Neither folic acid supplementation nor pregnancy affects the distribution of folate forms in the red blood cells of women

Abstract

It is not known whether folate metabolism is altered during pregnancy to support increased DNA and RNA biosynthesis. By using a state-of-the-art LC tandem mass spectrometry technique, the aim of this study was to investigate differences in RBC folate forms between pregnant and nonpregnant women and between nonpregnant women consuming different concentrations of supplemental folic acid. Forms of folate in RBCs were used to explore potential shifts in folate metabolism during early erythropoiesis. Total RBC folate and folate forms [tetrahydrofolate; 5-methyltetrahydrofolate (5-methyl-THF); 4α-hydroxy-5-methyl-tetrahydrofolate (an oxidation product of 5-methyl-THF); 5-formyl-tetrahydrofolate; and 5,10-methenyl-tetrahydrofolate] were measured in 4 groups of women (n = 26): pregnant women (PW) (30-36 wk of gestation) consuming 1 mg/d of folic acid, and nonpregnant women consuming 0 mg/d (NPW-0), 1 mg/d (NPW-1), and 5 mg/d (NPW-5) folic acid. The mean ± SD RBC folate concentration of the NPW-0 group (890 ± 530 nmol/L) was lower than the NPW-1 (1660 ± 350 nmol/L) and NPW-5 (1980 ± 570 nmol/L) groups as assessed by microbiologic assay (n = 26, P < 0.0022). No difference was found between the NPW-1 and NPW-5 groups. We detected 5-methyl-THF [limit of detection (LOD) = 0.06 nmol/L] in all groups and tetrahydrofolate (LOD = 0.2 nmol/L) in most women regardless of methylenetetrahydrofolate reductase genotype. Most women consuming folic acid supplements had detectable concentrations of 5,10-methenyl-tetrahydrofolate (LOD = 0.31 nmol/L). However, there was no difference in the relative distribution of 5-methyl-THF (83-84%), sum of non-methyl folates (0.6-3%), or individual non-methyl folate forms in RBCs across groups. We conclude that although folic acid supplementation in nonpregnant women increases RBC total folate and the concentration of individual folate forms, it does not alter the relative distribution of folate forms. Similarly, distribution of RBC folate forms did not differ between pregnant and nonpregnant women. This trial was registered at clinicaltrials.gov as NCT01741077.

FIGURE 1

Disposition of study participants and availability of samples for analysis. MTHFR, methylenetetrahydrofolate reductase; NPW-0, non-pregnant women not consuming a folic acid supplement; NPW-1, non-pregnant women consuming a 1 mg/d folic acid supplement; NPW-5, non-pregnant women consuming a 5 mg/d folic acid supplement; PW, pregnant women (consuming a 1 mg/d folic acid supplement).

FIGURE 2

RBC total folate concentrations in PW consuming 1 mg/d of folic acid and nonpregnant women consuming 0, 1, or 5 mg/d of folic acid as determined by microbiologic assay or LC-MS/MS. Women homozygous for the C677T MTHFR genotype were not included in these analyses. Values are means ± SDs; PW (n = 6), NPW-0 (n = 5), NPW-1 (n = 5), NPW-5 (n = 7). Within an assay, means without a common letter differ, P < 0.05. MTHFR, methylenetetrahydrofolate reductase; NPW-0, nonpregnant women not consuming a folic acid supplement; NPW-1, nonpregnant women consuming a 1-mg/d folic acid supplement; NPW-5, nonpregnant women consuming a 5-mg/d folic acid supplement; PW, pregnant women.