邓老凉茶颗粒的超高效液相色谱质谱联用指纹图谱研究

Fingerprint Analysis of Dengs Herbal Tea Granule by Ultra High Performance Liquid Chromatography Coupled with Mass Spectrometry

建立了适用于邓老凉茶颗粒质量控制的超高效液相色谱质谱联用指纹图谱分析方法。样品采用甲醇索氏萃取60 min,萃取液采用超高效液相色谱质谱法进行指纹图谱分析。色谱柱采用Waters ACQUITY HSST3 C18(150 mm×3.0 mm,1.8μm),以0.5%甲酸-乙腈为流动相进行梯度洗脱,流速为0.8 mL/min,柱温35℃。质谱采用负离子ESI模式,选择基峰离子流质量色谱图进行指纹图谱研究。32个共有峰在15 min内得到良好分离,其中15个共有峰通过对照品进行了确证。通过《中药色谱指纹图谱相似度评价系统2004A版》对邓老凉茶颗粒样品进行相似度分析,15个批次样品的相似度均达到0.960以上,表明邓老凉茶颗粒的产品质量稳定性很好。以32个共有峰的相对峰面积进行主成分分析,邓老凉茶颗粒样品之间的细微质量差异得到明显区分。该方法快速、高效、可靠,可有效地用于邓老凉茶颗粒的质量控制。

A fingerprint analysis method was developed for the quality control of Deng-s herbal tea granule（DHTG） by ultra high performance liquid chromatography coupled with mass spectrometry （UHPLC-MS）.The samples were extracted by Soxhlet extraction with methanol as solvent for 60 min.UHPLC-MS separation was performed on a Waters ACQUITY HSS T3 C18（150 mm×3.0 mm,1.8 μm） column by gradient elution with 0.5% formic acid-acetonitrile as mobile phase,at the temperature of 35 ℃ and a flow rate of 0.8 mL/min.The base peak chromatograms（BPC） obtained with negative-ion electrospray ionization（ESI） mass spectra were selected for the fingerprint analysis.A good separation of 32 common peaks was achieved in 15 min,and 15 of them were confirmed by reference substances.Similarity evaluation was performed by a professional software named Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine（Version 2004A）,and the similarity among 15 batches of samples was no less than 0.960,which confirmed that the quality of DHTG was stable.The principal component analysis（PCA） was performed by using the samples as variables and the relative peak areas of 32 common peaks as observations.A fine difference among DHTG samples was expressed in the PCA result.The developed method was fast and efficient,and was suitable for the quality control of DHTG.