ABSTRACT
In this study, high-performance liquid chromatography was used to determine four components of Shaoyao Gancao Decoction (SGD), and the effect of purification was evaluated using fingerprints, similarity analysis and cell experiments. An effective method for isolation and purification of SGD was established. The adsorption/desorption properties of SGD were evaluated using resin screening, isothermal analysis, adsorption kinetics, and dynamic adsorption-desorption experiments. It was shown that the Langmuir equation fitted the isotherm data well and that a pseudo-second-order model accurately described kinetic adsorption on AB-8 resin. Analysis of thermodynamic parameters showed that the adsorption process was exothermic. Under the optimal process conditions, the concentrations of albiflorin, paeoniflorin, liquiritin and ammonium glycyrrhizinate in the product were 73.05, 134.04, 45.04 and 75.00 mg/g, respectively. The yields of the four components were 71.89 %-86.19 %. Cell experiments showed that the purified SGD retained anti-inflammatory activity. This research lays the foundation for the separation and purification of SGD and subsequent preparation research.
Subject(s)
Drugs, Chinese Herbal , Glucosides , Monoterpenes , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/isolation & purification , Chromatography, High Pressure Liquid/methods , Glucosides/isolation & purification , Glucosides/chemistry , Monoterpenes/isolation & purification , Monoterpenes/chemistry , Adsorption , Flavanones/isolation & purification , Flavanones/chemistry , Flavanones/analysis , Animals , Glycyrrhizic Acid/isolation & purification , Glycyrrhizic Acid/chemistry , Glycyrrhizic Acid/analysis , Bridged-Ring Compounds/chemistry , Bridged-Ring Compounds/isolation & purification , Mice , Resins, Synthetic/chemistry , Anti-Inflammatory Agents/isolation & purification , Anti-Inflammatory Agents/chemistry , Humans , RAW 264.7 CellsABSTRACT
The development of new cultivars is essential for establishing a method of producing licorice in Japan. A suitable new cultivar for domestic licorice production, known as the interspecific hybrid strain C-18, was developed by crossbreeding Glycyrrhiza uralensis Fisch. (as the seed parent, possessing a high glycyrrhizin (GL) content, strain OMP-28) and Glycyrrhiza glabra L. (as the pollen parent, known for vigorous growth, strain OMP-10). Both G. uralensis and G. glabra are specified in the Pharmacopoeia of Japan (18th edition) as the source plants for Glycyrrhizae Radix. After 2 years of cultivation, strain C-18 exhibited robust growth, with a fresh weight of 148.8 g and a stem diameter of 0.89 mm. The GL content in the dry weight was measured at 3.61%. Seedlings cultivated from rhizomes in the field for 2 years showed a tap root fresh weight per plant of 120 ± 21 g, with an average GL content relative to the dry weight of 2.68% ± 0.38%. Although glabridin, a characteristic compound of G. glabra, was not detected, glycycoumarin, a characteristic compound of G. uralensis, was detected via HPLC analysis. Strain C-18 contained glycycoumarin as a characteristic compound of G. uralensis but lacked glabridin, a compound characteristic of G. glabra. Additionally, 2,3-dehydrokievitone (1) and parvisoflavone A (2) were identified as distinctive components of the interspecific hybrid (G. uralensis × G. glabra) C-18.
Subject(s)
Glycyrrhiza uralensis , Glycyrrhiza , Glycyrrhiza/chemistry , Glycyrrhiza uralensis/chemistry , Glycyrrhizic Acid/analysis , Glycyrrhizic Acid/chemistry , Japan , Plant Extracts/chemistry , Chromatography, High Pressure Liquid , Hybridization, Genetic , Plant Roots/chemistryABSTRACT
INTRODUCTION: Glycyrrhizin (GLY) and sennoside A (SA) are characteristic bioactive marker compounds of the Kampo medicine Daiokanzoto. Their accurate detection in blends of Rhei rhizoma and Glycyrrhizae radix of several species (4:1 or 4:2) is essential for quality control and to ensure therapeutic efficacy. A rapid, efficient assay can significantly facilitate their detection. OBJECTIVE: To establish a rapid qualitative assay for GLY and SA detection, a lateral flow immunoassay (LFA) was developed using specific monoclonal antibody (mAb) nanoparticles. METHODOLOGY: This assay harnesses the competitive binding of mAb nanoparticles to the immobilized analytes on test strips and free analytes in the samples. Two conjugates for detecting GLY and SA, GLY-bovine serum albumin and SA-human serum albumin, were separately immobilized on the test zones of LFA strips. The detection mechanism is reliant on the visual detection of color changes in the test zones. RESULTS: When GLY and SA were present in samples, they contended with the immobilized conjugates on the strip to bind with the mAb nanoparticles and produced distinct color patterns in the test zones. The limits of detection of the assay for GLY and SA were both 3.13 µg/mL. The capability of the LFA was substantiated using plant samples and Daiokanzoto, and its alignment with indirect competitive ELISA results was confirmed. CONCLUSION: The introduced LFA is a groundbreaking procedure that offers a rapid, straightforward, and sensitive method for simultaneously detecting GLY and SA in Daiokanzoto samples. It is instrumental in ensuring product quality.
