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 Table of Contents  
ORIGINAL ARTICLE
Year : 2018  |  Volume : 2  |  Issue : 4  |  Page : 265-268

Prospective study on role of folic acid and vitamin B12 in early pregnancy and spontaneous abortion


1 Department of Biochemistry, Pt. B.D. Sharma University of Health Sciences, Rohtak, Haryana, India
2 Department of Biochemistry, Obstetrics and Gynecology, Pt. B.D. Sharma PGIMS, Rohtak, Haryana, India

Date of Submission15-Jul-2018
Date of Decision05-Aug-2018
Date of Acceptance12-Aug-2018
Date of Web Publication11-Dec-2018

Correspondence Address:
Dr. Simmi Kharb
H. No. 1396, Sector-1, Rohtak, Haryana
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/bbrj.bbrj_85_18

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  Abstract 


Background: Vitamin B12 maintains normal folate metabolism during pregnancy. Maternal folate status has been associated with various adverse pregnancy outcomes. The aim of this study is to predict the risk of abortion in early pregnancy by studying the relation between folate, Vitamin B12 levels, and rate of abortion. This prospective, observational study was carried out on 100 pregnant women with singleton pregnancy of 6–12 weeks of gestation. Methods: The patients were divided into three groups, namely, Group 1 (control group) comprising of 30 normal pregnant women with no history of abortion; Group 2 (control group) comprising of 35 pregnant females with history of previous abortion; and Group 3 (study group) comprising of 35 pregnant women were coming with chief complaint of bleeding per vaginum. About 5 ml blood was taken and serum was separated using centrifugation. Serum folate and Vitamin B12 levels were estimated by chemiluminescence. Data so obtained were computed as mean + standard deviation and SPSS/ANOVA was applied. Results: Serum folate and Serum B12 levels were comparable in all the three groups and the difference was statistically significant. 26.66%, 37.13%, and 31.42% patients had folate deficiency in Groups 1, 2, and 3, respectively. Out of which, 3.33%, 5.71%, and 5.71% patients aborted from folate deficient Groups 1, 2, and 3, respectively. Thus, folate deficiency was not found to be associated with abortion (P = 0.944). Patients with Vitamin B12 deficiency were 73.33%, 82.85%, and 37.13% in Groups 1, 2, and 3, respectively; out of which 3.33% patients from Group 1, 8.57% from Group 2, and 2.85% patients from Group 3 were aborted. Correlation between maternal serum B12 deficiency and abortion was found to be insignificant (P = 0.551). Conclusion: A positive correlation was seen between serum folate and B12 levels. All females planning for conception should be given folate and Vitamin B12 supplementation.

Keywords: Abortion, folic acid, pregnancy, Vitamin B12


How to cite this article:
Kharb S, Singh A, Bala J, Gahlawat P, Nanda S. Prospective study on role of folic acid and vitamin B12 in early pregnancy and spontaneous abortion. Biomed Biotechnol Res J 2018;2:265-8

How to cite this URL:
Kharb S, Singh A, Bala J, Gahlawat P, Nanda S. Prospective study on role of folic acid and vitamin B12 in early pregnancy and spontaneous abortion. Biomed Biotechnol Res J [serial online] 2018 [cited 2019 Aug 19];2:265-8. Available from: http://www.bmbtrj.org/text.asp?2018/2/4/265/247246




  Introduction Top


Pregnancy is a period of rapid growth and organ differentiation. Women are prone to develop folate deficiency due to low dietary folate intake increased or metabolic requirement for folate. Furthermore, maternal folate status has been associated with other adverse pregnancy outcomes, namely, preeclampsia, congenital malformations (orofacial clefts and neural tube defects [NTDs]), preterm delivery, spontaneous abortion, fetal growth restriction, and fetal death.[1] Folic acid supplementation has been known to prevent megaloblastic anemia during pregnancy and NTDs in the newborn infant.[2],[3],[4]

