Ziziphi Spinosae Semen(ZSS),a traditional Chinese medicine,is used in clinics for the treatment of insomnia in China and other Asian countries.Herein,we described for the first time a comparative pharmacokinetics stud...Ziziphi Spinosae Semen(ZSS),a traditional Chinese medicine,is used in clinics for the treatment of insomnia in China and other Asian countries.Herein,we described for the first time a comparative pharmacokinetics study of the six major compounds of ZSS in normal control(NC)and para-chlorophenylalanine(PCPA)-induced insomnia model(IM)rats that were orally administered the aqueous extract of ZSS.An ultra-high-performance liquid chromatography coupled with quadrupole orbitrap mass(UHPLC-Q-Orbitrap-MS)method was developed and validated for the simultaneous determination of coclaurine,magnoflorine,spinosin,6000-feruloylspinosin,jujuboside A(JuA),and jujuboside B(JuB)in ZSS in rat plasma.The established approach was successfully applied to a comparative pharmacokinetic study.The systemic exposures of spinosin and 6000-feruloylspinosin were decreased in the IM group compared to the NC group,while plasma clearance(CL)was significantly increased.The Tmax values of JuA and JuB in IM rats were significantly lower than those in NC rats.The T1/2 of JuA in the IM group was significantly accelerated.The pharmacokinetic parameters of coclaurine and magnoflorine were not evidently affected between the two groups.These results indicate that the pathological state of insomnia altered the plasma pharmacokinetics of spinosin,6000-feruloylspinosin,JuA,and JuB in the ZSS aqueous extract,providing an experimental basis for the role of ZSS in insomnia treatment.The comparative pharmacokinetics-based UHPLC-Q-Orbitrap-MS using full-scan mode can therefore provide a reliable and suitable means for the screening of potentially effective substances applied as quality markers of ZSS.展开更多
Due to the scarcity of resources of Ziziphi spinosae semen (ZSS), many inferior goods and even adulterants are generally found in medicine markets. To strengthen the quality control, HPLC fingerprint common pattern ...Due to the scarcity of resources of Ziziphi spinosae semen (ZSS), many inferior goods and even adulterants are generally found in medicine markets. To strengthen the quality control, HPLC fingerprint common pattern established in this paper showed three main bioactive compounds in one chromatogram simultaneously. Principal component analysis based on DAD signals could discriminate adulterants and inferiorities. Principal component analysis indicated that all samples could be mainly regrouped into two main clusters according to the first principal component (PC1, redefined as Vicenin II) and the second principal component (PC2, redefined as zizyphusine). PC1 and PC2 could explain 91.42%of the variance. Content of zizyphusine fluctuated more greatly than that of spinosin, and this result was also confirmed by the HPTLC result. Samples with low content of jujubosides and two common adulterants could not be used equivalently with authenticated ones in clinic, while one reference standard extract could substitute the crude drug in pharmaceutical production. Giving special consideration to the well-known bioactive saponins but with low response by end absorption, a fast and cheap HPTLC method for quality control of ZSS was developed and the result obtained was commensurate well with that of HPLC analysis. Samples having similar fingerprints to HPTLC common pattern targeting at saponins could be regarded as authenticated ones. This work provided a faster and cheaper way for quality control of ZSS and laid foundation for establishing a more effective quality control method for ZSS.展开更多
A simple, reproducible and reliable HPLC fingerprint analysis method of Ziziphi Spinosae Semen(ZSS) and Ziziphi Mauritianae Semen(ZMS) was established, and then it was applied to analyze samples collected from differe...A simple, reproducible and reliable HPLC fingerprint analysis method of Ziziphi Spinosae Semen(ZSS) and Ziziphi Mauritianae Semen(ZMS) was established, and then it was applied to analyze samples collected from different locations. A total of 12 common fingerprint peaks were designated in ZSS and ZMS, respectively, and five of which were definitely identified. Of these common peaks, 10 peaks were presented in both ZSS and ZMS, whereas two were characteristic for ZSS and ZMS, respectively. Furthermore, the two characteristic peaks of ZMS were definitely assigned as 6′′′-sinapoylspinosin and frangufoline by HPLC guided purification and structural elucidation. The HPLC fingerprint analysis was proved to be useful in identifying and discriminating ZSS and ZMS, which was beneficial for quality control of ZSS and its products.展开更多
