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
[Objectives] In order to develop Triplostegia glandulifera Wall.ex DC. into foromula granules, a high performance liquid chromatography (HPLC) determination method and a TLC identification method for loganic acid in d...[Objectives] In order to develop Triplostegia glandulifera Wall.ex DC. into foromula granules, a high performance liquid chromatography (HPLC) determination method and a TLC identification method for loganic acid in decoction pieces, standard decoction and formula granules of T. glandulifera were established, and the quantity value transfer relationship of T. glandulifera pieces-standard decoction-formula granules was investigated.[Methods] An Agilent 1260 II high performance liquid chromatograph and a Waters Symmetry C 18 column (4.6 mm×250 mm, 5 μm) were used to perform gradient elution with acetonitrile-0.1%phosphoric acid solution as the mobile phase at a column temperature of 30 ℃ and a flow rate of 1 mL/min, and the detection wavelength was 240 nm. The TLC identification method of T. glandulifera was established using ethyl acetate-methanol-water (10 : 2 : 1) as the developer, and examination was carried out under a UV lamp (254 nm). The quantity value transfer law was analyzed by using the extract yield, the content of loganic acid and TLC chromatograms as the main evaluation indexes.[Results] The method for the determination and identification of loganic acid is stable, reproducible and reliable. The average yield, average loganic acid content and average loganic acid content transfer rate of 3 batches of T. glandulifera formula granules were, respectively, 18.7%, 41.1 mg/g and 43.1%, each of which was within corresponding range of mean±3SD of the 15 batches of standard decoction.[Conclusions] The content determination and TLC identification methods of loganic acid can be used to evaluate the quality of T. glandulifera formula granules. This study provides data basis for further research on T. glandulifera formula granules, and promotes the modernization of medicines for ethnic minorities.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
目的在遵循人用药品注册技术要求国际协调会(International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use,ICH)制剂稳定性指导原则的指导下,在采用超高效液相色谱-飞行...目的在遵循人用药品注册技术要求国际协调会(International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use,ICH)制剂稳定性指导原则的指导下,在采用超高效液相色谱-飞行时间质谱(UPLC-Q/TOF-MS)联用技术对中药传统水蜜丸制剂二至丸进行多成分定性定量研究的基础上,进行二至丸长期稳定性和加速稳定性研究,并考察了制剂影响因素对制剂稳定性的影响。方法采用Acquity UPLC BEH C18色谱柱(100 mm×2.1mm,1.7μm),流动相为0.1%甲酸水溶液-乙腈,梯度洗脱,体积流量0.2 m L/min,采用电喷雾电离源(ESI),正、负离子模式扫描。在遵循ICH制剂稳定性指导原则的指导下,进行了18个月的长期稳定性试验[温度(25±2)℃,相对湿度(RH)(60±5)%],加速稳定性试验[温度(40±2)℃,RH(75±5)%]和制剂影响因素试验(包装材料类型和密封条件)。结果采用以上方法共鉴定出二至丸中20个化学成分(红景天苷、蟛蜞菊内酯、10-羟基木犀榄苷二甲酯、木犀榄苷-11甲酯、马钱苷酸、松果菊苷、女贞苦苷、橄榄苦苷酸、毛蕊花苷/异毛蕊花苷、女贞苷、特女贞苷、ligustroflavone、特女贞苷异构体、阿富汗丁香苷F、异女贞苷、女贞酸、橄榄苦苷、女贞苷-G13、青藤碱、3-O-顺式-对-香豆酰委陵菜酸),并对其进行定量及归一化半定量研究。在长期稳定性研究中,20个成分均保持稳定;然而在加速稳定性试验条件下,11种成分(10-羟基木犀榄苷二甲酯、松果菊苷、女贞苦苷、橄榄苦苷酸、毛蕊花苷、女贞苷、特女贞苷、ligustroflavone、女贞酸、女贞苷-G13、青藤碱)的量明显下降。制剂影响因素试验结果显示采用钠钙玻璃包装材料替代现有市售聚酯包装,在密封或敞口条件下,制剂中各成分稳定性均显著提高。在采用相同包材时密封条件对制剂稳定性也存在显著性影响。结论通过较为系统的中药传统展开更多
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.
