An HPLC-DAD method combined with second-order calibration based on the alternating trilinear decomposition(ATLD)algorithm with the aid of region selection was developed to simultaneously and quantitatively characteriz...An HPLC-DAD method combined with second-order calibration based on the alternating trilinear decomposition(ATLD)algorithm with the aid of region selection was developed to simultaneously and quantitatively characterize the synergistic relationships and cumulative excretion of the four bioactive ingredients of Radix Gentianae Macrophyllae in vivo.Although the analytes spectra substantially overlapped with that of the biological matrix,the overlapping profiles between analytes and co-eluting interferences can be successfully separated and accurately quantified by the ATLD method on the basis of the strength of region selection.The proposed approach not only determined the content change but also revealed the synergistic relationships and the cumulative excretion in vivo of the four ingredients in urine and feces samples collected at different excretion time intervals.In addition,several statistical parameters were employed to evaluate the accuracy and precision of the method.Quantitative results were confirmed by HPLC-mass spectrometry.Satisfactory results indicated that the proposed approach can be utilized to investigate the pharmacokinetics of Radix Gentianae Macrophyllae excretion in vivo.展开更多
To develop a HPLC-DAD-ESI-TOF/MS analysis method for the determination of gentiopicroside and loganic acid in Radix gentianae samples and for the research of their fingerprints.The samples were extracted using ASE for...To develop a HPLC-DAD-ESI-TOF/MS analysis method for the determination of gentiopicroside and loganic acid in Radix gentianae samples and for the research of their fingerprints.The samples were extracted using ASE for 10 min under 100 ℃ and 9.65 MPa,and divided into water phase and chloroform phase and analyzed them with HPLC-DAD-ESI-TOF/MS method respectively.Based on this method,the HPLC fingerprints of Radix gentianae were established.Comparing the spectrogram and mass spectrum of the chromatogram peak with the reference value,three compounds in water phase were identified as gentiopicroside,asafetida acid and loganic acid.There is no report of the compounds in chloroform phase.The content of gentiopicroside and loganic acid in samples of different groups were determined,separately.The fingerprints were compared by the software of the similarity evaluation system for chromatographic fingerprint.The water phase fingerprint congruence coefficients of samples from six different areas were above 0.90,however,the chloroform phase fingerprint congruence coefficients were within 0.62-0.99.This method can be used for determination of potent component in Radix gentianae and its quality control.Radix gentianae from different producing areas have the largest diversities,and the diversities embodied in the content of chloroform phase compounds.展开更多
目的比较不同灭菌方法对龙胆和秦艽化学成分的影响和对微生物的杀灭效果。方法采用干热灭菌、湿热灭菌和钴60辐照灭菌3种方法对3批龙胆和3批秦艽药材粉末进行灭菌处理,采用液相指纹图谱方法分析龙胆、秦艽中的化学成分。按照《中国药典...目的比较不同灭菌方法对龙胆和秦艽化学成分的影响和对微生物的杀灭效果。方法采用干热灭菌、湿热灭菌和钴60辐照灭菌3种方法对3批龙胆和3批秦艽药材粉末进行灭菌处理,采用液相指纹图谱方法分析龙胆、秦艽中的化学成分。按照《中国药典》2015年版微生物限度检查方法检测了不同灭菌处理样品的需氧菌总数、霉菌酵母菌计数、耐胆盐革兰阴性菌、大肠埃希菌。结果主要成分龙胆苦苷对辐照敏感,3 k Gy的小剂量辐照即可使龙胆苦苷减少15%或18%;干热灭菌或湿热灭菌使龙胆和秦艽在7.8 min处新增1小峰(峰4);龙胆中峰1经辐照后有所增加,但该物质并不是辐照产生的特异成分,干热灭菌后该峰也有所增加。3种灭菌方法均呈现出随着灭菌强度的增大,发生变化的峰的数量和变化率都有所增加的规律。灭菌效果简单排序为辐照>湿热>干热。结论干热、湿热、辐照3种方式灭菌处理对龙胆、秦艽成分除龙胆苦苷外影响不大,部分成分有增加或减少,其中湿热(115℃、15 min)和辐照25 k Gy处理对成分的影响最大。辐照虽然灭菌效果良好,但是对主要成分龙胆苦苷的影响显著,故不适宜用于龙胆和秦艽的灭菌处理。展开更多
