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**Chapter 18**

**Antioxidant Role of Ascorbic Acid and**

José Luis Silencio Barrita and

http://dx.doi.org/10.5772/52181

for reducing inflammatory symptoms.

**1. Introduction**

of hydroxyproline.

María del Socorro Santiago Sánchez

Additional information is available at the end of the chapter

**His Protective Effects on Chronic Diseases**

Ascorbic acid (AA), commonly known as vitamin C, plays an important role in the human body, although its function at the cellular level is not yet clear. It is necessary for the synthe‐ sis of collagen, a protein that has many connective functions in the body. Among the sub‐ stances and structures that contain collagen are bone, cartilage and the surrounding material, as well as carrier substances and materials of union muscle, skin and other tissues. It also requires (AA) for the synthesis of hormones, neurotransmitters and in the metabolism of certain amino acids and vitamins. Participate in the liver for detoxification of toxic sub‐ stances and blood level for immunity. As an antioxidant reacts with histamine and peroxide

Its antioxidant capacity is associated with reduced incidence of cancer. The requirement for vitamin C for adults is well defined but they have not been uniform across different cul‐ tures, so their need has been defined as culture-specific. They have also defined other roles in cellular processes and reactions. Some epidemiological data mentioned its usefulness in reducing cold with increasing consumption of foods rich in vitamin, so people sometimes ingest an overdose of it. In most reports mention that discrete increases in blood levels of this vitamin reduces the risk of death in all conditions. Although there are many functions of vitamin C, his role in health is discussed mostly in relation to its role as an antioxidant and its effects on cancer, blood pressure, immunity, drug metabolism and urinary excretion

Antioxidants play important roles in cellular function and have been implicated in processes associated with aging, including vascular, inflammatory damage and cancer. In the case of

> © 2013 Barrita and Sánchez; licensee InTech. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

### **Chapter 18**

### **Antioxidant Role of Ascorbic Acid and His Protective Effects on Chronic Diseases**

José Luis Silencio Barrita and María del Socorro Santiago Sánchez

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/52181

### **1. Introduction**

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[176] Jin, X., Hidiroglou, N., Lok, E., Taylor, M., Kapal, K., et al. (2012). Dietary Selenium (Se) and Vitamin E (VE) Supplementation Modulated Methylmercury-Mediated Changes in Markers of Cardiovascular Diseases. *Rats Cardiovascular Toxicology*, 12,

[177] El-Desoky, G., Abdelreheem, M., AL-Othman, A., Alothman, Z., Mahmoud, M., & Yusuf, K. (2012). Potential hepatoprotective effects of vitamin E and selenium on hepatotoxicity induced by malathion in rats. *African Journal of Pharmacy and Pharma‐*

min E Cancer Prevention Trial) status. *Molecular Medicine*, 17, 134-143.

and meta-analysis. *Mayo Clinic Proceedings*, 83, 23-34.

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*Journal of American Medical Association*, 301, 39-51.

Controlled Trials. *Nutrition and Cancer*, 62, 719-727.

*Journal of Nutrition*, 137, 1338-1340.

10-24.

*cology*, 6, 806-813.

*ics*, 21, 1315-1330.

Ascorbic acid (AA), commonly known as vitamin C, plays an important role in the human body, although its function at the cellular level is not yet clear. It is necessary for the synthe‐ sis of collagen, a protein that has many connective functions in the body. Among the sub‐ stances and structures that contain collagen are bone, cartilage and the surrounding material, as well as carrier substances and materials of union muscle, skin and other tissues. It also requires (AA) for the synthesis of hormones, neurotransmitters and in the metabolism of certain amino acids and vitamins. Participate in the liver for detoxification of toxic sub‐ stances and blood level for immunity. As an antioxidant reacts with histamine and peroxide for reducing inflammatory symptoms.

Its antioxidant capacity is associated with reduced incidence of cancer. The requirement for vitamin C for adults is well defined but they have not been uniform across different cul‐ tures, so their need has been defined as culture-specific. They have also defined other roles in cellular processes and reactions. Some epidemiological data mentioned its usefulness in reducing cold with increasing consumption of foods rich in vitamin, so people sometimes ingest an overdose of it. In most reports mention that discrete increases in blood levels of this vitamin reduces the risk of death in all conditions. Although there are many functions of vitamin C, his role in health is discussed mostly in relation to its role as an antioxidant and its effects on cancer, blood pressure, immunity, drug metabolism and urinary excretion of hydroxyproline.

Antioxidants play important roles in cellular function and have been implicated in processes associated with aging, including vascular, inflammatory damage and cancer. In the case of

© 2013 Barrita and Sánchez; licensee InTech. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

AA its antioxidant role is useful since it contributes to the maintenance of the vascular sys‐ tem and the reduction of atherogenesis through regulation in collagen synthesis, production of prostacyclin and nitric oxide. In addition to this antioxidant role, the AA has actions at the molecular level because it acts as a cofactor of enzymes such as dopamine hydroxylase (EC <IP_ADDRESS>), influencing neurotransmitter concentration, improves lysosomal protein deg‐ radation and mediates consumer monosodium glutamate.

### **1.1. History**

Since the nineteenth and early twentieth century research on these compounds led to the discovery of vitamins.4 Since 1901, a publication of Wildiers first described the stimulating effect of small amounts of organic material in the growth of yeast; this effect was the subject of many publications and only after several years was universally accepted. Wilders gave the name "bios" to the substance or substances causing increased growth of yeast. In the years since it was shown that bios were multiple in nature, and changes fractionated as bios I, bios IIA, IIB and others.5

Name (IUPAC) systematic: (R) -3, 4-dihydroxy-5-((S) -1, 2-dihydroxyethyl) furan-2 (5H)-one;

Antioxidant Role of Ascorbic Acid and His Protective Effects on Chronic Diseases

http://dx.doi.org/10.5772/52181

451

Ascorbic acid contains several structural elements that contribute to their chemical behavior: the structure of the lactones and two enolic hydroxyl groups and a primary and secondary alcohol group. Enediol structure motivates their antioxidant properties, as can be oxidized easily enediols to diketones. Therefore, the carbonyl groups endioles neighbors are also

Ascorbic acid forms two bonds intermolecular hydrogen bonds (shown in red in the figure) that contribute substantially to the stability and with it the chemical qualities of the structure

Ascorbic acid is rapidly interconvert in two unstable diketone tautomers by proton transfer, though it is most stable in the enol form. The proton of the enol is lost, and again acquired by the electrons from the double bond to produce a diketone. This reaction is an enol. There

**Figure 1.** Structure of ascorbic acid and dehydroascorbic acid.

**2.2. Physical and chemical properties**

**Figure 2.** Hydrogen bridges formed by ascorbic acid

are two possible ways: 1, 2-diketone and 1, 3-diketone (figure 3).

called reductive.

endiol.

CAS Number 50-81-7: Formula: C6H8O6; mol wt. 176.13 g / mol

1747. - Lind cured scurvy in British sailors with oranges and lemons.

1907. - It is reproduced experimental scurvy in guinea pigs.

1928. - Eascott identified bios I as meso-inositol. Szent-Gyorgy and Glen published the isola‐ tion of vitamin C or hexuronic acid.

1933. - Allison, Hoover and Burk describe a compound that promotes the respiration and growth of Rhizobium, which designated "Coenzyme R". Then, it is defined molecular struc‐ ture and synthesis of vitamin C.

1937. - the Nobel Prize in Chemistry was awarded to Walter Haworth for his work in deter‐ mining the structure of ascorbic acid (shared with Paul Karrer for his work on vitamins) and the Nobel Prize for medicine was awarded to Albert von Szent-Györgyi Nagyrápolt for his studies on the biological functions of ascorbic acid.

### **2. Definition**

Vitamin C is defined as hexuronic acid, cevitáminic acid or xiloascórbic acid. The term vita‐ min C is generally used to describe all these compounds, although the representative of which is ascorbic acid.

#### **2.1. Structure, formula and chemical characteristics**

Ascorbic acid is the enolic form of one α-ketolactone. Ascorbic acid solution is easily oxi‐ dized to the diketo form referred to as dehydroascorbic acid, which can easily be converted into oxalic acid , diketogulonic acid or threonic acid.

**Figure 1.** Structure of ascorbic acid and dehydroascorbic acid.

Name (IUPAC) systematic: (R) -3, 4-dihydroxy-5-((S) -1, 2-dihydroxyethyl) furan-2 (5H)-one; CAS Number 50-81-7: Formula: C6H8O6; mol wt. 176.13 g / mol

### **2.2. Physical and chemical properties**

AA its antioxidant role is useful since it contributes to the maintenance of the vascular sys‐ tem and the reduction of atherogenesis through regulation in collagen synthesis, production of prostacyclin and nitric oxide. In addition to this antioxidant role, the AA has actions at the molecular level because it acts as a cofactor of enzymes such as dopamine hydroxylase (EC <IP_ADDRESS>), influencing neurotransmitter concentration, improves lysosomal protein deg‐

Since the nineteenth and early twentieth century research on these compounds led to the

effect of small amounts of organic material in the growth of yeast; this effect was the subject of many publications and only after several years was universally accepted. Wilders gave the name "bios" to the substance or substances causing increased growth of yeast. In the years since it was shown that bios were multiple in nature, and changes fractionated as bios

1928. - Eascott identified bios I as meso-inositol. Szent-Gyorgy and Glen published the isola‐

1933. - Allison, Hoover and Burk describe a compound that promotes the respiration and growth of Rhizobium, which designated "Coenzyme R". Then, it is defined molecular struc‐

1937. - the Nobel Prize in Chemistry was awarded to Walter Haworth for his work in deter‐ mining the structure of ascorbic acid (shared with Paul Karrer for his work on vitamins) and the Nobel Prize for medicine was awarded to Albert von Szent-Györgyi Nagyrápolt for his

Vitamin C is defined as hexuronic acid, cevitáminic acid or xiloascórbic acid. The term vita‐ min C is generally used to describe all these compounds, although the representative of

Ascorbic acid is the enolic form of one α-ketolactone. Ascorbic acid solution is easily oxi‐ dized to the diketo form referred to as dehydroascorbic acid, which can easily be converted

Since 1901, a publication of Wildiers first described the stimulating

radation and mediates consumer monosodium glutamate.

450 Oxidative Stress and Chronic Degenerative Diseases - A Role for Antioxidants

1747. - Lind cured scurvy in British sailors with oranges and lemons.

1907. - It is reproduced experimental scurvy in guinea pigs.

studies on the biological functions of ascorbic acid.

**2.1. Structure, formula and chemical characteristics**

into oxalic acid , diketogulonic acid or threonic acid.

**1.1. History**

discovery of vitamins.4

I, bios IIA, IIB and others.5

tion of vitamin C or hexuronic acid.

ture and synthesis of vitamin C.

**2. Definition**

which is ascorbic acid.

Ascorbic acid contains several structural elements that contribute to their chemical behavior: the structure of the lactones and two enolic hydroxyl groups and a primary and secondary alcohol group. Enediol structure motivates their antioxidant properties, as can be oxidized easily enediols to diketones. Therefore, the carbonyl groups endioles neighbors are also called reductive.

Ascorbic acid forms two bonds intermolecular hydrogen bonds (shown in red in the figure) that contribute substantially to the stability and with it the chemical qualities of the structure endiol.

**Figure 2.** Hydrogen bridges formed by ascorbic acid

Ascorbic acid is rapidly interconvert in two unstable diketone tautomers by proton transfer, though it is most stable in the enol form. The proton of the enol is lost, and again acquired by the electrons from the double bond to produce a diketone. This reaction is an enol. There are two possible ways: 1, 2-diketone and 1, 3-diketone (figure 3).

Chemical data: IUPAC Name: D-isoascorbic acid, CAS Number: [89-65-6] Molecular formu‐

Antioxidant Role of Ascorbic Acid and His Protective Effects on Chronic Diseases

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453

A short-term supplementation with physiological doses of antioxidant vitamins, carote‐ noids and trace elements during alcohol rehabilitation clearly improves micronutrient status indicators. Heavy smokers in particular seem to respond to vitamin C supplemen‐

In the liver accumulate micronutrients such as Vitamin A, E and iron, and therefore, in pa‐ tients with hepatic impairment is a deficit of them due to reduced intake, as well as intesti‐ nal transport and liver stores. The alteration of fat soluble vitamins is especially important in

Moreover, in the alcoholic patient levels water-soluble vitamins are low due to the effect of ethanol on its metabolism, producing pyridoxine deficiency, retinol, cobalamin, folate and niacin. In fact, in chronic alcoholism may develop Wernicke encephalopathy. It has also shown a direct relationship between oxidative stress and disease severity liver, requiring the micronutrients with antioxidant activity, being increased the needs of vitamin E and C.

