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1.
Article in Chinese | WPRIM | ID: wpr-846388

ABSTRACT

Objective: To establish an HPLC fingerprint of Qingxin Zishen Prescription Decoction (QZPD) and determine the contents of its multiple components, so as to provide a scientific basis for quality control. Methods: HPLC analysis was performed on a Phenomenex Kinetex C18 column (100 mm × 4.60 mm, 2.6 μm) for gradient elution with the mobile phase consisting of methanol, acetonitrile and 0.2% formic acid aqueous. The detection wavelength was set at 245 nm and 280 nm, and the column temperature was 40 ℃. Fingerprints of ten batches of QZPD were determined, and the similarities among fingerprints were evaluated. Attributive analysis and identification of common peaks were performed and the contents of 15 components were determined. Results: The fingerprint similarities of 10 batches of QZPD were ranged from 0.923 to 0.998 compared with the reference fingerprint, and 33 common peaks were identified in the fingerprint. Among them, seven peaks (P11, P14-P16, P24, P29, P30) were identified from Coptidis Rhizoma, two peaks (P7, P19) were identified from Nelumbinis Plumula, two peaks (P14, P21) were identified from Ziziphi Spinosae Semen, nine peaks (P4, P5, P10, P17, P18, P28, P31-P33) were identified from Salvia miltiorrhiza, nine peaks (P1-P3, P6, P8, P9, P12, P13, P20) were identified from Corni Fructus, while five peaks (P22, P23, P25-P27) cannot be originated and none of the common peaks was identified from Rehmanniae Radix, Uncariae Ramulus Cum Uncis and Triticum aestivum. By comparing with the chemical reference, fifteen components, including gallic acid (P2), 5-hydroxymethylfurfural (P3), danshensu (P4), protocatechuic aldehyde (P5), morroniside (P9), caffeic acid (P10), cornin (P12), loganin (P13), magnoflorine (P14), coptisine (P24), lithospermic acid (P28), berberine (P29), palmatine (P30), salvianolic acid B (P31) and salvianolic acid E (P33), were identified and quantified. The contents of the fifteen components were 158.3-248.2, 233.6-321.3, 45.9-166.0, 24.3-38.6, 800.7-1 263.6, 26.6-54.9, 44.5-108.2, 470.4-757.3, 85.6-178.6, 11.1-34.2, 56.2-106.4, 25.9-138.9, 21.0-59.2, 951.6-2 244.7 and 38.6-92.8 μg/g, respectively. Conclusion: The method established in this study is stable and highly reproducible, and can provide basis for quality control of QZPD.

2.
Article in Chinese | WPRIM | ID: wpr-846150

ABSTRACT

Objective: To reveal the molecular mechanism of Chaiyin Granules in treatment of coronavirus infection based on network pharmacology and molecular docking. Methods: The chemical constituents of Chaiyin Granules were collected by TCMSP database. SwissTargetPrediction database and GeneCards database were used to predict the potential targets of active ingredients and coronavirus. The potential active ingredients and its targets of Chaiyin Granules in the treatment of coronavirus infection were found through Venn diagram. The potential active compounds-targets network and the PPI network were visualized by Cytoscape 3.7.0. GO-enriched analysis and KEGG pathways analysis were constructed on STRING database. The molecular docking of potential active compounds and key targets was achieved by autodock vina 1.1.2. Results: Fifty-one potential active ingredients and 14 potential targets for Chaiyin Granules on treatment of coronavirus infection were obtained. KEGG pathways analysis showed that 44 metabolic pathways were involved to Chaiyin Granules effect on coronavirus infection, including MAPK signaling pathway, PI3K-Akt signaling pathway, mTOR signaling pathway, Fc epsilon RI signaling pathway and IL-17 signaling pathway. The results of molecular docking showed that baicalin, cubebin, coptisine, daidzein-4,7-diglucoside, linarin, liquiritin, luteolin and wogonin in Chaiyin Granules had strong affinity with NTRK2, PRKCα, TNF, VEGFA, GSK3β. Conclusion: This study elaborated that baicalin, cubebin and coptisine in Chaiyin Granules interacted with NTRK2, PRKCα, TNF, VEGFA, GSK3β and regulated PI3K-Akt/mTOR, ErbB/Ras and IL-17 signaling pathways to inhibit the invasion and replication of coronavirus and enhance immunity to battle against coronavirus infection. This study provides a research basis and theoretical basis for the application of Chaiyin Granules in the treatment of anti-coronavirus infection.

