复方蜘蛛香挥发油GC-MS指纹图谱的建立及多组分定量分析

Establishment of GC-MS fingerprint and quantitative analysis of multi-components of volatile oil from Compound Zhizhuxiang

目的建立复方蜘蛛香挥发油GC-MS指纹图谱及其10个指标成分的定量分析方法,为其质量控制及评价提供参考。方法提取15批复方蜘蛛香挥发油,采用GC-MS技术建立指纹图谱,使用中南大学“中药指纹图谱计算软件”计算相似度,并对共有峰进行归属。采用聚类分析(cluster analysis,CA)、主成分分析(principal component analysis,PCA)、正交偏最小二乘法-判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA)对指纹图谱进行评价。利用GC法测定挥发油中10个指标成分的含量。结果15批挥发油得量在4.4~6.8 mL;GC-MS指纹图谱有31个共有峰,其中峰1、11归属于蜘蛛香挥发油;峰31归属于木香挥发油;峰6~9、15、21归属于草果挥发油;峰2~5、10、18为蜘蛛香、草果挥发油共有;峰23、25、27、29、30为蜘蛛香、木香挥发油共有;峰12~14、16、17、19、20、22、24、26、28为蜘蛛香、木香、草果3者挥发油共有;15批样品指纹图谱与中位数法生成的指纹图谱共有模式的相似度在0.798~0.998;聚类分析可以将样品分为2类,其中S1、S2、S5、S6、S11、S14、S15为第1类;S3、S4、S7~S10、S12、S13为第2类;PCA及OPLS-DA可将样品分为4类,其中S1、S2、S5、S6为第1类,S3、S4、S7~S9为第2类,S10、S12、S13为第3类,S11、S14、S15为第4类。异戊酸(峰1)、α-蒎烯(峰2)、β-蒎烯(峰3)、1,8-桉叶素(峰7)、α-松油醇(峰13)、香叶醇(峰15)、β-榄香烯(峰23)、β-丁香烯(峰25)、反式-橙花叔醇(峰28)、石竹素(峰29)的质量浓度在19.49~43.24、2.90~13.15、7.47~17.19、182.94~248.19、12.83~20.48、21.82~57.63、1.54~2.83、3.07~6.50、12.06~19.02、2.94~5.43 mg/mL。结论建立的复方蜘蛛香挥发油指纹图谱及定量分析方法分离度好、准确性高,可为复方蜘蛛香挥发油质量控制及评价提供参考。

Objective To establish the GC-MS fingerprint of the volatile oil of Compound Zhizhuxiang(CZ)and quantitative analysis method of 10 index components,providing a reference for the quality control and evaluation of volatile oil of CZ.Methods A total of 15 batches of volatile oil from the CZ were extracted.The GC-MS technology was used to establish the fingerprint of volatile oils,then the“Chinese Medicine Fingerprint Calculation Software”of Central South University was used to evaluate the similarity,and the common peaks were assigned.Fingerprints were assessed by cluster analysis(CA),principal component analysis(PCA),and orthogonal partial least squares-discriminant analysis(OPLS-DA).The contents of 10 index components in CZ were determined by GC.Results The volatile oil yield of 15 batches of CZ was between 4.4 and 6.8 mL.31 Common peaks were identified in the GC-MS fingerprint.Among them,peaks 1,11 belonged to the volatile oil from Zhizhuxiang(Valerianae Jatamansi Rhizoma et Radix,VJRR);Peak 31 belonged to the volatile oil from Muxiang(Aucklandiae Radix,AR);Peaks 6—9,15,and 21belonged to the volatile oil from Caoguo(Tsaoko Fructus,TF);Peaks 2—5,10,18 were shared by the volatile oil from VJRR and TF;Peaks 23,25,27,29,30 was shared by the volatile oil from VJRR,and AR,peaks 12—14,16,17,19,20,22,24,26,28 shared by the volatile oil from VJRR,AR and TF.The similarity between the 15 batches of sample fingerprints and the median fingerprint was between 0.798 and 0.998.The samples could be divided into two categories by cluster analysis,among S1,S2,S5,S6,S11,S14,and S15 are the first category;S3,S4,S7—S10,S12,and S13 are the second category.The samples could be divided into four categories by PCA and OPLS-DA;among them,S1,S2,S5,and S6 are the first category;S3,S4,S7—S9 are the second category;S10,S12,and S13 are the third category;S11,S14,and S15 are the fourth category.The content of isovaleric acid(peak 1),α-pinene(peak 2),β-pinene(peak 3)1,8-cineole(peak 7),α-terpineol(peak 13),geraniol(peak 15),β-elemene(peak 23),β-caryophyllene(peak 25),nerolidol(peak 28),caryophyllene oxide(peak 29)was between 19.49—43.24,2.90—13.15,7.47—17.19,182.94—248.19,12.83—20.48,21.82—57.63,1.54—2.83,3.07—6.50,12.06—19.02,2.94—5.43 mg/mL.Conclusion The established GC-MS fingerprint and quantitative analysis method of volatile oil from CZ have good separation and high accuracy.It can provide a reference for the quality control and evaluation of the volatile oil from CZ.