高效液相色谱法分析芦丁在灌胃给药大鼠体内的组织分布

High performance liquid chromatography analysis of tissue distribution of Rutin in rats after intragastric administration

目的建立大鼠各组织中芦丁含量测定的高效液相色谱法(HPLC)。方法色谱条件:固定相为Agilent Zorbax SB-C_(18)柱(4.6 mm×250 mm,5μm),以黄酮类化合物山奈酚为内标,采用流动相为0.03%磷酸溶液-甲醇,以0.8 ml/min的流速进行梯度洗脱,检测波长为258 nm,柱温为30℃。芦丁溶于5%羧甲基纤维素钠(CMC-Na)制成混悬剂,大鼠按生药剂量200 mg/(kg·d)进行灌胃给药,采用HPLC法测定各组织中芦丁的含量。结果各组织中芦丁的线性范围不同,线性关系良好(r>0.990)。灌胃给药5次后,大鼠组织中的芦丁检出良好,在各组织含量分布中,以肾脏中含量最高,脾中含量最低。结论该方法灵敏度高、专属性强,结果可靠,使用范围广,测定方法简单,成本较低,适用于研究芦丁在大鼠中的组织分布。

Objective To establish an HPLC method for the determination of Rutin content in rats. Methods Rutin was analyzed on Agilent Zorbax SB-CI18 column （4. 6 mm×250 ram,5 ptm） with mobile phase consisting of phosphoric acid solution （0. 03% ） and methyl alcohol. The flow rate was 0. 8 ml/min for gradient elute. The column temperature was 30 ℃. The wavelength of UV detector was set at 258 nm. Rutin was soluble in 5% carboxymethyl cel- lulose sodium （CMC-Na） to make suspension. Rats were given the dose of 200 mg/（kg-d） by intragastric administration. The tissue of liver,heart, spleen,lung,kidney, muscle and brain were quickly taken after 5 times of intragastric administration. The concentrations of Rutin in tissue were determined by HPLC-UV method. Results The calibration curves for Rutin in tissue were obtained in the different concentration range with good linearity and recovery. The deviations of the intra-day and inter-day precision and accuracy were less than 15%. The recovery was greater than 80% ,and RSD was less than 15%. The sequence in the concentration of Rutin after 5 times of administration was ：kidney 〉 mus- cle 〉 lung 〉 liver 〉 heart 〉 spleen 〉 brain. Conclusion The method is simple, specific, sensitive, accurate and inexpensive. It can be used for the study of tissue distribution of Rutin in rats.