DESI-MSI在半夏泻心汤质量控制中的应用

Application of DESI-MSI in Quality Control of Banxia Xiexintang

目的:制备不同产地15批次半夏泻心汤物质基准及对应实物,分别应用超高效液相色谱-二极管阵列检测器检测法(UHPLC-DAD)与解吸电喷雾电离-敞开式离子化质谱成像技术(DESI-MSI技术)对不同产地15批次半夏泻心汤对应实物进行分析,考察DESI-MSI在经典名方质量控制中的优势。方法:以半夏泻心汤为研究模型,采用UHPLC-DAD建立半夏泻心汤物质基准及对应实物指纹图谱,并对指标成分含量、出膏率进行考察。同时,以半夏泻心汤对应实物冻干粉为研究载体,将其复溶后以5μL定量毛细管点样于定性滤纸,并固定于载玻片制成样本,以甲醇-甲酸(1000∶1)为喷雾溶剂,流量3μL·min-1,扫描范围100~1200 Da,空间分辨率300μm,喷雾电压3 k V,采样锥电压±40 V,喷雾器(N2)压力0.5 MPa,喷雾器入射角60度,收集角10度,离子源温度120℃,正、负离子全扫描样本。结果:DESI-MSI能检测到指标成分甘草苷、黄芩苷、汉黄芩苷,及共有峰芹糖基甘草苷、小檗碱、甘草酸等,并对其半定量分析,分析结果与UHPLC-DAD基本一致。同时,DESI-MSI还能检测到来自甘草、黄连、黄芩、大枣和人参的甘草皂苷G2,巴马汀,黄连碱,芦丁,人参皂苷Rg1等16种成分,并对其相对含量进行直观呈现,定性分析能力远强于UHPLC-DAD。结论:DESI-MSI技术无需进行复杂的样品处理过程,且灵敏度高、分析能力强,可以在无对照品情况下进行复杂样品的定性和相对含量分析,可作为经典名方物质基准、对应实物及配方颗粒样品的质控手段。

Objective:To prepare 15 batches of Banxia Xiexintang substance benchmark and lyophilized powder from different places,and the lyophilized powder was analyzed by ultra-high performance liquid chromatography with diode array detection(UHPLC-DAD)and desorption electrospray ionization-mass spectrometry imaging(DESI-MSI)in order to investigate the advantages of DESI-MSI in quality control of famous classical formulas.Method:Taking Banxia Xiexintang as the research model,fingerprints of the substance benchmark and lyophilized powder were established by UHPLC-DAD,and the content of index components and the yield of dry extract were also investigated.Meanwhile,as the research carrier,the lyophilized powder corresponding to Banxia Xiexintang was dissolved in methanol and dotted on qualitative filter paper with 5μL quantitative capillary,and fixed it on the slide to make samples.The samples were analyzed on a DESI-MSI system in positive and negative ion mode with methanol-formic acid(1000∶1,flow rate of 3μL·min-1)as spray solvent,N2 as spray gas(pressure of 0.5 MPa).The scanning range was 100-1200 Da,the spatial resolution was 300μm,the spray voltage was 3 k V,the sampling cone voltage was±40 V,incidence angle of sprayer was 60 degree,its collection angle was 10 degree,the ion source temperature was 120℃.Result:DESI-MSI could not only detect the index components of liquiritin,baicalin and wogonoside,as well as the common peaks of liquiritin apioside,berberine and glycyrrhizic acid,but also analyzed them semi-quantitatively,the analysis results were basically consistent with UHPLC-DAD.At the same time,DESI-MSI could detect 16 other components from Glycyrrhizae Radix et Rhizoma,Coptidis Rhizoma,Scutellariae Radix,Jujubae Fructus and Ginseng Radix et Rhizoma,such as licoricesaponin G2,palmatine,coptisine,rutin and ginsenoside Rg1,and present their relative content visually.The qualitative analysis ability of DESI-MSI was much better than UHPLC-DAD.Conclusion:DESI-MSI can be used as the quality control method for substance benchmark and lyophilized powder and dispensing granules of famous classical formulas with advantages of high sensitivity,strong analytical ability,no complex sample processing,qualitative and relative content analysis of complex samples without reference substance.