Yang Yue, Liu Ying, Feng Wei-Hong, Liu Xiao-Qian, Li Chun, Wang Zhi-Min
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
Zhongguo Zhong Yao Za Zhi. 2022 Jun;47(11):3000-3006. doi: 10.19540/j.cnki.cjcmm.20211227.202.
The purpose of this paper is to establish the HPLC fingerprint and content determination method for Crataegus pinnatifida seeds. The separation was developed on a Welch Ultimate XB C_(18) column(4.6 mm×250 mm, 5 μm) by gradient elution with acetonitrile-water containing 0.1% formic acid as the mobile phase at the flow rate of 1 mL·min(-1), the column temperature of 30 ℃, the injection volume of 10 μL, and the detection wavelength of 320 nm. Eighteen batches of samples were analyzed under the above chromatographic conditions to establish the fingerprint of C. pinnatifida seeds from different producing areas and a total of 24 common peaks were selected. The structures of three main chromatographic peaks were identified by comparison to reference substances, and the three compounds were simultaneously analyzed for content determination. They were identified as erythro-(7S,8R)-guaiacylglycerol-β-coniferyl aldehyde ether, threo-(7R,8R)-guaiacylglycerol-β-coniferyl aldehyde ether, and balanophonin, respectively. The relative similarity of fingerprints of 18 batches of samples and references ranged from 0.928 to 0.999, and the content of the three compounds was 0.055 1-0.182 7, 0.061 8-0.225 8, and 0.156 8-0.405 6 mg·g(-1), respectively. SPSS 17.0 and SIMCA 14.1 were used for cluster analysis, principal component analysis(PCA), and orthogonal partial least squares discriminant analysis(OPLS-DA) on the common peaks of the HPLC fingerprint of C. pinnatifida seeds. The results showed that there were significant differences between the two batches of samples from Liaoning province and the other samples, and the three compounds to be tested were the main components leading to the difference of C. pinnatifida seeds. The established method was simple and reliable and can be used for qualitative and quantitative analysis of C. pinnatifida seeds. The findings of this study are expected to provide a scientific basis for quality control of C. pinnatifida seeds.
本文旨在建立山楂籽的高效液相色谱指纹图谱及含量测定方法。采用Welch Ultimate XB C₁₈色谱柱（4.6 mm×250 mm，5 μm），以含0.1%甲酸的乙腈 - 水为流动相进行梯度洗脱，流速为1 mL·min⁻¹，柱温30℃，进样量10 μL，检测波长320 nm。在上述色谱条件下对18批样品进行分析，建立不同产地山楂籽的指纹图谱，共选出24个共有峰。通过与对照品比较鉴定了3个主要色谱峰的结构，并同时对这3种化合物进行含量测定。它们分别被鉴定为赤式 - (7S,8R)-愈创木基甘油 - β - 松柏醛醚、苏式 - (7R,8R)-愈创木基甘油 - β - 松柏醛醚和巴兰诺芬宁。18批样品指纹图谱与对照图谱的相对相似度在0.928至0.999之间，3种化合物的含量分别为0.055 1 - 0.182 7、0.061 8 - 0.225 8和0.156 8 - 0.405 6 mg·g⁻¹。使用SPSS 17.0和SIMCA 14.1对山楂籽高效液相色谱指纹图谱的共有峰进行聚类分析、主成分分析（PCA）和正交偏最小二乘法判别分析（OPLS - DA）。结果表明，辽宁省的两批样品与其他样品之间存在显著差异，所测的3种化合物是导致山楂籽差异的主要成分。所建立的方法简便可靠，可用于山楂籽的定性和定量分析。本研究结果有望为山楂籽的质量控制提供科学依据。