Subject(s)
Glycyrrhizic Acid , Sennosides , Glycyrrhizic Acid/analysis , Immunoassay/methods , Antibodies, Monoclonal , Humans , Nanoparticles/chemistry , Serum Albumin, Bovine/chemistry , Limit of Detection , Animals , Serum Albumin, Human/analysis , Drugs, Chinese Herbal/chemistryABSTRACT
This study focused on the separation, characterization, content determination, and antiviral efficacy research on colloidal particles with different sizes in Maxing Shigan Decoction(MXSG). The mixed colloidal phase of MXSG was initially separated into small colloidal particle segment(S), medium colloidal particle segment(M), and big colloidal particle segment(B) using ultrafiltration. Further fine separation was performed using size-exclusion chromatography. Dynamic light scattering(DLS) and transmission electron microscopy(TEM) were employed to characterize the size and morphology of the separated colloidal particles. UPLC-MS/MS was used to determine the content of ephedrine, amygdalin, glycyrrhizic acid, and the EDTA complexometric titration was used to measure the calcium(Ca~(2+)) content in different colloidal phases. Finally, a respiratory syncytial virus(RSV) infection mouse model was established using intranasal administration. The experimental groups included a blank group, a model group, a ribavirin group, an MXSG group, an S group, an M group, and a B group. Oral administration was given for treatment, and pathological changes in mouse lung tissue and organ indices were evaluated. The results of the study showed that the distribution of ephedrine, amygdalin, glycyrrhizic acid, and Ca~(2+) content was not uniform among different colloidal segments. Among them, the B segment had the highest proportions of the three components, except for Ca~(2+), accounting for 46.35%, 53.72%, and 92.36%, respectively. Size-exclusion chromatography separated colloidal particles with uniform morphology in the size range of 100-500 nm. Compared to the S and M segments, the B segment showed an increased lung index inhibition rate(38.31%), spleen index, and thymus index in RSV-infected mice, and it improved the infiltration of inflammatory cells and lung injury in the lung tissue of mice. The complex components in MXSG form colloidal particles of various sizes and morphologies through heating, and small-molecule active components such as ephedrine, amygdalin, glycyrrhizic acid, and Ca~(2+) participate in the assembly to varying degrees. The main material basis for the antiviral effect of MXSG is the colloidal particles with certain particle sizes formed by the assembly of active components during the heating process.
Subject(s)
Amygdalin , Drugs, Chinese Herbal , Mice , Animals , Amygdalin/chemistry , Drugs, Chinese Herbal/chemistry , Glycyrrhizic Acid/analysis , Ephedrine/analysis , Chromatography, Liquid , Tandem Mass Spectrometry , Antiviral Agents/pharmacologyABSTRACT
Dispensing granules of Chinese medicine (DGCM) have emerged as a more convenient alternative to traditional decoction (TD) of Chinese medicine, gaining popularity in recent years. However, the debate surrounding the consistency of DGCM compared to TD remains unresolved. In this study, three batches of Baishao and Gancao DGCM were obtained from manufacturers A, B, and C, and 15 batches of crude drugs were procured from hospital pharmacies for the preparation of dispensing granule decoction (DGD) and TD of Shaoyao-Gancao decoction (SGD). The HPLC-UV method was employed to determine the levels of gallic acid, paeoniflorin, albiflorin, liquiritin, liquiritin apioside, isoliquiritin apioside, isoliquiritin, glycyrrhizic acid, and isoliquiritigenin. The analgesic and antispasmodic effects were assessed using the hot plate and acetic acid writhing test in mice. To evaluate the consistency of chemical constituents and pharmacological effects between the two decoctions, the Criteria Importance Though Intercriteria Correlation (CRITIC) method combined with chemometrics was employed. Grey relation analysis (GRA) was used to assess the comprehensive quality consistency of the two decoctions. The CRITIC results revealed certain differences in chemical constituents and pharmacological effects between the selected DGCM and TD. Notably, DGD-A/C exhibited a significant difference from TD (p > 0.05), whereas DGD-B demonstrated no significant difference from TD (p > 0.05). The GRA analysis demonstrated that the overall quality consistency between DGD-B and TD was the highest among the three manufacturers. This study presents a method for evaluating the quality consistency of DGCM and TD of SGD, offering novel insights into the evaluation of consistency between DGCM and TD.