Pernicious anemia is a known cause of infertility,[5] and therefore, severe clinical deficiency of Vitamin B12 cannot coexist with pregnancy. Ball and Giles reported that 20% of women in mid-pregnancy had Vitamin B12 concentrations below the nonpregnant cutoff level.[6] While others have reported that 53% of the women had decreased Vitamin B12 as compared to nonpregnant individuals. Low Vitamin B12 has been associated with pregnancies affected by anencephaly.[7] Some studies had reported that maternal Vitamin B12 status was an independent risk factor for NTD.[8] It has been observed that low amniotic Vitamin B12 levels or elevated transcobalamin can occur during NTD-affected pregnancies.[9],[10] Reznikoff-Etiévant et al. in their study in patients with very early recurrent abortions (before 5 weeks of gestation) or early recurrent abortions (before12 weeks) observed that ten cases (9%) had Vitamin B12 levels lower than 180 pg/mL compared with controls (1%: women with one or more children and no history of abortion) and after Vitamin B12 treatment in 10 cases, 4 went on to normal full-term pregnancies.[11]

Medical Research Council study has shown that folic acid is protective against NTD. Many researchers found that folic acid supplements also reduced the risk of babies developing cleft lip or cleft palate.[4] Folate deficiency can result in spontaneous abortion, recurrent pregnancy loss, and stillbirth.[1] A Liverpool study in low-income women after histidine loading, it was noted that abnormal formiminoglutamic acid excretion was more frequent in younger gravidas and multiple pregnancies. They suggested that folate deficiency was closely associated with placental abruption and recurrent spontaneous abortion.[12] In a trial of periconceptional supplementation with folic acid, it was observed that folic acid decreased the risk of NTDs by causing abortion of the fetuses.[13]

In view of the possible association of maternal folate and Vitamin B12 levels with progress and outcome of pregnancy, the present study was designed to analyze Vitamin B12 and folate levels in early pregnancy (6–12 weeks) to predict the risk of abortion.


  Methods Top


This prospective observational study was carried out in 100 pregnant women with singleton pregnancy of 6–12 weeks of gestation admitted in labor room under emergency condition and attending the Outpatient Department of Obstetrics and Gynaecology, Pt. B.D. Sharma, PGIMS, Rohtak, for routine antenatal examination in collaboration with Department of Biochemistry. Women who were not sure of dates, known case of diabetes mellitus, hypertension, thyroid disorder, renal or liver disease, autoimmune disorder, and multiple pregnancies were excluded. Informed consent was obtained from these women, and the study was approved by ethical committee of the institutional postgraduate research board. The proposal was reviewed and approved by Research and Ethical committee of the department of Biochemistry, Obstetrics and Gynecology, Rohtak, Haryana, India. The Ethics Committee Approval number was IN. Har. Rohtak 2017, and date was 1.1. 2017-1.2-2018.

The patients were divided into three groups, namely, Group 1 (control group) comprising of 30 normal pregnant women with no history of abortion; Group 2 (control group) comprising of 35 pregnant females with history of previous abortion; and Group 3 (study group) comprising of 35 pregnant women were coming with chief complaint of bleeding per vaginum. A detailed history and thorough general physical examination was carried out in all the pregnant women. Routine antenatal investigations were done at the time of first antenatal visit. Five milliliters blood was drawn aseptically and serum was separated by centrifugation. Routine investigations and serum folate and Vitamin B12 levels were estimated in maternal blood by chemiluminescence. Reference range for serum folate was 4.5–20 ng/ml and for serum B12 was 118–435 pg/ml. All the patients were followed every 4 weekly till 20 weeks and the outcome was noted in terms of anembryonic pregnancy, missed abortion, spontaneous abortion, or continuation of pregnancy.

Statistical consideration

Data thus collected were presented as mean ± standard deviation and Student's t-test was done using SPSS (Statistical Package for the Social Sciences, version 17.0, SPSS Inc, Chicago, Illinois, USA).


