使用液相色谱串联质谱法对植绒拭子宫颈阴道分泌物中的依非韦伦进行定量分析的方法的验证及临床应用

Validation and clinical application of a method to quantify efavirenz in cervicovaginal secretions from flocked swabs using liquid chromatography tandem mass spectrometry.

作者信息

Olagunju Adeniyi, Nwogu Jacinta, Eniayewu Oluwasegun, Atoyebi Shakir, Amara Alieu, Kpamor John, Bolaji Oluseye, Adejuyigbe Ebunoluwa, Owen Andrew, Khoo Saye

机构信息

Department of Pharmacology and Therapeutics, University of Liverpool, Liverpool, UK.

Department of Pharmaceutical Chemistry, Obafemi Awolowo University, Ile-Ife, Nigeria.

出版信息

Wellcome Open Res. 2022 Apr 7;6:246. doi: 10.12688/wellcomeopenres.17202.3. eCollection 2021.

DOI:10.12688/wellcomeopenres.17202.3

PMID:36034058

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9379332/

Abstract

A liquid chromatography tandem mass spectrometry method to quantify drugs in dried cervicovaginal secretions from flocked swabs was developed and validated using the antiretroviral efavirenz as an example. Cervicovaginal swabs (CVS) were prepared by submerging flocked swabs in efavirenz-spiked plasma matrix. Time to full saturation, weight uniformity, recovery and room temperature stability were evaluated. Chromatographic separation was on a reverse-phase C18 column by gradient elution using 1mM ammonium acetate in water/acetonitrile at 400 µL/min. Detection and quantification were on a TSQ Quantum Access triple quadrupole mass spectrometer operated in negative ionisation mode. The method was used to quantify efavirenz in CVS samples from human immunodeficiency virus (HIV)-positive women in the VADICT study (NCT03284645). A total of 98 samples (35 paired intensive CVS and DBS pharmacokinetic samples, 14 paired sparse CVS and DBS samples) from 19 participants were available for this analysis. Swabs were fully saturated within 15 seconds, absorbing 128 µL of plasma matrix with coefficient of variation (%CV) below 1.3%. The method was linear with a weighting factor (1/X) in the range of 25-10000 ng/mL with inter- and intra-day precision (% CV) of 7.69-14.9%, and accuracy (% bias) of 99.1-105.3%. Mean recovery of efavirenz from CVS was 83.8% (%CV, 11.2) with no significant matrix effect. Efavirenz remained stable in swabs for at least 35 days after drying and storage at room temperature. Median (range) CVS efavirenz AUC was 16370 ng*h/mL (5803-22088), C was 1618 ng/mL (610-2438) at a T of 8.0 h (8.0-12), and C was 399 ng/mL (110-981). Efavirenz CVS:plasma AUC ratio was 0.41 (0.20-0.59). Further application of this method will improve our understanding of the pharmacology of other therapeutics in the female genital tract, including in low- and middle-income countries.

摘要

以抗逆转录病毒药物依非韦伦为例，开发并验证了一种用于定量植绒拭子干燥宫颈阴道分泌物中药物的液相色谱串联质谱法。通过将植绒拭子浸入添加依非韦伦的血浆基质中来制备宫颈阴道拭子（CVS）。评估达到完全饱和的时间、重量均匀性、回收率和室温稳定性。色谱分离在反相C18柱上进行，采用梯度洗脱，流动相为含1mM醋酸铵的水/乙腈溶液，流速为400µL/min。检测和定量在TSQ Quantum Access三重四极杆质谱仪上进行，采用负离子模式运行。该方法用于定量VADICT研究（NCT03284645）中人类免疫缺陷病毒（HIV）阳性女性的CVS样本中的依非韦伦。共有来自19名参与者的98个样本（35对密集CVS和DBS药代动力学样本，14对稀疏CVS和DBS样本）可用于该分析。拭子在15秒内完全饱和，吸收128µL血浆基质，变异系数（%CV）低于1.3%。该方法在25-10000 ng/mL范围内呈线性，加权因子为（1/X），日间和日内精密度（%CV）为7.69-14.9%，准确度（%偏差）为99.1-105.3%。依非韦伦从CVS中的平均回收率为83.8%（%CV，11.2），无明显基质效应。依非韦伦在干燥并室温储存后，在拭子中至少稳定35天。在T为8.0小时（8.0-12）时，CVS中依非韦伦的AUC中位数（范围）为16370 ng*h/mL（5803-22088），C为1618 ng/mL（610-2438），C为399 ng/mL（110-981）。依非韦伦CVS：血浆AUC比值为0.41（0.20-0.59）。该方法的进一步应用将增进我们对包括低收入和中等收入国家在内的女性生殖道中其他治疗药物药理学的理解。