基金Supported by Yunnan Provincial Science and Technology Major Project(202102AA310027).
文摘[Objectives] In order to develop Triplostegia glandulifera Wall.ex DC. into foromula granules, a high performance liquid chromatography (HPLC) determination method and a TLC identification method for loganic acid in decoction pieces, standard decoction and formula granules of T. glandulifera were established, and the quantity value transfer relationship of T. glandulifera pieces-standard decoction-formula granules was investigated.[Methods] An Agilent 1260 II high performance liquid chromatograph and a Waters Symmetry C 18 column (4.6 mm×250 mm, 5 μm) were used to perform gradient elution with acetonitrile-0.1%phosphoric acid solution as the mobile phase at a column temperature of 30 ℃ and a flow rate of 1 mL/min, and the detection wavelength was 240 nm. The TLC identification method of T. glandulifera was established using ethyl acetate-methanol-water (10 : 2 : 1) as the developer, and examination was carried out under a UV lamp (254 nm). The quantity value transfer law was analyzed by using the extract yield, the content of loganic acid and TLC chromatograms as the main evaluation indexes.[Results] The method for the determination and identification of loganic acid is stable, reproducible and reliable. The average yield, average loganic acid content and average loganic acid content transfer rate of 3 batches of T. glandulifera formula granules were, respectively, 18.7%, 41.1 mg/g and 43.1%, each of which was within corresponding range of mean±3SD of the 15 batches of standard decoction.[Conclusions] The content determination and TLC identification methods of loganic acid can be used to evaluate the quality of T. glandulifera formula granules. This study provides data basis for further research on T. glandulifera formula granules, and promotes the modernization of medicines for ethnic minorities.
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.
文摘目的在遵循人用药品注册技术要求国际协调会(International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use,ICH)制剂稳定性指导原则的指导下,在采用超高效液相色谱-飞行时间质谱(UPLC-Q/TOF-MS)联用技术对中药传统水蜜丸制剂二至丸进行多成分定性定量研究的基础上,进行二至丸长期稳定性和加速稳定性研究,并考察了制剂影响因素对制剂稳定性的影响。方法采用Acquity UPLC BEH C18色谱柱(100 mm×2.1mm,1.7μm),流动相为0.1%甲酸水溶液-乙腈,梯度洗脱,体积流量0.2 m L/min,采用电喷雾电离源(ESI),正、负离子模式扫描。在遵循ICH制剂稳定性指导原则的指导下,进行了18个月的长期稳定性试验[温度(25±2)℃,相对湿度(RH)(60±5)%],加速稳定性试验[温度(40±2)℃,RH(75±5)%]和制剂影响因素试验(包装材料类型和密封条件)。结果采用以上方法共鉴定出二至丸中20个化学成分(红景天苷、蟛蜞菊内酯、10-羟基木犀榄苷二甲酯、木犀榄苷-11甲酯、马钱苷酸、松果菊苷、女贞苦苷、橄榄苦苷酸、毛蕊花苷/异毛蕊花苷、女贞苷、特女贞苷、ligustroflavone、特女贞苷异构体、阿富汗丁香苷F、异女贞苷、女贞酸、橄榄苦苷、女贞苷-G13、青藤碱、3-O-顺式-对-香豆酰委陵菜酸),并对其进行定量及归一化半定量研究。在长期稳定性研究中,20个成分均保持稳定;然而在加速稳定性试验条件下,11种成分(10-羟基木犀榄苷二甲酯、松果菊苷、女贞苦苷、橄榄苦苷酸、毛蕊花苷、女贞苷、特女贞苷、ligustroflavone、女贞酸、女贞苷-G13、青藤碱)的量明显下降。制剂影响因素试验结果显示采用钠钙玻璃包装材料替代现有市售聚酯包装,在密封或敞口条件下,制剂中各成分稳定性均显著提高。在采用相同包材时密封条件对制剂稳定性也存在显著性影响。结论通过较为系统的中药传统
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.