There are few data on vitamin needs and trace elements in burned patients. the large tissue loss, decreased gastrointestinal absorption, increased urinary losses, changes in distribution, and a high degree catabolism, that are made increased the needs of vitamins and trace ele‐ ments. Clinical guidelines recommend giving also established daily requirements and addi‐

Also, increase the intake of vitamin C (1,000 mg/day) as it promotes the healing process, and vitamin A (10,000 IU/day) for its effect immune and protective skin and mucous mem‐ branes. Also, is required additional vitamin D due to high risk of fractures in these group

Zinc supplement is suitable dose of 220 mg/day, as is involved in protein synthesis and tis‐ sue regeneration. Furthermore, Chan et al indicate that in the week post-injury, there are high losses exuding of copper, being necessary to increase their requirements (4.5 mg/day of

The increased production of oxygen species reactive in this clinical situation requires admin‐ istration of antioxidants (ascorbic acid, glutathione, carotenoids, vitamin A and E) been shown to reduce mortality, protecting the micro vascular circulation, reducing the peroxida‐ tion tissue lipid. According to some authors, surgical stress may necessitate supplementa‐ tion of ascorbic acid, alpha tocopherol and trace elements, associating too low preoperative levels of vitamin A (<0.77 mol/L) with an increase of postoperative infection and mortality. At present, it is unknown whether supplementation micronutrient for a short period of time

patients but have not yet been established Daily exact requirements.

la: C6H8O6, molar mass: 176.13 g/mol; Melting point: 169-172 ° C

**3. Relationship with other nutrients**

patients with steatorrhea or cholestasis.

tional doses of certain micronutrients.

copper sulfate).

**3.1. Vitamin. A and E**

tation (Gueguen, 2003)

**Figure 3.** Nucleophilic attack on the proton ascorbic enol to give 1,3 diketone.

#### **2.3. Vitamers or vitameric forms**

The vitamer of a particular vitamin is any chemical compound which generally has the same molecular structure and each shows a different vitamin activity in a biological system which is deficient of the vitamin.

The vitamin activity of multiple vitamers is due to the ability (sometimes limited) of the body to convert one or many vitamers in another vitamer for the same enzymatic cofactor which is active in the body as the most important form of the vitamin. As part of the defini‐ tion of the vitamin, the body can not completely synthesize an optimal amount of vitamin activity of foodstuffs simple, without a certain minimum amount of vitamer as base. Not all vitamers have the same vitamin power by mass or weight. This is due to differences in the absorption and the variable interconversion several vitamers in the vitamin.

For ascorbic acid per se, may be mentioned the following vitamers: dehydroascorbic acid, erythorbic acid (figure 4) and the following salts: sodium ascorbate, calcium ascorbate, and others. (Rogur, 2010)

**Figure 4.** Structure of erythorbic acid, isoascorbic or Arabian-ascorbic

Chemical data: IUPAC Name: D-isoascorbic acid, CAS Number: [89-65-6] Molecular formu‐ la: C6H8O6, molar mass: 176.13 g/mol; Melting point: 169-172 ° C

### **3. Relationship with other nutrients**

### **3.1. Vitamin. A and E**

**Figure 3.** Nucleophilic attack on the proton ascorbic enol to give 1,3 diketone.

452 Oxidative Stress and Chronic Degenerative Diseases - A Role for Antioxidants

The vitamer of a particular vitamin is any chemical compound which generally has the same molecular structure and each shows a different vitamin activity in a biological system which

The vitamin activity of multiple vitamers is due to the ability (sometimes limited) of the body to convert one or many vitamers in another vitamer for the same enzymatic cofactor which is active in the body as the most important form of the vitamin. As part of the defini‐ tion of the vitamin, the body can not completely synthesize an optimal amount of vitamin activity of foodstuffs simple, without a certain minimum amount of vitamer as base. Not all vitamers have the same vitamin power by mass or weight. This is due to differences in the

For ascorbic acid per se, may be mentioned the following vitamers: dehydroascorbic acid, erythorbic acid (figure 4) and the following salts: sodium ascorbate, calcium ascorbate, and

absorption and the variable interconversion several vitamers in the vitamin.

**Figure 4.** Structure of erythorbic acid, isoascorbic or Arabian-ascorbic

**2.3. Vitamers or vitameric forms**

is deficient of the vitamin.

others. (Rogur, 2010)

A short-term supplementation with physiological doses of antioxidant vitamins, carote‐ noids and trace elements during alcohol rehabilitation clearly improves micronutrient status indicators. Heavy smokers in particular seem to respond to vitamin C supplemen‐ tation (Gueguen, 2003)

In the liver accumulate micronutrients such as Vitamin A, E and iron, and therefore, in pa‐ tients with hepatic impairment is a deficit of them due to reduced intake, as well as intesti‐ nal transport and liver stores. The alteration of fat soluble vitamins is especially important in patients with steatorrhea or cholestasis.

Moreover, in the alcoholic patient levels water-soluble vitamins are low due to the effect of ethanol on its metabolism, producing pyridoxine deficiency, retinol, cobalamin, folate and niacin. In fact, in chronic alcoholism may develop Wernicke encephalopathy. It has also shown a direct relationship between oxidative stress and disease severity liver, requiring the micronutrients with antioxidant activity, being increased the needs of vitamin E and C.

There are few data on vitamin needs and trace elements in burned patients. the large tissue loss, decreased gastrointestinal absorption, increased urinary losses, changes in distribution, and a high degree catabolism, that are made increased the needs of vitamins and trace ele‐ ments. Clinical guidelines recommend giving also established daily requirements and addi‐ tional doses of certain micronutrients.

Also, increase the intake of vitamin C (1,000 mg/day) as it promotes the healing process, and vitamin A (10,000 IU/day) for its effect immune and protective skin and mucous mem‐ branes. Also, is required additional vitamin D due to high risk of fractures in these group patients but have not yet been established Daily exact requirements.

Zinc supplement is suitable dose of 220 mg/day, as is involved in protein synthesis and tis‐ sue regeneration. Furthermore, Chan et al indicate that in the week post-injury, there are high losses exuding of copper, being necessary to increase their requirements (4.5 mg/day of copper sulfate).

The increased production of oxygen species reactive in this clinical situation requires admin‐ istration of antioxidants (ascorbic acid, glutathione, carotenoids, vitamin A and E) been shown to reduce mortality, protecting the micro vascular circulation, reducing the peroxida‐ tion tissue lipid. According to some authors, surgical stress may necessitate supplementa‐ tion of ascorbic acid, alpha tocopherol and trace elements, associating too low preoperative levels of vitamin A (<0.77 mol/L) with an increase of postoperative infection and mortality. At present, it is unknown whether supplementation micronutrient for a short period of time could restore plasma antioxidant levels after surgery. Some authors suggest that antioxi‐ dants could lead to improved metabolism and ventricular function after cardiac surgery. Al‐ so state that patient's major surgery may benefit from selenium, even before surgery, to action at the level of oxidative stress. The ESPEN recommended in these surgical patients treated with parenteral nutrition, supplement micronutrient recommended daily doses, the vitamin supplementation is unnecessary if the patient is on concomitant enteral nutrition, oral or parenteral.

require thiamine, niacin, vitamin A, E and C, B complex, zinc (15-20 mg/day and 10 mg/L

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455

Consumption of 0.6 mg equivalents of alpha tocopherol/g linoleic acid is suitable for human adults. The minimum requirement of vitamin E related to the consumption of fatty acids with a high degree of unsaturation can be calculated with a specific formula that must take into account the peroxibility of polyunsaturated fatty acids is based on the results of animal experiments. But still no convincing evidence of how much vitamin E is required in relation

Studies so far show that the effects of supplementation with EPA and DHA lipid peroxidiza‐ cion increase even when the amounts of vitamin E present are adequate in relation to the oxida‐ tive potential of these fatty acids. On the other hand the calculation of the requirement for vitamin E using current data from recent consumption, show that a reduction in total fat intake with a concomitant increase in consumption of polyunsaturated fatty acids, including EPA and DHA results in an increased need intake of vitamin E. In fact the methods used to investigate the requirements of vitamin E and polyunsaturated fatty acid intake (erythrocyte hemolysis) and the techniques used to assess lipid peroxidation (malondialdehyde analysis, MDA) may

Therefore, further studies are needed to establish the requirement of vitamin E when intake of unsaturated fatty acids of longer chain increases. For this purpose it is necessary to use functional techniques based on the measurement of in vivo lipid peroxidation. Until then or until the available data suggest using the index of 0.6 mg of alpha tocopherol per gram of ingested PUFA. However it is likely that higher levels are necessary for vitamin fats are rich

The diet of our ancestors was less dense in calories, being higher in fiber, rich in fruits, vege‐ tables, lean meat and fish. As a result, the diet was significantly lower in total fat and satu‐ rated fat, but containing equal amounts of n-6 essential fatty acids and n-3. Linoleic acid (LA) is the major n-6 fatty acid and alpha-linolenic acid (ALA) is the major n-3 fatty acid.

In the body, LA is metabolized to arachidonic acid (ARA), and ALA is metabolized to eico‐ sapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The essential fatty acid ratio n-6: n-3 is 1 to 2:1, with higher levels of long chain polyunsaturated fatty acids (PUFA), such as EPA, DHA and ARA, today's diet. Today this ratio is about 10 to 1 or 20 and 25 to 1, indicat‐ ing that Western diets are deficient in n-3 fatty acids compared with the diet that humans

The n-3 and n-6 are not interconvertible in the human body and are important components of practically all cell membranes. The n-6 fatty acids and n-3 influence eicosanoid metabo‐ lism, gene expression, and intercellular communication cell to cell. The polyunsaturated fat‐ ty acid composition of cell membranes is largely dependent on food ingestion. Therefore, appropriate amounts of n-6 fatty acids and n-3 in the diet should be considered in making

to consumption of polyunsaturated fatty acids: EPA (20:5 n-3) and DHA (22:6 n-3).

be inappropriate for measuring a quantitative relationship between the two loads.

in fatty acids containing more than two double bonds (Valk, 2000).

evolved and established patterns genetic.

dietary recommendations.

intestinal leaks) and selenium (rise up to 120 ug/day) in patients with sepsis.

**3.3. Polyunsaturated fatty acids**

### **3.2. Minerals such as selenium, iron and zinc**

Recent studies have shown that in patients with inflammatory bowel disease there is a cor‐ relation between the level of some antioxidants such as selenium, vitamin C and E and clini‐ cal improvement and reduction in serum levels of TNF-α and decrease in steroid dose to 65%. Low serum zinc levels have been correlated with increased blood pressure, disease cor‐ onary type II diabetes mellitus and hyperlipidemia. Also, high intake of magnesium (> 500-1000 mg/day) can lower high blood pressure, and be effective in acute myocardial in‐ farction and atherosclerosis. Houston recommended to prevent the emergence and develop‐ ment of hypertension, administration of additional vitamins and trace elements. Finally, oxidative stress plays an important role in the initiation and maintenance of the pathogene‐ sis of cardiovascular disease and its complications. For some authors, antioxidants such as vitamins E and C, beta-carotene, selenium and zinc, may act by reducing the cardiovascular risk, although evidence is limited.

Malnutrition is a common feature of inflammatory intestinal diseases, being frequent the deficit of vitamins B12, A, D, E and K, steatorrhea, ileal resection or extensive lesions in the intestine. GI bleeding contributing to iron losses, and through diarrhea and fistulas is loss of electrolytes and trace elements (copper, magnesium, selenium and zinc).

The role of antioxidant micronutrients in the clinical and functional improvement has been described by different authors. Thus, a low intake of selenium, beta-carotene and vitamins E and C, can reduce the body's natural defenses and increase inflammation of the airways, and therefore the selenium (100-200 ug/d) been associated with improved lung function, es‐ pecially in smokers. Gazdik et al indicate that supplementation of 200 ug/d of selenium in asthmatic patients produced a statistically significant decrease in the use of corticosteroids. Loannidis and McClave et al indicate that antioxidants such as selenium, vitamin A, vitamin C and vitamin E reduce pancreatic inflammation and pain, and prevent the occurrence of exacerbations. Recently in a double blind study in patients with chronic pancreatitis were given daily supplements of 600 ug of selenium, 9,000 IU of beta-carotene, 540 mg of vitamin C, 270 IU of vitamin E and 2,000 mg methionine, the pain was reduced significantly, as well as stress markers oxidative normalized plasma concentrations of antioxidants.

For some authors, parenteral administration of ascorbic acid can lower the morbidity and mortality of these patients in a randomized, double-blind placebo-controlled; we observed that mortality at day 28 decreased in the group of patients who received ascorbate and vita‐ min E by intravenous infusion. Some authors recommend increasing the contribution of an‐ tioxidants such as vitamin C, retinol, vitamin E, beta-carotene and selenium. Also appear to require thiamine, niacin, vitamin A, E and C, B complex, zinc (15-20 mg/day and 10 mg/L intestinal leaks) and selenium (rise up to 120 ug/day) in patients with sepsis.

