毛细管电色谱测定复方丹参注射液中原儿茶醛和原儿茶酸的含量

Quantitative determination of protocatechuic aldehyde and protocatechuic acid in compound Danshen injection by capillary electrochromatography

目的 建立毛细管电色谱分离测定复方丹参注射液中原儿茶醛(PAH)和原儿茶酸(PA)的内标定量测定方法。方法 用ODS柱为固定相,甲醇和2 mmol·L-1磷酸缓冲液(pH 3.0)为流动相,电压为-8kV,流速为10μL·min-1,压力为6766.59kPa,紫外检测波长为255 nm,4-羟基苯甲酸为内标。结果 原儿茶醛和原儿茶酸的线性范围分别是0.2-2.5ng和0.042-0.52 ng;相关系数分别为0.999 96和0.999 97;加样回收率分别为98.02%(RSD=3.42％)和100.0％(RSD)=0.93％);检测限分别为0.085 ng和0.036 ng;精密度实验RSD分别为0.52％和0.33％;重现性实验RSD分别为0.27％和0.34％;稳定性实验RSD分别为0.37％和0.32％:柱效分别为113246和257901;PA与PAH的分离度为4.854,PAH与4-羟基苯甲酸的分离度为4.620。结论 该方法简便,快速,准确,灵敏度高,重现性好,样品用量少,适用于丹参及含丹参制剂的样品中原儿茶醛和原儿茶酸的分离测定。

OBJECTIVE: To develop a method for the quantitative determination of protocatechuic aldehyde (PAH) and protocatechuic acid (PA) in Compound Danshen Injection by capillary electrochromatography. METHODS: Stationary phase was ODS. The mobile phase was composed of methanol and 2 mmol&middotL-1 H3 PO4 buffer solution at pH 3.0. The voltage was at -8 kV. The flow rate was 10 μL&middotmin-1. Pressure was 6 766.59 kPa and UV detection wavelength was 255 nm. The internal standard was 4-hydrobenzoic acid. RESULTS: The linearity ranges of PAH and PA were 0.2 - 2.5 ng and 0.042 - 0.52 ng respectively. The correlation coefficients were 0. 999 96 and 0.999 97 respectively. The average recoveries of spiked sample were 98.02% (RSD = 3.34%) and 99.94% (RSD = 0.91%) respectively. The detection limits were 0.085 ng and 0.036 ng respectively. The RSD (n = 5) of intra-batch variation were 0.52% and 0.33% respectively. The RSD (n = 5) of inter-batch variation were 0.27% and 0.34%. The RSD of stability were 0.37% and 0.32% respectively. The column efficiencies per meter were 113 246 and 257 901 respectively. The resolution between PA and PAH was 4.854 and the resolution between PAH and 4-hydrobenzoic acid was 4.620. CONCLUSION: The method was simple, rapid, accurate, sensitive and reproducible with small sample consuming. It is suitable for the determination of PAH and PA in Danshen and Danshen preparations.