高效液相色谱法同时测定补阳还五汤灌胃大鼠血清中阿魏酸和芍药苷的含量

Determination of erulic acid and paeoniflorin in serum of rats after intragastric administration of Buyang Huanwu decoction by HPLC

目的建立高效液相色谱法(HPLC)同时测定大鼠灌胃补阳还五汤后血清中阿魏酸和芍药苷的含量测定方法。方法12只大鼠采用随机数字表法分为空白对照组和补阳还五汤组,每组各6只,制备含药血清,采用HPLC分析,Phenomsil C_(18)柱(4.6 mm×250 mm,5μm),流动相为0.3%乙酸水和乙腈,紫外检测波长为230 nm,流速为1 mL·min^(-1),柱温30℃,每次进样量为10μL,梯度洗脱。结果阿魏酸和芍药苷的线性回归方程分别为■=1.567 3X-7.332 1、■=1.320 1X+2.091 1,r值分别为0.999 8、0.999 7,线性范围分别为0.52~16.64μg·L^(-1)、0.21~3.36μg·L^(-1);精密度、专属性、稳定性、基质效应和提取回收率考查均符合要求。结论 HPLC可以用于同时测定大鼠灌胃补阳还五汤后血清中阿魏酸和芍药苷的含量。

Objective To establish a method of dtermining of paeoniflorin and erulic acid in serum of rats after intragastric administration of Buyang Huanwu decoction by high performance liquid chromatography（ HPLC）. Methods 12 rats were randomly by random number table method divided into control group and Buyang Huanwu decoction group and 6 rats in each group. Preparation of Medicated serum were prepared and analyzed by HPLC,with C18 column（ 4. 6 mm × 150 mm,5 μm）. The mobile phase were 0. 3% acetic acid and acetonitrile. The wavelength of detection,flow rate,column temperature and sample size were 230 nm,1 m L·min-1,30 ℃and 10 μL respectively. Results Linear regression equation,r value and linear range of erulic acid were ■ = 1. 567 3 X-7. 332 1,0. 999 8 and 0. 52 ~ 16. 64 μg·L^-1 respectively,meanwhile,Linear regression equation,r value and linear range of paeoniflorin were ■ = 1. 320 1 X ＋ 2. 091 1,0. 999 7 and 0. 21 ~ 3. 36 μg·L^-1 respectively. The investigation of precision,specificity,stability and recovery rate of extraction were all qualified. Conclusions HPLC can determine the levels of paeoniflorin and erulic acid in serum of rats after intragastric administration of Buyang Huanwu decoction.