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目的建立一种便捷、准确测定宫颈癌化疗患者血浆中顺铂(Cisplatin,DDP)浓度的高效液相色谱测定法(HPLC),为实时监测血浆顺铂浓度及制定化疗方案提供实验依据。方法取500μL待检者血浆,先将血浆样本经甲醇沉淀蛋白,取上清液经二乙基二硫代氨基甲酸钠(DDTC)衍生,再由三氯甲烷萃取。色谱柱为Ultimate Acclaim120C18(5μm,4.6mm×250mm),流动相为甲醇∶水=75∶25;流速为1.0mL/min;紫外检测波长为254nm;柱温35℃;进样量20μL。结果血浆顺铂浓度检测的线性范围为0.2~10mg/L(r=0.9995),最低检测限为0.057mg/L(S/N=3),相对回收率为100.8%±3.6%,日内和日间RSD均<10%。结论该方法处理过程简便、快速,重复性好,可用于宫颈癌化疗患者血浆顺铂浓度测定和顺铂药物动力学研究。
Objective To establish a convenient and accurate HPLC method for the determination of Cisplatin (DDP) in the plasma of patients with cervical cancer and to provide an experimental basis for real-time monitoring of plasma Cisplatin concentration and chemotherapy regimen. Methods 500μL of plasma were collected from the plasma samples. The plasma samples were first precipitated with methanol and the supernatant was derivatized with DDTC and extracted with chloroform. The column was Ultimate Acclaim120C18 (5μm, 4.6mm × 250mm). The mobile phase was methanol: water = 75:25, the flow rate was 1.0mL / min, the UV detection wavelength was 254nm, the column temperature was 35 ℃ and the injection volume was 20μL. Results The linear range of plasma cisplatin concentration was 0.2-10 mg / L (r = 0.9995), the lowest detection limit was 0.057 mg / L (S / N = 3), and the relative recovery was 100.8% ± 3.6% Between RSD <10%. Conclusion The method is simple, rapid and reproducible. It can be used to determine the concentration of cisplatin in plasma and the pharmacokinetics of cisplatin in patients with cervical cancer.