Subject(s)
Drugs, Chinese Herbal , Glycyrrhiza , Mice , Animals , Drugs, Chinese Herbal/chemistry , Glycyrrhiza/chemistry , Glycyrrhizic Acid/pharmacology , Glycyrrhizic Acid/analysis , Chromatography, High Pressure Liquid/methodsABSTRACT
Qinggusan is the 69th prescription in the first batch of "Catalogue of Ancient Chinese Classic Formulas". In modern clinical practiceï¼ Qinggusan is mainly used to treat noninfectious fever. Howeverï¼ because few studies on Qinggusan reference samples and their quality value transfer are availableï¼ the development and promotion of its compound preparations are restricted. Thereforeï¼ establishing an accurate and comprehensive quality control method to clarify the critical quality attributes of Qinggusan reference samples is of great importance. In this studyï¼ 15 batches of Qinggusan reference samples were processed to determine the range of their dry extract ratios. Quantitative high-performance liquid chromatography ï¼HPLCï¼ fingerprint analysis was performed using a Waters Symmetry Shield RP18 column ï¼250 mm×4.6 mmï¼ 5 µmï¼ with acetonitrile-0.1% ï¼v/vï¼ formic acid aqueous solution as the mobile phase in gradient elution mode. The flow rate was 1.0 mL/minï¼ the column temperature was 30 âï¼ and the detection wavelength was 254 nm. The HPLC fingerprints of the Qinggusan reference samples were established under these conditions to evaluate their similarity. The established method was systematically validated and found to demonstrate good precisionï¼ repeatabilityï¼ and sample stability. Subsequentlyï¼ characteristic peaks were identified and attributed by HPLC-quadrupole-time-of-flight-mass spectrometry ï¼HPLC-Q-TOF-MSï¼ analysis. MS was performed in electrospray ionization modeï¼ the data were collected in both positive- and negative-ion modesï¼ and the detection range was m/z 50-2000. The contents and transfer rate ranges of the index componentsï¼ namelyï¼ gentiopicrinï¼ mangiferinï¼ picroside â ¡ï¼ picroside â ï¼ and glycyrrhizic acidï¼ were determined to analyze the quality value transfer of the samples. The results demonstrated that the dry extract rate of the 15 batches of Qinggusan reference samples ranged from 24.10% to 26.88% and that their fingerprint similarities were generally greater than 0.95. Twelve common peaks were identified by reference identificationï¼ literature comparisonï¼ and high-resolution MS analysis. Twelve compoundsï¼ including six iridoid glycosidesï¼ two flavonoidsï¼ one alkaloidï¼ one triterpenoid saponinï¼ and two others. Among themï¼ L-piceinï¼ androsinï¼ picroside â £ï¼ picroside â ¡ and picroside â were from Picrorhizae Rhizomaï¼ loganin acidï¼ swertiamarin and gentiopicrin were from Gentianae Macrophyllae Radixï¼ neomangiferin and mangiferin were from Anemarrhenae Rhizomaï¼ dichotomine B was from Stellariae Radixï¼ and glycyrrhizic acid was from Glycyrrhizae Radix et Rhizoma. The five key components presented good linear relationships in their respective linear rangesï¼ and all correlation coefficients were higher than 0.999. The relative standard deviations ï¼RSDsï¼ of precisionï¼ stabilityï¼ and repeatability were less than 1.3%. The average recoveries varied between 95.92% and 102.5%ï¼ with RSDs less than 3.9%ï¼ these values meet the requirements defined in the 2020 edition of the Chinese Pharmacopoeia. The contents of gentiopicrinï¼ mangiferinï¼ picroside â ¡ï¼ picroside â ï¼ and glycyrrhizic acid in the 15 batches of reference samples were in the range of 17.92-27.55ï¼ 1.83-4.42ï¼ 23.08-36.44ï¼ 8.43-15.04ï¼ and 0.94-2.39 mg/gï¼ respectivelyï¼ and their transfer rates from the decoction pieces to the reference samples were 47.91%-63.95%ï¼ 22.96%-59.39%ï¼ 60.82%-77.82%ï¼ 64.25%-99.53%ï¼ and 15.30%-39.30%ï¼ respectively. In this studyï¼ the chemical components of Qinggusan reference samples were comprehensively identified and their quality value transfer was studied through the combination of HPLC fingerprinting and MS. Clarification of the critical quality attributes of Qinggusan reference samples could provide a basis for the quality control of Qinggusan compound preparations.
Subject(s)
Drugs, Chinese Herbal , Glycyrrhizic Acid , Glycyrrhizic Acid/analysis , Drugs, Chinese Herbal/analysis , Plant Extracts , Quality Control , Chromatography, High Pressure LiquidABSTRACT
Quality consistency of Glycyrrhiza formula granules is essential for guaranteeing clinical efficacy. However, a suitable method to accurately and conveniently evaluate the consistency of the clinical efficacy of Glycyrrhiza formula granules is currently not available. This study established a method for the simultaneous determination of 12 active components in Glycyrrhiza formula granules using ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry. The rate of inhibition of cyclooxygenase-2 by different batches of Glycyrrhiza formula granules was determined. Near-infrared spectra were collected for different batches of Glycyrrhiza formula granules to detect their biological activity in the inhibition of cyclooxygenase-2. The quality consistency of the 11 batches of Glycyrrhiza formula granules was evaluated using principal component and correlation analyses. The results showed significant differences in the formula granules of Glycyrrhiza uralensis produced by the different manufacturers. Some differences were also observed among batches of formula granules produced by the same manufacturer. Correlation analysis of the chemical components and cyclooxygenase-2 activity showed that glycyrrhizic acid, liquiritin, and isoliquiritin were the main active components of Glycyrrhiza. Correlation analysis of the near-infrared spectra and cyclooxygenase-2 inhibition activity showed a high correlation between the active components and three characteristic bands: 3383-3995, 4227-4651, and 5315-5878 cm-1 . In this study, the main active anti-inflammatory components of Glycyrrhiza granules were screened. Thus, the near-infrared spectrum and characteristic active band of multi-index active components can be used to quickly detect the quality consistency of Glycyrrhiza formula granules, thereby improving the ability to control the quality and consistency of these granules.
