Diverse associations observed between pregnancy complications and RBC or plasma folates determined by an in-house developed LC-MS/MS method

Abstract

Background

As folates are essential for embryonic development and growth, it is necessary to accurately determine the levels of folates in plasma and red blood cells (RBCs) for clinical intervention. The aims of this study were to develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantitation of folates in plasma and RBCs and to examine the association between plasma and RBC folate concentrations and gestational diabetes mellitus (GDM), gestational hypertension (GH) and preeclampsia (PE).

Methods

With the in-house developed LC-MS/MS, a retrospective cross-sectional study was conducted. The healthy pregnant women of first- (n = 147), second- (n = 84) and third-trimester (n = 141) or the women diagnosed with GDM (n = 84), GH (n = 58) or PE (n = 23), that were aged between 22 and 46 years old and registered at our institute, were subjected for measurement of folic acid (FA) and 5-methyltetrahydrofolate (5-MTHF), followed by appropriate statistical association analysis.

Results

The assay for simultaneous quantitation of FA and 5-MTHF in plasma and RBCs was linear, stable, with imprecision less than 15% and recoveries within ±10%. The lower limits of quantification for FA and 5-MTHF measurement in whole blood were 0.57 and 1.09 nmol/L, and in plasma were 0.5 and 1 nmol/L, respectively. In the association analysis, the patients with lower RBC folate level (<906 nmol/L) presented higher risks of PE development (OR 4.861 [95% CI 1.411–16.505]) by logistic regression and restricted cubic spline (RCS) regression in a nonlinear fashion. In addition, higher level of plasma folates in pregnancy was significantly associated with GH risk but may be protective for the development of GDM.

Conclusions

The in-house developed LC-MS/MS method for folates and metabolites in plasma or RBC showed satisfactory analytical performance for clinical application. Further, the levels of folates and metabolites were diversely associated with GDM, GH and PE development.

Keywords:

• Folate
• plasma
• RBC
• LC-MS/MS
• gestational diabetes mellitus
• gestational hypertension
• preeclampsia

Disclosure statement

The authors declare that they have no conflict of interest.

Author contributions

All authors have certified the author list and the contribution description. All authors have read and approved the submitted manuscript and any substantially modified version of the manuscript. Contribution to work: Zhengwen Xu, Yanhong Zhai, Guanghui Li and Zheng Cao were involved in study conception and design; Zhengwen Xu, Lin Zhang, Jing Wang, Lanlan Meng, Lin Liu and Hongyuan Zhu were involved in patient recruitment; Youran Li, Ying Liu, Shengmin Liu, Jingru Sun, Lijun Shao and Lin Li were involved in the LC-MS/MS assay development and validation; Zhengwen Xu and Shaofei Su contributed to the statistical analysis and figure preparation; Lanlan Meng, Yanhong Zhai, Guanghui Li and Zheng Cao drafted the article and critically reviewed and approved the final article.

Data availability statement

The derived data supporting the findings of this study are included within the article and its supplemental materials. The raw data were generated at Beijing Obstetrics and Gynecology Hospital, Capital Medical University and are available upon reasonable request from the corresponding authors.

Additional information

Funding

This work was supported by the Training Fund for Open Projects at Clinical Institutes and Departments of Capital Medical University (CCMU2022ZKYXZ006), the Beijing Municipal Administration of Hospitals Incubating Program (No. PX2022058) and High-level Construction Project of Public Health Technical Personnel in Beijing Municipal Health System (No. Lingjunrencai-02-02). The funding bodies did not take part in the design of the study, the collection, analysis and interpretation of the data, or manuscript writing.