Ziziphi Spinosae Semen(ZSS) has been used for treatment of insomnia in China for centuries. To reveal the influence of insomnia on the levels of the neurotransmitters including serotonin(5-HT), glutamic acid(Glu),γ-a...Ziziphi Spinosae Semen(ZSS) has been used for treatment of insomnia in China for centuries. To reveal the influence of insomnia on the levels of the neurotransmitters including serotonin(5-HT), glutamic acid(Glu),γ-aminobutyric acid(GABA),noradrenaline(NE) and dopamine(DA), and to study the role of ZSS aqueous extract in the treatment of insomnia, an UPLC-ESIMS/MS method was developed and validated for simultaneous determination of five neurotransmitters in the rat brain. The brain samples were pretreated by one-step direct protein precipitation with acetonitrile. The analytes were detected in positive mode with multiple reaction monitoring(MRM) and the procedure was completed in less than 10 min. The method showed a good linearity(R^2 >0.9967) with the other validation parameters were within acceptance range. The results indicated that the concentration of 5-HT, GABA and DA is significantly lower(P < 0.01) in para-chlorophenylalanine(PCPA)-induced insomnia rat model group, while Glu and NE significantly higher than those in control group(P < 0.01). Treatment with ZSS aqueous extract(4 or 8 g·kg^1·d^-1 for seven days) could ameliorate the symptoms of insomnia by significantly changing the levels of the neurotransmitter parameters mentioned above. The data obtained in this study demonstrate that ZSS aqueous extract could ameliorate the symptoms of insomnia by modulating the levels of monoamines and amino acid neurotransmitters in the brain.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
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
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
Conventional quantity determination of Chinese herbal medicines by High-Performance Thin-Layer Chromatography (HPTLC) must be carried out on a TLC scanner for measuring the marker component on the HPTLC plate. General...Conventional quantity determination of Chinese herbal medicines by High-Performance Thin-Layer Chromatography (HPTLC) must be carried out on a TLC scanner for measuring the marker component on the HPTLC plate. Generally the HPTLC image of the herbal drug always takes the photograph as the permanent visual document. Considering the photo of the HPTLC image preserving all the visual quality-related parameters, there might be an alternative cost/effective approach worth studying. It would be presumably applicable to use the exclusively developed software as a virtual scanner instead of the TLC Scanner instrument to do digital scanning, integrating and calculating target components in the photo of the HPTLC image. It means digitally scanning the photo of the HPTLC image solely with software does the same thing as the instrumental scanner does. Based on this protocol, we primarily conducted the assay of the contents of the major saponins, jujuboside a and jujuboside b, in Ziziphi Spinosae Semen (Suan-Zao-Ren). Using home-developed software “digiscan” for acquiring the raw data and integrating the peaks area of the target components with “Origin Pro”, the contents of jujuboside a and jujuboside b can then be calculated respectively. The primary investigated experiment results were well compatible with official HPLC-ELSD method in Chinese Pharmacopoeia. The methodology validation and comparative analysis with HPLC confirmed that the unprecedented method is acceptable. The advantage of the recommended approach is far more simple, flexible and cost/effective than the conventional method, and it can be done at anytime and anywhere once only if the high quality HPTLC image photo is available. There would be a bright perspective for application to other herbal medicines. And the one-stop software for merging digital scanning, integration and calculation together is in progress.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
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 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supported by the Natural Science Foundation of State(Nos.81603289 and 81603251)the Base Program of the Joint Training Talents of Shanxi Province(No.2017JD01)the Key Laboratory of Effective Substances Research and Utilization in TCM of Shanxi province(No.201605D111004)
文摘Ziziphi Spinosae Semen(ZSS) has been used for treatment of insomnia in China for centuries. To reveal the influence of insomnia on the levels of the neurotransmitters including serotonin(5-HT), glutamic acid(Glu),γ-aminobutyric acid(GABA),noradrenaline(NE) and dopamine(DA), and to study the role of ZSS aqueous extract in the treatment of insomnia, an UPLC-ESIMS/MS method was developed and validated for simultaneous determination of five neurotransmitters in the rat brain. The brain samples were pretreated by one-step direct protein precipitation with acetonitrile. The analytes were detected in positive mode with multiple reaction monitoring(MRM) and the procedure was completed in less than 10 min. The method showed a good linearity(R^2 >0.9967) with the other validation parameters were within acceptance range. The results indicated that the concentration of 5-HT, GABA and DA is significantly lower(P < 0.01) in para-chlorophenylalanine(PCPA)-induced insomnia rat model group, while Glu and NE significantly higher than those in control group(P < 0.01). Treatment with ZSS aqueous extract(4 or 8 g·kg^1·d^-1 for seven days) could ameliorate the symptoms of insomnia by significantly changing the levels of the neurotransmitter parameters mentioned above. The data obtained in this study demonstrate that ZSS aqueous extract could ameliorate the symptoms of insomnia by modulating the levels of monoamines and amino acid neurotransmitters in the brain.
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.
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.
文摘Conventional quantity determination of Chinese herbal medicines by High-Performance Thin-Layer Chromatography (HPTLC) must be carried out on a TLC scanner for measuring the marker component on the HPTLC plate. Generally the HPTLC image of the herbal drug always takes the photograph as the permanent visual document. Considering the photo of the HPTLC image preserving all the visual quality-related parameters, there might be an alternative cost/effective approach worth studying. It would be presumably applicable to use the exclusively developed software as a virtual scanner instead of the TLC Scanner instrument to do digital scanning, integrating and calculating target components in the photo of the HPTLC image. It means digitally scanning the photo of the HPTLC image solely with software does the same thing as the instrumental scanner does. Based on this protocol, we primarily conducted the assay of the contents of the major saponins, jujuboside a and jujuboside b, in Ziziphi Spinosae Semen (Suan-Zao-Ren). Using home-developed software “digiscan” for acquiring the raw data and integrating the peaks area of the target components with “Origin Pro”, the contents of jujuboside a and jujuboside b can then be calculated respectively. The primary investigated experiment results were well compatible with official HPLC-ELSD method in Chinese Pharmacopoeia. The methodology validation and comparative analysis with HPLC confirmed that the unprecedented method is acceptable. The advantage of the recommended approach is far more simple, flexible and cost/effective than the conventional method, and it can be done at anytime and anywhere once only if the high quality HPTLC image photo is available. There would be a bright perspective for application to other herbal medicines. And the one-stop software for merging digital scanning, integration and calculation together is in progress.
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
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.
