左归饮高效液相色谱指纹图谱研究及5种成分含量测定

Study on the HPLC Fingerprint of Zuogui Drink and Content Determination of Five Components

目的为左归饮的质量控制提供依据。方法采用高效液相色谱(HPLC)法建立15批左归饮样品的指纹图谱,色谱柱为Agilent 5 HC-C_(18)柱(250 mm×4.6 mm,5μm),流动相为0.2%甲酸水溶液-乙腈(梯度洗脱),流速为1.0 mL/min,检测波长为250 nm,柱温为30℃,进样量为20μL。采用中药色谱指纹图谱相似度评价系统(2012.1307023版)进行相似度评价,确定共有峰及指认成分,并测定其含量。结果15批样品指纹图谱与对照指纹图谱的相似度均在0.975~0.999之间。标定共有峰33个,指认出其中5个,分别为5-羟甲基糠醛、莫诺苷、马钱苷、甘草苷、甘草酸。5种成分质量浓度分别在0.017~0.266 mg/mL、0.049~0.784 mg/mL、0.034~0.539 mg/mL、0.016~0.250 mg/mL、0.009~0.150 mg/mL范围内与峰面积线性关系良好(R^(2)>0.9990,n=5);精密度、稳定性、重复性试验结果的RSD均小于3.0%;平均加样回收率分别为101.40%,98.72%,98.00%,101.53%,99.13%,RSD分别为2.06%,2.09%,1.65%,1.56%,0.40%(n=6);样品含量分别为49.838~140.933μg/g、131.060~340.103μg/g、68.081~178.313μg/g、12.108~47.326μg/g、1.190~55.483μg/g。结论所建立的方法可用于左归饮HPLC指纹图谱的建立及其中5个成分的含量测定,可更全面地为左归饮的质量评价提供依据。

Objective To provide a basis for the quality control of Zuogui Drink.Methods Fingerprint of 15 batches of Zuogui Drink samples was established by the high-performance liquid chromatography(HPLC)method.The chromatographic column was the Agilent 5 HC-C_(18) column(250 mm×4.6 mm,5μm),the mobile phase was 0.2%formic acid aqueous solution-acetonitrile(gradient elution),the flow rate was 1.0 mL/min,the detection wavelength was 250 nm,the column temperature was 30℃,and the injection volume was 20μL.The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(Version 2012.1307023)was used for similarity evaluation,the common peaks were marked,the components were identified,and the contents were determined.Results The similarity between the fingerprint of 15 batches of samples and the reference fingerprint was in the range of 0.975-0.999.A total of 33 common peaks were marked,and five of them were identified,namely 5-hydroxymethylfurfural(5-HMF),morroniside,loganin,liquiritin and glycyrrhizic acid.The linear ranges of the above five components were 0.017-0.266 mg/mL,0.049-0.784 mg/mL,0.034-0.539 mg/mL,0.016-0.250 mg/mL,and 0.009-0.150 mg/mL(R^(2)>0.9990,n=5)respectively.The RSDs of precision,stability and repeatability tests were all lower than 3.0%.The average recovery rates of the above five components were 101.40%,98.72%,98.00%,101.53%,99.13%,with RSDs of 2.06%,2.09%,1.65%,1.56%,0.40%(n=6)respectively.The contents of 5-HMF,morroniside,loganin,liquiritin and glycyrrhizic acid in samples were in the range of 49.838-140.933μg/g,131.060-340.103μg/g,68.081-178.313μg/g,12.108-47.326μg/g,and 1.190-55.483μg/g,respectively.Conclusion The established method can be used for the establishment of HPLC fingerprint of Zuogui Drink and the contents determination of the five components,which can provide a basis for the comprehensive quality evaluation of Zuogui Drink.