Subject(s)
Drugs, Chinese Herbal , Glycyrrhiza uralensis , Glycyrrhiza , Drugs, Chinese Herbal/analysis , Cyclooxygenase 2 , Glycyrrhiza/chemistry , Glycyrrhiza uralensis/chemistry , Glycyrrhizic Acid/analysis , Chromatography, High Pressure Liquid/methodsABSTRACT
This study focused on the separation, characterization, content determination, and antiviral efficacy research on colloidal particles with different sizes in Maxing Shigan Decoction(MXSG). The mixed colloidal phase of MXSG was initially separated into small colloidal particle segment(S), medium colloidal particle segment(M), and big colloidal particle segment(B) using ultrafiltration. Further fine separation was performed using size-exclusion chromatography. Dynamic light scattering(DLS) and transmission electron microscopy(TEM) were employed to characterize the size and morphology of the separated colloidal particles. UPLC-MS/MS was used to determine the content of ephedrine, amygdalin, glycyrrhizic acid, and the EDTA complexometric titration was used to measure the calcium(Ca~(2+)) content in different colloidal phases. Finally, a respiratory syncytial virus(RSV) infection mouse model was established using intranasal administration. The experimental groups included a blank group, a model group, a ribavirin group, an MXSG group, an S group, an M group, and a B group. Oral administration was given for treatment, and pathological changes in mouse lung tissue and organ indices were evaluated. The results of the study showed that the distribution of ephedrine, amygdalin, glycyrrhizic acid, and Ca~(2+) content was not uniform among different colloidal segments. Among them, the B segment had the highest proportions of the three components, except for Ca~(2+), accounting for 46.35%, 53.72%, and 92.36%, respectively. Size-exclusion chromatography separated colloidal particles with uniform morphology in the size range of 100-500 nm. Compared to the S and M segments, the B segment showed an increased lung index inhibition rate(38.31%), spleen index, and thymus index in RSV-infected mice, and it improved the infiltration of inflammatory cells and lung injury in the lung tissue of mice. The complex components in MXSG form colloidal particles of various sizes and morphologies through heating, and small-molecule active components such as ephedrine, amygdalin, glycyrrhizic acid, and Ca~(2+) participate in the assembly to varying degrees. The main material basis for the antiviral effect of MXSG is the colloidal particles with certain particle sizes formed by the assembly of active components during the heating process.
Subject(s)
Mice , Animals , Amygdalin/chemistry , Drugs, Chinese Herbal/chemistry , Glycyrrhizic Acid/analysis , Ephedrine/analysis , Chromatography, Liquid , Tandem Mass Spectrometry , Antiviral Agents/pharmacologyABSTRACT
The decoction turns into a complex multiphase system following exposure to high temperature and a complex chemical environment. However, the differences in the concentration of key active ingredients in different phase states and the release of drugs in sedimentary phase have yet to be elucidated. A simple ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the simultaneous quantitative determination of brucine, strychnine, liquiritin, isoliquiritin, isoliquiritigenin and glycyrrhizic acid concentrations and it was applied to compare the content of different phases and measure the release characteristics of the sedimentary phase in "Glycyrrhiza glabra-Nux vomica" decoction (NGD). The results show that the method's selectivity, precision (intraday and interday ≤ 2%), matrix effect (101-108%), recovery and stability results were acceptable according to the guidelines. The method is sensitive and reliable. The content determination results show that the most toxic strychnine in the sedimentary phase accounted for 75.70% of the total components. The different components exhibited differential release in different media, and its components were released in the artificial intestinal fluid up to 81.02% in 12 h. Several components conformed to the primary kinetic model and the Ritger-Peppas model, and the most toxic compound exhibited slow release, thus conforming to the Ritger-Peppas model. This study provides a standard of reference for studies investigating reduction in toxicity of the combination of Glycyrrhiza glabra (Glycyrrhiza glabra L.) and Nux vomica (Strychnos nux-vomica L.).
Subject(s)
Strychnos nux-vomica , Chromatography, High Pressure Liquid , Chromatography, Liquid , Glycyrrhizic Acid/analysis , Seeds/chemistry , Strychnine/chemistry , Strychnos nux-vomica/chemistry , Tandem Mass SpectrometryABSTRACT
Tojapride is composed of Caulis Perillae, Rhizoma Cyperi, Radix Glycyrrhizae, Citrus aurantium L., Coptis chinensis Franch, Pericarpium Citri Reticulatae, Reynoutria japonica Houtt, Tetradium ruticarpum, and Cleistocactus sepium. It has the effects of inhibiting gastric acid and relieving pain. It is clinically used for treating gastroesophageal reflux disease. To further study the pharmacodynamic properties of Tojapride, the systematic characterization of the chemical constituents in Tojapride was investigated using ultra-performance liquid chromatography with Q-Exactive Orbitrap mass spectrometry combined with parallel reaction monitoring for the first time. Eventually, a total of 222 compounds, including flavonoids, alkaloids, and glycyrrhizic acid derivatives, were identified based on the chromatographic retention times, MS/MS2 information, and bibliography data; a total of 218 of these were reported for the first time as being present in Tojapride. This newly developed approach provides a powerful tool for extending our understanding of chemical constituents of Tojapride, which can be further extended to other TCMP composition research.
