UPLC-TOF结合多元数学统计方法建立五加参蛤蚧精中蛤蚧的鉴别方法

UPLC-TOF Combined with Multivariate Mathematical Statistics to Establish the Identification Method of Gecko in WuJiaShen GeJieJing

目的建立五加参蛤蚧精中蛤蚧的定性鉴别方法,为判定五加参蛤蚧精中是否投料正品蛤蚧提供参考。方法取样品至1.5 mL离心管中,离心(12000 r·min^(-1),5 min),取上清液,即得;色谱分析采用Waters BEH C_(18)色谱柱(2.1×50 mm,1.7μm),柱温35℃,流速为0.5 mL·min^(-1),流动相为0.1%甲酸水溶液(A)-乙腈(B),梯度洗脱。质谱测定采用电喷雾离子源(ESI)的飞行时间质谱(TOF/MS);正负离子模式下采集MSE数据;质量扫描范围m/z100~1200。结果分别对8个不同厂家的五加参蛤蚧精中的蛤蚧定性离子用多元数学统计方法进行比较、拟合,最终决定对样品采取未酶解的前处理方式,采集了ESI(正和负)的MSE数据,将162.1136、482.3709、508.3879这3个正离子特征离子和514.2794、353.2299、407.2765这3个负离子特征离子做为蛤蚧鉴别的6个特征离子。8个厂家中有一个厂家未检出这6个特征离子,怀疑为蛤蚧伪品投料。结论所建立的蛤蚧鉴别方法可作为规范五加参蛤蚧精中蛤蚧真伪判定的质量控制手段。

OBJECTIVE To establish the qualitative identification method of Gecko in the essence of WuJiaShen GeJieJing,and to provide a reference for determining whether the essence of WuJiaShen GeJieJing contains genuine Gecko.METHODS The sample was put into a 1.5 mL centrifuge tube,centrifuged(12000 r·min^(-1),5 min),and the supernatant was taken;Waters BEH C_(18)(2.1×50 mm,1.7μm)chromatographic column was used,Column temperature was 35℃,Flow rate was 0.5 mL·min^(-1),Mobile phase was 0.1%formic acid(A)-acetonitrile(B),gradient elution.Time of flight mass spectrometry(TOF/MS)of electric spray ion source(ESI)was used for mass spectrometry;Collect MSE data in positive and negative ion mode;The mass scanning range was m/z 100~1200.RESULTS The qualitative ions of Gekko in WuJiaShen GeJieJing from eight different manufacturers were compared and fitted with multiple mathematical statistical methods.Finally,sample was decided to adopt the pretreatment method without enzymolysis,and the MSE data of ESI(positive and negative)were collected.The three positive ion characteristic ions of 162.1136,482.3709 and 508.3879 and the three negative ion characteristic ions of 514.2794,353.2299 and 407.2765 were used as the six characteristic ions for Gecko identification.One of the eight manufacturers did not detect these six characteristic ions,and it was suspected that they were fed for gecko counterfeits.CONCLUSION The established gecko identification method can be used as a quality control method to standardize the determination of gecko in the WuJiaShen GeJieJing.