BackgroundDried blood spots (DBS) from fingertip prick blood can enable high throughput fatty acid profiling but may be prone to lipid peroxidation during storage. The use of butylated hydroxytoluene (BHT) on chromatography paper can prevent polyunsaturated fatty acid (PUFA) loss but examinations on the length of storage times possible are not comprehensive.MethodIn the first study, venous whole blood was saturated on paper strips pre-soaked with 0, 2.5 or 5.0 mg/mL BHT and exposed to air for up to 28 days. In a second study, the effect of sealing DBS on 5.0 mg/mL BHT-soaked chromatography strips in capped test tubes or vacuum sealed polypropylene bags with and without nitrogen purging was examined over eight weeks. The fatty acid composition of the DBS were determined by gas chromatography and the effect of sample storage on omega-3 biomarkers were examined.ResultsPUFA and omega-3 biomarkers in DBS stored without BHT were dramatically reduced by day 3. In general, BHT delayed decreases in eicosapentaenoic + docosahexaenoic acid from baseline (3.2 ± 0.2 wt%) to 28 days (2.6 ± 0.03 wt%) of storage. In the % n-3 highly unsaturated fatty acids (HUFA) in total HUFA biomarker, BHT was more effective at preventing changes, particularly with 5.0 mg/mL BHT where no differences were detected up to 28 days. Sealed storage with BHT tended to increase the stability of the PUFA in DBS and nitrogen purging did not appear to provide additional benefits. The % n-3 HUFA in total HUFA biomarker also appeared to be more stable in the sealed storage study.ConclusionsThe storage of DBS in sealed containers with BHT may prevent PUFA degradation for up to 8 weeks. The % n-3 HUFA in total HUFA biomarker appears to provide a more consistent assessment of omega-3 status throughout storage as compared with other omega-3 blood biomarkers.
An increased dietary intake of n-3 highly unsaturated fatty acids (HUFA; >or=20 carbons, >or=3 carbon-carbon double bonds), particularly eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3), is associated with the decreased risk and incidence of several morbidities afflicting the elderly, including cognitive decline, dementia, rheumatoid arthritis, and macular degeneration. In this study, the dietary intake and blood levels of fatty acids were directly determined in residents of a retirement home or assisted living phase of a continuum of care facility for Canadian seniors. Finger-tip-prick blood samples, 3-day food duplicates, and 3-day food records were collected. The fatty acid composition of food duplicates and blood was determined by gas chromatography. Fifteen participants (7 male, 8 female; 87.1 +/- 4.8 years of age) completed the protocol. The daily intake of EPA and DHA combined, determined directly, was 70 mg (95% CI, 41-119) or 0.036% of total energy (95% CI, 0.022-0.058). In finger-tip-prick blood, the percent of n-3 HUFA in total HUFA of whole blood, a biomarker of n-3 polyunsaturated fatty acid status, was 28.8 +/- 5.2%. Correlations between daily n-3 HUFA intake and n-3 HUFA in blood were not significant (r = 0.14; n = 15), but became significant after the removal of 2 participants who appeared to consume fish irregularly (r = 0.59; n = 13). The n-3 HUFA intake and corresponding n-3 HUFA blood levels of Canadian long-term care residents are lower than levels estimated to prevent several morbidities associated with aging.
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