Sensitive and simultaneous quantitation of 6β‐hydroxycortisol and cortisol in human plasma by LC‐MS/MS coupled with stable isotope dilution method

Hirano Ryohei; Yokokawa Akitomo; Furuta Takashi; Shibasaki Hiromi

摘要

Abstract > CYP3A phenotyping provides a means for personalized drug therapy. We focused our attention on the plasma 6β‐hydroxycortisol (6β‐OHF) to cortisol ratio as an index for CYP3A phenotyping. In the present study, we developed a sensitive and reliable method for the simultaneous determination of 6β‐OHF and cortisol in human plasma using high‐performance liquid chromatography/tandem mass spectrometry together with picolinylester derivatization or nonderivatization methods and 6β‐[9,11,12,12‐ 2 H <sub>4</sub> ]hydroxycortisol and [1,2,4,19‐ 13 C <sub>4</sub> ]cortisol as internal standards for in vivo CYP3A phenotyping in humans. The lower limits of quantification were 38.513?pg/mL for 6β‐OHF and 38.100?pg/mL for cortisol. The relative error and relative standard deviation of the lower limits of quantification were?5% for both methods. The intra‐day and inter‐day assay reproducibilities of the determined 6β‐OHF and cortisol concentrations were consistent with the actual amounts added as relative errors and relative standard deviations for both methods, which were 5.4% and 3.9%, respectively. Both methods were applied for the quantification of plasma 6β‐OHF and cortisol concentrations in healthy subjects taking oral contraceptives. The absolute concentrations and time course of 6β‐OHF and cortisol were found to be consistent when measured using the 2 methods. The ratio as an index for in vivo CYP3A activity decreased after 21?days of taking oral contraceptives for both methods. To the best of our knowledge, this is the first study reporting the detailed investigation of accuracy and precision in the simultaneous measurement of 6β‐OHF and cortisol in human plasma using liquid chromatography/tandem mass spe </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> <div class="translation abstracttxt"> <span class="zhankaihshouqi fivelineshidden" id="abstract"> <span>机译：</span><Abstract XMLNS =“http://www.wiley.com/namespaces/wiley”type =“main”xml：lang =“en”> <title type =“main”>抽象</ title> > cyp3a表型提供个性化药物治疗的手段。我们将注意力集中在血浆6β-羟基（6β-OHF）上以皮质醇的比例作为CYP3A表型的指标。在本研究中，我们使用高效液相色谱/串联质谱法同时测定人血浆中6β-OHF和皮质醇的敏感性和可靠的方法，以及吡啶基衍生物衍生化或非衍生化方法和6β-[9,11， 12,12- 2 </ sup> h <sub> 4 </ sub>]羟基硅藻和[1,2,4,19- 13 </ sup> c <sub> 4 </ sub> ]皮质醇作为人类体内CYP3A表型的内部标准。 6β-OHF的定量率下限为38.513〜OHF和38.100〜100〜100μl的皮质醇。两种方法的较低定量限制的相对误差和相对标准偏差为Δ≤≤2个方法。确定的6β-OHF和皮质醇浓度的日期和日间测定再现能量与作为相对误差和两种方法的相对标准偏差加入的实际量，它们分别为5.5％，分别。在服用口服避孕药的健康受试者中施用两种方法以定量血浆6β-OHF和皮质醇浓度。发现6β-OHF和皮质醇的绝对浓度和时间过程在使用2种方法测量时是一致的。作为在两种方法服用口服避孕药的21天后的体内CYP3A活性的比例降低。据我们所知，这是第一项研究报告使用液相色谱/串联质量SPE在人血浆同时测量6β-OHF和皮质醇中的准确性和精度的详细研究 </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> </div> <div class="record"> <h2 class="all_title" id="enpatent33" >著录项</h2> <ul> <li> <span class="lefttit">来源</span> <div style="width: 86%;vertical-align: text-top;display: inline-block;"> <a href='/journal-foreign-34906/'>《Journal of mass spectrometry: JMS》</a> <b style="margin: 0 2px;">|</b><span>2018年第8期</span><b style="margin: 0 2px;">|</b><span>共10页</span> </div> </li> <li> <div class="author"> <span class="lefttit">作者</span> <p id="fAuthorthree" class="threelineshidden zhankaihshouqi"> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Hirano Ryohei&option=202" target="_blank" rel="nofollow">Hirano Ryohei;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Yokokawa Akitomo&option=202" target="_blank" rel="nofollow">Yokokawa Akitomo;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Furuta Takashi&option=202" target="_blank" rel="nofollow">Furuta Takashi;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Shibasaki Hiromi&option=202" target="_blank" rel="nofollow">Shibasaki Hiromi;</a> </p> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zkzz" style="display: none;">展开▼</span> </div> </li> <li> <div style="display: flex;"> <span class="lefttit">作者单位</span> <div style="position: relative;margin-left: 3px;max-width: 639px;"> <div class="threelineshidden zhankaihshouqi" id="fOrgthree"> <p>Department of Medicinal Chemistry and Clinical Pharmacy School of PharmacyTokyo University of Pharmacy and Life SciencesTokyo Japan;</p> <p>Department of Medicinal Chemistry and Clinical Pharmacy School of PharmacyTokyo University of Pharmacy and Life SciencesTokyo Japan;</p> <p>Department of Medicinal Chemistry and Clinical Pharmacy School of PharmacyTokyo University of Pharmacy and Life SciencesTokyo Japan;</p> <p>Department of Medicinal Chemistry and Clinical Pharmacy School of PharmacyTokyo University of Pharmacy and Life SciencesTokyo Japan;</p> </div> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zhdw" style="display: none;">展开▼</span> </div> </div> </li> <li > <span class="lefttit">收录信息</span> <span style="width: 86%;vertical-align: text-top;display: inline-block;"></span> </li> <li> <span class="lefttit">原文格式</span> <span>PDF</span> </li> <li> <span class="lefttit">正文语种</span> <span>eng</span> </li> <li> <span class="lefttit">中图分类</span> <span><a href="https://www.zhangqiaokeyan.com/clc/1187.html" title="分析化学">分析化学;</a></span> </li> <li class="antistop"> <span class="lefttit">关键词</span> <p style="width: 86%;vertical-align: text-top;"> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=derivatization&option=203" rel="nofollow">derivatization;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=LC‐MS/MS&option=203" rel="nofollow">LC‐MS/MS;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=nonderivatization&option=203" rel="nofollow">nonderivatization;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=plasma 6β‐hydroxycortisol&option=203" rel="nofollow">plasma 6β‐hydroxycortisol;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=plasma cortisol&option=203" rel="nofollow">plasma cortisol;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=stable isotope‐labeled compound&option=203" rel="nofollow">stable isotope‐labeled compound;</a> </p> <div class="translation"> 机译：衍生化;LC-MS / MS;非立体化;血浆6β-羟基硅藻;血浆皮质醇;稳定同位素标记的化合物; 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