3.
Article in Chinese | WPRIM | ID: wpr-873325

ABSTRACT

Objective::To investigate in vivo and in vitro metabolites of coptisine and their metabolic pathways. Method::SD rats were given coptisine by single gavage (dose of 25 mg·kg-1). Urine and feces from 0 h to 48 h, bile from 0 h to 24 h, and plasma and brain tissue samples at 0.25, 1, 2 h after administration were collected.In vitro metabolism was incubated with rat liver microsomes and intestinal flora.The metabolites were analyzed and identified by the high-resolution HPLC-MS/MS technique.The liquid chromatography separation was carried out on ZORBAX SB-C18 column (4.6 mm×150 mm, 5 μm) with acetonitrile-0.1% formic acid solution as the mobile phase for gradient elution, the flow rate was 1.0 mL·min-1, and column temperature was 25 ℃.The mass spectra were obtained in positive and negative ion mode with electrospray ionization (ESI), the scanning range was m/z 50-1 200.The relative molecular weight was determined according to the quasi-molecular ion peaks.The structures of metabolites were elucidated by comparing the data with literature data, including main ion peaks, UV spectrum and HPLC retention time information. Result::A total of 17 metabolites were identified in each sample, including 11 phase Ⅰ metabolites and 6 phase Ⅱ metabolites.The pathways to these metabolites were hydroxylation, demethylation, dehydrogenation, sulfation and glucuronide conjugation. Conclusion::Coptisine can produce metabolic reaction of phase Ⅰ and phase Ⅱ in rat, and metabolites are predominantly present in urine, and the main metabolic site is liver.Coptisine is poorly absorbed and rarely metabolized in gastrointestinal tract, so it is mostly excreted through feces by prototype.This experiment can provide material basis for the pharmacodynamics and pharmacology of coptisine.

4.
Article in Chinese | WPRIM | ID: wpr-872804

ABSTRACT

Objective::To determine the relationship between the characters and the main components of Chelidonii Herba pieces. Method::The main components of the samples were determined by HPLC, and the characters of Chelidonii Herba Pieces were evaluated by sensory evaluation. The correlation between the characters and components was calculated by correlation formula. Result::The content of 6 components, such as chelidonine, was determined by HPLC. Based on the result of sensory evaluation, there was a certain correlation between the characters and the components. Character descriptions that " some have visible white powder, some have white pubescence, and sometimes they have visible yellow florets" had a very low similarity with total alkali, indicating that these characters and total alkali content was not related. The similarity between the description of " hollow stem" had a high similarity with total alkali content, indicating that the amount of stem was related to the total alkali content. The character description of " more broken leaves" was negatively correlated with total alkaloids in the similarity, which indicated that the content of total alkaloids was less when there were more leaves(or more broken leaves), otherwise, the content of total alkaloids was relatively higher. Conclusion::The established HPLC method is simple and feasible. This study objectively quantifies the descriptions of Chelidonii Herba pieces characters, correlates them with the main components of Chelidonii Herba pieces, and then preliminarily judges the quality of Chelidonii Herba pieces according to the appearance of the characters, which provides a theoretical basis for the identification of Chelidonii Herba pieces in the market by experience, and ideas for the study of the characters of other traditional Chinese medicines.

