Holleran Julianne L, Parise Robert A, Guo Jianxia, Kiesel Brian F, Taylor Sarah E, Ivy S Percy, Chu Edward, Beumer Jan H
Cancer Therapeutics Program, UPMC Hillman Cancer Center, Pittsburgh, PA, United States.
Cancer Therapeutics Program, UPMC Hillman Cancer Center, Pittsburgh, PA, United States; Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, United States.
J Pharm Biomed Anal. 2020 Sep 10;189:113464. doi: 10.1016/j.jpba.2020.113464. Epub 2020 Jul 7.
We developed a high-performance liquid chromatography mass spectrometry method for quantitating iohexol in 50 μL human plasma. After acetonitrile protein precipitation, chromatographic separation was achieved with a Shodex Asahipak NH2P-50 2D (5 μm, 2 × 150 mm) column and a gradient of 0.1 % formic acid in acetonitrile and 0.1 % formic acid in water over a 10 min run time. Mass spectrometric detection was performed on a Micromass Quatromicro triple-stage bench-top mass spectrometer with electrospray, positive-mode ionization. The assay was linear from 1 to 500 μg/mL for iohexol, proved to be accurate (101.3-102.1 %) and precise (<3.4 %CV), and fulfilled Food and Drug Administration (FDA) criteria for bioanalytical method validation. Recovery from plasma was 53.1-64.2 % and matrix effect was trivial (-3.4 to -1.3 %). Plasma freeze thaw stability (97.4-99.4 %), stability for 5 months at -80 °C (95.5-103.3 %), and stability for 4 h at room temperature (100.6-103.3 %) were all acceptable. This validated assay using a deuterated internal standard will be an important tool in measuring iohexol clearance and determining glomerular filtration rate (GFR) in patients.
我们开发了一种高效液相色谱质谱法，用于定量测定50μL人血浆中的碘海醇。经乙腈蛋白沉淀后，使用Shodex Asahipak NH2P - 50 2D（5μm，2×150mm）色谱柱，在10分钟的运行时间内，以乙腈中的0.1%甲酸和水中的0.1%甲酸进行梯度洗脱，实现色谱分离。质谱检测在配备电喷雾正模式电离的Micromass Quatromicro台式三级质谱仪上进行。该测定法对碘海醇的线性范围为1至500μg/mL，经证明准确（101.3 - 102.1%）且精密（<3.4%CV），并符合美国食品药品监督管理局（FDA）生物分析方法验证标准。从血浆中的回收率为53.1 - 64.2%，基质效应微不足道（-3.4至-1.3%）。血浆冻融稳定性（97.4 - 99.4%）、在-80°C下5个月的稳定性（95.5 - 103.3%）以及在室温下4小时的稳定性（100.6 - 103.3%）均可接受。这种使用氘代内标的经过验证的测定法将成为测量患者碘海醇清除率和确定肾小球滤过率（GFR）的重要工具。