### **3.3. Polyunsaturated fatty acids**

could restore plasma antioxidant levels after surgery. Some authors suggest that antioxi‐ dants could lead to improved metabolism and ventricular function after cardiac surgery. Al‐ so state that patient's major surgery may benefit from selenium, even before surgery, to action at the level of oxidative stress. The ESPEN recommended in these surgical patients treated with parenteral nutrition, supplement micronutrient recommended daily doses, the vitamin supplementation is unnecessary if the patient is on concomitant enteral nutrition,

Recent studies have shown that in patients with inflammatory bowel disease there is a cor‐ relation between the level of some antioxidants such as selenium, vitamin C and E and clini‐ cal improvement and reduction in serum levels of TNF-α and decrease in steroid dose to 65%. Low serum zinc levels have been correlated with increased blood pressure, disease cor‐ onary type II diabetes mellitus and hyperlipidemia. Also, high intake of magnesium (> 500-1000 mg/day) can lower high blood pressure, and be effective in acute myocardial in‐ farction and atherosclerosis. Houston recommended to prevent the emergence and develop‐ ment of hypertension, administration of additional vitamins and trace elements. Finally, oxidative stress plays an important role in the initiation and maintenance of the pathogene‐ sis of cardiovascular disease and its complications. For some authors, antioxidants such as vitamins E and C, beta-carotene, selenium and zinc, may act by reducing the cardiovascular

Malnutrition is a common feature of inflammatory intestinal diseases, being frequent the deficit of vitamins B12, A, D, E and K, steatorrhea, ileal resection or extensive lesions in the intestine. GI bleeding contributing to iron losses, and through diarrhea and fistulas is loss of

The role of antioxidant micronutrients in the clinical and functional improvement has been described by different authors. Thus, a low intake of selenium, beta-carotene and vitamins E and C, can reduce the body's natural defenses and increase inflammation of the airways, and therefore the selenium (100-200 ug/d) been associated with improved lung function, es‐ pecially in smokers. Gazdik et al indicate that supplementation of 200 ug/d of selenium in asthmatic patients produced a statistically significant decrease in the use of corticosteroids. Loannidis and McClave et al indicate that antioxidants such as selenium, vitamin A, vitamin C and vitamin E reduce pancreatic inflammation and pain, and prevent the occurrence of exacerbations. Recently in a double blind study in patients with chronic pancreatitis were given daily supplements of 600 ug of selenium, 9,000 IU of beta-carotene, 540 mg of vitamin C, 270 IU of vitamin E and 2,000 mg methionine, the pain was reduced significantly, as well

For some authors, parenteral administration of ascorbic acid can lower the morbidity and mortality of these patients in a randomized, double-blind placebo-controlled; we observed that mortality at day 28 decreased in the group of patients who received ascorbate and vita‐ min E by intravenous infusion. Some authors recommend increasing the contribution of an‐ tioxidants such as vitamin C, retinol, vitamin E, beta-carotene and selenium. Also appear to

electrolytes and trace elements (copper, magnesium, selenium and zinc).

as stress markers oxidative normalized plasma concentrations of antioxidants.

oral or parenteral.

**3.2. Minerals such as selenium, iron and zinc**

454 Oxidative Stress and Chronic Degenerative Diseases - A Role for Antioxidants

risk, although evidence is limited.

Consumption of 0.6 mg equivalents of alpha tocopherol/g linoleic acid is suitable for human adults. The minimum requirement of vitamin E related to the consumption of fatty acids with a high degree of unsaturation can be calculated with a specific formula that must take into account the peroxibility of polyunsaturated fatty acids is based on the results of animal experiments. But still no convincing evidence of how much vitamin E is required in relation to consumption of polyunsaturated fatty acids: EPA (20:5 n-3) and DHA (22:6 n-3).

Studies so far show that the effects of supplementation with EPA and DHA lipid peroxidiza‐ cion increase even when the amounts of vitamin E present are adequate in relation to the oxida‐ tive potential of these fatty acids. On the other hand the calculation of the requirement for vitamin E using current data from recent consumption, show that a reduction in total fat intake with a concomitant increase in consumption of polyunsaturated fatty acids, including EPA and DHA results in an increased need intake of vitamin E. In fact the methods used to investigate the requirements of vitamin E and polyunsaturated fatty acid intake (erythrocyte hemolysis) and the techniques used to assess lipid peroxidation (malondialdehyde analysis, MDA) may be inappropriate for measuring a quantitative relationship between the two loads.

Therefore, further studies are needed to establish the requirement of vitamin E when intake of unsaturated fatty acids of longer chain increases. For this purpose it is necessary to use functional techniques based on the measurement of in vivo lipid peroxidation. Until then or until the available data suggest using the index of 0.6 mg of alpha tocopherol per gram of ingested PUFA. However it is likely that higher levels are necessary for vitamin fats are rich in fatty acids containing more than two double bonds (Valk, 2000).

The diet of our ancestors was less dense in calories, being higher in fiber, rich in fruits, vege‐ tables, lean meat and fish. As a result, the diet was significantly lower in total fat and satu‐ rated fat, but containing equal amounts of n-6 essential fatty acids and n-3. Linoleic acid (LA) is the major n-6 fatty acid and alpha-linolenic acid (ALA) is the major n-3 fatty acid.

In the body, LA is metabolized to arachidonic acid (ARA), and ALA is metabolized to eico‐ sapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The essential fatty acid ratio n-6: n-3 is 1 to 2:1, with higher levels of long chain polyunsaturated fatty acids (PUFA), such as EPA, DHA and ARA, today's diet. Today this ratio is about 10 to 1 or 20 and 25 to 1, indicat‐ ing that Western diets are deficient in n-3 fatty acids compared with the diet that humans evolved and established patterns genetic.

The n-3 and n-6 are not interconvertible in the human body and are important components of practically all cell membranes. The n-6 fatty acids and n-3 influence eicosanoid metabo‐ lism, gene expression, and intercellular communication cell to cell. The polyunsaturated fat‐ ty acid composition of cell membranes is largely dependent on food ingestion. Therefore, appropriate amounts of n-6 fatty acids and n-3 in the diet should be considered in making dietary recommendations.

These two classes of polyunsaturated fatty acids must be distinguished because they are metabolically and functionally distinct and have opposing physiological functions, balance is important for maintaining homeostasis and normal development. Studies with nonhuman primates and human newborns indicate that DHA is essential for normal functional devel‐ opment of the retina and the brain, especially in premature babies. A balanced n-6/n-3 ratio in the diet is essential for normal growth and development and should lead to reduced car‐ diovascular disease and other chronic diseases and improve mental health. Although a rec‐ ommended dietary allowance for essential fatty acids does not exist, an adequate intake (AI) has been estimated for essential fatty acids n-6 and n-3 by an international scientific working group. The final recommendations are for Western societies, reduce the consumption of n-6 fatty acids and increased intake of n-3 fatty acids. (Simopoulus, 2000)

**4.2. Fruits and vegetables**

Lemon juice Orange juice, canned Col common, dehydrated Peppers, sweet, ripe, red, raw Currants, black, European, raw

parsley, dry Orange juice green radish, raw Grape juice Orange peel, raw Pokeroot fruit Mustard seeds

leaves of kale, boiled, drained

Leaves and stems of cress, raw

Brussels sprouts, cooked in water

Broccoli cooked Brussels sprouts cooked

Cauliflower, raw

Cabbage, red, raw Strawberries Raw

Lamb crude

to quantify when using colorimetric methods.

Vitamin C is a major constituent of fruits and vegetables, which also contain citric acid, oxa‐ lates and substances such as anthocyanins, coloring agents and carotenoids that are difficult

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Currently there is great interest in relation to consumption of natural foods and mainly on the content of nutrients in fruits, vegetables and vitamin C. This interest is due in part to vitamin C is probably one of the most widely used nutrients in the food and pharmaceutical industry. Used as a supplement, additive, preservative, as an antioxidant in processed

> > 76 61 59

foods. Table 2 shows the main foods that are good sources of vitamin C.

Acerola 1743 Coriander leaves dry 567 red peppers, spicy, ripe, raw 369 Orange juice, dehydrated 359 Grape juice, dehydrated 350 common Guavas, whole, raw 242 Dried Tomato juice 239 red pepper, spicy, immature, raw 235

Papaya 56

**Table 2.** Acid content of ascorbic acid in different foods and different presentations

**FOOD CONCETRATION (mg/100g)**

### **4. Vitamin C sources**

#### **4.1. Food sources in Mexico**

The main sources of ascorbic acid are presented in Table 1. The results of vitamin C are shown as the mean and correspond to the official tables of composition of Mexican foods.

**Table 1.** Main sources of ascorbic acid in México

### **4.2. Fruits and vegetables**

These two classes of polyunsaturated fatty acids must be distinguished because they are metabolically and functionally distinct and have opposing physiological functions, balance is important for maintaining homeostasis and normal development. Studies with nonhuman primates and human newborns indicate that DHA is essential for normal functional devel‐ opment of the retina and the brain, especially in premature babies. A balanced n-6/n-3 ratio in the diet is essential for normal growth and development and should lead to reduced car‐ diovascular disease and other chronic diseases and improve mental health. Although a rec‐ ommended dietary allowance for essential fatty acids does not exist, an adequate intake (AI) has been estimated for essential fatty acids n-6 and n-3 by an international scientific working group. The final recommendations are for Western societies, reduce the consumption of n-6

The main sources of ascorbic acid are presented in Table 1. The results of vitamin C are shown as the mean and correspond to the official tables of composition of Mexican foods.

fatty acids and increased intake of n-3 fatty acids. (Simopoulus, 2000)

456 Oxidative Stress and Chronic Degenerative Diseases - A Role for Antioxidants

**FOOD CONCENTRATION (mg/100g EP)**

poblan chili 364 "trompito" chili 320 yucca flower 273 guava 199 Marañon 167 cauliflower 127 red bell pepper 160 guajillo chili 100 garlic 99 Nanche fruit 86 orange 76 manila mango 76 serran chili 65 pumpkin 58 watercress 51 beet 20 cucumber 13

**4. Vitamin C sources**

**4.1. Food sources in Mexico**

EP = edible portion

**Table 1.** Main sources of ascorbic acid in México

Vitamin C is a major constituent of fruits and vegetables, which also contain citric acid, oxa‐ lates and substances such as anthocyanins, coloring agents and carotenoids that are difficult to quantify when using colorimetric methods.

Currently there is great interest in relation to consumption of natural foods and mainly on the content of nutrients in fruits, vegetables and vitamin C. This interest is due in part to vitamin C is probably one of the most widely used nutrients in the food and pharmaceutical industry. Used as a supplement, additive, preservative, as an antioxidant in processed foods. Table 2 shows the main foods that are good sources of vitamin C.

**Table 2.** Acid content of ascorbic acid in different foods and different presentations

In Table 3 shows the analysis of vitamin C in different parts of the food such as edi‐ ble portion thereof, the seed or plant center and the shell and stalks that are normally discarded.

**Food Concentration**

**Table 4.** Content of ascorbic acid in Mexican fruits and vegetables

rose apple and apple are the best examples.

The shell or skin of many fruits and vegetables is usually discarded, but these same wastes have significant amounts of ascorbic acid, which is shown in Table 5, where the lime peel,

**(mg / 100 g EP)**

Antioxidant Role of Ascorbic Acid and His Protective Effects on Chronic Diseases

squash 7.2±0.9 94.2 spinach 8.5±0.2 91.6 potatoes 74.6±6.7 79.7 cucumber 93.0±7.1 96.0 green tomato 222.8±10.7 90.9 poblano chile 191.0±7.8 91.9 green pepper 195.5±9.5 94.3 nopales 268.9±30.1 95.2 cambray onion 17.0±0.7 93.0 carrot 50.4±5.6 87.2 white cabbage 184.7±17.2 93.2 grapefruit 261.3±10.7 87.8 mango 319.6±5.3 84.6 watermelon 56.2±8.9 92.5 banana 333.7±6.3 76.5 orange 279.8±39.4 84.0 mamey 31.6±1.2 66.8 plum 331.1±17.9 88.1 grape 66.1±13.6 84.3 apple 458.1±17.8 84.5 beet 70.8±6.8 85.3 lemon 39.4±2.5 88.3 avocado 256.2±61.9 84.3 sweet lime 306.8±23.4 89.8

**Moisture (%)**

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**Table 3.** Distribution of ascorbic acid (mg/100g) in some fruits and vegetables produced in México

Below the edible portion the moisture (%) is indicated in parenthesis. The data shown in Tables 3 through 6 are original and have not been published yet. EP = Edible portion

Table 4 shows the concentration of ascorbic acid in the main fruits and vegetables con‐ sumed in Mexico. Data are presented as mean and standard deviation. It shows that even the concentration of vitamin C is lower in the edible portion in the shells report‐ ed in Table 5.

