An enantiomeric quantitative LC-MS/MS method for tolvaptan and itsmonohydroxylates in human plasma using a reversed-phase separation procedure
An enantiomeric quantitative LC-MS/MS method for tolvaptan and itsmonohydroxylates in human plasma using a reversed-phase separation procedure
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使用反相分离程序对人血浆中托伐普坦及其单羟基化物进行对映体定量 LC-MS/MS 方法
DOI：
10.1016/j.jpba.2019.113061
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发表时间：
2020
影响因子：
3.4
通讯作者：
Junichi Kawakami
中科院分区：
文献类型：
--
作者：
Shunta Akutsu;Takafumi Naito;Kohei Hoshikawa;Masao Saotome;Yuichiro Maekawa;Junichi Kawakami
具有不对称碳的外消旋托伐普坦通过CYP 3A代谢为三对具有V2受体拮抗活性的二醇形式的单羟基化对映体。本研究旨在建立人血浆中5 R-和5S-托伐普坦及其单羟基化对映体的同时定量液相色谱-串联质谱法，并将其应用于患者样本。采用多糖衍生物手性柱，以反相洗脱模式分离去蛋白血浆样品。质谱仪以正离子电喷雾电离模式运行。通过5 R-和5S-托伐普坦的重组CYP 3A 4/5消化鉴别托伐普坦单羟基化物对映异构体的色谱峰。5 R-和5S-托伐普坦的校准曲线范围为0.25-125 ng/mL，4 R5 R-和4S 5S-二醇的校准曲线范围为0.025-12.5 ng/mL，4S 5 R-、4 R5 S-、3S 5 R-和3R 5S-二醇的校准曲线范围为0.025-38.15 ng/mL，差异较大。其人血浆中的处理前回收率和基质因子分别为85.2- 112.9%和86.9- 113.1%。所有分析物的日内和日间准确度和不精密度分别为92.3- 113.8%和3.5- 14.6%。采用5倍稀释程序，心力衰竭患者中5 R-和5S-托伐普坦、4 R5 R-、4S 5S-、4S 5 R-、4 R5 S-、3S 5 R-和3R 5S-二醇的血浆浓度范围为0.634-28.4、2.37-131、0.525-15.4、0.0970-4.08、6.82-108、0.271-6.49、0.394-4.18和4.81-39.8 ng/mL。总之，本方法具有可接受的分析性能水平，可有助于心力衰竭患者血浆5 R-和5S-托伐普坦及其单羟基化对映体的表征。
Racemic tolvaptan possessing an asymmetric carbon is metabolized to three pairs of monohydroxylate enantiomers of diol form with V2receptor antagonistic activity via CYP3A. This study aimed to develop a simultaneous quantitative liquid chromatography-tandem mass spectrometry method for 5R- and 5S-tolvaptan and their monohydroxylate enantiomers in human plasma and to apply it to patient samples. Deproteinized plasma specimens were separated using a polysaccharide derivative chiral column in a reversed-phase elution mode. The mass spectrometer was run in positive ion electrospray ionization mode. The chromatographic peaks of tolvaptan monohydroxylate enantiomers were identified by the recombinant CYP3A4/5 digestion of 5R- and 5S-tolvaptan. The calibration curves ranged over the plasma concentrations of 0.25–125 ng/mL for 5R- and 5S-tolvaptan, 0.025–12.5 ng/mL for 4R5R- and 4S5S-diols, and 0.025–38.15 ng/mL for 4S5R-, 4R5S-, 3S5R-, and 3R5S-diols with a large variation. Their pre-treatment recovery rates and matrix factors in human plasma were 85.2–112.9 % and 86.9–113.1 %, respectively. The intra- and inter-day accuracy and imprecision were 92.3–113.8 % and 3.5–14.6 % for all analytes, respectively. The plasma concentration ranges of 5R- and 5S-tolvaptan, 4R5R-, 4S5S-, 4S5R-, 4R5S-, 3S5R-, and 3R5S-diols in heart failure patients with a 5-fold dilution procedure were 0.634–28.4, 2.37–131, 0.525–15.4, 0.0970–4.08, 6.82–108, 0.271–6.49, 0.394–4.18, and 4.81–39.8 ng/mL, respectively. In conclusion, the present method has an acceptable analytical performance level and can be helpful for characterization of the plasma 5R- and 5S-tolvaptan and their monohydroxylate enantiomers in heart failure patients.