5-氟尿嘧啶-N-琥珀酰壳聚糖纳米粒在荷瘤小鼠体内组织分布分析方法

HPLC determination of 5-fluorouracil loaded N-Succinyl-Chitosan (Suc-Chi) nanoparticles in sarcoma1 80-bearing mice

目的：建立5-氟尿嘧啶-N-琥珀酰壳聚糖纳米粒（5-FU-Sur-Chi／NPs）在荷瘤小鼠血浆和组织中高效液相色谱（HPLC）分析方法。方法：生物样品经甲醇沉淀蛋白质并提取药物，采用HPLC法测定药物含量。色谱柱：Hypersil ODS C18（4．6mm×200mm，5μm）；流动相：水-甲醇（99：1）以磷酸调pH值6．2；流速：0．8mL·min^-1；紫外检测波长：266nm；柱温：室温；进样量：20μL；内标：利巴韦林。结果：该方法线性范围为0．1～10mg·L^-1，线性关系良好（r^2=0．9932～0．9979），方法回收率、日内和日间精密度的RSD均小于15％。结论：该分析方法灵敏性、准确性较高，适用于5-FU-Suc-Chi／NPs荷瘤小鼠肿瘤靶向性及药动学研究。

OBJECTIVE To establish an HPLC method for analysis of pharmacokinetics and tissue distribution of 5-Fu-Suc- Chi-NPs in Sarcomal80-bearing mice. METHODS The biological sample were diluted by methanol which also precipitated the protein in the plasma and tissues. The Hypersil ODS C18 column（4. 6 mm × 200 mm, 5μm）was used as analytical column. The column temperature was 25℃. The mobile phase consisted of methanol water-（99：1, v/v）at a flow rate of 0. 8 mL·min^-1. The UV detection wavelength was 226 nm. RESULTS The calibration curves were linear in the range of 0. 1 - 10 mg·L^-1 ,and it has fine linear correlation（correlation coefficient,r^2 = 0. 993 2-0. 997 9）. The recovery rate of the method, and the intra- and inter-day precisions expressed as the relative standard deviation were less than 15 % respectively. CONCLUSION The method had high sensitivity and precision. It is suitable for determination of 5-Fu-Suc-Chi-NPs in Sarcoma180-bearing mice.