**Table 4.** Content of ascorbic acid in Mexican fruits and vegetables

In Table 3 shows the analysis of vitamin C in different parts of the food such as edi‐ ble portion thereof, the seed or plant center and the shell and stalks that are normally

> 848.9±11.2 (81.8)

> > 19.9±0.5 (94.2)

1916.5±186.7 (69.5)

623.0±35.1 (82.5)

> 82.1±4.9 (81.8)

115.8±1.2 (85.8)

> 4.2±1.0 (85.8)

Below the edible portion the moisture (%) is indicated in parenthesis. The data shown in

Table 4 shows the concentration of ascorbic acid in the main fruits and vegetables con‐ sumed in Mexico. Data are presented as mean and standard deviation. It shows that even the concentration of vitamin C is lower in the edible portion in the shells report‐

Tables 3 through 6 are original and have not been published yet. EP = Edible portion

**Table 3.** Distribution of ascorbic acid (mg/100g) in some fruits and vegetables produced in México

(93.0)

266.6±6.7 (90.5)

1044.5±50.9 (65.7)

**Food Edible portion Shell Seed Stem**

discarded.

Apple 458.1±17.8

chayote 7.2±0.9

onion 17.0±0.7

lime 256.2±17.2

pomarrosa 531.7±36.4

guanábana 140.7±3.6

beat 70.8±6.8

Potato 74.6±6.7

ed in Table 5.

(84.5)

458 Oxidative Stress and Chronic Degenerative Diseases - A Role for Antioxidants

(94.2)

(87.6)

(87.1)

(83.6)

(80.0)

(85.3)

(80.3)

The shell or skin of many fruits and vegetables is usually discarded, but these same wastes have significant amounts of ascorbic acid, which is shown in Table 5, where the lime peel, rose apple and apple are the best examples.

**Food-industrialized**

**Use as a preservative**

**5. Deficiencies**

gulated lymph (20-53 ug/108

ents on the label of the final product

acids (palmitate and ascorbyl stearate).

**5.1. Primary deficiency – scurvy. Signs and symptoms**

Vitamin C (ascorbic acid) is water soluble and sensitive to oxygen. For this reason, it may partially destroyed in foods during processing, if exposed to air during storage or if treated with water. Manufacturers can protect them from oxidation by adding vitamin C. The addi‐ tion of ascorbic acid as an antioxidant should be appropriately marked in the list of ingredi‐

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Ascorbic acid and its salts are practically insoluble in lipids (fats), for this reason that is often used in the food industry as an antioxidant and preservative greasy foods, in order to avoid rancid. Their salts are usually used with a solubilizing agent (usually a monoglyceride) to improve its implementation. Also this often used in the processing industry of cosmetics products. Sodium ascorbate is a sodium salt of ascorbic acid (vitamin C) and formula C6H7NaO6. This form is used in the food industry for their functions antiseptic, antioxidants, and preservatives. Ascorbyl palmitate is an ester formed by ascorbic acid (vitamin C) and palmitic acid creating a liposoluble form of vitamin C. It is used in the food industry as an

Is usually used as a food preservative and as antioxidant in the food industry, a typical case is found as a bread improver additive. In industry collecting fruit prevents the color oxida‐ tive change known browning. Is often added to foods treated with nitrite in order to reduce the generation of nitrosamines (a carcinogen), so commonly found in sausages and cold

Ascorbic acid and its sodium, potassium and calcium salts are used widely as antioxidants and additives. These compounds are soluble in water, so that fats do not protect against oxi‐ dation. For this purpose may be used ascorbic acid esters with fat soluble long chain fatty

A frank deficiency of vitamin C causes scurvy, a disease characterized by multiple hemorrhag‐ es. Scurvy in adults is manifested by lassitude, weakness, irritability, vague muscle pain, joint pain and weight loss. Early signs objectives are as bleeding gums, tooth loss and gingivitis.

The diagnosis of scurvy, is achieved by testing plasma ascorbic acid, low concentration indicates low levels in tissues. It is generally accepted that ascorbic acid concentration in the layer of coa‐

garding vitamin C and its concentration in tissues. In the most extreme cases scurvy shows: bleeding gums and skin, perifollicular bleeding and ocular petechiae, salivary and lachrymal glands "drier", functional neuropathy, lower limb edema, psychological disturbances, anemia

and poor healing of wounds. The consumption of snuff lowers blood levels of vitamin C.

leukocytes) is the most reliable indicator of nutritional status re‐

cuts. In the same way is generally used in the food industry as an acidity regulator.

antioxidant (code E 304). It is wrong to think that is a natural antioxidant

**Table 5.** Content of ascorbic acid in the shell of some fruits and vegetables

Table 6 shows the values of ascorbic acid in some plant species used as flavoring for Mexi‐ can dishes. In most cases the amount used for the preparation of food is very low and some‐ times do not amount to more than 2% by weight of the end plate. However their presence in cooked food gives organoleptic properties suitable for the acceptance of it and especially the potential of the flavors of food.

**Table 6.** Content of ascorbic acid in some food used as spices

### **Food-industrialized**

**Shell of concentration**

460 Oxidative Stress and Chronic Degenerative Diseases - A Role for Antioxidants

**Table 5.** Content of ascorbic acid in the shell of some fruits and vegetables

**spices concentration**

**Table 6.** Content of ascorbic acid in some food used as spices

potential of the flavors of food.

**(mg/100g)**

Table 6 shows the values of ascorbic acid in some plant species used as flavoring for Mexi‐ can dishes. In most cases the amount used for the preparation of food is very low and some‐ times do not amount to more than 2% by weight of the end plate. However their presence in cooked food gives organoleptic properties suitable for the acceptance of it and especially the

**(mg /100g EP)**

coriander 56.0±7.3 85.3 yerbabuena 226.9±21.9 86.8 epazote leaves 18.3±1.1 86.3 chard leaves 8.5±0.9 92.7 chard stem 15.7±0.9 95.0 parsley leaves 243.9±17.2 84.5 parsley stem 182.1±11.0 90.2 celery leaves 5.6±0.2 88.4 celery stem 7.8±0.9 90.3 corn grain 35.3±6.0 70.7 corn "hair" 55.9±1.2 85.4

CARROT 129.1±10.1 88.5 APPLE 848.9±11.2 81.8 BITTER LEMON 39.4±2.5 71.7 AVOCADO 187.1±27.2 82.0 BEET 115.8±1.2 85.8 SMALL POTATO 4.2±1.0 85.5 BIG POTATO 2.1±0.1 80.3 POMARROSA 622.9±35.1 82.5 LIME 1916.4±186.7 69.4 CHAYOTE 19.9±0.5 94.2

**moisture (%)**

**moisture (%)**

Vitamin C (ascorbic acid) is water soluble and sensitive to oxygen. For this reason, it may partially destroyed in foods during processing, if exposed to air during storage or if treated with water. Manufacturers can protect them from oxidation by adding vitamin C. The addi‐ tion of ascorbic acid as an antioxidant should be appropriately marked in the list of ingredi‐ ents on the label of the final product

Ascorbic acid and its salts are practically insoluble in lipids (fats), for this reason that is often used in the food industry as an antioxidant and preservative greasy foods, in order to avoid rancid. Their salts are usually used with a solubilizing agent (usually a monoglyceride) to improve its implementation. Also this often used in the processing industry of cosmetics products. Sodium ascorbate is a sodium salt of ascorbic acid (vitamin C) and formula C6H7NaO6. This form is used in the food industry for their functions antiseptic, antioxidants, and preservatives. Ascorbyl palmitate is an ester formed by ascorbic acid (vitamin C) and palmitic acid creating a liposoluble form of vitamin C. It is used in the food industry as an antioxidant (code E 304). It is wrong to think that is a natural antioxidant

#### **Use as a preservative**

Is usually used as a food preservative and as antioxidant in the food industry, a typical case is found as a bread improver additive. In industry collecting fruit prevents the color oxida‐ tive change known browning. Is often added to foods treated with nitrite in order to reduce the generation of nitrosamines (a carcinogen), so commonly found in sausages and cold cuts. In the same way is generally used in the food industry as an acidity regulator.

Ascorbic acid and its sodium, potassium and calcium salts are used widely as antioxidants and additives. These compounds are soluble in water, so that fats do not protect against oxi‐ dation. For this purpose may be used ascorbic acid esters with fat soluble long chain fatty acids (palmitate and ascorbyl stearate).

### **5. Deficiencies**

### **5.1. Primary deficiency – scurvy. Signs and symptoms**

A frank deficiency of vitamin C causes scurvy, a disease characterized by multiple hemorrhag‐ es. Scurvy in adults is manifested by lassitude, weakness, irritability, vague muscle pain, joint pain and weight loss. Early signs objectives are as bleeding gums, tooth loss and gingivitis.

The diagnosis of scurvy, is achieved by testing plasma ascorbic acid, low concentration indicates low levels in tissues. It is generally accepted that ascorbic acid concentration in the layer of coa‐ gulated lymph (20-53 ug/108 leukocytes) is the most reliable indicator of nutritional status re‐ garding vitamin C and its concentration in tissues. In the most extreme cases scurvy shows: bleeding gums and skin, perifollicular bleeding and ocular petechiae, salivary and lachrymal glands "drier", functional neuropathy, lower limb edema, psychological disturbances, anemia and poor healing of wounds. The consumption of snuff lowers blood levels of vitamin C.

### **Clinical manifestations of Vitamin C deficiency**

Clinical manifestations of deficiency have been described at several levels: a) mesenchymal. - By the presence of petechiae, ecchymosis, curly hair, peri-follicular hemorrhages, bleeding gums, swollen, hyperkeratosis, Sjôgren syndrome, dyspnea, arthralgia, edema and poor heal‐ ing, b) systemics, characterized by fatigue, weakness and lassitude c) psychological and neuro‐ logical, by the presence of depression, hysteria, hypochondriasis and vasomotor instability.

wound healing. • Dry and separated strands in the hair. • Tendency to bruising. • Gingivi‐ tis (gum inflammation). • nosebleeds. • Possible weight gain because of slowed metabolism.

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**6. Laboratory methods for its measurement in foods and biological fluid**

In the analysis of vitamin C, for the methods commonly used consume many time and therefore overestimates the concentration, due to other oxydizable species different of vita‐ min C; the determination by liquid chromatography of high resolution (HPLC), with electro‐ chemical detection, for example, they require equipment not always available in smalls laboratories and also, is very expensive. However this method quantifies all the forms of the vitamin C present in the sample, and even it detects an epimer of ascorbic acid, the eritorbic acid or isoascorbic acid. The samples of vitamin C saturation are used to establish the defi‐ ciency of ascorbate in tissue and are useful to confirm the diagnostic of scurvy when the pa‐ tient has a normal absorption (Engelfried, 1944). It has been described 3 types of tests to determine the tissue saturation, the first 2 are easy to make but they don't cover the problem on totally, the third test is complicated and it's only useful in research work listed below:

**•** The vitamin C in the plasma is not found doing a metabolic function; it is rather in a

**•** from one tissue to another. Its lost or decrease does not indicates the intracellular sta‐ tus of this vitamin. A well-nourish adult with a free acid ascorbic diet decreases his serum levels of acid to cero in about 6 weeks; however only after many weeks of more deprivation the scurvy symptoms appears. So that for this reason the scurvy patients have low levels of ascorbic acid in the plasma. The vitamin C determination in plasma after a charge dose generally reflects the vitamin proportion, which has been stored by the tissues, however is tough to do a completely quantitative techni‐ que, because in high doses of this vitamin, the plasmatic concentration exceeds the kidney threshold which causes the lost of this vitamin in urine. For a specific meas‐ ure most be given multiple small doses, to avoid an excess in the blood levels above

**b.** Measurement of kidney excretion with and without sample dose: The most important problems in this measurement are those concerning to the collection of urine, more than the vitamin measurement. The excretion of the vitamin has been correlated with the cre‐ atinine excretion. This is because the creatinine is used as a real and simple indicator of

**c.** Tissue Measurement This is the one of the 3 techniques which gives a real representa‐ tion of desaturation of the vitamin. This measurement is difficult because of problems in the sampling of tissue. In the case of ascorbic acid is recommended two methods to measure tissue levels. In the first method is measured its concentration in the buffy coat

• Rough, dry and scaly. • Pain and swelling of the joints. • Weakened tooth enamel.

**a.** Measurement of blood levels with and without a test sample:

transit

the renal threshold.

glomerular filtration.

### **5.2. Deficiencies secondary and association with other diseases**

Severe deficiency of vitamin C leads to Scurvy. This rarely occurs but can be observed deficien‐ cies in those who consume a diet without vegetables and fruits, alcoholism, in older people with limited diets, severely ill patients with chronic stress and in infants fed cow's milk.

