沉香高效液相特征图谱

HPLC Characteristic Chromatogram of Agarwood

【目的】对《中国药典》(2015)高效液相色谱法进行改进和完善,建立既适用于野生沉香也适用于人工沉香的高效液相特征图谱,为沉香的科学鉴别和质量评价提供科学依据。【方法】采用Phenomenex luna C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%甲酸溶液,梯度洗脱,流速0.7 mL·min^(-1),柱温31℃,检测波长252 nm。以29批次的参考样品和验证样品为研究对象,优化沉香高效液相色谱分离条件,考察所建高效液相特征图谱的精密度、稳定性和重复性。利用《中药色谱指纹图谱相似度评价系统》确定沉香样品共有的高效液相特征峰。【结果】方法学考察结果表明,所建高效液相特征图谱的精密度、稳定性和重复性良好。选定野生沉香和人工沉香高效液相图谱特征峰相对保留时间的RSD均小于2%,具有较好的保留重现性。验证结果表明,所建高效液相特征图谱对15批次验证样品可以有效、准确鉴别,其中,5批次伪品沉香无法与特征图谱匹配,6批次野生沉香、4批次人工沉香验证样品均符合沉香高效液相特征图谱。【结论】29批次样品兼顾野生沉香和人工沉香的特点,能够更全面、准确地反映沉香质量。所建高效液相特征图谱对野生沉香和人工沉香具有普适性,且符合方法学验证要求,可用于沉香的质量控制。

【Objective】To provide scientific basis for the identification and quality evaluation of agarwood,HPLC condition listed in＂Chinese Pharmacopoeia＂（2015） was improved,and the characteristic chromatogram for both wild agarwood and artificial agarwood was established. 【Method】 The analysis was carried out on a Phenomenex luna C18 column（250 mm × 4. 6 mm,5 μm） with a gradient mobile phase of acetonitrile-0. 1% formic acid at a flow rate of0. 7 mL·min^（-1）. The detection wavelength was set at 252 nm and the column temperature was 31 ℃. 29 batches of samples including both reference and validation ones were investigated. The chromatographic conditions were optimized and the precision,stability and repeatability of the method were investigated. The common characteristic peaks were analyzed with ＂Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica＂.【Result】 The results show that the precision,stability and repeatability of the method are good. The RSD of relative retention time of the characteristic peaks selected for either wild agarwood or cultivated agarwood is less than 2%. The results of 15 batches of validation samples show that the HPLC characteristic chromatogram of agarwood is accurate and effective for the identification of agarwood. 5 batches of adulterated agarwood among them cannot meet with the characteristic chromatogram,but the HPLC characteristic chromatogram of both 6 batches of wild agarwood and 4 batches of cultivated agarwood samples are consistent with those of agarwood. 【Conclusion】 29 batches of samples contain both wild and cultivated agarwood,and the developed HPLC method shows good applicable to both wild and cultivated agarwood,which conforms to the requirements of method ology validation and can be used for the quality control of agarwood.