HPCE测定附子活性成分的血浆蛋白结合率

Study on the Plasma Protein Combine Rate of the Active Compounds of Aconiti Lateralis Radix Praeparata by HPCE

目的建立适用于附子生物碱类化学成分血浆蛋白结合率的方法。方法通过考察进样方式、电泳缓冲液组成等因素确定色谱条件,并采用该条件测定苯甲酰次乌头原碱、苯甲酰新乌头原碱和苯甲酰乌头原碱的血浆蛋白结合率。结果经考察确定色谱条件为:电压进样,缓冲液为100 mmol·L^(-1)硼砂-硼酸缓冲溶液(pH=9.0)∶甲醇=7∶3,分离电压为15 kV,检测波长为230 nm。在1,2,5μg·mL^(-1)3个剂量下,附子中苯甲酰次乌头原碱的蛋白结合率分别为(43.81±3.2)%,(42.51±3.6)%,(42.09±2.3)%;苯甲酰新乌头原碱的蛋白结合率分别为(39.59±4.0)%,(34.58±3.7)%,(37.02±3.0)%;苯甲酰乌头原碱的蛋白结合率分别为(48.23±4.6)%,(33.26±3.9)%,(32.60±3.8)%。结论超滤法结合HPCE技术可作为一种新型药物蛋白结合率测定的方法。

OBJECTIVE To establish a method for evaluating the plasma protein binding rate of alkaloid in Aconiti Lateralis Radix Praeparata. METHODS The chromatography condition such as sample injection condition and the ingredient of electrophoretic buffer solution were evaluated. And the plasma protein binding rate of benzoylhypaconine, benzoylaconine and benzoylmesaconine were analyzed. RESULTS The chromatography condition was as followes: voltage injection, electrophoretic buffer solution [100 mmol·L-1 borate saline buffer(pH=9.0)∶menthol=7∶3], separation voltage was 15 kV, the detection wavelength was 230 nm. Within the level of 1, 2, 5 μg·mL-1, the plasma protein binding rates of benzoylhypaconine were(43.81±3.2)%,(42.51±3.6)%,(42.09±2.3)%; the plasma protein binding rates of benzoylaconine were(39.59±4.0)%,(34.58±3.7)%,(37.02±3.0)%; and the plasma protein binding rates of benzoylmesaconine were(48.23±4.6)%,(33.26±3.9)%,(32.60±3.8)%. CONCLUSION Ultrafiltration combined with HPCE technology can be used as a new method for the analysis of plasma protein binding rate.