Symptoms of scurvy are follicular hyperkeratosis, gingival swelling and inflammation (in gums), bleeding gums, loose teeth, dry mouth and eyes, hair loss and dry skin, among other symptoms that can lead to death.

By deficiency of collagen, the wounds do not heal scars and wounds of previous rupture and may lead to secondary infections. Neurotic disorders are common, consisting of hyste‐ ria and depression, followed by decreased psychomotor activity. It is not safe indiscriminate administration of ascorbic acid, since as the body becomes saturated, decreased absorption, and giving large doses, abruptly deleted. So if you continue with diet low in vitamin, may appear "rebound scurvy". In addition to "rebound scurvy," gastric intolerance and kidney, its use decreases the cobalamin (vitamin B12), a substance synthesized by the body.

Eating a balanced and varied diet high in fruits and vegetables, the minimum dose of vita‐ min C, this completely covered. The daily requirement in an adult male is 90 mg/d and a woman of 75 mg/d (mg/day), although there are always situations where it is necessary to increase the dose of vitamin A through supplementation. Such circumstances or situations are:• Pregnancy and Lactation, • Alcoholics and smokers, • diabetics, • Allergic and asth‐ matic, • People who take daily medications or medications such as oral contraceptives, cor‐ tisone, antibiotics, etc.

#### **Anemia by Vitamin C Deficiency**

Anemia of vitamin C Deficiency is a rare type of anemia that is caused by a severe and very pro‐ longed lack of vitamin C. In this type of anemia, the bone marrow produces small red blood cells (microcytosis). This deficiency is diagnosed by measuring the values of vitamin C in white blood cells. One tablet of vitamin C per day corrects the deficiency and anemia cure.

#### **Associated symptoms**

Associated deficiency or lack of vitamin C (ascorbic acid) can produce or be reflected by: Swollen and bleeding gums, dry, rough skin, spontaneous bruising, Impaired wound heal‐ ing, bleeding nose, joint pain and swelling, anemia and dental enamel weakened. Very small amounts of vitamin C may be associated with signs and symptoms of deficiency, including: • Anemia, • Bleeding gums, • Decreased, ability to fight infection. • Decrease the rate of wound healing. • Dry and separated strands in the hair. • Tendency to bruising. • Gingivi‐ tis (gum inflammation). • nosebleeds. • Possible weight gain because of slowed metabolism. • Rough, dry and scaly. • Pain and swelling of the joints. • Weakened tooth enamel.

### **6. Laboratory methods for its measurement in foods and biological fluid**

In the analysis of vitamin C, for the methods commonly used consume many time and therefore overestimates the concentration, due to other oxydizable species different of vita‐ min C; the determination by liquid chromatography of high resolution (HPLC), with electro‐ chemical detection, for example, they require equipment not always available in smalls laboratories and also, is very expensive. However this method quantifies all the forms of the vitamin C present in the sample, and even it detects an epimer of ascorbic acid, the eritorbic acid or isoascorbic acid. The samples of vitamin C saturation are used to establish the defi‐ ciency of ascorbate in tissue and are useful to confirm the diagnostic of scurvy when the pa‐ tient has a normal absorption (Engelfried, 1944). It has been described 3 types of tests to determine the tissue saturation, the first 2 are easy to make but they don't cover the problem on totally, the third test is complicated and it's only useful in research work listed below:

**a.** Measurement of blood levels with and without a test sample:

**Clinical manifestations of Vitamin C deficiency**

462 Oxidative Stress and Chronic Degenerative Diseases - A Role for Antioxidants

symptoms that can lead to death.

tisone, antibiotics, etc.

**Associated symptoms**

**Anemia by Vitamin C Deficiency**

**5.2. Deficiencies secondary and association with other diseases**

Clinical manifestations of deficiency have been described at several levels: a) mesenchymal. - By the presence of petechiae, ecchymosis, curly hair, peri-follicular hemorrhages, bleeding gums, swollen, hyperkeratosis, Sjôgren syndrome, dyspnea, arthralgia, edema and poor heal‐ ing, b) systemics, characterized by fatigue, weakness and lassitude c) psychological and neuro‐ logical, by the presence of depression, hysteria, hypochondriasis and vasomotor instability.

Severe deficiency of vitamin C leads to Scurvy. This rarely occurs but can be observed deficien‐ cies in those who consume a diet without vegetables and fruits, alcoholism, in older people

Symptoms of scurvy are follicular hyperkeratosis, gingival swelling and inflammation (in gums), bleeding gums, loose teeth, dry mouth and eyes, hair loss and dry skin, among other

By deficiency of collagen, the wounds do not heal scars and wounds of previous rupture and may lead to secondary infections. Neurotic disorders are common, consisting of hyste‐ ria and depression, followed by decreased psychomotor activity. It is not safe indiscriminate administration of ascorbic acid, since as the body becomes saturated, decreased absorption, and giving large doses, abruptly deleted. So if you continue with diet low in vitamin, may appear "rebound scurvy". In addition to "rebound scurvy," gastric intolerance and kidney,

Eating a balanced and varied diet high in fruits and vegetables, the minimum dose of vita‐ min C, this completely covered. The daily requirement in an adult male is 90 mg/d and a woman of 75 mg/d (mg/day), although there are always situations where it is necessary to increase the dose of vitamin A through supplementation. Such circumstances or situations are:• Pregnancy and Lactation, • Alcoholics and smokers, • diabetics, • Allergic and asth‐ matic, • People who take daily medications or medications such as oral contraceptives, cor‐

Anemia of vitamin C Deficiency is a rare type of anemia that is caused by a severe and very pro‐ longed lack of vitamin C. In this type of anemia, the bone marrow produces small red blood cells (microcytosis). This deficiency is diagnosed by measuring the values of vitamin C in white

Associated deficiency or lack of vitamin C (ascorbic acid) can produce or be reflected by: Swollen and bleeding gums, dry, rough skin, spontaneous bruising, Impaired wound heal‐ ing, bleeding nose, joint pain and swelling, anemia and dental enamel weakened. Very small amounts of vitamin C may be associated with signs and symptoms of deficiency, including: • Anemia, • Bleeding gums, • Decreased, ability to fight infection. • Decrease the rate of

blood cells. One tablet of vitamin C per day corrects the deficiency and anemia cure.

with limited diets, severely ill patients with chronic stress and in infants fed cow's milk.

its use decreases the cobalamin (vitamin B12), a substance synthesized by the body.

and platelets which correlates good with the first signs of scurvy, making the most rec‐ ommended technique. The second method determines the tissue saturation grade for an intradermal test, using dichlorophenol indophenol, which depends of skin reductor substances, which made this nonspecific.

tain the greatest sensitivity and resolution in the chromatographic signal. These tests are

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The test is performed with the standard addition curve and calibration curve AA patterns between 1.0 to 25.0 mg/L phosphoric acid, 0.05 N, to determine the effect matrix in the meth‐ od of quantification. The proposal is a column Hypersil ODS C18, 5µm x 4.0 mm x 250 mm.

Ascorbic acid or Vitamin C (C6H8O6) can be determined by means of an iodometric titration. Vitamin C is a mild reducing agent that reacts rapidly with tri-iodide ion, this reaction is generated in a known excess of tri-iodide ion (I3-) by reacting iodate iodide, is allowed to react and then the excess is titrated by I3-back with a solution of thiosulphate. -The method

The recommended daily ingestion (intake) is of 60 to 100 mg to avoid the appearance of disease symptoms that are produced by deficiencies of this vitamin. The infants require a little more of 100mg/day, although there is controversy over the minimum amount of this vitamin. We must take into account that this vitamin is very labile at heat and oxy‐ gen presence. The ascorbic acid is specific in the treatment of scurvy; the required dose could be better measured by the urinary excretion after a saturation dose. Depending of the required saturation velocity is the daily dose recommended which varies between 0.2 and 2.0 g/d. In the vitamin C deficiency, the tissue saturation is obtained with 3 daily

performed with the following mobile phases:

Water-methanol-acetonitrile (74.4: 25.0: 0.6)

**6.3. Other methods reported**

is based on the following reactions:

<sup>8</sup>*<sup>I</sup>* <sup>−</sup> <sup>+</sup> *IO*<sup>3</sup> <sup>−</sup> <sup>+</sup> <sup>6</sup>*<sup>H</sup>* <sup>+</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>→</sup>3*H*2*<sup>O</sup>* <sup>+</sup> <sup>3</sup>*I*<sup>3</sup>

*<sup>I</sup>* <sup>3</sup><sup>−</sup> <sup>+</sup> 2(*S*2*O*3)<sup>−</sup><sup>2</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>→</sup><sup>3</sup> *<sup>I</sup>* <sup>−</sup> <sup>+</sup> (*S*4*O*6)<sup>−</sup><sup>2</sup> thiosulfate tetrathionate

Ascorbic acid dehydroascorbic acid

<sup>−</sup> <sup>+</sup> *<sup>H</sup>*2*<sup>O</sup>* <sup>+</sup> *<sup>I</sup>*<sup>3</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>→</sup>*C*6*H*8*O*<sup>6</sup> <sup>+</sup> <sup>2</sup>*<sup>H</sup>* <sup>+</sup> <sup>+</sup> <sup>3</sup>*<sup>I</sup>*

**7. Requirements and recommendations in Mexico**

Methanol-buffer solution: 0.03 M KH2PO4, pH = 2.7, (99:1)

2% KH2PO4, pH = 2.3

1% NaH2PO4, pH = 2.7

Iodometric titration

*C*6*H*8*O*<sup>6</sup>

Acetonitrile-Water (70:30)

### **6.1. Spectrophotometric method**

Another proposed method is highly sensitive colorimetric determination of ascorbic acid with 2,4-dinitrophenylhydrazine.

The method can be applied in virtually any laboratory is simple to perform and requires little complicated equipment compared to HPLC. Samples can be from serum to food. In foods the determination may be affected if the food contains natural dyes interfering reading the wavelength of detection. However, it is an easy to implement, since it is in‐ expensive and sensitive.

The use of UV-Visible spectroscopy (UV-Vis), for the determination of AA, is widely used in research with food, since this acid has strong electronic transitions in the UV region, facili‐ tating their identification and quantification by this technique.

Vitamin C is easily affected by such factors as moisture, light, air, heat, metal ions such as iron and copper, oxygen and the alkaline medium. After decomposition under these condi‐ tions is easily transformed into various compounds such as: oxalic acid, L-threonic acid, Lxylonic acid and L-dehydroascorbic acid, and in turn the latter are irreversibly transformed into acid-diketo 2.3 L gluconic acid, which is its main degradation product.

Within the official methods described for the analysis of vitamin C in tablet, one of the most widely used is the direct titration with 2,6-dichlorophenol indophenol by simple and rapid result. The method is valid if it is known that the composition of the sample no interfering substances and the concentration of dehydroascorbic acid is negligible, therefore, can be ap‐ plied to a freshly prepared sample, but not useful in stability studies of vitamin C.

### **6.2. Chromatographic method (HPLC)**

However, high performance liquid chromatography (HPLC) ensures detection and quantita‐ tion limits lower, which also facilitate the elimination of the effects caused by the matrix (in‐ terference to other methods of analysis); this technique used as an essential tool in detailed kinetic studies.

Quantification of Ascorbic Acid by HPLC.

Using a standard solution of 50 mg AA/L phosphoric acid, 0.05 N and taking into account the various reports, on the conditions for the quantification of AA in samples, it optimizes the mobile phase and the wavelength of the detector (scanning from 200 to 320 nm) to ob‐ tain the greatest sensitivity and resolution in the chromatographic signal. These tests are performed with the following mobile phases:

2% KH2PO4, pH = 2.3

and platelets which correlates good with the first signs of scurvy, making the most rec‐ ommended technique. The second method determines the tissue saturation grade for an intradermal test, using dichlorophenol indophenol, which depends of skin reductor

Another proposed method is highly sensitive colorimetric determination of ascorbic acid

The method can be applied in virtually any laboratory is simple to perform and requires little complicated equipment compared to HPLC. Samples can be from serum to food. In foods the determination may be affected if the food contains natural dyes interfering reading the wavelength of detection. However, it is an easy to implement, since it is in‐

The use of UV-Visible spectroscopy (UV-Vis), for the determination of AA, is widely used in research with food, since this acid has strong electronic transitions in the UV region, facili‐

Vitamin C is easily affected by such factors as moisture, light, air, heat, metal ions such as iron and copper, oxygen and the alkaline medium. After decomposition under these condi‐ tions is easily transformed into various compounds such as: oxalic acid, L-threonic acid, Lxylonic acid and L-dehydroascorbic acid, and in turn the latter are irreversibly transformed

Within the official methods described for the analysis of vitamin C in tablet, one of the most widely used is the direct titration with 2,6-dichlorophenol indophenol by simple and rapid result. The method is valid if it is known that the composition of the sample no interfering substances and the concentration of dehydroascorbic acid is negligible, therefore, can be ap‐

However, high performance liquid chromatography (HPLC) ensures detection and quantita‐ tion limits lower, which also facilitate the elimination of the effects caused by the matrix (in‐ terference to other methods of analysis); this technique used as an essential tool in detailed

Using a standard solution of 50 mg AA/L phosphoric acid, 0.05 N and taking into account the various reports, on the conditions for the quantification of AA in samples, it optimizes the mobile phase and the wavelength of the detector (scanning from 200 to 320 nm) to ob‐

substances, which made this nonspecific.

