苦豆子总碱GC-MS分析及幽门螺杆菌抑菌活性实验研究  

GC-MS Analysis of Total Alkaloids in Kudouzi(Sophora alopecuroides L.)and Experimental Study on Antibacterial Activity of Helicobacter pylori

作　　者：王维 戴碧珊 梁玉梅 WANG Wei;DAI Bishan;LIANG Yumei(The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong,China;Zhujiang Hospital of Southern Medical University,Guangzhou 510000,Guangdong,China)

机构地区：[1]广州中医药大学第一附属医院,广东广州510405 [2]南方医科大学珠江医院,广东广州510000

出　　处：《辽宁中医杂志》2024年第12期136-141,共6页Liaoning Journal of Traditional Chinese Medicine

基　　金：广东省中医药局科研项目(20231098);广东省医学科研基金项目(A2017178)。

摘　　要：目的基于气相色谱-质谱联用技术(gaschromatography-mass spectrometry,GC-MS)分析对苦豆子中的生物碱进行定性、定量分析,获得含量较高的5种苦豆子生物碱探究其与苦豆子总碱对幽门螺杆菌(Helicobacter pylori,Hp)临床耐药株的体外抑菌功效。方法采用732强酸性阳离子交换树脂法、Hydromatrix为基质的固相萃取法和有机溶剂液液萃取法制备新疆苦豆子总碱,并用GC-MS法分析总碱中生物碱的组分及相对含量。收集上消化道疾病患者胃黏膜标本,分离、鉴定出52株Hp临床菌株,应用E-试验方法检测对甲硝唑、克拉霉素、阿莫西林的耐药性,对32株临床耐药株进行筛选;应用液体倍比稀释法检测经GC-MS法分析中含量较高的5种苦豆子生物碱与苦豆子总碱对32株临床耐药株的体外抑菌效果。结果GC-MS法检测苦豆子含有13种已知生物碱,其中槐定碱、槐果碱、苦参碱、异槐果碱、野啶碱、lamprolobine和槐胺碱为主要生物碱,占所检出13种生物碱中的相对百分含量分别为25.82%、5.81%、13.28%、8.02%、7.34%、4.88%和3.15%。药敏实验显示,Hp对甲硝唑、克拉霉素耐药率高,对阿莫西林敏感。GC-MS法分析含量较高的5种苦豆子生物碱与苦豆子总碱对临床耐药株体外抑菌效果显著,以苦豆子总碱最为显著,其抗Hp作用较强;苦参碱与槐果碱也存在一定的抗Hp作用;槐定碱、异槐果碱、野啶碱的抑菌效果相对较弱。结论GC-MS法可快速分析苦参碱、槐定碱等生物碱,在苦豆子、苦参及其生物碱相关药物的质量控制方面具有一定的应用价值。Hp菌株对甲硝唑耐药率高,因此不选择甲硝唑作为根除Hp感染的一线药物。在多种有效成分中苦豆子总碱在体外抗Hp效果较好,可研发成为治疗Hp感染的中药单剂或复方制剂。Objective Based on gaschromatography-mass spectrometry(GC-MS)analysis,to qualitatively and quantitatively analyze the alkaloids in Kudouzi(Sophora alopecuroides L.),and to obtain 5 high alkaloids contents of Kudouzi(Sophora alopecuroides L.)to explore their in vitro antibacterial efficacy against clinical resistant strains of Helicobacter pylori(Hp)with total alkaloids from Kudouzi(Sophora alopecuroides L.).Methods The total alkaloids of Xinjiang Kudouzi(Sophora alopecuroides L.)were prepared using 732 strong acidic cation exchange resin method,solid-phase extraction with Hydromatrix as matrix,and organic solvent liquid-liquid extraction method.The components and relative contents of alkaloids in the total alkaloids were analyzed by GC-MS.The gastric mucosa samples were collected from patients with upper gastrointestinal diseases,52 clinical strains of Hp were isolated and identified.E-test method was applied to drug resistance of metronidazole,clarithromycin and amoxicillin,and 32 clinical drug resistant strains were screened.The liquid dilution method was used to detect the in vitro antibacterial effect of 5 alkaloids and total alkaloids of Kudouzi(Sophora alopecuroides L.)with high content analyzed by GC-MS on 32 clinically resistant strains.Results The GC-MS method detected that Kudouzi(Sophora alopecuroides L.)contained 13 known alkaloids,among which sophoridine,sophocarpine,matrine,isosophoridine,noritine,lamprolobine and sophoramine were the main alkaloids,accounting for 25.82%,15.81%,13.28%,8.02%,7.34%,4.88%and 3.15%of the 13 alkaloids detected,respectively.According to drug sensitivity experiments,Hp had a high resistance rate to metronidazole and clarithromycin,and was sensitive to amoxicillin.GC-MS analysis showed the 5 alkaloids and total alkaloids of Kudouzi(Sophora alopecuroides L.)with higher contents had significant in vitro antibacterial activity against clinically resistant strains,with the total alkaloids of Kudouzi(Sophora alopecuroides L.)being the most significant and having a strong anti Hp

关 键 词：苦豆子总碱 GC-MS分析 幽门螺杆菌 抑菌活性实验 

分 类 号：R285[医药卫生—中药学]