Suzuki Yosuke, Sasamoto Yuri, Yoshijima Chisato, Tanaka Ryota, Ono Hiroyuki, Ando Tadasuke, Shin Toshitaka, Mimata Hiromitsu, Itoh Hiroki, Ohno Keiko
Department of Medication Use Analysis and Clinical Research, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo, 204-8588, Japan.
Department of Medication Use Analysis and Clinical Research, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo, 204-8588, Japan.
J Pharm Biomed Anal. 2020 May 30;184:113202. doi: 10.1016/j.jpba.2020.113202. Epub 2020 Feb 24.
In chronic kidney disease (CKD), organic anion transporting polypeptide (OATP)1B activity is reduced by mechanisms involving 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF), a uremic toxin. Coproporphyrin-I (CP-I) is a sensitive and specific endogenous probe for phenotyping OATP1B activity and a potentially useful tool to individualize the dosage of OATP1B substrates. In this study, we developed and validated an assay for simultaneous quantification of CP-I and CMPF in human plasma using ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS). The samples were prepared by solid phase extraction, and then subjected to UHPLC-MS/MS quantification. The assay fulfilled the requirements of the US Food and Drug Administration (FDA) guideline for assay validation, with a lower limit of quantification of 0.1 for CP-I and 50 ng/mL for CMPF. Recovery rates from human plasma ranged from 97.3%-109.8% for CP-I, and 94.1%-113.3% for CMPF. Matrix effects corrected by internal standards varied between 107.2 % and 119.3 % for CP-I, and between 90.4 % and 107.4 % for CMPF. The validated assay was applied to measurement of plasma CP-I and CMPF concentrations in 10 healthy volunteers, 14 stage 3-5 CKD patients, and 14 stage 5D CKD patients. The concentrations measured in all samples were within the calibration ranges. Our novel method may be clinically useful for simultaneous measurement of plasma CP-I and CMPF concentrations in human samples, and contribute to reveal the in vivo relationship of OATB1B activity with accumulation of CMPF in CKD patients.
在慢性肾脏病（CKD）中，有机阴离子转运多肽（OATP）1B活性通过涉及3-羧基-4-甲基-5-丙基-2-呋喃丙酸（CMPF，一种尿毒症毒素）的机制而降低。粪卟啉-I（CP-I）是用于OATP1B活性表型分析的灵敏且特异的内源性探针，也是使OATP1B底物剂量个体化的潜在有用工具。在本研究中，我们开发并验证了一种使用超高效液相色谱-串联质谱法（UHPLC-MS/MS）同时定量人血浆中CP-I和CMPF的分析方法。样品通过固相萃取制备，然后进行UHPLC-MS/MS定量分析。该分析方法符合美国食品药品监督管理局（FDA）分析方法验证指南的要求，CP-I的定量下限为0.1，CMPF为50 ng/mL。人血浆中的回收率CP-I为97.3%-109.8%，CMPF为94.1%-113.3%。经内标校正的基质效应CP-I在107.2%至119.3%之间，CMPF在90.4%至107.4%之间。经验证的分析方法应用于测量10名健康志愿者、14名3-5期CKD患者和14名5D期CKD患者的血浆CP-I和CMPF浓度。所有样品中测得的浓度均在校准范围内。我们的新方法可能在临床上有助于同时测量人样品中的血浆CP-I和CMPF浓度，并有助于揭示CKD患者中OATB1B活性与CMPF蓄积的体内关系。