Subject(s)
Alkaloids , Drugs, Chinese Herbal , Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacology , Flavonoids/analysis , Glycyrrhizic Acid/analysis , Phytochemicals/pharmacology , Tandem Mass Spectrometry/methodsABSTRACT
Licorice, the root of Glycyrrhiza spp., is used in a large number of herbal medicines, such as traditional Chinese medicines, Japanese Kampo medicines, and therapeutic drugs. Since glycyrrhizin (GL) is among the main components in licorice and exhibits numerous beneficial pharmacological activities, the content of GL directly affects biological activity. The quality control based on GL content is an important factor in ensuring biological activity; however, the content of GL in licorice varies depending on plant cultivation environment, genetic factors, and species type. Previously, we prepared an anti-GL monoclonal antibody (anti-GL mAb) and employed it in various immunochemical assays for quality control of licorice and licorice-based products. In this study, we employed the anti-GL mAb in chemiluminescence enzyme immunostaining (CLEIS) to develop a very simple, rapid, specific, and sensitive quality control assay for licorice products, with a limit of detection of 3.9 ng. Furthermore, the CLEIS assay enabled semiquantitative analysis of GL in Kampo medicines. Our results showed that multiple samples can be simultaneously analyzed using CLEIS, and it is a useful tool for determining GL content, as well as ensuring chemical quality control of licorice-containing products and herbal medicines.
Subject(s)
Glycyrrhiza , Plants, Medicinal , Antibodies, Monoclonal , Glycyrrhiza/chemistry , Glycyrrhizic Acid/analysis , Glycyrrhizic Acid/pharmacology , Luminescence , Medicine, Kampo , Plant Extracts/pharmacologyABSTRACT
Shengjiang Xiexin decoction, a traditional Chinese medical formula, has been utilized to alleviate the delayed-onset diarrhea induced by irino tecan. However, the chemical constituents of this formula and the activities of its constituents remain unclear. In this study, ultrahigh-performance liquid chromatography-quadrupole/orbitrap high-resolution mass spectrometry was employed to comprehensively analyze the chemical constituents of Shengjiang Xiexin decoction. A total of 270 components, including flavonoids, coumarins, triterpenoids, alkaloids, diarylheptanoids and others, were identified or characterized. Multidrug resistance-associated protein 2 is an efflux transporter responsible for regulating drug absorption. A total of 20 characteristic components from the formula were selected to evaluate their effects on the function of multidrug resistance-associated protein 2 using the vesicular transport assay. Glycyrrhizic acid and glycyrrhetinic acid were identified as potential multidrug resistance-associated protein 2 inhibitors, while 9 flavonoid aglycones increased the uptake of the substrate [3 H]-estradiol 17-ß-glucuronide in the vesicles. This was the first systematic investigation of the chemical constituents from Shengjiang Xiexin decoction and the effect of its characteristic components on the transporter. The results offered a basis for further exploring the detoxification mechanisms of this formula and its interactions with other drugs.
Subject(s)
Alkaloids , Drugs, Chinese Herbal , Glycyrrhetinic Acid , Chromatography, High Pressure Liquid/methods , Chromatography, Liquid , Coumarins/analysis , Diarylheptanoids , Drugs, Chinese Herbal/chemistry , Estradiol , Flavonoids/analysis , Glucuronides , Glycyrrhizic Acid/analysis , Mass Spectrometry/methods , Multidrug Resistance-Associated Protein 2ABSTRACT
The present study established specific chromatograms and a method for determining multiple primary components in Yinqiao Powder decoctions and compared the change rules of chemical composition in powder and piece decocting processes of Yinqiao Powder to provide a scientific basis for the modern research of preparations of Yinqiao Powder. Powder and piece decoctions of Yinqiao Powder were prepared. The specific chromatograms were determined and the content of 17 primary components was measured by high-performance liquid chromatography(HPLC), including adoxosidic acid, neochlorogenic acid, forsythoside E, loganic acid, chlorogenic acid, cryptochlorogenic acid, sweroside, forsythoside â , forsythoside H, forsythoside A, isochlorogenic acid B, E-aldosecologanin, hesperidin, phillyrin, arctiin, liquiritigenin, and dipotassium glycyrrhizinate. The effect of decocting time on the chemical composition in powder and piece decoctions of Yinqiao Powder was investigated. As a result, the specific chromatogram similarities of powder decoctions of Yinqiao Powder with different decocting time were high, which indicated that their chemical compositions were similar, while the similarities of piece decoctions were low, suggesting similar chemical compositions with big differences. In powder decoctions, the concentrations of neochlorogenic acid, cryptochlorogenic acid, forsytherin H, and isochlorogenic acid B increased with the prolongation of decocting time, and those of adoxosidic acid, forsythoside E, forsythoside â , E-aldosecologanin, phillyrin, dipotassium glycyrrhizinate, loganic acid, arctiin, sweroside, and liquiritigenin increased firstly and tended to be stable, while those of forsythoside A, chlorogenic acid, and hesperidin increased firstly and then decreased. In piece decoctions, the