HPLC法测定荷瘤小鼠血浆中华卟啉钠含量及药动学研究

Determination of sinoporphyrin sodium in tumor-bearing mouse plasma by HPLC method

目的建立HPLC法测定荷瘤小鼠体内华卟啉钠的含量,并分析其在荷瘤小鼠体内的药动学情况。方法采用Waters XBridge C_(18)(3.0 mm×100 mm,3.5μm)色谱柱,流动相为A:乙腈-甲醇(20∶80),B:水(1%乙酸、0.1%三乙胺),梯度洗脱,流速0.7 ml/min,检测波长380 nm。荷瘤小鼠尾静脉给药后在规定时间点采血,分离得到血浆,乙腈沉淀蛋白后按照上述方法进行样品检测,采用DAS 2.0软件进行统计矩计算和分析。结果华卟啉钠浓度在70.8~14 160 ng/ml范围内线性关系良好(r=0.999 8)。华卟啉钠主要药动学参数:c_(max)=(24 127.59±1 415.23)ng/ml,t_(max)=0.083 h,t_(1/2)=(9.59±1.25)h,MRT_(0-∞)=(11.77±1.73)h,AUC_(0-∞)=(34 775.83±6 185.43)h·ng/ml。结论该方法充分考虑光敏剂华卟啉钠的样品稳定性对检测准确度的影响,具有灵敏、快速、准确、样品处理简单等特点,适用于华卟啉钠临床前荷瘤小鼠血浆样本的分析和研究。

Objective To establish a HPLC method for the assay of sinoporphyrin sodium （DVDMS） in tumor-bearing mouse plasma and to study its pharmacokinetics.Methods The column was Waters XBridge C18 （3.0 mm×100 mm, 3.5 μm）. Gradient elution was applied with mobile phase A as the mixture of acetonitrile-methanol （20：80） and B as the aqueous solution of 1% acetic acid and 0.1% triethylamine at flow rate 0.7 ml/min. The detection wavelength was 380 nm. DVDMS was administrated to tumor-bearing mice by tail vein injection. The blood samples were collected at designated time and centrifuged for plasma. DVDMS in plasma samples were extracted by protein precipitation and analyzed by the HPLC method mentioned above. Pharmacokinetic parameters were calculated by DAS 2.0 with statistical moment analysis.Results DVDMS showed good linearity within the ranges of 70.8-14 160 ng/ml （r=0.9998）. The main pharmacokinetic parameters were calculated as follows：cmax=（24 127.59±1 415.23） ng/ml, tmax=0.083 h, t1/2=（9.59±1.25） h, MRT0-∞=（11.77±1.73） h, AUC0-∞=（34 775.83±6 185.43） h·ng/ml. Conclusion This HPLC method is sensitive, rapid and accurate, which can be used for analysis and research of DVDMS in plasma samples of tumor-bearing mice.