Simultaneous quantitation of four androgens and 17‐hydroxyprogesterone in polycystic ovarian syndrome patients by LC‐MS/MS

Cao, Zheng; Lu, Yifan; Cong, Yang; Liu, Ying; Li, Youran; Wang, Husheng; Zhang, Chenglong; Huang, Weijian; Liu, Jingrui; Dong, Ying; Tang, Guodong; Luo, Yiqi Ruben; Yin, Chenghong; Zhai, Yanhong

doi:10.1002/jcla.23539

Cited by 14 publications

(12 citation statements)

References 45 publications

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“…The method in the present study requires an additional derivatization step compared to the direct measurement. However, compared to these reports, the LLOQ of DHT (14.5 pg/mL) in the present study showed higher sensitivity than hydroxylamine hydrochloride derivatization [18] or direct measurement [19,23] by using LLE after PA derivatization. Also, methods using solid-phase extraction have been developed for the detection of steroids, but these methods are not efficient in time and cost -effective compared with liquid-liquid extraction [16,24,25].…”

Section: Discussioncontrasting

confidence: 89%

“…Similar to the acylation reaction of fluorinated anhydrides and the hydroxyl group of the seventeenth carbon position in the steroid reaction, derivatization using PA showed a higher sensitivity in the detection of 17-OH steroids, such as corticosteroids, in ESI-LC/MS [ 17 ]. Recently, LC-MS based quantification methods for androgens such as DHT that utilize various sample sources were developed, and 5AR inhibition studies were conducted [ 16 , 18 , 19 , 20 , 21 , 22 ]. The method in the present study requires an additional derivatization step compared to the direct measurement.…”

Section: Discussionmentioning

confidence: 99%

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Development of a Liquid Chromatography/Mass Spectrometry-Based Inhibition Assay for the Screening of Steroid 5-α Reductase in Human and Fish Cell Lines

et al. 2021

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Steroid 5-α reductase (5AR) is responsible for the reduction of steroids to 5-α reduced metabolites, such as the reduction of testosterone to 5-α dihydrotestosterone (DHT). A new adverse outcome pathway (AOP) for 5AR inhibition to reduce female reproduction in fish (AOP 289) is under development to clarify the antiestrogenic effects of 5AR inhibitors in female fish. A sensitive method for the DHT analysis using chemical derivatization and liquid chromatography–tandem mass spectrometry was developed. A cell-based 5AR inhibition assay that utilizes human cell lines, a transient overexpression system, and fish cell lines was developed. The measured IC50 values of two well-known 5AR inhibitors, finasteride and dutasteride, were comparable in the different systems. However, the IC50 of dutasteride in the fish cell lines was lower than that in the human cell lines. Finasteride showed a higher IC50 against the RTG-2 cell line. These results demonstrated that 5ARs inhibition could differ in terms of structural characteristics among species. The assay has high sensitivity and reproducibility and is suitable for the application in 5AR inhibition screening for various endocrine disruption chemicals (EDCs). Future studies will continue to evaluate the quantitative inhibition of 5AR by EDCs to compare the endocrine-disrupting pathway in different species.

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Section: Discussioncontrasting

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Development of a Liquid Chromatography/Mass Spectrometry-Based Inhibition Assay for the Screening of Steroid 5-α Reductase in Human and Fish Cell Lines

et al. 2021

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“…The LLOQs was calculated by analysing the serially diluted QC specimens spiked with IS over 5 days. The LLOQ was defined as the average concentration at which the S/N ratio >10, CV <20% and bias were within ±20% [ 19 ].…”

Section: Methodsmentioning

confidence: 99%

“…The precision was evaluated by intra-assay precision ( n = 10 batches) and inter-assay precision ( n = 25 batches from three separate days) estimation with QC materials. The accuracy was assessed by the recovery studies, in which the recovery of each androgen was calculated at high-, medium-, and low-level QCs by comparing the IS peak area ratio of extracted QC samples to the IS peak area ratio of non-extracted standard solutions at the same concentration [ 19 ]. The stability of the analytes in plasma was assessed by evaluating plasma samples kept at 4 and 21 °C (room temperature) for 6 days.…”

Section: Methodsmentioning

confidence: 99%

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

Meng

et al. 2022

Annals of Medicine

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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 R 2 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.

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