concentration of chlorogenic acid increased firstly and then decreased with the prolongation of decocting time, while those of the remaining 16 components showed an upward trend. The concentrations of adoxosidic acid, forsythoside E, forsythoside â , E-aldosecologanin, phillyrin, dipotassium glycyrrhizinate, forsythoside A, forsythoside H, and chlorogenic acid in powder decoctions were higher than those in piece decoctions. The concentrations of hesperidin, loganic acid, phillyrin, sweroside, liquiritigenin, neochlorogenic acid, and cryptochlorogenic acid in powder decoctions were higher than those in piece decoctions within 40 min of decocting. The concentration of isochlorogenic acid B in powder decoctions was lower than that in piece decoction 10 min after decocting. The results showed that the decocting time and particle size of raw medicinal materials had certain effects on the content of chemical components in decoctions of Yinqiao Powder. Compared with the piece decocting, the powder decocting could achieve faster resolution of chemical components and higher concentrations, which confirmed the scientific evidence of the traditional powder decocting method of Yinqiao Powder. For the piece decocting of prescriptions of Yinqiao Powder, extraction time should be prolonged and extraction times should be increased to achieve the same effect as the powder decocting.
Subject(s)
Drugs, Chinese Herbal , Hesperidin , Neoplasms, Multiple Primary , Chlorogenic Acid/analysis , Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/chemistry , Glycyrrhizic Acid/analysis , Humans , PowdersABSTRACT
Glycyrrhiza uralensis Fisch is a medicinal plant widely used to treat multiple diseases in Europe and Asia, and its efficacy largely depends on liquiritin and glycyrrhizic acid. The regulatory pattern responsible for the difference in efficacy between wild and cultivated G. uralensis remains largely undetermined. Here, we collected roots and rhizosphere soils from wild (WT) G. uralensis as well as those farmed for 1 year (C1) and 3 years (C3), generated metabolite and transcript data for roots, microbiota data for rhizospheres and conducted comprehensive multi-omics analyses. We updated gene structures for all 40 091 genes in G. uralensis, and based on 52 differentially expressed genes, we charted the route-map of both liquiritin and glycyrrhizic acid biosynthesis, with genes BAS, CYP72A154 and CYP88D6 critical for glycyrrhizic acid biosynthesis being significantly expressed higher in wild G. uralensis than in cultivated G. uralensis. Additionally, multi-omics network analysis identified that Lysobacter was strongly associated with CYP72A154, which was required for glycyrrhizic acid biosynthesis. Finally, we developed a holistic multi-omics regulation model that confirmed the importance of rhizosphere microbial community structure in liquiritin accumulation. This study thoroughly decoded the key regulatory mechanisms of liquiritin and glycyrrhizic acid, and provided new insights into the interactions of the plant's key metabolites with its transcriptome, rhizosphere microbes and environment, which would guide future cultivation of G. uralensis.
Subject(s)
Glycyrrhiza uralensis , Plants, Medicinal , Glycyrrhiza uralensis/chemistry , Glycyrrhiza uralensis/genetics , Glycyrrhiza uralensis/metabolism , Glycyrrhizic Acid/analysis , Glycyrrhizic Acid/metabolism , Plant Roots/metabolism , Plants, Medicinal/genetics , Plants, Medicinal/metabolism , SoilABSTRACT
A quantitative analysis of multi-components by single marker(QAMS) method was established for the simultaneous determination of ephedrine hydrochloride, protocatechuic acid, 5-caffeoylquinic acid, 4-hydroxybenzoic acid, naringin, neohesperidin, glycyrrhizic acid, and praeruptorin A in Jizhi Syrup by high performance liquid chromatography(HPLC) with ultraviolet multi-wavelength detection system, and its feasibility in quality evaluation of Jizhi syrup was verified. With naringin as the internal reference substance, the relative correction factors and chromatographic peak localization methods of other seven components were respectively established at 210, 254, 280, and 320 nm. The method reproducibility was validated, and the result of QAMS were compared with those obtained by the external standard method(ESM) to verify the accuracy and feasibility of the method. The relative correction factors of ephedrine hydrochloride, protocatechuic acid, 5-caffeoylquinic acid, 4-hydroxybenzoic acid, neohesperidin, glycyrrhizic acid, and praeruptorin A with naringin as reference were 0.846, 0.582, 0.608, 0.293, 0.913, 2.207, and 0.940, respectively, which presented excellent reproducibility under different experimental conditions. Furthermore, QAMS and ESM showed no significant difference in the results for 15 batches of samples. Except protocatechuic acid and 5-caffeoylquinic acid, other six compounds were the exclusive components of single medicinal materials. In addition, glycyrrhizic acid and praeruptorin A were identified in the Jizhi Syrup for the first time, filling up the blank of no component detected in Glycyrrhizae Radix et Rhizoma and Peucedani Radix. The method established in this study is convenient, efficient, specific, accurate, and reliable, which can comprehensively and effectively evaluate the quality of Jizhi Syrup to ensure the safety and efficacy of this drug in clinical application.
