高效液相色谱法测定腹腔热灌注化疗后腹腔液及血浆的顺铂浓度

目的：建立高效液相色谱法测定腹腔热灌注化疗后腹腔液及血浆顺铂浓度。方法：采用高效液相色谱法测定腹腔热灌注化疗完成后血浆和腹腔液中的顺铂浓度，即经二乙基二硫代氨基甲酸钠络合衍生化后用流动相溶解的产物Pt（DDTC）₂的浓度。色谱柱为Hypersil-ODS₂（250 mm×4.6 mm，5 μm），流动相为水-乙腈（30∶70），流速1.0 mL·min^-1，检测波长254 nm，柱温25℃。结果：顺铂的血浆检测线性范围为0.1~1.5 μg·mL^-1（r²=0.999 7），平均回收率99.6%（n=6）；顺铂的腹腔液检测线性范围为2~25 μg·mL^-1（r²=0.998 8），平均回收率100.7%（n=6），最低检测浓度均为0.1 μg·mL^-1。利用已建立的方法，本研究还对5位行腹腔热灌注化疗的患者进行了血浆及腹腔液顺铂药物浓度检测。发现腹腔热灌注化疗结束后顺铂平均吸收率98.38%。结论：本法是一种准确有效的检测方法，适用于腹腔热灌注化疗后血浆及腹腔液顺铂药物浓度监测和药动学研究，为后续设计针对化疗药物浓度监测的临床实验提供了技术基础。

Objective: To develop an HPLC methodss for determining human plasma and ascites concentrations of cisplatin(DDP)after hyperthermic intraperitoneal perfusion chemotherapy(HIPEC). Methods: DDP was extracted from the human plasma and ascites DDP and reacted with sodium diethyldithioearbamate(DDTC). The product Pt(DDTC)₂ extracted by acetonitrile ether was determined using high-performance liquid chromatography(HPLC)with the mobile phase of water and acetonitrile at the ratio of 30:70 and the flow rate of 1.0 mL·min^-1.The separation was performed on a Hypersil-ODS₂ column(250 mm×4.6 mm,5 μm)with column temperature at 25℃. The derivatives of DDP were detected at the wavelength of 254 nm. Results: The linear ranges of DDP were 0.1-1.5 μg·mL^-1 for plasma(r²=0.999 7)and 2-25 μg·mL^-1 for ascites(r²=0.998 8). The average recoveries of DDP were 99.6% for plasma(n=6)and 100.7% for ascites(n=6). The minimum concentration detectable was 0.1 μg·mL^-1. Using the established methods,plasma and peritoneal fluid concentrations of cisplatin were detected in 5 patients undergoing HIPEC. It was found that the average absorption rate of cisplatin was 98.38% after HIPEC. Conclusion: The method is accurate and effective for determining plasma and ascites DDP levels after HIPEC and it can be used in pharmacokinetic study and concentration monitoring of DDP. It provided a technical basis for the subsequent design of clinical trials for monitoring the concentration of DDP.

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