Zhou Lei, Kang Jie, Fan Li, Ma Xiao-Chi, Zhao Hai-Yu, Han Jian, Wang Bao-rong, Guo De-An
The State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, No. 38 Xueyuan Road, Beijing 100083, PR China.
J Pharm Biomed Anal. 2008 May 12;47(1):39-46. doi: 10.1016/j.jpba.2007.12.009. Epub 2007 Dec 15.
A high-performance liquid chromatography-diode array UV detection-electrospray ionization tandem mass spectrometry (HPLC-DAD-ESI-MS) method was developed and validated for the simultaneous analysis of seven major constituents in Cortex Fraxini, including esculin, esculetin, fraxetin, fraxin, escuside, oleuropein and ligustroside. The contents of the seven constituents were determined by using HPLC-DAD, and the chemical structures of these constituents were identified by using HPLC-DAD-ESI-MS method. The separation was performed on an Agilent Zorbax SB-C18 column (250 mm x 4.6 mm i.d., 5 microm) with gradient elution of acetonitrile and 0.3% aqueous acetic acid within 40 min. Detection was performed at 254 nm. The calibration curves showed good linearity (r2>0.9992). The limits of detection (LOD) ranged from 1.07 to 3.19 ng/ml and limits of quantification (LOQ) ranged from 2.79 to 12.75 ng/ml, respectively. The intra- and inter-day precision was less than 5% and the accuracy was ranging from 96.49% to 103.55%. The recovery of the assay was in the range of 97.61-104.36%. The method was successfully applied to the quantification of the seven constituents in different samples of Cortex Fraxini. The results indicated that the developed method could be considered to be a simple, rapid and reliable method for the quality evaluation of Cortex Fraxini.
建立了一种高效液相色谱 - 二极管阵列紫外检测 - 电喷雾电离串联质谱（HPLC - DAD - ESI - MS）方法，并对其进行了验证，用于同时分析秦皮中的七种主要成分，包括七叶苷、七叶内酯、秦皮素、秦皮苷、秦皮甲素、橄榄苦苷和女贞苷。采用HPLC - DAD测定这七种成分的含量，并采用HPLC - DAD - ESI - MS方法鉴定这些成分的化学结构。分离在Agilent Zorbax SB - C18柱（250 mm×4.6 mm内径，5μm）上进行，用乙腈和0.3%的乙酸水溶液在40分钟内进行梯度洗脱。检测波长为254 nm。校准曲线显示出良好的线性（r2>0.9992）。检测限（LOD）范围为1.07至3.19 ng/ml，定量限（LOQ）范围分别为2.79至12.75 ng/ml。日内和日间精密度均小于5%，准确度范围为96.49%至103.55%。该方法的回收率在97.61 - 104.36%范围内。该方法成功应用于不同秦皮样品中七种成分的定量分析。结果表明，所建立的方法可被认为是一种用于秦皮质量评价的简单、快速且可靠的方法。
