Premium
Effect of Antioxidants on Vitamins A, E and Carotenoid Stability in Dried Blood Spots
Author(s) -
Arredondo Francisco X,
Craft Neal E
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.892.7
Subject(s) - chemistry , tocopherol , vitamin e , butylated hydroxyanisole , antioxidant , chromatography , carotenoid , vitamin , food science , sodium ascorbate , whole blood , vitamin c , retinol , ascorbic acid , biochemistry , medicine , surgery
Dry blood spot (DBS) collection offers a more practical alternative to serum collection, particularly in remote areas where cold storage is not possible. Unfortunately, many analytes, such as retinol and α‐tocopherol, are sensitive to rapid oxidation as the DBS ages on the card resulting in inaccurate measurements. One report indicated that, from day one post‐collection, retinol values can drop up to 25% and α‐tocopherol up to 50%. (Granado, et al. Quimica Clinica 23:121, 2004). Pretreatment of DBS cards with 0.5% sodium ascorbate as an antioxidant improved the recovery of α‐tocopherol during a single determination in several individuals. (Montalvo‐Velarde et al. Rev. Centro de Inv. (Mex) 7:33, 2006). Objective To investigate the use of DBS cards pretreated with non‐phenolic and phenolic antioxidants on the stability of vitamin A, α‐tocopherol, and carotenoids eluted at different times. Methods Sodium ascorbate (SA) and ascorbyl palmitate (AP) were the non‐phenolic antioxidants; butylated hydroxyanisole (BHA), 2–6, diterbutyl‐4‐methylphenol (BHT) and terbutyl hydroquinone (TBHQ) were used as phenolic antioxidants. Concentrations ranged from 0.005 to 0.5 % in the treatment solutions. What man 903 Protein‐Saver cards were saturated with the indicated antioxidants and allowed to dry overnight. Cards were spotted with 65 μL of blood collected in sodium heparin from a healthy volunteer. DBS retinol, α‐tocopherol, and carotenoids were estimated via reversed‐phase HPLC at 11, 24, 31, and 64 days post‐collection and storage at −20°C. Matching frozen plasma was processed for reference at each time the DBS were examined. Results DBS retinol had stabilized by day 11 with recoveries from 60 to 73 % compared to the liquid plasma values. BHA in the pretreatment solution at 0.05% provided the highest retinol recovery at 73.6% and remained stable through day 64. By day‐11, α‐tocopherol values were between 65–80% of those found in liquid plasma. TBHQ at 0.5% in the pretreatment solutions limited oxidation to a 12% drop from day‐11 compared with day 64. This represents a substantial improvement compared to the 50% drop reported by Granado et. al. Lute in was 72–93% of plasma content by Day‐11 and still above 76% in 0.5% TBHQ by Day‐64. β‐carotene had declined to 33–52% of plasma levels by Day‐11. The highest recovery was 28.4% in 0.5% TBHQ. Interestingly, the least % decrease from Day‐11 to Day‐64 for all analytes was observed for AP. Retinol, α‐tocopherol, and carotenoids in liquid plasma remained stable throughout the study. Conclusions The results indicate that incorporation of antioxidants into the DBS collection cards improved the recovery of retinol, α‐tocopherol and carotenoids. TBHQ was the most effective antioxidant overall, but AP minimized the % losses from Day‐11 to Day‐64. It is probable that antioxidant treatment of collection cards would be beneficial for other analytes.