Delivery prediction by quantitative analysis of four steroid metabolites with liquid chromatography tandem mass spectrometry in asymptomatic pregnant women

Abstract

Background

Prediction of delivery is important for assessing due dates, providing adequate prenatal care, and suggesting appropriate interventions in preterm and post-term pregnancies. Recent metabolomic findings suggested that the temporal abundance information of metabolome can be used to predict delivery timing with high accuracy in a cohort of healthy women. However, a targeted and quantitative assay is required to further validate the clinical performance and utility of this group of metabolomic candidates in delivery prediction with a larger and independent cohort.

Method

LC-MS/MS quantitative assays were applied to determine the plasma concentrations of four steroid metabolites, including oestriol-16-glucuronide (E3-16-Gluc), 17-alpha-hydroxyprogesterone (17-OHP), tetrahydrodeoxycorticosterone (THDOC), and androstane-3,17-diol (A-3,17-Diol) in asymptomatic women of singleton pregnancies (≥30^th gestational weeks). Subsequent statistical analysis was conducted to assess the performance of the above candidates in delivery prediction.

Result

Using LC-MS/MS, four steroids were separated and quantified in 5.5 min. The coefficients of variation (CVs) of the four analytes at the lower limit of quantification ranged from 7.9% to 14.6%, with the R² values greater than 0.990 in the calibration curves. Of the 585 recruited pregnant women who ended up with spontaneous delivery, 17.1% and 82.9% of the subjects delivered within and after 7 days since plasma collection, respectively. In the receiver operator curve analysis, the gestational age-adjusted area under the curve of the combined measurements of E3-16-Gluc and 17-OHP was 0.69 (95% CI: 0.60–0.76), with the sensitivity of 87.0% (95% CI: 78.8%–92.9%) and specificity of 60.2% (95% CI: 55.7%–64.6%). Moreover, the positive and the negative predictive values were 28.3%–34.0% and 93.1%–97.4% respectively for this combined panel.

Conclusion

We performed analytical and clinical validation of a quantitation LC-MS/MS panel for the four steroids in the plasma of pregnant women. The steroid metabolites panel of E3-16-Gluc and 17-OHP was potentially useful for predicting delivery within one week in asymptomatic women of singleton pregnancies.

Key messages

• A quantitative LC-MS/MS assay for determining the plasma levels of 17-OHP, THDOC, A-3,17-Diol and E3-16-Gluc was developed and validated, in order to evaluate their predictive performance in asymptomatic delivery of singleton pregnancy. The levels of E3-16-Gluc and 17-OHP were found to be significantly elevated at the time of sampling in women that delivered within one week and their combinational testing may be potentially useful in delivery prediction.

Keywords:

• Delivery prediction
• metabolism
• LC-MS/MS
• steroid
• assay validation

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: Lanlan Meng, Lin Li, Yanhong Zhai, Zhen Zhao and Zheng Cao were involved in study conception and design; Lanlan Meng, Zhengwen Xu, Lin Liu, Yuanyuan Zheng and Xiaowei Liu were involved in patient recruitment; Lanlan Meng, Shengmin Liu, Youran Li, Ying Liu, Qixin He and Yuting Cong were involved in the LC-MS/MS assay development and validation; Lanlan Meng, Shaofei Su and Yifan Lu contributed to the statistical analysis and figure preparation; Lanlan Meng, Zhen Zhao and Zheng Cao drafted the article and critically reviewed and approved the final article.

Disclosure statement

The authors declare that they have no conflict of interest.

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article and its Supplemental materials.

Additional information

Funding

This work was supported by the Capital Medical University (No. PYZ20056) and the Beijing Municipal Administration of Hospitals Incubating Program (No. PX2020060). The funding body did not take part in the design of the study, the collection, analysis and interpretation of the data, or manuscript writing.