HPLC法同时测定云连药材中6种生物碱的含量

摘要

OBJECTIVE:To develop a method for simultaneous determination of 6 alkaloids in Coptis teeta.METHODS:The determination was performed on XtimateTM C18 with mobile phase consisted of 30 mmol/L ammonium bicarbonate [containing 0.1％ triethylamine and 0.7％ ammonia-acetonitrile (gradient elution)] at the flow rate of 1.0 mL/min.The detection wavelength was set at 270 nm,the column temperature was 30 ℃,and the sample size was 10 μL.RESULTS:The linear ranges of jateorrhizine,columbamine,epiberberine,coptisine,palmatine hydrochloride and berberine hydrochloride were 0.85-16.96 mg/L (r=0.999 9),1.25-24.96 mg/L(r=0.999 8),2.05-40.96 mg/L(r=0.999 9),3.65-72.96 mg/L(r=0.999 9),2.88-57.60 mg/L(r=0.999 9) and 13.25-264.96 mg/L(r=0.999 9),respectively.RSDs of precision,stability and reproducibility tests were all lower than 3.0％.Recoveries were 97.14％-102.14％ (RSD=1.93％,n=6),97.00％-102.00％ (RSD=2.06％,n=6),98.18％-101.82 ％ (RSD=1.79％,n=6),96.15％-101.28％ (RSD=2.06％,n=6),96.88％-101.88％ (RSD=1.87％,n=6),99.31％-103.76％ (RSD=1.89％,n=6),respectively.CONCLUSIONS:The method is simple,accurate,stability and reproducible,and can be used for simultaneous determination of 6 alkaloids in Coptis teeta.%目的:建立同时测定云连药材中6种生物碱含量的方法.方法:色谱柱为XtimateTMC18,流动相为30 mmol/L碳酸氢铵溶液(含0.1％三乙胺和0.7％氨水)-乙腈(梯度洗脱),流速为1.0 mL/min,检测波长为270 nm,柱温为30℃,进样量为10μL.结果:药根碱、非洲防己碱、表小檗碱、黄连碱、盐酸巴马汀、盐酸小檗碱的检测质量浓度线性范围分别为0.85～16.96 mg/L(r=0.999 9)、1.25～24.96 mg/L (r=0.999 8)、2.05～40.96 mg/L (r=0.999 9)、3.65～72.96 mg/L (r=0.999 9)、2.88～57.60 mg/L(r=0.999 9)、13.25～264.96 mg/L(r=0.999 9);精密度、稳定性、重复性试验的RSD＜3.0％;加样回收率分别为97.14％～102.14％ (RSD=1.93％,n=6)、97.00％～102.00％ (RSD=2.06％,n=6)、98.18％～101.82％ (RSD=1.79％,n=6)、96.15％～101.28％ (RSD=2.06％,n=6)、96.88％～101.88％(RSD=1.87％,n=6)、99.31％～103.76％ (RSD=1.89％,n=6).结论:该方法操作简便,精密度、稳定性、重复性好,可用于云连药材中6种生物碱含量的同时测定.