464 Oxidative Stress and Chronic Degenerative Diseases - A Role for Antioxidants

tating their identification and quantification by this technique.

**6.2. Chromatographic method (HPLC)**

Quantification of Ascorbic Acid by HPLC.

into acid-diketo 2.3 L gluconic acid, which is its main degradation product.

plied to a freshly prepared sample, but not useful in stability studies of vitamin C.

**6.1. Spectrophotometric method**

with 2,4-dinitrophenylhydrazine.

expensive and sensitive.

kinetic studies.

Acetonitrile-Water (70:30)

1% NaH2PO4, pH = 2.7

Methanol-buffer solution: 0.03 M KH2PO4, pH = 2.7, (99:1)

Water-methanol-acetonitrile (74.4: 25.0: 0.6)

The test is performed with the standard addition curve and calibration curve AA patterns between 1.0 to 25.0 mg/L phosphoric acid, 0.05 N, to determine the effect matrix in the meth‐ od of quantification. The proposal is a column Hypersil ODS C18, 5µm x 4.0 mm x 250 mm.

### **6.3. Other methods reported**

Iodometric titration

Ascorbic acid or Vitamin C (C6H8O6) can be determined by means of an iodometric titration. Vitamin C is a mild reducing agent that reacts rapidly with tri-iodide ion, this reaction is generated in a known excess of tri-iodide ion (I3-) by reacting iodate iodide, is allowed to react and then the excess is titrated by I3-back with a solution of thiosulphate. -The method is based on the following reactions:

<sup>8</sup>*<sup>I</sup>* <sup>−</sup> <sup>+</sup> *IO*<sup>3</sup> <sup>−</sup> <sup>+</sup> <sup>6</sup>*<sup>H</sup>* <sup>+</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>→</sup>3*H*2*<sup>O</sup>* <sup>+</sup> <sup>3</sup>*I*<sup>3</sup> *C*6*H*8*O*<sup>6</sup> <sup>−</sup> <sup>+</sup> *<sup>H</sup>*2*<sup>O</sup>* <sup>+</sup> *<sup>I</sup>*<sup>3</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>→</sup>*C*6*H*8*O*<sup>6</sup> <sup>+</sup> <sup>2</sup>*<sup>H</sup>* <sup>+</sup> <sup>+</sup> <sup>3</sup>*<sup>I</sup>* Ascorbic acid dehydroascorbic acid *<sup>I</sup>* <sup>3</sup><sup>−</sup> <sup>+</sup> 2(*S*2*O*3)<sup>−</sup><sup>2</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>−</sup> <sup>→</sup><sup>3</sup> *<sup>I</sup>* <sup>−</sup> <sup>+</sup> (*S*4*O*6)<sup>−</sup><sup>2</sup> thiosulfate tetrathionate

### **7. Requirements and recommendations in Mexico**

The recommended daily ingestion (intake) is of 60 to 100 mg to avoid the appearance of disease symptoms that are produced by deficiencies of this vitamin. The infants require a little more of 100mg/day, although there is controversy over the minimum amount of this vitamin. We must take into account that this vitamin is very labile at heat and oxy‐ gen presence. The ascorbic acid is specific in the treatment of scurvy; the required dose could be better measured by the urinary excretion after a saturation dose. Depending of the required saturation velocity is the daily dose recommended which varies between 0.2 and 2.0 g/d. In the vitamin C deficiency, the tissue saturation is obtained with 3 daily doses of 700 mg each one for 3 days. Harris and cols. defined the tissue saturation as a sufficient storage of ascorbic acid where occurs a excretion of 50 mg or even more in a period of 4 to 5 hours after 1 dose of 700 mg/d.

**9.2. Interaction with ROS**

substrate of the oxidative damage16.

Vitamin C quickly debug the reactive species of oxygen and nitrogen just as superoxide, hi‐ droperoxile radicals, aqueous peroxyl radicals, singlet oxygen, ozone, peroxynitrite, nitro‐ gen dioxide, nitroxide radicals and hypochlorous acid thereby protecting in fact other

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Although the AA (ascorbic acid) reacts quickly with the hydroxyl radicals (constant speed > 109 Lmol-1s-1) is clumsy to debug this radical preferentially over other substrates. This is be‐ cause hydroxyl radicals are very reactive and they will combine immediately with nearest substratum in their environment at a limited speed because of its diffusion. Vitamin C can also act as a co-antioxidant when regenerate the α-tocopherol (vitamin E) from the α-toco‐ pheroxil radical produced when this is debugged from the lipid-soluble radicals just made. This is a function potentially important because in the *in vitro* experiments have shown that α-tocopherol can act as a pro-oxidant in absentia of co-oxidants just as vitamin C. However

AA can regenerate urates, glutathione and β-carotene *in vitro* from their respective oxidation products with one unpaired electron (urate radicals, glutathionil radicals and cations of β-caro‐ tene radicals). Two important properties of vitamin C make it an ideal antioxidant. The first one is its low potential reduction of ascorbate (282 mV) and its oxidation product with an elec‐ tron, the ascorbile radical (2174 mV), which is derivates from its functional group en-diol in the molecule. This low potential of reduction of the ascorbate and the ascorbile radical makes them potentially appropriate for oxidation-reduction reactions and that why the vitamin acts as a soluble antioxidant terminal molecule. The second property which makes the vitamin an ideal antioxidant is the stability and the low reactivity of the just made ascorbile radical when the as‐

The ascorbyl radical disproportionate rapidly to form ascorbate and dehydroascorbic acid (equations 1 and 2), or it is retro-reduced to ascorbate by an enzyme semi-dehydroascorbate reductase dependent of NADH. The oxidation product of two AA electrons, dehydroascor‐ bic acid, can be reduced by itself to ascorbate because of the glutathione, by enzymes de‐ pendents of glutathione: glutaredoxin (dehydroascorbate oxidoreductase [glutathione dehydrogenase (ascorbate)]) or by an enzyme dependent of selenium (seleno-enzyme): the tioredoxine reductase. Alternatively, the dehydroascorbic acid gets quickly and irreversibly

Where in equation 1 shows the reversible oxidation of 2 ascorbate electrons (AH2) to the as‐ corbile radical (A•2) and dehydroascorbic acid (A) respectively; the equation 2 shows the

« «• **AH A A 2 2** (1)

• • +®+ **A A AH A 22 2** (2)

**A ADCG** ® ® oxalate, threonate, etc. (3)

the relevance *in vivo* of the interaction of both vitamins it's not that clear yet.

corbate debug the reactive species of oxygen and nitrogen. (equation 1).

hydrolyzed to 2,3-dicetogulonic acid (ADCG) (equation 3):

The decreased levels in smokers are basically explained because they consume fewer sour‐ ces of the vitamin. In this kind of population will be required a 50% more of the recom‐ mended dose of the vitamin. Because of the daily recommendation (RDA) it is defined as the daily ingestion average of food which is enough to cover the nutriments required for healthy people in a group of the population, it's necessary to continually assess these recom‐ mendations for vitamin C. The totalities of the reviewed information suggest that a con‐ sumption of 90-100 mg of this vitamin is enough for the optimum reduction of chronic disease risk in non-smoking men and women. Although some reports are suggested amounts up to 120 mg/day.

### **8. Toxicity and hypersensitivity**

High doses of the vitamin (5-15 g/day), may cause osmotic diarrhea because it is ingested more vitamin of which can be absorbed. Also ascorbic acid can provoke intestinal cramps and acidification of the urine, leading to the formation of oxalate stones in the kidney of uri‐ nary tract. An exaggerated complementation during pregnancy may high the fetal require‐ ment and result in the presence of scurvy in the newborn. It is also credited with the destruction of vitamin B12 of food during the ingestion.

Since the oxalic acid is a metabolite of the catabolism of the ascorbic acid is likely to be the formation of oxalate crystals in kidney in patient's susceptible to nucleation and therefore the formation of crystals or "kidney stones" when it is consumed excess of the vitamin. This relation however does not extend to subjects which are not susceptible to the formation of these kidney stones.

### **9. Biochemical functions**

#### **9.1. Paper as an antioxidant**

Vitamin C is a soluble antioxidant important in biological fluids. An antioxidant is defined as "any substance which, when present in lower concentrations compared with the oxidable substrates (for example, proteins, lipids and carbohydrates and even nucleic acids) avoids or prevent significantly the oxidation of this substratum". The definition also given by the Food Nutrition Board is "A dietary antioxidant is a substance present in food which decreases sig‐ nificantly the adverse effects of the reactive species of oxygen (ROS), reactive species of ni‐ trogen (RNS), or both for the normal physiology function in humans.

### **9.2. Interaction with ROS**

doses of 700 mg each one for 3 days. Harris and cols. defined the tissue saturation as a sufficient storage of ascorbic acid where occurs a excretion of 50 mg or even more in a

The decreased levels in smokers are basically explained because they consume fewer sour‐ ces of the vitamin. In this kind of population will be required a 50% more of the recom‐ mended dose of the vitamin. Because of the daily recommendation (RDA) it is defined as the daily ingestion average of food which is enough to cover the nutriments required for healthy people in a group of the population, it's necessary to continually assess these recom‐ mendations for vitamin C. The totalities of the reviewed information suggest that a con‐ sumption of 90-100 mg of this vitamin is enough for the optimum reduction of chronic disease risk in non-smoking men and women. Although some reports are suggested

High doses of the vitamin (5-15 g/day), may cause osmotic diarrhea because it is ingested more vitamin of which can be absorbed. Also ascorbic acid can provoke intestinal cramps and acidification of the urine, leading to the formation of oxalate stones in the kidney of uri‐ nary tract. An exaggerated complementation during pregnancy may high the fetal require‐ ment and result in the presence of scurvy in the newborn. It is also credited with the

Since the oxalic acid is a metabolite of the catabolism of the ascorbic acid is likely to be the formation of oxalate crystals in kidney in patient's susceptible to nucleation and therefore the formation of crystals or "kidney stones" when it is consumed excess of the vitamin. This relation however does not extend to subjects which are not susceptible to the formation of

Vitamin C is a soluble antioxidant important in biological fluids. An antioxidant is defined as "any substance which, when present in lower concentrations compared with the oxidable substrates (for example, proteins, lipids and carbohydrates and even nucleic acids) avoids or prevent significantly the oxidation of this substratum". The definition also given by the Food Nutrition Board is "A dietary antioxidant is a substance present in food which decreases sig‐ nificantly the adverse effects of the reactive species of oxygen (ROS), reactive species of ni‐

trogen (RNS), or both for the normal physiology function in humans.

period of 4 to 5 hours after 1 dose of 700 mg/d.

466 Oxidative Stress and Chronic Degenerative Diseases - A Role for Antioxidants

amounts up to 120 mg/day.

these kidney stones.

**9. Biochemical functions**

**9.1. Paper as an antioxidant**

**8. Toxicity and hypersensitivity**

destruction of vitamin B12 of food during the ingestion.

Vitamin C quickly debug the reactive species of oxygen and nitrogen just as superoxide, hi‐ droperoxile radicals, aqueous peroxyl radicals, singlet oxygen, ozone, peroxynitrite, nitro‐ gen dioxide, nitroxide radicals and hypochlorous acid thereby protecting in fact other substrate of the oxidative damage16.

Although the AA (ascorbic acid) reacts quickly with the hydroxyl radicals (constant speed > 109 Lmol-1s-1) is clumsy to debug this radical preferentially over other substrates. This is be‐ cause hydroxyl radicals are very reactive and they will combine immediately with nearest substratum in their environment at a limited speed because of its diffusion. Vitamin C can also act as a co-antioxidant when regenerate the α-tocopherol (vitamin E) from the α-toco‐ pheroxil radical produced when this is debugged from the lipid-soluble radicals just made. This is a function potentially important because in the *in vitro* experiments have shown that α-tocopherol can act as a pro-oxidant in absentia of co-oxidants just as vitamin C. However the relevance *in vivo* of the interaction of both vitamins it's not that clear yet.