基金support from the National Natural Science Foundation of China(Grants Nos.21776321,21576297,21205145,21575039)Key Projects of Technological Innovation of Hubei Province(2016ACA138)The Open Research Program(Grant Nos.2015ZD001,2015ZD002 and 2015ZY006)。
文摘An HPLC-DAD method combined with second-order calibration based on the alternating trilinear decomposition(ATLD)algorithm with the aid of region selection was developed to simultaneously and quantitatively characterize the synergistic relationships and cumulative excretion of the four bioactive ingredients of Radix Gentianae Macrophyllae in vivo.Although the analytes spectra substantially overlapped with that of the biological matrix,the overlapping profiles between analytes and co-eluting interferences can be successfully separated and accurately quantified by the ATLD method on the basis of the strength of region selection.The proposed approach not only determined the content change but also revealed the synergistic relationships and the cumulative excretion in vivo of the four ingredients in urine and feces samples collected at different excretion time intervals.In addition,several statistical parameters were employed to evaluate the accuracy and precision of the method.Quantitative results were confirmed by HPLC-mass spectrometry.Satisfactory results indicated that the proposed approach can be utilized to investigate the pharmacokinetics of Radix Gentianae Macrophyllae excretion in vivo.
文摘To develop a HPLC-DAD-ESI-TOF/MS analysis method for the determination of gentiopicroside and loganic acid in Radix gentianae samples and for the research of their fingerprints.The samples were extracted using ASE for 10 min under 100 ℃ and 9.65 MPa,and divided into water phase and chloroform phase and analyzed them with HPLC-DAD-ESI-TOF/MS method respectively.Based on this method,the HPLC fingerprints of Radix gentianae were established.Comparing the spectrogram and mass spectrum of the chromatogram peak with the reference value,three compounds in water phase were identified as gentiopicroside,asafetida acid and loganic acid.There is no report of the compounds in chloroform phase.The content of gentiopicroside and loganic acid in samples of different groups were determined,separately.The fingerprints were compared by the software of the similarity evaluation system for chromatographic fingerprint.The water phase fingerprint congruence coefficients of samples from six different areas were above 0.90,however,the chloroform phase fingerprint congruence coefficients were within 0.62-0.99.This method can be used for determination of potent component in Radix gentianae and its quality control.Radix gentianae from different producing areas have the largest diversities,and the diversities embodied in the content of chloroform phase compounds.
文摘目的比较不同灭菌方法对龙胆和秦艽化学成分的影响和对微生物的杀灭效果。方法采用干热灭菌、湿热灭菌和钴60辐照灭菌3种方法对3批龙胆和3批秦艽药材粉末进行灭菌处理,采用液相指纹图谱方法分析龙胆、秦艽中的化学成分。按照《中国药典》2015年版微生物限度检查方法检测了不同灭菌处理样品的需氧菌总数、霉菌酵母菌计数、耐胆盐革兰阴性菌、大肠埃希菌。结果主要成分龙胆苦苷对辐照敏感,3 k Gy的小剂量辐照即可使龙胆苦苷减少15%或18%;干热灭菌或湿热灭菌使龙胆和秦艽在7.8 min处新增1小峰(峰4);龙胆中峰1经辐照后有所增加,但该物质并不是辐照产生的特异成分,干热灭菌后该峰也有所增加。3种灭菌方法均呈现出随着灭菌强度的增大,发生变化的峰的数量和变化率都有所增加的规律。灭菌效果简单排序为辐照>湿热>干热。结论干热、湿热、辐照3种方式灭菌处理对龙胆、秦艽成分除龙胆苦苷外影响不大,部分成分有增加或减少,其中湿热(115℃、15 min)和辐照25 k Gy处理对成分的影响最大。辐照虽然灭菌效果良好,但是对主要成分龙胆苦苷的影响显著,故不适宜用于龙胆和秦艽的灭菌处理。
