柱前衍生-气相色谱-质谱联用法同时测定茯苓中的可溶性糖与三萜酸  

Simultaneous determination of soluble sugars and triterpene acids in Poria cocos by gas chromatography-mass spectrometry with pre-column derivatization

作　　者：陈菲[1] 李惠华[1] 陈淳[1] 吴承文 明艳林[1,3] CHEN Fei;LI Huihua;CHEN Chun;WU Chengwen;MING Yanlin(Fujian Key Laboratory of Subtropical Plant Physiology and Biochemistry,Fujian Institute of Subtropical Botany,Xiamen 361006,China;Forestry Bureau of Youxi County,Youxi 365100,China;National Plant Introduction&Quarantine Base(Xiamen),Xiamen Overseas Chinese Subtropical Plant Introduction Garden,Plant Introduction&Quarantine and Plant Product Key Laboratory of Xiamen City,Xiamen 361002,China)

机构地区：[1]福建省亚热带植物研究所,福建省亚热带植物生理生化重点实验室,福建厦门361006 [2]尤溪县林业局,福建尤溪365100 [3]国家农作物国外引种隔离检疫基地(厦门),厦门华侨亚热带植物引种园,厦门市植物引种检疫与植物源产物重点实验室,福建厦门361002

出　　处：《厦门大学学报（自然科学版）》2024年第3期532-540,共9页Journal of Xiamen University：Natural Science

基　　金：厦门市重大科技计划(3502Z20211006)。

摘　　要：[目的]通过建立柱前衍生-气相色谱-质谱(GC-MS)法实现茯苓(Poria cocos)各成分的快速高效分离检测,为茯苓有效成分的分析提供新方法并为茯苓原料品质判断提供参考.[方法]采用超声辅助提取法,以反油酸为内标物,甲氧胺(MOX)和N-甲基-N-(三甲基硅烷)三氟乙酰胺[含1%(质量分数)三甲基氯硅烷](MSTFA+1%TMCS)为肟化-硅烷化衍生试剂,采用GC-MS同时检测茯苓中的果糖、葡萄糖、海藻糖、阿拉伯糖醇、甘露糖醇、山梨糖醇、麦芽糖醇、茯苓酸、茯苓新酸A和茯苓新酸B.[结果]优化后的衍生条件为:200μL茯苓的甲醇提取液氮气吹干后,加入50μL MOX,于40℃反应2 h,然后加入250μL(MSTFA+1%TMCS),于40℃下反应30 min.衍生后的目标化合物在40 min内完成分离分析,线性关系良好,相关系数均在0.9935以上,该方法的定量限为0.008~1.000μg/mL,检测限为0.003~0.300μg/mL.茯苓样品加标回收率为82.0%~122.4%,相对标准偏差为1.05%~4.43%.[结论]本方法样品用量少,结果稳定且灵敏度高,可用于检测茯苓样品中可溶性糖和微量的三萜酸类化合物,同时也为检测其他样品中的三萜酸类化合物提供更多的选择.[Objective]Poria cocos has long been used in China as a medicinal and food mushroom due to its sedative,tranquilizing,and diuretic properties.Previous research has confirmed that soluble sugars and triterpene acids are its major nutritional and functional components,and the contents are substantially affected by factors such as the planting area,harvesting period,and processing methods.It is vital to comparatively identify and quantify soluble sugars and triterpene acids rapidly and accurately for the further development of fermented mycelia.[Methods]P.cocos was extracted using ultrasonic extraction with methanol,followed by derivatization with methoxyamine(MOX)and N-methyl-N-(trimethylsilyl)trifluoroacetamide(MSTFA)containing 1%(mass fraction)trimethylchlorosilane(TMCS).The effects of various variables(e.g.,volume ratio of menthanol,extraction time,volume of MSTFA,volume of MOX,methoximation time,and silylation time)on the extraction and derivation processes were investigated through single factor experiments.The derivatives were analyzed using gas chromatography-mass spectrometry(GC-MS),qualitatively in full scan mode with a scan range of m/z 40800,and quantitatively in selected ion scan mode.Derivatives were identified based on their retention time and electron impact(EI)-MS library match using commercial National Institute of Standards and Technology(NIST)library and standards.[Results]Methanol was selected as the extraction solvent,due to the highest extraction yields of pachymic acid(PA),poricoic acid A(PAA),and poricoic acid B(PAB),as well as shorter nitrogen blow-dry time.The extraction yields of derivatives increased sharply with increasing extraction time from 30 min to 60 min,then plateaued.Considering extraction efficiency,the extraction time was determined to be 60 min.For the volume of MOX,the yields of reducing sugars derivatives increased and reached the peak at 50μL.The efficiency of further silylation derivations would be reduced by the presence of pyridine in the methoximation reagent.For redu

关 键 词：衍生 气相色谱-质谱法 可溶性糖 三萜酸 茯苓 

分 类 号：O657.63[理学—分析化学]