AA can regenerate urates, glutathione and β-carotene *in vitro* from their respective oxidation products with one unpaired electron (urate radicals, glutathionil radicals and cations of β-caro‐ tene radicals). Two important properties of vitamin C make it an ideal antioxidant. The first one is its low potential reduction of ascorbate (282 mV) and its oxidation product with an elec‐ tron, the ascorbile radical (2174 mV), which is derivates from its functional group en-diol in the molecule. This low potential of reduction of the ascorbate and the ascorbile radical makes them potentially appropriate for oxidation-reduction reactions and that why the vitamin acts as a soluble antioxidant terminal molecule. The second property which makes the vitamin an ideal antioxidant is the stability and the low reactivity of the just made ascorbile radical when the as‐ corbate debug the reactive species of oxygen and nitrogen. (equation 1).

The ascorbyl radical disproportionate rapidly to form ascorbate and dehydroascorbic acid (equations 1 and 2), or it is retro-reduced to ascorbate by an enzyme semi-dehydroascorbate reductase dependent of NADH. The oxidation product of two AA electrons, dehydroascor‐ bic acid, can be reduced by itself to ascorbate because of the glutathione, by enzymes de‐ pendents of glutathione: glutaredoxin (dehydroascorbate oxidoreductase [glutathione dehydrogenase (ascorbate)]) or by an enzyme dependent of selenium (seleno-enzyme): the tioredoxine reductase. Alternatively, the dehydroascorbic acid gets quickly and irreversibly hydrolyzed to 2,3-dicetogulonic acid (ADCG) (equation 3):

$$\mathbf{^\star A}\mathbf{H}\_2 \leftrightarrow \mathbf{A} \bullet \mathbf{^\star}\_2 \leftrightarrow \mathbf{A} \tag{1}$$

$$\mathbf{A}\bullet\_{2} + \mathbf{A}\bullet\_{2} \to \mathbf{A}\mathbf{H}\_{2} + \mathbf{A} \tag{2}$$

$$\mathbf{A} \rightarrow \mathbf{A} \mathbf{DCG} \rightarrow \text{oxalate}, \text{ threeone}, \text{etc.} \tag{3}$$

Where in equation 1 shows the reversible oxidation of 2 ascorbate electrons (AH2) to the as‐ corbile radical (A•2) and dehydroascorbic acid (A) respectively; the equation 2 shows the dismutation of the ascorbile radical to transform ascorbate and dehydroascorbic acid; and equation 3 show the hydrolysis of the dehydroascorbic acid to ADCG, which decomposes to oxalate, treonate y many other products. Vitamin C has been recognized and accepted by the FDA as one of 4 dietary antioxidants, the other 3 are vitamin E, β-carotene precursor of vitamin A and selenium as an essential component of antioxidant enzymes glutathione per‐ oxidase (GPx) and thioredoxin reductase.

Although there is substantial scientific evidence of the role of antioxidant vitamin C and its effects on human health are needed more research that guarantee the role of vitamin both *in vivo* and *in vitro*, particularly because the AA is a redox active compound, which may act not only as an antioxidant but also as pro-oxidant in the presence of ion redox active transition metal.

The reduction of metallic ions like iron and copper for the vitamin C *in vitro* (equation 4) results in the formation of hydroxyl radicals highly reactive way to the reaction of this ions with hydrogen peroxide, a process known as the Fenton chemistry (equation 5), The lipid hydroperoxides can also "break" because of reduced metallic ions, forming lipid alkoxy radi‐ cals (equation 6) which can begin and spread chain reactions of the lipidic peroxidation. However the shown mechanism in the equation 5 requires the availability of free ions, redox active metallic ions and a low index vitamin C/metallic ion, conditions unlikely under nor‐ mal conditions *in vivo*. Although has been shown that in biological fluids like plasma, the vitamin C acts like an antioxidant towards the lipids even in presence of free active ions.

$$\mathbf{A}\mathbf{H}\_2 + \mathbf{M}(n+1) \to \mathbf{A}\bullet\_2 + \mathbf{M}n + \mathbf{H} + \tag{4}$$

**9.3. Molecular mechanisms intracellular**

quired for the synthesis of catecholamine.

**10. Ascorbic acid in cancer**

The molecular mechanisms of the anti-scurvy effect of vitamin C are very broad and so low studied. Also Vitamin C is a cofactor of many involved enzymes in the collagen biosynthe‐ sis, carnitine and neurotransmitters. The pro-collagen dioxygenase (proline hydroxylase) and the procolagene-lisine-5-dioxigenese are two enzymes involved in the synthesis of the collagen which needs ascorbic acids for maximum activity. The posttranslational hydroxyla‐ tion of the lysine and proline residues of these enzymes are indispensable for the synthesis and formation of the stable helix which forms the collagen. So that the difference of the vita‐ min leads to the formation of weak structures causing lost of teeth, pain in joints, disorders

Antioxidant Role of Ascorbic Acid and His Protective Effects on Chronic Diseases

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469

Two dioxigenases involved in the carnitine synthesis also require vitamin C for its activity. Carnitine is essential for the transport of long chain fatty acids to the mitochondria so one deficiency of vitamin C will bring consequences just as fatigue and lethargy which are late symptoms of scurvy. Besides that the vitamin C is a cofactor for the synthesis of catechola‐ mines, in particular for the conversion of dopamine to norepinephrine catalyzed by the en‐ zyme dopamine-β-monooxygenase. Depression, hypochondriasis, and behavioral changes

Other kind of enzymes where vitamin C acts as a cofactor are the ones involved in the pepti‐ des amidations and in the tyrosine metabolism (this are also of the mono and dioxygenases kind). It is also implicated in the cholesterol metabolism to bile acids, way of 7-α-monooxy‐ genase and in the adrenal steroids metabolism. The hydroxylation of aromatic drugs and

The role of vitamin C, due to its redox potential is to reduce metal ions present in the active sites of enzymes mono and dioxygenases. Ascorbate for instance acts as a co substrate in these reactions, not as a coenzyme. The reduction of iron, involved by the presence of vita‐ min improves the intestinal absorption of dietary non heme iron. Other proposals include the maintenance of the thiol groups of proteins, keeping in its reduced form of glutathione addition, a cellular antioxidant and enzyme cofactor, and tetrahydrofolate as a cofactor re‐

The recommended daily allowance (RDA) for ascorbic acid varies from 100 to 120 mg/day for adults. Have been attributed many benefits just like its antioxidant power, antiathero‐ genic, anticarcinogenic, immunomodulatory and anti-cold. However these benefits have been subject of debate and controversies because of the danger in the use of mega doses of‐ ten used and its prooxidant effects and antioxidants. Discussed even if ascorbic acid cause cancer or promote or interfere with cancer therapy, the experts panels of dietary antioxi‐

carcinogens by cythocrome P-450, gets better also by reducing agents like vitamin C.

of the connective tissue, and poor healing, characteristic signs of scurvy.

are common in scurvy as a result of deficient dopamine hydroxylation.

*9.3.1. As an enzyme cofactor*

$$\bullet \text{ H}\_2\text{O}\_2 + \text{Mn} \rightarrow \bullet \text{OH}^\cdot + \text{OH}\_2 + \text{M} \,(n+1) \tag{5}$$

$$\bullet \text{ LOOH} + \text{Mn} \rightarrow \text{LO} \\ \bullet \text{ } + \text{OH}\_2 + \text{M} \\ \text{(}n+\text{1)} \tag{6}$$

Where in equation 4 shows the reduction of metallic active redox ions M(*n*+1)] because of the ascorbate to form de ascorbile radical and the reduced metal (M*n),* the equation 5 shows the productions of hydroxyl radicals highly reactive (•OH) of the reaction of the hydrogen peroxide (H2O2) with the reduced metallic ions and the equation 6 shows the reaction of the lipid hydroperoxides (LOOH) with the reduced metallic ions to form alkoxy radicals (LO•). Although there no convincing evidence of a prooxidant effect of vitamin C on humans, exist a substantial evidence of its antioxidant activity. Interestingly its antioxidant activity does not correlate directly in its anti-curvy effect.

Because of this, the experts considerate that if the antioxidant activity of Vitamin C is accept‐ ed *in vivo* and that if this is relevant for human health, then scurvy should not be considerate as the only criterion for the nutritional fitness or for determine the ideal quantity or required of the vitamin.

### **9.3. Molecular mechanisms intracellular**

#### *9.3.1. As an enzyme cofactor*

dismutation of the ascorbile radical to transform ascorbate and dehydroascorbic acid; and equation 3 show the hydrolysis of the dehydroascorbic acid to ADCG, which decomposes to oxalate, treonate y many other products. Vitamin C has been recognized and accepted by the FDA as one of 4 dietary antioxidants, the other 3 are vitamin E, β-carotene precursor of vitamin A and selenium as an essential component of antioxidant enzymes glutathione per‐

Although there is substantial scientific evidence of the role of antioxidant vitamin C and its effects on human health are needed more research that guarantee the role of vitamin both *in vivo* and *in vitro*, particularly because the AA is a redox active compound, which may act not only as an antioxidant but also as pro-oxidant in the presence of ion redox

The reduction of metallic ions like iron and copper for the vitamin C *in vitro* (equation 4) results in the formation of hydroxyl radicals highly reactive way to the reaction of this ions with hydrogen peroxide, a process known as the Fenton chemistry (equation 5), The lipid hydroperoxides can also "break" because of reduced metallic ions, forming lipid alkoxy radi‐ cals (equation 6) which can begin and spread chain reactions of the lipidic peroxidation. However the shown mechanism in the equation 5 requires the availability of free ions, redox active metallic ions and a low index vitamin C/metallic ion, conditions unlikely under nor‐ mal conditions *in vivo*. Although has been shown that in biological fluids like plasma, the vitamin C acts like an antioxidant towards the lipids even in presence of free active ions.

Where in equation 4 shows the reduction of metallic active redox ions M(*n*+1)] because of the ascorbate to form de ascorbile radical and the reduced metal (M*n),* the equation 5 shows the productions of hydroxyl radicals highly reactive (•OH) of the reaction of the hydrogen peroxide (H2O2) with the reduced metallic ions and the equation 6 shows the reaction of the lipid hydroperoxides (LOOH) with the reduced metallic ions to form alkoxy radicals (LO•). Although there no convincing evidence of a prooxidant effect of vitamin C on humans, exist a substantial evidence of its antioxidant activity. Interestingly its antioxidant activity does

Because of this, the experts considerate that if the antioxidant activity of Vitamin C is accept‐ ed *in vivo* and that if this is relevant for human health, then scurvy should not be considerate as the only criterion for the nutritional fitness or for determine the ideal quantity or required

+ +® + + + • ( ) **AH M 1 A M H 2 2** *n n* (4)

+® + + + • ( ) **H O M OH OH M 1 2 2 <sup>2</sup>** *n n* (5)

• +® + + + ( ) **<sup>2</sup> LOOH M LO OH M 1** *n n* (6)

oxidase (GPx) and thioredoxin reductase.

468 Oxidative Stress and Chronic Degenerative Diseases - A Role for Antioxidants

not correlate directly in its anti-curvy effect.

of the vitamin.

active transition metal.

The molecular mechanisms of the anti-scurvy effect of vitamin C are very broad and so low studied. Also Vitamin C is a cofactor of many involved enzymes in the collagen biosynthe‐ sis, carnitine and neurotransmitters. The pro-collagen dioxygenase (proline hydroxylase) and the procolagene-lisine-5-dioxigenese are two enzymes involved in the synthesis of the collagen which needs ascorbic acids for maximum activity. The posttranslational hydroxyla‐ tion of the lysine and proline residues of these enzymes are indispensable for the synthesis and formation of the stable helix which forms the collagen. So that the difference of the vita‐ min leads to the formation of weak structures causing lost of teeth, pain in joints, disorders of the connective tissue, and poor healing, characteristic signs of scurvy.

Two dioxigenases involved in the carnitine synthesis also require vitamin C for its activity. Carnitine is essential for the transport of long chain fatty acids to the mitochondria so one deficiency of vitamin C will bring consequences just as fatigue and lethargy which are late symptoms of scurvy. Besides that the vitamin C is a cofactor for the synthesis of catechola‐ mines, in particular for the conversion of dopamine to norepinephrine catalyzed by the en‐ zyme dopamine-β-monooxygenase. Depression, hypochondriasis, and behavioral changes are common in scurvy as a result of deficient dopamine hydroxylation.

Other kind of enzymes where vitamin C acts as a cofactor are the ones involved in the pepti‐ des amidations and in the tyrosine metabolism (this are also of the mono and dioxygenases kind). It is also implicated in the cholesterol metabolism to bile acids, way of 7-α-monooxy‐ genase and in the adrenal steroids metabolism. The hydroxylation of aromatic drugs and carcinogens by cythocrome P-450, gets better also by reducing agents like vitamin C.