Subject(s)
Drugs, Chinese Herbal , Glycyrrhizic Acid , Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/chemistry , Ephedrine , Glycyrrhizic Acid/analysis , Reproducibility of ResultsABSTRACT
According to the polarity of different components in Sanpian Decoction, two fingerprints were established. Then the substance benchmark freeze-dried powder of 15 batches of Sanpian Decoction was prepared, followed by the determination of the fingerprints, index component content, and dry extract rates, the identification of attribution of characteristic peaks, and the calculation of similarities between these fingerprints and the reference(R), the content and transfer rate ranges of ferulic acid, sinapine thiocyanate, liquiritin, and glycyrrhizic acid, and the dry extract rate range. The results showed that the similarities of 15 batches of the substance benchmark fingerprints with R were all greater than 0.900.Further summarization of the characteristic peaks revealed that there were a total of 20 characteristic peaks in fingerprint 1, among which, eight were from Sinapis Semen, four from Paeoniae Radix Alba, six from Chuanxiong Rhizoma, and two from Glycyrrhizae Radix et Rhizoma. A total of 16 characteristic peaks were observed in fingerprint 2, including one from Sinapis Semen, three from Paeoniae Radix Alba, eight from Chuanxiong Rhizoma, and four from Glycyrrhizae Radix et Rhizoma. The average dry extract rate of 15 batches of substance benchmarks was 18.25%, with a dry extract rate range of 16.28%-20.76%. The index component content and transfer rate ranges were listed as follows: 0.15%-0.18% and 38.81%-58.05% for ferulic acid; 0.26%-0.42% and 36.51%-51.02% for sinapine thiocyanate; 0.09%-0.15% and 48.80%-76.61% for liquiritin; 0.13%-0.24% and 23.45%-35.61% for glycyrrhizic acid. The fingerprint, dry extract rate, and index component content determination was combined for analyzing the quality value transfer of substance benchmarks in the classic prescription Sanpian Decoction.The established quality evaluation method for the substance benchmarks was stable and feasible, which has provided a basis for the quality control of Sanpian Decoction and the follow-up development of related preparations.
Subject(s)
Drugs, Chinese Herbal , Paeonia , Benchmarking , Chromatography, High Pressure Liquid , Glycyrrhizic Acid/analysis , Quality Control , ThiocyanatesABSTRACT
The methods for determining the characteristic chromatogram and index components content of Xuanfu Daizhe Decoction were established to provide a scientific basis for the quality evaluation of substance benchmarks and preparations. Eighteen batches of Xuanfu Daizhe Decoction were prepared with the decoction pieces of different batches and of the same batch were prepared respectively, and the HPLC characteristic chromatograms of these samples were established. The similarities of the chromatographic fingerprints were analyzed. With liquiritin, glycyrrhizic acid, 6-gingerol, ginsenoside Rg_1, and ginsenoside Re as index components, the high performance liquid chromatography was established for content determination with no more than 70%-130% of the mass average as the limit. The results showed that there were 19 characteristic peaks corresponding to the characteristic chromatograms of 18 batches of Xuanfu Daizhe Decoction, including 8 peaks representing liquiritin, 1,5-O-dicaffeoylqunic acid, ginsenoside Rg_1, ginsenoside Re, 1-O-acetyl britannilactone, ginsenoside Rb_1, glycyrrhizic acid, and 6-gingerol, and the fingerprint similarity was greater than 0.97. The contents of liquiritin, glycyrrhizic acid, 6-gingerol, and ginsenosides Rg_1 + Re in the prepared Xuanfu Daizhe Decoction samples were 0.53%-0.86%, 0.61%-1.2%, 0.023%-0.068%, and 0.33%-0.66%, respectively. Except for several batches, most batches of Xuanfu Daizhe Decoction showed stable contents of index components, with no discrete values. The characteristic chromatograms and index components content characterized the information of Inulae Flos, Ginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma, and Zingiberis Rhizoma Recens in Xuanfu Daizhe Decoction. This study provides a scientific basis for the further research on the key chemical properties of substance benchmark and preparations of Xuanfu Daizhe Decoction.