A sensitive,fast and comprehensive method for the quality assessment of Semen Ziziphi Spinosae(SZS)standard decoction with characterization of its chemical components was developed and validated.UPLC-Q/TOF-MS/MS syste...A sensitive,fast and comprehensive method for the quality assessment of Semen Ziziphi Spinosae(SZS)standard decoction with characterization of its chemical components was developed and validated.UPLC-Q/TOF-MS/MS system was used to identify thirty-six chemical components of SZS standard decoction which included nucleosides,phenolic acids,alkaloids,and flavonoids.Furthermore,a UPLC-PDA method was validated to simultaneously determine adenosine,protocatechuic acid,magnoflorine,catechin,protocatechin,vicenin II,spinosin,kaempferol-3-rutinoside,and 6'''-feruloylspinosin which represent four species of characteristic compounds.The qualitative method had been validated according to Chinese Pharmacopoeia(2015 edition)in terms of lineary,repeatability,recovery and stability for all analytes,with the results showing good precision,accuracy and stability.In conclusion,the method using UPLC combined with MS and PDA provided a novel way for the standardization and identification of SZS standard decoction,and also offered a basis for qualitative analysis and quality assessment of the preparations for SZS standard decoction.展开更多
Background:Sleep is essential for maintaining human health,and insomnia is a widespread problem.Traditional Chinese medicine(TCM)has been used for centuries to treat sleep disorders,with fewer reported side effects co...Background:Sleep is essential for maintaining human health,and insomnia is a widespread problem.Traditional Chinese medicine(TCM)has been used for centuries to treat sleep disorders,with fewer reported side effects compared to conventional treatments.Objective:This study seeks to investigate the sleep-promoting effects of the GSZ formula,which comprisesγ-aminobutyric acid(GABA),Schisandrae Chinensis Fructus(Wuweizi in Chinese),and Ziziphi Spinosae Semen(Suanzaoren in Chinese).In addition,this study aims to explore the active ingredients and potential mechanisms underlying the sleep-enhancing effects of the formula.Methods:The impact of GSZ on sleep was evaluated using two models,the complete sleep model and the sub-threshold sleep model.Mice were randomly divided into five groups and orally administered GSZ solution(0.33 g/kg/day or 0.99 g/kg/day),positive drug diazepam(2.50 mg/kg)or a control solution for 30 days.Hypno-sis model was established in mice using pentobarbital sodium.Sleep duration and incidence were measured by recording when the righting reflex of mice disappeared for more than 1 min.GABA and dopamine(DA)levels in mouse brain tissue were measured using ELISA kits.The ingredients of the GSZ formula were identified using mass spectrometry,and the targets of these ingredients and disease-related genes were retrieved from public databases.A network medicine approach was used to calculate the shortest path between ingredient targets and disease-related proteins.The expression levels of potential proteins,such as Akt,p-Akt,GSK-3β,and p-GSK-3β,were analyzed using Western blotting based on the predicted results.Results:GSZ significantly prolonged sleep duration and enhanced the sleep rate in mice(P<0.05).Furthermore,it elevated GABA levels and reduced DA levels in the mouse brain(P<0.05).Network medicine analysis suggested that GABA,stearic acid,genistin,and coumestrol may be the most crucial active ingredients for sleep improve-ment.Western blotting analysis demonstrated that GSZ modulated the protein expression levels of p-Akt/Akt and p-GSK-3β/GSK-3β(P<0.05).Conclusion:Our study demonstrated that the GSZ formula could improve sleep,with key ingredients likely being GABA,stearic acid,genistin,and coumestrol.The mechanism might involve the regulation of the Akt/GSK-3βpathway,as revealed by the network medicine analysis and experimental validation.Our current new findings shed light on the potential mechanisms underlying the sleep-enhancing effects of the GSZ formula,which could provide experimental evidence to develop innovative treatments for insomnia.展开更多
The holistic characterization and quality control of all the medicinal herbs of proprietary Chinese medicines(PCMs)are of great significance to ensure their safety,efficacy,and consistency.Thin-layer chromatography(TL...The holistic characterization and quality control of all the medicinal herbs of proprietary Chinese medicines(PCMs)are of great significance to ensure their safety,efficacy,and consistency.Thin-layer chromatography(TLC),a simple and classic approach for qualitatively characterizing and examining quality markers of natural products,has been widely used in the characterization and quality control of traditional Chinese medicines.Zaoren Anshen(ZRAS)capsule,prepared from three medicinal herbs of fried Ziziphi Spinosae Semen,Salvia Miltiorrhiza Radix et Rhizoma,and vinegar-processed Schisandrae Chinensis Fructus,is a famous PCM in China for the treatment of insomnia,amnesia,and dizziness in clinical practice.However,no effective method is available so far for simultaneous identification and examination of all the three medicinal herbs of ZRAS capsule.In the present study,we developed a TLC method via twice-development and visualization by UV light or chromogenic agent,which could be used for simultaneous qualitative identification of all the three medicinal herbs of ZRAS capsule in one plate.Moreover,the sample preparation method was optimized.The developed TLC method was rapid,simple,low-cost,and effective,and thus it could be used for quality control of ZRAS capsule.展开更多
基金This work was supported by grants from the National Natural Science Foundation of China(No:81603289,81603251)Key Laboratory of Effective Substances Research and Utilization in TCM of Shanxi province(No:201605D111004)and Key Technology Research Zhen Dong Special Project from Shanxi Science and Technology Department(No:2016ZD0105).
