A methodology for examining the association between plasma volume and micronutrient biomarker mass and concentration in healthy eumenorrheic women

Background. Accurate estimation and interpretation of nutritional biomarker con- centrations are important in nutritional research, clinical care, and public health surveillance. Plasma volume (PV) may affect the interpretation of plasma biomarkers but is rarely measured. We aimed to examine the association between plasma volume (PV) and micronutrient biomarker concentrations and mass as part of pilot work to develop methods. Methods. Nine healthy women with regular menstrual cycles provided fasting blood samples to measure micronutrient biomarkers. Indocyanine green was injected, and five timed blood draws were taken from 2 to 5 min to measure PV. Visits were scheduled around menstrual cycle day 2. Retinol, 25-hydroxyvitamin D, riboflavin, alpha-tocopherol, zinc, copper, magnesium, manganese, cobalt, iron, and ferritin concentrations were measured in serum. Total circulating micronutrient biomarker mass was calculated from PV and concentration. Results. The mean PV was 2067± 470 mL. PV correlated positively with concentration of iron (r =0:87, P = 0.005); other correlations were weaker with p > 0.05. PV and total mass of retinol (r =0:90), 25(OH)D (r =0:75), zinc (r =0:88), copper (r =0:83), magnesium (r = 0:93), manganese (r = 0:72), and iron (r = 0:92) were strongly correlated (all p < 0.05). PV was positively correlated with circulating micronutrient mass for most biomarkers, implying that concentrations are maintained at different volumes of plasma. Larger studies are needed to further examine these relationships. Conclusion. Though there appear to be some association between micronutrient biomarker mass and plasma volume, we are unable to draw a firm conclusion about any relationship from these results because of the small sample size. We consider these findings as a preliminary analysis to establish methods for future studies.