UPLC-PDA法测定杨树花口服液中非法添加物黄芩苷

Determination of Baicalin in Yangshuhua Koufuye by UPLC-PDA

为了建立超高效液相色谱-二极管阵列检测法(UPLC-PDA法)测定杨树花口服液中非法添加物黄芩苷,取适量黄芩苷对照品、杨树花口服液阴性样品、阳性添加样品、抽检样品经50%甲醇溶解,超声提取,滤液过滤并定量稀释后,作为供试品溶液,采用UPLC-PDA法检测。色谱分离采用ACQUITY UPLCTMHSS T3色谱柱为分离柱,柱温:35℃;流动相体系:A项为乙腈,B项为0.4%磷酸水溶液,进行梯度洗脱;流速:0.35 m L/min;进样量:10μL;选择二极管阵列检测器,检测波长为278 nm,上机测定。通过保留时间、光谱图、峰纯度等参数对黄芩苷进行定性定量检测。结果表明,黄芩苷在0.5~100μg/m L的范围内线性关系良好(R2=0.999);杨树花口服液中添加黄芩苷0.05 mg/m L,信噪比(S/N)>3,确定为方法的检测限;添加0.1 mg/m L,信噪比(S/N)>10,确定为方法的定量限;杨树花口服液在0.1、0.2、0.4、0.6 mg/m L黄芩苷添加水平的回收率为95%~105%,批内批间变异系数均小于10%,准确度和精密度良好,完全满足检测需求。而且峰纯度角与阈值符合要求。本方法经济快速、灵敏、重现性好,适用于杨树花口服液中非法添加黄芩苷的定性定量检测。

To establish a UPLC-PDA method for determination and confirmation of baicalin added illegally in Yangshuhua Koufuye. Methods： The amount of baicalin control products, Yangshuhua Koufuye negative samples, the positive samples added baicalin, sampling appropriate amount dissolved in 50% methanol, with ultrasonic extraction,filtration and quantitative diluted and then detected by UPLC-PDA. An ACQUITY UPLCTM HSS T3 column was used with gradient elution of acetonitrile and 0.4% phosphoric acid as the mobile fhase （0.35 mL/min） and with column temperature 35℃; sample size： 10p.L; The signals were detected by PDA detector, and the detection wavelength was 278 nm. By comparing the retention time, spectra, peak purity and other parameters to identify Baicalin. Baicalin in the range of 0.5 - 100 pLg/mL of the linear relationship is good （R2= 0.999） ; The detection limit was 0.05 mg/mL（ s/n〉3）, and the quantitation limit was 0.1 mg/mL（ s/n〉 10）. Added at the level of 0.1,0.2,0.4,0.6 mg/mL recovery rate was 95%- 105%, the intra and inter assay coefficients of variation were less than 10%, with good accuracy and precision, fully meet the detection requirements. But the peak purity and threshold requirements. The mothod was rapid, reliable, sensitive and reproducibility, adapts to the determination of baicalin in yangshuhua koufuye, and can be used as a qualitative, quantitative determination method.