Kasper S C, Mattiuz E L, Swanson S P, Chiu J A, Johnson J T, Garner C O
Lilly Research Laboratories, Department of Drug Disposition, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN 46285, USA.
J Chromatogr B Biomed Sci Appl. 1999 Apr 16;726(1-2):203-9. doi: 10.1016/s0378-4347(99)00017-1.
A reversed-phase high-performance liquid chromatographic-electrochemical assay was developed and validated for the quantification of olanzapine in human breast milk. The assay involved a solid-phase extraction (SPE) of olanzapine and its internal standard on a Bond Elut Certify LRC mixed-mode cartridge. After conditioning of the SPE cartridge, human milk (1 ml) was passed through the cartridge. The cartridge was washed with five separate washing steps to remove endogenous compounds, and the analytes were eluted with ethyl acetate-ammonium hydroxide (98:2, v/v) solution. The eluate was evaporated to dryness (gentle stream of nitrogen at 40 degrees C), and the residue was dissolved in mobile phase. The extract was injected onto a YMC basic column (150 mmx4.6 mm I.D., 5 microm particle size) at a flow-rate of 1 ml/min. A mixture of 75 mM phosphate buffer, pH 7.0-acetonitrile-methanol (48:26:26, v/v/v) was used as the mobile phase. Standard curves with a lower limit of quantitation of 0.25 ng/ml of olanzapine were linear (r2> or =0.9992) over a range of 0.25-100 ng/ml. Based on the analysis of quality control (QC) samples, the average inter-day accuracy (RE) was 99.0% with an average precision (CV) of 6.64% over the entire range. The stability of olanzapine in human milk was established after three freeze-thaw-heat cycles and storage at -70 degrees C for 10 months. The validated method was used to measure olanzapine concentrations in human milk during a clinical trial.
建立了一种反相高效液相色谱 - 电化学分析法，并验证其用于定量测定人母乳中奥氮平的含量。该分析方法包括在Bond Elut Certify LRC混合模式小柱上对奥氮平及其内标进行固相萃取（SPE）。在对SPE小柱进行预处理后，将人乳（1 ml）通过小柱。通过五个单独的洗涤步骤清洗小柱以去除内源性化合物，然后用乙酸乙酯 - 氢氧化铵（98:2，v/v）溶液洗脱分析物。将洗脱液蒸发至干（在40℃下用氮气缓流），残余物溶解于流动相中。提取物以1 ml/min的流速注入YMC碱性柱（150 mm×4.6 mm内径，5μm粒径）。使用75 mM磷酸盐缓冲液（pH 7.0）-乙腈-甲醇（48:26:26，v/v/v）的混合物作为流动相。奥氮平定量下限为0.25 ng/ml的标准曲线在0.25 - 100 ng/ml范围内呈线性（r2≥0.9992）。基于质量控制（QC）样品的分析，在整个范围内平均日间准确度（RE）为99.0%，平均精密度（CV）为6.64%。在经过三次冻融 - 加热循环以及在 - 70℃下储存10个月后，确定了奥氮平在人乳中的稳定性。在一项临床试验中，使用经过验证的方法测量人乳中奥氮平的浓度。
