HPLC法定性定量分析苦豆草药材中多个生物碱成分

HPLC analysis of multiple alkaloids in the whole herbs of Sophora alopecuroides L

目的:建立苦豆草药材中多个生物碱同时定性定量分析的高效液相色谱法。方法:采用DiamonsilTM C18柱(5μm,250mm×4.6mm),以0.05mol.L-1磷酸二氢钠水溶液-乙腈-甲醇-高氯酸钠(940mL:50mL:10mL:25g;pH4.5)为流动相,流速1mL.min-1,检测波长210nm,柱温40℃(0～39min)→50℃(40～49min)→40℃(50～55min)。结果:本方法在55min内定性分析了宁夏市售苦豆草中9个生物碱,并对其中6个进行了定量测定。苦参碱、槐定碱、槐果碱、苦豆碱、氧化槐果碱、氧化苦参碱的浓度均在0.002～2.00mg.mL-1范围内与峰面积呈良好线性关系,平均回收率(n=3)分别为97.4%～98.3%(RSD≤1.3%),97.5%～98.3%(RSD≤1.2%),96.8%～98.0%(RSD≤1.2%),97.1%～98.0%(RSD≤1.0%),97.9%～98.3%(RSD≤1.1%),97.1%～98.7%(RSD≤0.8%)。结论:本法灵敏、准确,专属性好,适用于苦豆草药材中生物碱成分的定性鉴别和定量分析。

Objective：To establish an HPLC method for the simultaneous qualitative and quantitative determination of multiple alkaloids in the whole herbs of Sophora alopecuroides.Methods：The method was conducted on a DiamonsilTM C18 column（5 μm,250 mm×4.6 mm）with 0.05 mol·L-1 sodium dihydrogen phosphate-acetonitrile-methyl alcohol-sodium perchlorate（940 mL：50 mL：10 mL：25 g;pH 4.5）as mobile phase,flow rate of 1 mL· min-1,UV detection at 210 nm and column temperature at 40 ℃（0-39 min）→50 ℃（40-49 min）→40 ℃（50-55 min）.Results：Nine alkaloids in S.alopecuroides from Ningxia herbs market were identified and six of them were simultaneously determinated by the developed method in 55 min.Matrine,sophoridine,sophocarpine,aloperine,oxysophocarpine and oxymatrine all showed good linearity in the range of 0.002-2.00 mg·mL-1;The average recoveries（n=3）were 97.4%-98.3%（RSD≤1.3%）,97.5%-98.3%（RSD≤1.2%）,96.8%-98.0%（RSD≤1.2%）,97.1%-98.0%（RSD≤1.0%）,97.9%-98.3%（RSD≤1.1%）,and 97.1%-98.7%（RSD≤0.8%）.Conclusion：The developed method is sensitive,accurate and specific for the identification and determination of the alkaloids in the whole herbs of S.alopecuroides.