  Results Top


In the present study, serum B12 levels were significantly higher in the study group as compared to controls [Table 1]. The mean values of serum Vitamin B12 in Group 1 were higher in Group 2 as compared to Group 1 and lower than Group 3 [Table 1]. The difference between the mean values of serum B12 of women in three groups was statistically significant.
Table 1: Biochemical investigations in three groups

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On ccorrelating folate levels with abortion, it was observed that in Group 1, a total of 26.66% (n = 8) women had folate deficiency, of these women 23.33% (n = 7) had normal pregnancy and 3.33% (n = 1) resulted in abortion during the first trimester. Seventy-three percent of the women had normal folate levels; out of which 70% (n = 21) had normal pregnancy, while 3.33% (n = 1) women aborted spontaneously. The difference between the women with folate levels <4.5 ng/ml and those with >4.5 ng/ml was statistically insignificant (P = 0.439). Similarly, in women with Group 2, a total of 37.13% (n = 13) had folate deficiency; out of which 31.42% (n = 11) had normal pregnancy, while 5.71% (n = 2) resulted in abortion. 62.85% (n = 22) women had normal folate levels out of which 57.14% (n = 20) had normal pregnancy and 5.71% (n = 2) women aborted. The difference between the women with folate levels <4.5 ng/ml and those with >4.5 ng/ml was found to be statistically insignificant (P = 0.571).

In Group 3, a total of 31.42% (n = 11) women had folate deficiency; out of which 25.71% (n = 9) had normal pregnancy, while 5.71% (n = 2) aborted. Women with normal folate levels were 68.56% (n = 24) out of which 54.28% (n = 19) had normal pregnancy and 14.28% (n = 5) women aborted. The difference between the women with folate levels <4.5 ng/ml and those with >4.5 ng/ml was found to be statistically insignificant (P = 0.855, [Table 2]).
Table 2: Correlation of serum folate levels with abortion

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Serum Vitamin B12 was correlated with abortion and it was noted that in Group 1, a total of 73.33% (n = 22) women had Vitamin B12 deficiency out of which 70% (n = 21) had normal pregnancy, while 3.33% (n = 1) resulted in abortion. 26.66% women had normal B12 levels out of which 23.33% (n = 7) had normal pregnancy and 3.33% (n = 1) women aborted. The difference between the women with serum B12 levels <118 pg/mL and those with >118 pg/mL was found to be statistically insignificant (P = 0.439). Furthermore, in Group 2, a total of 82.85% women were found with Vitamin B12 deficiency out of which 74.28% (n = 26) had normal pregnancy and 8.57% (n = 3) aborted. Women with normal serum B12 levels were 17.13% (n = 6); out of which 14.28% (n = 5) had normal pregnancy and 2.85% (n = 1) women aborted. The difference between the women with serum B12 levels <118 pg/mL and those with >118 pg/mL was found to be statistically insignificant (P = 0.657, [Table 3]).
Table 3: Correlation of serum Vitamin B12 levels with abortion

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In Group 3, a total 37.13% (n = 13) women had Vitamin B12 deficiency out of which 31.42% (n = 11) had normal pregnancy and 5.71% (n = 2) resulted in abortion. 62.85% (n = 22) women had normal serum B12 levels out of which 48.57% (n = 17) had normal pregnancy and 14.28% (n = 5) women aborted. The difference between the women with serum B12 levels <118 pg/mL and those with >118 pg/mL was found to be statistically insignificant (P = 0.599).


  Discussion Top


In the present study, mean serum folate levels were within normal range in all the three groups. The difference between the mean value of serum folate of women in three groups was statistically significant (P = 0.01). Reference range for serum folate was 4.5–20 ng/ml. Group 2 and Group 3 showed serum folate levels on lower limits of normal range. Lower limits of normal folate values could be due to increased demand during pregnancy, poor nutritional status, malabsorption, noncompliance of folate supplementation during periconceptional period, less interpregnancy interval, or nausea-vomiting occurring in the first trimester of pregnancy and vegetarian status as all the women were vegetarian in the present study.