文摘Ziziphi Spinosae Semen(ZSS),a traditional Chinese medicine,is used in clinics for the treatment of insomnia in China and other Asian countries.Herein,we described for the first time a comparative pharmacokinetics study of the six major compounds of ZSS in normal control(NC)and para-chlorophenylalanine(PCPA)-induced insomnia model(IM)rats that were orally administered the aqueous extract of ZSS.An ultra-high-performance liquid chromatography coupled with quadrupole orbitrap mass(UHPLC-Q-Orbitrap-MS)method was developed and validated for the simultaneous determination of coclaurine,magnoflorine,spinosin,6000-feruloylspinosin,jujuboside A(JuA),and jujuboside B(JuB)in ZSS in rat plasma.The established approach was successfully applied to a comparative pharmacokinetic study.The systemic exposures of spinosin and 6000-feruloylspinosin were decreased in the IM group compared to the NC group,while plasma clearance(CL)was significantly increased.The Tmax values of JuA and JuB in IM rats were significantly lower than those in NC rats.The T1/2 of JuA in the IM group was significantly accelerated.The pharmacokinetic parameters of coclaurine and magnoflorine were not evidently affected between the two groups.These results indicate that the pathological state of insomnia altered the plasma pharmacokinetics of spinosin,6000-feruloylspinosin,JuA,and JuB in the ZSS aqueous extract,providing an experimental basis for the role of ZSS in insomnia treatment.The comparative pharmacokinetics-based UHPLC-Q-Orbitrap-MS using full-scan mode can therefore provide a reliable and suitable means for the screening of potentially effective substances applied as quality markers of ZSS.
文摘Due to the scarcity of resources of Ziziphi spinosae semen (ZSS), many inferior goods and even adulterants are generally found in medicine markets. To strengthen the quality control, HPLC fingerprint common pattern established in this paper showed three main bioactive compounds in one chromatogram simultaneously. Principal component analysis based on DAD signals could discriminate adulterants and inferiorities. Principal component analysis indicated that all samples could be mainly regrouped into two main clusters according to the first principal component (PC1, redefined as Vicenin II) and the second principal component (PC2, redefined as zizyphusine). PC1 and PC2 could explain 91.42%of the variance. Content of zizyphusine fluctuated more greatly than that of spinosin, and this result was also confirmed by the HPTLC result. Samples with low content of jujubosides and two common adulterants could not be used equivalently with authenticated ones in clinic, while one reference standard extract could substitute the crude drug in pharmaceutical production. Giving special consideration to the well-known bioactive saponins but with low response by end absorption, a fast and cheap HPTLC method for quality control of ZSS was developed and the result obtained was commensurate well with that of HPLC analysis. Samples having similar fingerprints to HPTLC common pattern targeting at saponins could be regarded as authenticated ones. This work provided a faster and cheaper way for quality control of ZSS and laid foundation for establishing a more effective quality control method for ZSS.
基金National Project for Standardization of Chinese Material Medica(Grant No.ZYBZH-Y-JIN-34).
文摘A simple, reproducible and reliable HPLC fingerprint analysis method of Ziziphi Spinosae Semen(ZSS) and Ziziphi Mauritianae Semen(ZMS) was established, and then it was applied to analyze samples collected from different locations. A total of 12 common fingerprint peaks were designated in ZSS and ZMS, respectively, and five of which were definitely identified. Of these common peaks, 10 peaks were presented in both ZSS and ZMS, whereas two were characteristic for ZSS and ZMS, respectively. Furthermore, the two characteristic peaks of ZMS were definitely assigned as 6′′′-sinapoylspinosin and frangufoline by HPLC guided purification and structural elucidation. The HPLC fingerprint analysis was proved to be useful in identifying and discriminating ZSS and ZMS, which was beneficial for quality control of ZSS and its products.
