2.6. Blood Collection and Analysis of Systemic Biomarkers
TG, TNF-α
BLOOD
Participants were instructed to fast at least eight hours prior to their scheduled blood draw for the biochemical assessments. Blood samples (10 mL) were collected from each participant’s forearm vein using a 23-gauge needle and vacutainer tubes containing EDTA pre- and post-interventions. The collected samples were promptly centrifuged to separate plasma and stored in multiple aliquots at −80 °C for the analysis of relative biomarkers. Triglyceride (TG) levels were determined using a triglyceride colorimetric assay kit (Cayman Chemical Company, Ann Arbor, MI, USA). IL-6 and TNF-α were assessed using commercially available ELISA kits (Raybiotek, Atlanta, GA, USA). Malondialdehyde (MDA) levels were assessed using a colorimetric assay kit (Northwest Life Science Specialties, Vancouver, WA, USA). 8-hydroxy-2′-deoxyguanosine (8-OHdG) was analyzed using a DNA/RNA oxidative damage ELISA kit (Cayman Chemical Company, Ann Arbor, MI, USA). All samples were analyzed in duplicate.