The role of vitamin C, due to its redox potential is to reduce metal ions present in the active sites of enzymes mono and dioxygenases. Ascorbate for instance acts as a co substrate in these reactions, not as a coenzyme. The reduction of iron, involved by the presence of vita‐ min improves the intestinal absorption of dietary non heme iron. Other proposals include the maintenance of the thiol groups of proteins, keeping in its reduced form of glutathione addition, a cellular antioxidant and enzyme cofactor, and tetrahydrofolate as a cofactor re‐ quired for the synthesis of catecholamine.

### **10. Ascorbic acid in cancer**

The recommended daily allowance (RDA) for ascorbic acid varies from 100 to 120 mg/day for adults. Have been attributed many benefits just like its antioxidant power, antiathero‐ genic, anticarcinogenic, immunomodulatory and anti-cold. However these benefits have been subject of debate and controversies because of the danger in the use of mega doses of‐ ten used and its prooxidant effects and antioxidants. Discussed even if ascorbic acid cause cancer or promote or interfere with cancer therapy, the experts panels of dietary antioxi‐ dants and related compounds have been concluded that the data *in vivo* does not shows clearly a direct relation between the excess ingestion and the formation of kidney stones, the prooxidant effects and the excess absorption of iron.

that the cytotoxicity induced by vitamin C is mediated primarily by the formation of hy‐ drogen peroxide in the cell surface. The cytotoxic activity may also be mediated by the presence of cupric ions (Cu2+) in malignant melanoma cells that react with vitamin C to form free radicals in solution. Vitamin C also invests into cells, transforming them chem‐

Antioxidant Role of Ascorbic Acid and His Protective Effects on Chronic Diseases

http://dx.doi.org/10.5772/52181

471

Studies of cell surface and ultrastructure suggest that cancer cells after administration of vi‐ tamin C had cytolysis, cell membrane damage, mitochondrial changes, nuclear and nucleo‐ lar reduction and an increase in the formation of phagolysosomes. Changes in cell surface as cytolysis showed predominantly increased synthesis of collagen and disruption of the cell

The quantitative estimation of cellular organelles shown that vitamin C affects the intracel‐ lular distribution of the organelles, event that plays an important role in the citodifferentia‐ tion of the carcinogenic cell and this is the shared effect that not only vitamin C has, but also vitamin A and E. Changes in the Golgi complex and apoptotic activity and autophagic addi‐ tion to changes in cell surface and in some cases even the reversal of transformed cells to their normal cell types are needed in the possible reduction in incidence of various cancers.

Have also been associated changes in the protein synthesis, DNA and ARA with the dif‐ ferentiation and proliferation cell. But these mechanisms are not clear yet. It have been mentioned that many of this metabolic effects are mediated by the transcription and translation at genomic level. This vitamins modulate the DNA synthesis and the genetic expression in a similarly to hormones and steroids.Their effects can affect the chemical mutagenicity and the cell status. The vitamins can control the cell replication affecting the DNA, RNA and proteins in specific places which are target of electrophyles, promot‐ ing the rearrangement of codons in the altered cells and the translocation of specific genes or carcinogens. In this way, vitamins A, E and C affects directly the DNA, RNA

The vitamin C administration decrease the DNA synthesis in the core, the RNA synthesis in the nucleolus and the protein in the cytoplasm of these cells. This inhibition is accompanied

Have been proposed many mechanisms of the vitamin C activity in the prevention and

**3.** Inhibition of the hyaluronidase, keeping the substances around the tumor intact avoid‐

**1.** Would improve the immune system by increasing the lymphocytes production.

**5.** Correction of a likely ascorbate deficiency, seen in patients with cancer

by ultrastructural changes mentioned which decreases the cancer progression.

ically to a normal phenotype fine.

membrane with increased phagocytic activity and apoptotic.

and the protein synthesis in the carcinogen cells.

**2.** Stimulation in the collagen synthesis.

**Mechanism of action:**

treatment of cancer:

ing metastasis.

**4.** Inhibition of carcinogen virus

The epidemiological and clinic study does not shows conclusive benefic effects in many kinds of cancer, with the exception of stomach cancer. Recently it has tested several deriva‐ tives of ascorbic acid on cancer cells as ascorbic acid spheres. The ascorbyl stearate is a com‐ pound which inhibits the human carcinogenic cell proliferation, by interfering with the progression of the cellular cycle and inducing apoptosis by modulation of signal transduc‐ tion pathways. The cancer is a global public health problem with increasing levels of mortal‐ ity. Although exists a great variety and types of cancer, we can remark the role of vitamin C and its effects in this suffering. AA is effective protecting against the oxidative damage in tissue and also suppressing the carcinogens formation like nitrosamines.

Although vitamin C is a cytotoxic agent for tumor cells and non toxic for normal cells, in modern medicine and conventional favors more the use of powerful toxic chemotherapeutic agents. A great amount of studies have shown that the consumption of vitamin C is inverse‐ ly related with cancer with protective effects in cancer of lung, pancreas, stomach, cervix, rectum and oral cavity.

The guanine oxidation, a DNA purine, gets reduced significantly after the vitamin C supple‐ mentation, but the adenine oxidation, another purine, it's up high which suggest the antioxi‐ dant role of the vitamin. Other extensive studies both *in vivo* and *in vitro* have shown its ability to prevent, reduce or increase the adverse effects of chemotherapy. The combination of vitamin C and vitamin K already given in the chemotherapy increases the survival and the effects of various chemotherapeutic agents in a tumor-ascitic-murine model. The vitamin C has shown be safe even with the radiotherapy. The co administration of vitamin A, β-caro‐ tene, E and C can reduce the incidence and delay the progression of several cancers, such as skin, colon, stomach, esophagus, mammary gland and matrix.

Epidemiologic studies have revealed an inverse relation between the consumption of vi‐ tamin A, β-carotene, E and C and the incidence of several human cancers. There are a decrease in the risk and incidence of cancer in populations with high content of vitamins in plasma. The carcinogenesis is related with the cell differentiation, progression, metabo‐ lism and synthesis of collagen. The basic mechanism for the carcinogenesis is the cell dif‐ ferentiation because the cancer develops when a lost in this differentiation exists. And here is where the mentioned vitamins have a wide influence over de cell growth and its differentiation. Vitamin C is a strong antioxidant that acts synergistically with vitamin E in the purification of free radicals which are carcinogenic. The AA as sodium ascorbate exerts marked cytotoxic effects over many human cell lines when they are cultured. These effects are dose dependent.

Lupulescu reported that vitamin C (up to 200 ug/mL) did not cause any morphological change in mouse melanoma, neuroblastoma, and mouse and rat gliomas but is lethal for neuroblastoma cells. Cytotoxic effects are dependent cell also because they are stronger in human melanoma cells compared to mouse melanoma. It has even been suggested that the cytotoxicity induced by vitamin C is mediated primarily by the formation of hy‐ drogen peroxide in the cell surface. The cytotoxic activity may also be mediated by the presence of cupric ions (Cu2+) in malignant melanoma cells that react with vitamin C to form free radicals in solution. Vitamin C also invests into cells, transforming them chem‐ ically to a normal phenotype fine.

Studies of cell surface and ultrastructure suggest that cancer cells after administration of vi‐ tamin C had cytolysis, cell membrane damage, mitochondrial changes, nuclear and nucleo‐ lar reduction and an increase in the formation of phagolysosomes. Changes in cell surface as cytolysis showed predominantly increased synthesis of collagen and disruption of the cell membrane with increased phagocytic activity and apoptotic.

The quantitative estimation of cellular organelles shown that vitamin C affects the intracel‐ lular distribution of the organelles, event that plays an important role in the citodifferentia‐ tion of the carcinogenic cell and this is the shared effect that not only vitamin C has, but also vitamin A and E. Changes in the Golgi complex and apoptotic activity and autophagic addi‐ tion to changes in cell surface and in some cases even the reversal of transformed cells to their normal cell types are needed in the possible reduction in incidence of various cancers.

Have also been associated changes in the protein synthesis, DNA and ARA with the dif‐ ferentiation and proliferation cell. But these mechanisms are not clear yet. It have been mentioned that many of this metabolic effects are mediated by the transcription and translation at genomic level. This vitamins modulate the DNA synthesis and the genetic expression in a similarly to hormones and steroids.Their effects can affect the chemical mutagenicity and the cell status. The vitamins can control the cell replication affecting the DNA, RNA and proteins in specific places which are target of electrophyles, promot‐ ing the rearrangement of codons in the altered cells and the translocation of specific genes or carcinogens. In this way, vitamins A, E and C affects directly the DNA, RNA and the protein synthesis in the carcinogen cells.

The vitamin C administration decrease the DNA synthesis in the core, the RNA synthesis in the nucleolus and the protein in the cytoplasm of these cells. This inhibition is accompanied by ultrastructural changes mentioned which decreases the cancer progression.

#### **Mechanism of action:**

dants and related compounds have been concluded that the data *in vivo* does not shows clearly a direct relation between the excess ingestion and the formation of kidney stones, the

The epidemiological and clinic study does not shows conclusive benefic effects in many kinds of cancer, with the exception of stomach cancer. Recently it has tested several deriva‐ tives of ascorbic acid on cancer cells as ascorbic acid spheres. The ascorbyl stearate is a com‐ pound which inhibits the human carcinogenic cell proliferation, by interfering with the progression of the cellular cycle and inducing apoptosis by modulation of signal transduc‐ tion pathways. The cancer is a global public health problem with increasing levels of mortal‐ ity. Although exists a great variety and types of cancer, we can remark the role of vitamin C and its effects in this suffering. AA is effective protecting against the oxidative damage in

Although vitamin C is a cytotoxic agent for tumor cells and non toxic for normal cells, in modern medicine and conventional favors more the use of powerful toxic chemotherapeutic agents. A great amount of studies have shown that the consumption of vitamin C is inverse‐ ly related with cancer with protective effects in cancer of lung, pancreas, stomach, cervix,

The guanine oxidation, a DNA purine, gets reduced significantly after the vitamin C supple‐ mentation, but the adenine oxidation, another purine, it's up high which suggest the antioxi‐ dant role of the vitamin. Other extensive studies both *in vivo* and *in vitro* have shown its ability to prevent, reduce or increase the adverse effects of chemotherapy. The combination of vitamin C and vitamin K already given in the chemotherapy increases the survival and the effects of various chemotherapeutic agents in a tumor-ascitic-murine model. The vitamin C has shown be safe even with the radiotherapy. The co administration of vitamin A, β-caro‐ tene, E and C can reduce the incidence and delay the progression of several cancers, such as

Epidemiologic studies have revealed an inverse relation between the consumption of vi‐ tamin A, β-carotene, E and C and the incidence of several human cancers. There are a decrease in the risk and incidence of cancer in populations with high content of vitamins in plasma. The carcinogenesis is related with the cell differentiation, progression, metabo‐ lism and synthesis of collagen. The basic mechanism for the carcinogenesis is the cell dif‐ ferentiation because the cancer develops when a lost in this differentiation exists. And here is where the mentioned vitamins have a wide influence over de cell growth and its differentiation. Vitamin C is a strong antioxidant that acts synergistically with vitamin E in the purification of free radicals which are carcinogenic. The AA as sodium ascorbate exerts marked cytotoxic effects over many human cell lines when they are cultured.

Lupulescu reported that vitamin C (up to 200 ug/mL) did not cause any morphological change in mouse melanoma, neuroblastoma, and mouse and rat gliomas but is lethal for neuroblastoma cells. Cytotoxic effects are dependent cell also because they are stronger in human melanoma cells compared to mouse melanoma. It has even been suggested

tissue and also suppressing the carcinogens formation like nitrosamines.

skin, colon, stomach, esophagus, mammary gland and matrix.

prooxidant effects and the excess absorption of iron.

470 Oxidative Stress and Chronic Degenerative Diseases - A Role for Antioxidants

rectum and oral cavity.

These effects are dose dependent.

Have been proposed many mechanisms of the vitamin C activity in the prevention and treatment of cancer:

quired to maintain the normal properties of collagen. The decrease in AA concentration in plasma and Prolasa activity in diabetes can be normalized by the inhibitor of aldose reduc‐ tase. We conclude that in diabetic animals there is a deficiency of AA, which may be respon‐

Antioxidant Role of Ascorbic Acid and His Protective Effects on Chronic Diseases

http://dx.doi.org/10.5772/52181

473

The decrease in plasma ascorbic acid in diabetes plays an important role in the abnormali‐ ties of collagen and proteoglycans. These are the 2 major constituents of the extracellular matrix and its abnormalities are associated with the pathogenesis and complications of dia‐ betes. The structural similarity between glucose metabolism and AA and can interact at the

Ascorbic acid enhances the collagen and proteoglycans in fibroblast culture media. This stimulatory action is inhibited by high concentrations of glucose (25 mM). This effect how‐ ever, is not mediated AA consumption of fibroblasts. Insulin removes the inhibitory effect of glucose on the production of collagen, but the mechanism is not yet known. Thus high con‐ centrations of glucose in diabetes damage the action of ascorbic acid at the cellular level.