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
HPLC fingerprinting and quantification of gentiopicroside（GPS） and loganic acid（LA） in Gentianae Macrophyllae Radix（GMR） crude drugs were developed in this study.The samples were separated on Zorbax SB-C_（18） ...HPLC fingerprinting and quantification of gentiopicroside（GPS） and loganic acid（LA） in Gentianae Macrophyllae Radix（GMR） crude drugs were developed in this study.The samples were separated on Zorbax SB-C_（18） column（250 mm×4.6 mm, 5μm） with a linear gradient of acetonitrile and 0.04%phosphoric acid.The HPLC flow rate was 1.0 mL/min and a UV absorption was measured at 230 nm.An orthogonal L9（3^4） test was applied for the optimization of sample extraction conditions,and an aliquot of GMR sample（g） was extracted with 15-fold of 50%ethanol（mL） for 30 min by sonication.Quantitative analysis showed that the content of GPS（14.05 mg/g-74.61 mg/g） in all samples was obviously higher than that of LA（1.13 mg/g-40.46 mg/g）. Based on the content ratio of GPS over LA（1.8-11.4）,samples originated from Gentiana macrophylla（with content ratio of GPS over LA≤4.3） could be distinguished from those from G.dahurica and G.dahurica var.gracilipes（with content ratio of GPS over LA≥4.8）.The principle components analysis of the HPLC fingerprints showed that samples originated from G.macrophylla and G.dahurica（including G.dahurica var.gracilipes） could be divided into two groups.This established HPLC-DAD method could be efficiently used for the species identification and quality control of GMR crude drugs.展开更多
Gentianae Radix et Rhizoma(also called "Longdan" in Chinese)is commonly used for eliminating damp-heat and quenching the fire of liver and gall bladder in traditional Chinese medicine.In this study,a novel a...Gentianae Radix et Rhizoma(also called "Longdan" in Chinese)is commonly used for eliminating damp-heat and quenching the fire of liver and gall bladder in traditional Chinese medicine.In this study,a novel and reliable method using high-performance liquid chromatography(HPLC)was developed both for quantitative analysis of four bioactive compounds(loganic acid,swertiamarin,gentiopicroside and sweroside)and chemical fingerprint analysis of "Longdan".In quantitative analysis,four compounds showed good regressions(R^(2)>40.9987)within the test ranges and the recovery of the method was in the range 97.61-102.49%.In fingerprint analysis,ten characteristic peaks were selected to evaluate the similarities of the crude drugs,and the HPLC chromatograms of twenty samples from different regions of China showed similar patterns.The results demonstrated that the combination of the quantitative and chromatographic fingerprint analyses offered an efficient way to evaluate the quality consistency of Gentianae Radix et Rhizoma.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
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.
基金Research Program of Science Technology of Traditional Chinese Medicine(TCM) Sponsored by the State Administration of TCM of China(Grant No.04-05ZL01)
文摘HPLC fingerprinting and quantification of gentiopicroside（GPS） and loganic acid（LA） in Gentianae Macrophyllae Radix（GMR） crude drugs were developed in this study.The samples were separated on Zorbax SB-C_（18） column（250 mm×4.6 mm, 5μm） with a linear gradient of acetonitrile and 0.04%phosphoric acid.The HPLC flow rate was 1.0 mL/min and a UV absorption was measured at 230 nm.An orthogonal L9（3^4） test was applied for the optimization of sample extraction conditions,and an aliquot of GMR sample（g） was extracted with 15-fold of 50%ethanol（mL） for 30 min by sonication.Quantitative analysis showed that the content of GPS（14.05 mg/g-74.61 mg/g） in all samples was obviously higher than that of LA（1.13 mg/g-40.46 mg/g）. Based on the content ratio of GPS over LA（1.8-11.4）,samples originated from Gentiana macrophylla（with content ratio of GPS over LA≤4.3） could be distinguished from those from G.dahurica and G.dahurica var.gracilipes（with content ratio of GPS over LA≥4.8）.The principle components analysis of the HPLC fingerprints showed that samples originated from G.macrophylla and G.dahurica（including G.dahurica var.gracilipes） could be divided into two groups.This established HPLC-DAD method could be efficiently used for the species identification and quality control of GMR crude drugs.
基金supported by Key National Science Foundation of China(81130069)Beijing Tongrentang Co.Ltd.(D08080203640901)the State Key Laboratory of Traditional Chinese Medicine Resources Research and Development in Chendu University of TCM。
文摘Gentianae Radix et Rhizoma(also called "Longdan" in Chinese)is commonly used for eliminating damp-heat and quenching the fire of liver and gall bladder in traditional Chinese medicine.In this study,a novel and reliable method using high-performance liquid chromatography(HPLC)was developed both for quantitative analysis of four bioactive compounds(loganic acid,swertiamarin,gentiopicroside and sweroside)and chemical fingerprint analysis of "Longdan".In quantitative analysis,four compounds showed good regressions(R^(2)>40.9987)within the test ranges and the recovery of the method was in the range 97.61-102.49%.In fingerprint analysis,ten characteristic peaks were selected to evaluate the similarities of the crude drugs,and the HPLC chromatograms of twenty samples from different regions of China showed similar patterns.The results demonstrated that the combination of the quantitative and chromatographic fingerprint analyses offered an efficient way to evaluate the quality consistency of Gentianae Radix et Rhizoma.
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.
