UPLC法测定苏木中巴西苏木素和原苏木素B的含量

Determination of brazilin and protosappanin B in Sappan Lignum by UPLC

目的建立同时测定苏木中巴西苏木素和原苏木素B的质量分数的超高效液相（UPLC）法。方法采用UPLC-PDA方法,色谱柱为ACQUICTY UPLC HSS T3（2.1 mm×100 mm,1.8μm）,流动相为甲醇-质量分数0.2%的冰醋酸水溶液,梯度洗脱,洗脱时间7.5 min,柱温40℃,流量为0.3 m L/min,检测波长为280nm。进样量为10μL。结果巴西苏木素和原苏木素B均达到基线分离,质量浓度在0.195~200μg/m L范围内与峰面积有良好的线性关系,R2均为0.999 8;平均回收率分别为103.89%、99.83%,RSD为1.26%、1.87%。结论相比HPLC法,UPLC法明显提高了同时检测苏木中巴西苏木素和原苏木素B质量分数的分析速度,并且灵敏度更高,可为以后建立苏木质量检测方法新标准提供参考。

Objective To develop an UPLC method to determine brazilin and protosappanin B in aqueous extract of Sappan Lignum. Method Brazilin and protosappanin B were analyzed by UPLC. An ACQUICTY UPLC HSS T3（ 2. 1 mm × 100 mm,1. 8 μm） column was used with the mobile phase of methanol-water（ 0.2% glacial acetic acid） in gradient elution mode and it cost 7.5 minutes throughout the general courses.The column temperature was 40 ℃ with a flow rate of 0.3 m L / min and the detective wavelength was 280 nm.Each sample was injected by 10 μL. Results The results showed that the peaks of brazilin and protosappanin B were separated commendably at the same time. The chromatographic peak areas also had good linearity in a beamy range of 0.195-200 μg / m L（ R2= 0.999 8）. The average recoveries of brazilin and protosappanin B were 103. 89%（ RSD = 1. 26%） and 99. 83%（ RSD = 1. 87%）, respectively. Conclusions Compared to HPLC,it apparently promoted efficiency by UPLC in analyzing brazilin and protosappanin B of Sappan Lignum with higher sensitivity, which could provide a reference for the establishment of new determination standards of Sappan Lignum.