基金financially supported by the National Key Scientific Project for China New Drug Discovery and Twelfth Five-Year Plan-preclinical study on Suanzaoren granules(No.2014ZX09304306)the Key Technologies of Common Quality Evaluation of New Drugs(Grant no.2015010201)
文摘A sensitive,fast and comprehensive method for the quality assessment of Semen Ziziphi Spinosae(SZS)standard decoction with characterization of its chemical components was developed and validated.UPLC-Q/TOF-MS/MS system was used to identify thirty-six chemical components of SZS standard decoction which included nucleosides,phenolic acids,alkaloids,and flavonoids.Furthermore,a UPLC-PDA method was validated to simultaneously determine adenosine,protocatechuic acid,magnoflorine,catechin,protocatechin,vicenin II,spinosin,kaempferol-3-rutinoside,and 6'''-feruloylspinosin which represent four species of characteristic compounds.The qualitative method had been validated according to Chinese Pharmacopoeia(2015 edition)in terms of lineary,repeatability,recovery and stability for all analytes,with the results showing good precision,accuracy and stability.In conclusion,the method using UPLC combined with MS and PDA provided a novel way for the standardization and identification of SZS standard decoction,and also offered a basis for qualitative analysis and quality assessment of the preparations for SZS standard decoction.
基金supported by grants from the PhD Start-up Fund of Zunyi Medical University(F-ZH-012)Zunyi Medical University In-novation and Entrepreneurship Training Program(ZHCX2022078)for College Students.
文摘Background:Sleep is essential for maintaining human health,and insomnia is a widespread problem.Traditional Chinese medicine(TCM)has been used for centuries to treat sleep disorders,with fewer reported side effects compared to conventional treatments.Objective:This study seeks to investigate the sleep-promoting effects of the GSZ formula,which comprisesγ-aminobutyric acid(GABA),Schisandrae Chinensis Fructus(Wuweizi in Chinese),and Ziziphi Spinosae Semen(Suanzaoren in Chinese).In addition,this study aims to explore the active ingredients and potential mechanisms underlying the sleep-enhancing effects of the formula.Methods:The impact of GSZ on sleep was evaluated using two models,the complete sleep model and the sub-threshold sleep model.Mice were randomly divided into five groups and orally administered GSZ solution(0.33 g/kg/day or 0.99 g/kg/day),positive drug diazepam(2.50 mg/kg)or a control solution for 30 days.Hypno-sis model was established in mice using pentobarbital sodium.Sleep duration and incidence were measured by recording when the righting reflex of mice disappeared for more than 1 min.GABA and dopamine(DA)levels in mouse brain tissue were measured using ELISA kits.The ingredients of the GSZ formula were identified using mass spectrometry,and the targets of these ingredients and disease-related genes were retrieved from public databases.A network medicine approach was used to calculate the shortest path between ingredient targets and disease-related proteins.The expression levels of potential proteins,such as Akt,p-Akt,GSK-3β,and p-GSK-3β,were analyzed using Western blotting based on the predicted results.Results:GSZ significantly prolonged sleep duration and enhanced the sleep rate in mice(P<0.05).Furthermore,it elevated GABA levels and reduced DA levels in the mouse brain(P<0.05).Network medicine analysis suggested that GABA,stearic acid,genistin,and coumestrol may be the most crucial active ingredients for sleep improve-ment.Western blotting analysis demonstrated that GSZ modulated the protein expression levels of p-Akt/Akt and p-GSK-3β/GSK-3β(P<0.05).Conclusion:Our study demonstrated that the GSZ formula could improve sleep,with key ingredients likely being GABA,stearic acid,genistin,and coumestrol.The mechanism might involve the regulation of the Akt/GSK-3βpathway,as revealed by the network medicine analysis and experimental validation.Our current new findings shed light on the potential mechanisms underlying the sleep-enhancing effects of the GSZ formula,which could provide experimental evidence to develop innovative treatments for insomnia.
基金National Key Research and Development Program of China(Grant No.2018YFC1707300)。
文摘The holistic characterization and quality control of all the medicinal herbs of proprietary Chinese medicines(PCMs)are of great significance to ensure their safety,efficacy,and consistency.Thin-layer chromatography(TLC),a simple and classic approach for qualitatively characterizing and examining quality markers of natural products,has been widely used in the characterization and quality control of traditional Chinese medicines.Zaoren Anshen(ZRAS)capsule,prepared from three medicinal herbs of fried Ziziphi Spinosae Semen,Salvia Miltiorrhiza Radix et Rhizoma,and vinegar-processed Schisandrae Chinensis Fructus,is a famous PCM in China for the treatment of insomnia,amnesia,and dizziness in clinical practice.However,no effective method is available so far for simultaneous identification and examination of all the three medicinal herbs of ZRAS capsule.In the present study,we developed a TLC method via twice-development and visualization by UV light or chromogenic agent,which could be used for simultaneous qualitative identification of all the three medicinal herbs of ZRAS capsule in one plate.Moreover,the sample preparation method was optimized.The developed TLC method was rapid,simple,low-cost,and effective,and thus it could be used for quality control of ZRAS capsule.