肝郁脾虚证患者口服加味逍遥散后阿魏酸的血药浓度HPLC测定法

HPLC Determination of Concentration of Ferulic Acid in Blood of Patients with Liver Depression and Spleen Deficiency

目的 建立加味逍遥散汤剂中阿魏酸 (FA)在肝郁脾虚证患者体内血药浓度测定的HPLC法。方法 应用乙腈提取法预处理血样 ,RP -HPLC法测定 10例肝郁脾虚证患者在口服加味逍遥散汤剂后 8个时间点血清样本中的FA浓度。色谱条件 :KromasilC18柱 ( 2 5 0mm× 4.6mm ,5 μm)为固定相 ;甲醇 -水 -冰醋酸 ( 40∶5 9.6∶0 .4,V/V)为流动相 ,流速 :1.0mL/min ;检测波长λ :32 2nm。FA用内标 (对 -羟基苯甲醛 )鉴定。结果 FA的最低检测限为 0 .46 2ng(S/N =3) ,血清最低检测限浓度 4.6 92ng/mL ;线性范围为 7.82～ 195 .5ng/mL(r =0 .9971)。结论 本法简便、快速、灵敏、准确而专一 ,适用于肝郁脾虚证患者口服加味逍遥散汤剂后血清FA的测定。

OBJECTIVE To establish the HPLC method for determination of the concentration of ferulic acid in the blood of the patients with liver depression and spleen deficiency who take Modified Xiaoyao Powder. METHOD Acetonitrile extraction was used to pre-treat the blood sample. RP-HPLC method was used to determine the concentration of ferulic acid of ten patients at eight different times, under the following conditions: Kromasil C18 colume (250 mm × 4.6 mm × 5 μm) as the fixed phase; methanol - water - glacial acetic acid (40∶59.6∶0.4, V/V) as the mobile phase; flow rate: 1.0 mL/min; determination wavelength λ:322nm. Ferulic acid was identified with the internal standard (parahydroxybenzaldehyde). RESULT The minimum ferulic acid determination limit was 0.462 ng (S/N = 3), the minimum serum determination limit was4.692 ng/mL, and the linear range was within 7.82～195.5 ng/mL (r= 0.997 1). CONCLUSION This method is simple, speedy, sensitive and accurate, suitable for the determination of the concentration of ferulic acid in the serum of the patients with liver depression and spleen deficiency who take Modified Xiaoyao Powder.