Subject(s)
Drugs, Chinese Herbal , Ginsenosides , Benchmarking , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/chemistry , Ginsenosides/analysis , Glycyrrhizic Acid/analysis , Quality ControlABSTRACT
With reference to the production process documented in Chinese Pharmacopoeia, this paper prepared the calibrator samples of Xiaochaihu Granules from multiple batches and established a method for fingerprint analysis and content determination that could be used to evaluate Xiaochaihu Granules available in market. Multiple batches of Chinese herbal pieces contained in Xiaochaihu Granules were collected for preparing the calibrator samples according to the process in Chinese Pharmacopoeia. Following the establishment of fingerprints for calibrator samples by UHPLC, the method for determining the contents of saikosaponin B2, saikosaponin B1, baicalin, wogonoside, baicalein, liquiritin, glycyrrhizin G2 and glycyrrhizic acid in Xiaochaihu Granules was established. The experimental results showed that the fingerprints of calibrator samples had 26 common peaks, covering the chemical compounds of main herbs Bupleuri Radix, Scutellariae Radix, Changii Radix, Glycyrrhizae Radix et Rhizoma, and Rhizoma Zingiberis Recens. The similarity of fingerprints for 47 batches of Xiaochaihu Granules from 31 companies with the calibrator sample fingerprint ranged from 0.74 to 0.99, indicating good applicability of the established fingerprint. The contents of main components baicalin, saikosaponin B2, and glycyrrhizic acid in Xiaochaihu Granules were within the ranges of 22.917-49.108 mg per bag(RSD 19%), 0.28-2.19 mg per bag(RSD 62%), and 0.897-6.541 mg per bag(RSD 41%), respectively. The quality difference in saikosaponin B2, and glycyrrhizic acid among different manufacturers was significant. The fingerprint analysis and content determination method for calibrator samples of Xiaochaihu Granules prepared according to the production process in Chinese Pharmacopoeia has been proved suitable for evaluating the quality of Xiaochaihu Granules from different manufacturers. Saikosaponin B2, glycyrrhizic acid, and liquiritin should be added as content control indicators for Xiaochaihu Granules, aiming to further improve the product quality.
Subject(s)
Drugs, Chinese Herbal , Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/chemistry , Glycyrrhizic Acid/analysis , Rhizome/chemistry , Scutellaria baicalensisABSTRACT
A total of 18 batches of Zhuru Decoction samples were prepared. Chromatographic fingerprints were established for Zhuru Decoction and single decoction pieces, the content of which was then determined. The extraction rate ranges, content, and transfer rate ranges of puerarin, liquiritin, and glycyrrhizic acid, together with the common peaks and the similarity range of the fingerprints, were determined to clarify key quality attributes of Zhuru Decoction. The 18 batches of Zhuru Decoction samples had 25 common peaks and the fingerprint similarity higher than 0.95. Puerariae Lobatae Radix, Glycyrrhizae Radix et Rhizoma, and Zingiberis Rhizoma Recens had 21, 3, and 1 characteristic peaks, respectively. The 18 batches of samples showed the extraction rates within the range of 18.45%-25.29%. Puerarin had the content of 2.20%-3.07% and the transfer rate of 38.5%-45.9%; liquiritin had the content of 0.24%-0.85% and the transfer rate of 15.9%-37.5%; glycyrrhizic acid had the content of 0.39%-1.87% and the transfer rate of 16.2%-32.8%. In this paper, the quality value transmitting of substance benchmarks of Zhuru Decoction was analyzed based on chromatographic fingerprints, extraction rate, and the content of index components. A scientific and stable method was preliminarily established, which provided a scientific basis for the quality control and formulation development of Zhuru Decoction.
Subject(s)
Drugs, Chinese Herbal , Quality Control , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/standards , Glycyrrhizic Acid/analysis , Rhizome/chemistryABSTRACT
With reference to the production process documented in Chinese Pharmacopoeia, this paper prepared the calibrator samples of Xiaochaihu Granules from multiple batches and established a method for fingerprint analysis and content determination that could be used to evaluate Xiaochaihu Granules available in market. Multiple batches of Chinese herbal pieces contained in Xiaochaihu Granules were collected for preparing the calibrator samples according to the process in Chinese Pharmacopoeia. Following the establishment of fingerprints for calibrator samples by UHPLC, the method for determining the contents of saikosaponin B2, saikosaponin B1, baicalin, wogonoside, baicalein, liquiritin, glycyrrhizin G2 and glycyrrhizic acid in Xiaochaihu Granules was established. The experimental results showed that the fingerprints of calibrator samples had 26 common peaks, covering the chemical compounds of main herbs Bupleuri Radix, Scutellariae Radix, Changii Radix, Glycyrrhizae Radix et Rhizoma, and Rhizoma Zingiberis Recens. The similarity of fingerprints for 47 batches of Xiaochaihu Granules from 31 companies with the calibrator sample fingerprint ranged from 0.74 to 0.99, indicating good applicability of the established fingerprint. The contents of main components baicalin, saikosaponin B2, and glycyrrhizic acid in Xiaochaihu Granules were within the ranges of 22.917-49.108 mg per bag(RSD 19%), 0.28-2.19 mg per bag(RSD 62%), and 0.897-6.541 mg per bag(RSD 41%), respectively. The quality difference in saikosaponin B2, and glycyrrhizic acid among different manufacturers was significant. The fingerprint analysis and content determination method for calibrator samples of Xiaochaihu Granules prepared according to the production process in Chinese Pharmacopoeia has been proved suitable for evaluating the quality of Xiaochaihu Granules from different manufacturers. Saikosaponin B2, glycyrrhizic acid, and liquiritin should be added as content control indicators for Xiaochaihu Granules, aiming to further improve the product quality.