In the study, 26.66% (n = 8), 37.13% (n = 13), and 31.42% (n = 11) patients had folate deficiency in Groups 1, 2, and 3, respectively. Out of which 3.33% (n = 1), 5.71% (n = 2), and 5.71% (n = 2) women aborted from Group 1, 2, and 3, respectively. Hence, folate deficiency was not found to be associated with abortion (P > 0.05) (odds ratio [OR] = 0.84; 95% confidence interval [CI] 0.13–5.22). These findings were comparable with the Hungarian randomized trial and they had also concluded that folate deficiency has no significant effects on birth weight, miscarriage, and stillbirth after periconceptional multivitamin supplementation.[3] George et al. reported that low blood folate was a significant independent risk factor for spontaneous abortion (OR = 1.47; 95% CI 1.01–2.14) after adjustment for a number of known risk factors and other possible confounding variables but high folate status was associated with a nonsignificant decrease in risk (OR = 0.74; 95% CI, 0.47–1.10).[14]

In the present study, difference between the mean values of serum Vitamin B12 of women in three groups was statistically significant (P < 0.001). Seventy-three percent in Group 1, 83% in Group 2, and 37% in Group 3 had Vitamin B12 levels below cutoff (<118 pg/ml), out of which 3.33% (n = 1) women from Group 1, 8.57% (n = 3) from Group 2, and 2.85% (n = 2) women from Group 3, respectively, were aborted. Hence, Vitamin B12 deficiency was not significantly associated with abortion (P > 0.05) (OR = 0.61; 95% CI 0.10–3.76). Reference range for serum Vitamin B12 was 118–435 pg/ml. Hibbard and Spencer conducted a study on pregnant females and found that 53% of the women had Vitamin B12 levels below the normal range for nonpregnant individuals (150 pg/mL). Furthermore, 14% of the women had levels <50 pg/mL. However, in a 3-year postnatal follow-up of these women, only four women remained deficient after 6 months. They concluded that Vitamin B12 deficiency was common during pregnancy and in most of the cases, it was not associated with latent pernicious anemia. This was in accordance with the present study.[15]

In contrast, another study in pregnant females between 20 and 30 weeks of gestation, attending high-risk pregnancy clinic, a below-normal value of serum B12 was found in 22% of patients tested.[16] They agreed with the previous work by Ball and Giles.[6] These findings were in contrast with the study reported that women who had taken lower limit of Vitamin B12 as 180 pg/ml instead of 118 pg/ml in their study. Bennett followed the obstetric histories of women with recurrent severe Vitamin B12 deficiency (<180 pg/mL) and observed that 78.5% (n = 11) had frequent fetal loss. They suggested that Vitamin B12 deficiency leading to raised homocysteine levels and hypercoagulability could be the cause for the same.[17] Reznikoff-Etiévant et al. recruited women with very early recurrent abortions (VERA – before 5 weeks of gestation), or early recurrent abortions (ERA – before 12 weeks) were as cases and women with one or more children and no history of abortion as controls. 9% (n = 10) of the cases had Vitamin B12 levels lower than 180 pg/mL compared with 1% (n = 1) in the control group. Of the 10 cases, 4 went on to normal full-term pregnancies after Vitamin B12 treatment.[11]

In Group 1, a positive but nonsignificant correlation was seen between serum folate and Vitamin B12 levels (r = 0.124, P > 0.05), while a significant positive correlation was seen between serum folate and Vitamin B12 levels in both Groups 2 and 3. A much stronger correlation was seen between the two variables in Group 2 compared to Group 3 (r = 0.816 vs. 0.468). Both Vitamin B12 and folate had mean value in the lower limit of normal range in all the three groups. This might be related to poor nutritional intake, increased maternal and fetal demands, hemodilution, increased maternal catabolism, and renal clearance from plasma.[18]


  Conclusion Top


In the present study, difference between the mean values of serum Vitamin B12 and folate levels of women in three groups was statistically significant. On correlating serum folate and Vitamin B12 levels with abortion, the relation was found to be insignificant. Thus, folate and Vitamin B12 supplementation in periconceptional period reduces the risk of NTD, but their role in abortion needs to be studied on a larger scale. A positive correlation was seen between serum folate and Vitamin B12 levels. All females planning for conception should be given folate and Vitamin B12 supplementation.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Tamura T, Picciano MF. Folate and human reproduction. Am J Clin Nutr 2006;83:993-1016.  Back to cited text no. 1
    
