HPLC法同时测定不同等级黄柏饮片中多指标成分的含量

Simultaneous Content Determination of Multiple Indicator Ingredients in Diefferent Grades of Phellodendron amurense Decoction Piece by HPLC

目的：建立同时测定黄柏饮片中盐酸小檗碱、盐酸黄柏碱和木兰花碱含量的方法,并比较不同等级黄柏饮片中3种成分的含量。方法：采用高效液相色谱法。色谱柱为Phenomenex Luna C18,流动相为乙腈-0.05 mol/L磷酸二氢钾（梯度洗脱）,流速为1 ml/min,检测波长为280 nm,柱温为30℃,进样量为5μl。结果：盐酸小檗碱、盐酸黄柏碱、木兰花碱的检测进样量线性范围分别为0.387 0-7.740、0.044 4-0.888 0、0.048 0-0.960 0μg（r=0.999 9、0.999 8、0.999 9）;精密度、稳定性、重复性试验的RSD〈3%;加样回收率分别为95.61%-103.22%、96.18%-102.80%、97.93%-102.78%,RSD分别为2.80%、1.84%、1.84%（n=6）。结论：该方法操作简单、结果准确,适用于测定黄柏饮片中盐酸小檗碱、盐酸量柏碱和木兰花碱的含量;一级饮片中盐酸小檗碱和盐酸黄柏碱的含量高于二级饮片,但两个等级饮片中木兰花碱的含量无显著性差异。

OBJECTIVE：To establish a method for the simultaneous content determination of berberine hydrochloride,phellodendrine hydrochloride and magnoflorine in Phellodendron amurense decoction piece,and to campare the contents of the 3 ingredients in different grades of P. amurense decoction piece. METHODS：HPLC was performed on the column of Phenomenex Luna C18 with mobile phase of acetonitrile-0.05 mol/L KH2PO4（gradient elution）at a flow rate of 1 ml/min,the detection wavelength was280 nm,the column temperature was 30 ℃,and injection volume was 5 μl. RESULTS：The linear ranges were 0.387 0-7.740 μg for berberine hydrochloride（r=0.999 9）,0.044 4-0.888 0 μg for phellodendrine hydrochloride（r=0.999 8）and 0.048 0-0.960 0 μg for magnoflorine（r=0.999 9）;RSDs of precision,stability and reproducibility tests were lower than 3%,recoveries were95.61%-103.22%（RSD=2.80%,n=6）,96.18%-102.80%（RSD=1.84%,n=6）and 97.93%-102.78%（RSD=1.84%,n=6）.CONCLUSIONS：The method is simple and accurate,and can be used for the contents determination of berberine hydrochloride,phellodendrine hydrochloride and magnoflonine in P. amurense. The contents of berbenine hydrochloride and phellodendrine hydrochloride in the first-grade decation piece are higher than those in the second-grade decoction piece,and the content of magnoflorine in both decoction pieces shows no discernible differences.