应用超临界流体萃取分离-气相色谱-质谱分析法建立中药温郁金的指纹图谱及其系统聚类分析

收集生长于浙江省瑞安市8个产地的温郁金样品14批次,分别经清洗、冷冻干燥及碾磨粉碎,得到通过0.425mm筛孔药筛的粉末状样品14个。称取此样品各30g,分别按试验选定的最佳条件萃取温度35℃,萃取压力15MPa,萃取时间(静态30min,动态90min)]进行超临界流体萃取分离,所得萃取物收集于甲醇5mL中,并进行气相色谱-质谱法(GC-MS)分析。色谱分离采用HP-5ms石英毛细管色谱柱和在50~220℃之间程序升温模式;质谱测定采用电子轰击离子源(EI),在质荷比(m/z)50~650内进行扫描。所得总离子流色谱图中共有50多个色谱峰,选择保留时间在40min以内的共有峰26个作为考察对象,选择保留时间为17.12min的莪术二酮为参考峰,计算共有峰的相对保留时间α和峰面积百分比S以及各样品与S3的S值的最大差值ΔS(Max)%,并建立了14个样品的指纹图谱。根据GC-MS分析所得结果并与NIST 14谱库比对,对共有峰作出鉴定。通过SPSS 16.0软件,采用系统聚类分析法对所分析的14批次温郁金样品的指纹图谱进行分类比较。应用指纹图谱对2个市售温郁金产品作了组分鉴定和产地认定。

Institution of Fingerprints of the TCMWenyujin by GC-MS with Separation by SFE and Study on Their Categorization by the Systematic Clustering Analysis

Samples of the TCMwenjujin(WYJ)from 14 different batches grown in 8 cultural areas of the Rui′an city in Zhejiang Province were collected and pretreated by washing,freeze-drying and pulverizing to pass through the medicinal sieve with bore diameter of 0.425 mm,thus to otbain 14 analytical samples in powdery form.30 g each of the 14 samples were taken and extracted separately by SFE technique under the optimized conditions of temperature at 35 ℃,of pressured under 15 MPa and with times of extraction of 30 min(static)and 90 min(dynamic).The extract was collected in 5 mL of CH3OH and analyzed by GC-MS.Quartz capillary chromatographic column(HP-5 ms)and programmed temperature elevation mode in the temperature interval of 50℃ to 220℃ were adopted in GC separation,and EI with scanning in the range of m/z from 50 to 650 was chosen for MS determination.As shown in the TIC chromatogram,more than 50 chromatographic peaks were obtained,among which 26 peaks in common with their retention times within 40 min were selected for further examination,using the peak of the compound numbered 16 with retention time of 17.12 min as the reference peak.Values of relative retention timesα,percentages of peak areas S%andΔS(Max)of the 26 peaks in common were calculated and their fingerprints were instituted.Based on the results obtained by GC-MS,and through comparing with the data given by the data base of NIST-14,the compounds giving the 26 common peaks were characterized.The fingerprints of the 14 samples from 8 cultural areas were studied comparatively by the systematic clustering analysis using the software of SPSS 16.0,and categorization based on the cultural areas of the Wenyujin was realized.Two Wenyujin samples obtained from the market were analyzed by the proposed method,with their components identified and their cultural areas recognized.