摘要
目的:针对经典名方芍药甘草汤药渣,开展小分子资源性成分提取及定性、定量研究,明确该药渣中残留小分子化学成分情况,探讨其可资源化利用途径。方法:采用超高效液相色谱-四极杆-飞行时间串联质谱法(UPLC-QTOF-MS/MS)定性分析芍药甘草汤药渣中残留的小分子化学物质,使用Agilent C18色谱柱(3.0 mm×100 mm,2.7μm),流速0.4 mL·min^(-1),进样量5μL,流动相0.05%甲酸水溶液(A)-乙腈(B)梯度洗脱(0~1 min,14%~17.5%B;1~3 min,17.5%~19%B;3~4 min,19%~20%B;4~5 min,20%B;5~6 min,20%~21%B;6~9 min,21%B;9~22 min,21%~36%B;22~23 min,36%B;23~32 min,36%~43%B),电喷雾离子源(ESI),负离子模式扫描,扫描范围m/z 50~1 200;对其主要成分建立高效液相色谱法(HPLC)定量分析方法,使用Agilent C18反相色谱柱(4.6 mm×150 mm,5μm),检测波长235 nm,流速0.8 mL·min^(-1),流动相0.05%磷酸水溶液(A)-乙腈(B)梯度洗脱(0~1 min,14%~19%B;1~4 min,19%B;4~18 min,19%~50%B);比较有机溶剂提取、酶法提取2种不同技术处理前后芍药甘草汤药渣中主要成分含量变化。结果:定性分析了芍药甘草汤药渣中16个化学成分,定量分析发现药渣中化学成分残留较多,其中芍药苷、甘草苷等6个指标成分残留量达到原饮片中的70%以上;药渣经纤维素酶酶解后,甘草苷可转化为甘草素;酶法提取药渣中粗多糖含量是空白组的6倍,质量分数高达12%。结论:芍药甘草汤药渣仍存在较多的小分子资源成分,有较大开发潜力,可利用有机溶剂对药渣中目标成分进行再提取,亦可采用生物酶解技术将甘草苷转化为甘草素,还可通过酶法提取药渣中粗多糖后开发动物饲料。该研究可为芍药甘草汤制剂及配方颗粒等生产带来的药渣提供资源再利用参考依据和途径,以实现芍药甘草汤药渣的高值化利用。
Objective:Aiming at the residue of Shaoyao Gancaotang,the extraction,qualitative and quantitative study of the small molecule resource components were carried out to clarify the residual small molecule chemical components in the residue and explore the ways of its resource utilization.Method:The ultra-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-QTOFMS/MS)was used to qualitatively identify the residual small molecule substances in the dregs of Shaoyao Gancaotang.Agilent C18 reversed-phase chromatographic column(3.0 mm×100 mm,2.7 μm)was used at the flow rate of 0.4 mL·min^(-1),the injection volume was 5 μL,and the mobile phase was gradient eluted with 0.05%formic acid aqueous solution(A)-acetonitrile(B)(0-1 min,14%-17.5%B;1-3 min,17.5%-19%B;3-4 min,19%-20%B;4-5 min,20%B;5-6 min,20%-21%B;6-9 min,21%B;9-22 min,21%-36%B;22-23 min,36%B;23-32 min,36%-43%B),electrospray ionization(ESI)was employed with negative ion mode scanning and scanning range of m/z 50-1 200.A high performance liquid chromatography(HPLC)was established for the quantitative analysis of its main components with Agilent C18 reversed-phase chromatographic column(4.6 mm×150 mm,5 μm),the detection wavelength was set at 235 nm,the flow rate was 0.8 mL·min^(-1),and the injection volume was 5 μL.Mobile phase was 0.05% phosphoric acid(A)-acetonitrile(B)for gradient elution(0-1 min,14%-19%B;1-4 min,19%B;4-18 min,19%-50%B).The content changes of main components in the residue of Shaoyao Gancaotang were compared before and after two different techniques of organic solvent extraction and enzymatic extraction.Result:A total of 16 chemical components in the residue of Shaoyao Gancaotang were qualitatively analyzed,and quantitative analysis found that there were many chemical components in the residue,among which the residues of 6 index components such as paeoniflorin and liquiritin reached more than70% in the original decoction piece.After enzymolysis by cellulase,liquiritin in the residue could be converted int
作者
陈彭月
刘艳
陈莎
刘安
李海鹰
CHEN Peng-yue;LIU Yan;CHEN Sha;LIU An;LI Hai-ying(Hebei Key Laboratory of Pharmaceutical Quality Analysis and Control,College of Pharmaceutical Sciences,Hebei University,Baoding 071002,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2021年第24期171-177,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家重点研发计划项目(2019YFC1906601)
国家“重大新药创制”科技重大项目(2019ZX09201005-002)。
