高效液相色谱法测定不同产地功劳木中药根碱与巴马汀及小檗碱的含量

摘要

Objective To establish a liquid chromatography method for determination of Jatrorrhizine,Palmatine and Berberine in Mahonia from different areas by high pressure liquid chromatography.Methods The sample was treated by ultrasonic extraction with HCl-MeOH (1∶ 100,by volume).The analytical column was the Agilent ZORBAX SB-C18Column (150mm × 2.1mm,5μm).The mobile phase consisted of Methanol-0.1％ Formic acid,the flow rate was 0.4 ml/min,the column temperature was 40 ℃ and the detection wavelength was 345 nm.Results The calibrated linear curves of jatrorrhizine,palmatine and berberine concentrations were within the range of 0.005 00-0.500 00,0.005 00-0.750 00 and 0.000 55-0.220 00μg; the correlation coefficients (r) were 0.999 9,0.999 8 and 0.999 5,respectively.The recovery rates were 98.47％ (RSD =2.84％),98.58％ (RSD =3.34％) and 99.58％ (RSD =3.76％).There was a significant difference between the total amounts of alkaloids in Mahonia from different areas.They were 7.42,13.20,16.18 and 20.36 mg/g respectively.The maximum was 3 times higher than that of the minimum.The content of Jatrorrhizine was 3.91,8.06,3.78,7.91 mg/g,respectively.The Berberine was 1.42,7.64,8.61 and 10.55 mg/g; the Palmatine was 2.10,0.48,0.80 and 1.90 mg/g.The contents of the 3 alkaloids were related to the original plant variety,temperature,sunshine intensity and producing area.Conclusions The method is simple and fast.It can be used for simultaneous determination of Jatrorrhizine,Palmatine,Berberine in Mahonia.%目的 建立药根碱、巴马汀和小檗碱的含量测定的高效液相色谱法,并测定不同产地功劳木中药根碱、巴马汀和小檗碱含量.方法 样品用盐酸-甲醇(体积比1∶100)混昆合溶液超声提取,采用高效液相色谱法,色谱柱为Agilent ZORBAX SB-C18(150mm×2.1 mm,5μm),以甲醇-0.1％甲酸为流动相,流速为0.4 ml/min,柱温为40℃,检测波长为345 nm.配制一系列质量浓度(X)梯度的药根碱溶液(1.00、5.00、25.00、50.00、75.00、100.00 mg/L)、小檗碱溶液(l.00、2.50、5.00、25.00、75.00、100.00 mg/L)和巴马汀溶液(0.11、0.55、2.20、11.00、22.00、44.00 mg/L).分别取5μl注入高效液相色谱仪并记录药根碱、小檗碱和巴马汀的峰面积(Y).以浓度(X)为横坐标,色谱峰面积(y)为纵坐标绘制标准曲线,计算回归方程.取庆元百山祖、五凤垟麻宅村、泽雅源口、瓯海泽雅4个不同产地的功劳木药材,测定药根碱、小檗碱、巴马汀峰面积,每个样品测定3次.结果 上述色谱条件下,药根碱、巴马汀和小檗碱可以较好地分离,线性范围分别是0.005 00 ～0.500 00μg(r =0.999 9)、0.00500～ 0.750 00μg(r =0.999 8)、0.000 55 ～0.22000μg(r =0.999 5),平均回收率分别为98.47％[相对标准偏差(RSD)=2.84％]、98.58％ (RSD=3.34％)、99.58％(RSD=3.76％).不同产地功劳木3种生物碱总量有差异,庆元百山祖阔叶功劳木、五凤垟麻宅村功劳木、泽雅源口阔叶功劳木、瓯海泽雅阔叶功劳木的药根碱、巴马汀和小檗碱的总含量分别为7.42、16.18、13.20、20.36 mg/g,最高相差3倍;药根碱含量分别为3.91、8.06、3.78、7.91 mg/g,小檗碱含量分别为1.42、7.64、8.61、10.55 mg/g,巴马汀含量分别为2.10、0.48、0.80、1.90 mg/g.3种生物碱含量高低与原植物品种、温度、日照强度及产地有关.结论 高效液相色谱法测定药根碱、巴马汀和小檗碱的含量简便、快速、重现性好,适用于同时测定功劳木中药根碱、巴马汀和小檗碱的含量.