2.
Pardo J, Peled Y, Bar J, Hod M, Sela BA, Rafael ZB, et al. Evaluation of low serum Vitamin B (12) in the non-anaemic pregnant patient. Hum Reprod 2000;15:224-6.  Back to cited text no. 2
    
3.
Yang T, Gu Y, Wei X, Liang X, Chen J, Liu Y, et al. Periconceptional folic acid supplementation and Vitamin B12 status in a cohort of Chinese early pregnancy women with the risk of adverse pregnancy outcomes. J Clin Biochem Nutr 2017;60:136-42.  Back to cited text no. 3
    
4.
Prevention of neural tube defects: Results of the medical research council vitamin study. MRC vitamin study research group. Lancet 1991;338:131-7.  Back to cited text no. 4
    
5.
Jackson IM, Doig WB, McDonald G. Pernicious anaemia as a cause of infertility. Lancet 1967;2:1159-60.  Back to cited text no. 5
    
6.
Ball EW, Giles C. Folic acid and Vitamin B12 levels in pregnancy and their relation to megaloblastic anaemia. J Clin Pathol 1964;17:165-74.  Back to cited text no. 6
    
7.
Thompson MD, Cole DE, Ray JG. Vitamin B-12 and neural tube defects: The Canadian experience. Am J Clin Nutr 2009;89:697S-701S.  Back to cited text no. 7
    
8.
Molloy AM, Kirke PN, Brody LC, Scott JM, Mills JL. Effects of folate and Vitamin B12 deficiencies during pregnancy on fetal, infant, and child development. Food Nutr Bull 2008;29:S101-11.  Back to cited text no. 8
    
9.
Gardiki-Kouidou P, Seller MJ. Amniotic fluid folate, Vitamin B12 and transcobalamins in neural tube defects. Clin Genet 1988;33:441-8.  Back to cited text no. 9
    
10.
Afman LA, Van Der Put NM, Thomas CM, Trijbels JM, Blom HJ. Reduced Vitamin B12 binding by transcobalamin II increases the risk of neural tube defects. QJM 2001;94:159-66.  Back to cited text no. 10
    
11.
Reznikoff-Etiévant MF, Zittoun J, Vaylet C, Pernet P, Milliez J. Low Vitamin B(12) level as a risk factor for very early recurrent abortion. Eur J Obstet Gynecol Reprod Biol 2002;104:156-9.  Back to cited text no. 11
    
12.
Hook EB, Czeizel AE. Can terathanasia explain the protective effect of folic-acid supplementation on birth defects? Lancet 1997;350:513-5.  Back to cited text no. 12
    
13.
Greenberg JA, Bell SJ, Guan Y, Yu YH. Folic acid supplementation and pregnancy: More than just neural tube defect prevention. Rev Obstet Gynecol 2011;4:52-9.  Back to cited text no. 13
    
14.
George L, Mills JL, Johansson AL, Nordmark A, Olander B, Granath F, et al. Plasma folate levels and risk of spontaneous abortion. JAMA 2002;288:1867-73.  Back to cited text no. 14
    
15.
Hibbard ED, Spencer WJ. Low serum B12 levels and latent addisonian anaemia in pregnancy. J Obstet Gynaecol Br Commonw 1970;77:52-7.  Back to cited text no. 15
    
16.
Pardo J, Peled Y, Bar J, Hod M, Sela BA, Rafael ZB, et al. Evaluation of low serum Vitamin B (12) in the non-anaemic pregnant patient. Hum Reprod 2000;15:224-6.  Back to cited text no. 16
    
17.
Bennett M. Vitamin B12 deficiency, infertility and recurrent fetal loss. J Reprod Med 2001;46:209-12.  Back to cited text no. 17
    
18.
Steen MT, Boddie AM, Fisher AJ, Macmahon W, Saxe D, Sullivan KM, et al. Neural-tube defects are associated with low concentrations of cobalamin (Vitamin B12) in amniotic fluid. Prenat Diagn 1998;18:545-55.  Back to cited text no. 18
    



 
 
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