5.
Article in Chinese | WPRIM | ID: wpr-851132

ABSTRACT

Objective: To establish a spectrum-effect relationship betweent anti-inflammation effects and extracts of Corydalis yanhusuo, in order to provide ideas and methods for study of material basis of efficacy. Methods UPLC-Q-TOF/MS was used to establishe fingerprints of different extracts of C. yanhusuo, and the flurescent enzyme was used as a marker to perform the anti-inflammation activity test. Finally, the relationships between characteristic peaks and anti-inflammation activity was established by partial least squares regression analysis (PLSR) and gray relational analysis (GRA). The anti-inflammatory component obtained by spectral effect analysis was predicted by molecular docking technology, and its anti-inflammatory mechanism was preliminarily studied. Results:The 95% ethanol extract had significant anti-inflammatory activity. The characteristic peaks of No. 5 and 8-11 were significantly affected in PLSR and GRA. Molecular docking results showed that C. yanhusuo exerted anti-inflammatory effects by acting on PKC, ERK2, IKKβ, JAK1, PI3K-α, PI3K-γ, TNF-α, affecting the transmission of inflammatory signals. Conclusion: The anti-inflammatory effect of C. yanhusuo is the result of the combination of various components. The main anti-inflammatory components are coptisine, berberine, palmatine, dihydrogenine, and dehydrocryptine, which exert anti-inflammatory effects by affecting PI3K, JAK, PKC, ERK, IKKβ, and TNF-α signaling pathways.

6.
Article in Chinese | WPRIM | ID: wpr-850910

ABSTRACT

Objective: To establish a method for the determination of 13 active components (phellodendrine, chlorogenic acid, magnoflorine, geniposide, coptisine, epiberberine, jatrorrhizine, berberine, palmatine, baicalin, wogonoside, wogonin, and oroxylin A) in Huanglian Jiedu Decoction (HJD) by HPLC, and screen the effective fractions of HJD by the content and ratio of active components. Methods: The freeze-dried powder of HJD water decoction was extracted by petroleum ether, ethyl acetate, n-butanol and pure water by soxhlet extractor combined system solvent extraction method. The different polar fractions of HJD were obtained after drying, and the residue was used as the precipitation fraction. Subsequently, 13 active ingredients in HJD were detected by Agilent 1260 high performance liquid chromatography. Furthermore, the content of active components in HJD and its different polar fractions were detected, and the effective fraction of HJD were screened. Results: A stable and reliable HPLC method was successfully established, and the content of active components in HJD from high to low were: geniposide, berberine, palmatine, baicalin, wogonoside, coptisine, jatrorrhizine, epiberberine, magnoflorine, phellodendrine, wogonin, chlorogenic acid, and oroxylin A. Compared with the fractions of petroleum ether, ethyl acetate, purified water, and precipitation, the total content of active components in the fraction of n-butanol of HJD was the highest and its proportion is the closest to that of HJD, which was the effective fraction of HJD. Conclusion: In this study, a method for the determination of 13 active ingredients in HJD by HPLC was successfully established, and the n-butanol fraction of HJD was selected as the effective fraction of HJD based on the content of active components.

7.
Article in Chinese | WPRIM | ID: wpr-850821

ABSTRACT

Objective: To evaluate the anti-rheumatism effect, and predict the mechanism and Q-marker of YAO medicine compound containing Cissus pteroclada (Sifangteng in Chinese, SFT) in the treatment of rheumatoid arthritis (RA) in rats. Methods: SD rats were randomly divided into control group, model group, tripterygium glycosides group and SFT high/low dose (28.7, 7.2 g/kg) groups with eight rats in each group. Except the control group, the RA models in rats induced by Collange II collagen were established. The SFT group and the tripterygium glycosides group were given corresponding drugs by intragastric administration during the modeling period, while the other two groups were given the same volume of saline once daily for 28 d. The degree of foot swelling was measured and scored during the experiment. The levels of TNF-α and IL-1β in serum were measured by ELISA at the end of the experiment. SFT chemical components and predicting targets were searched and screened through TCMSP and Drugbank databases. The target of RA disease was searched by TTD database. The protein interaction network was constructed and visualized by String database and Cytoscape software, cluster analysis was analyzed by MCODE. GO and KEGG enrichment analysis was carried out using String database. Finally, combined with the validity and measurability of chemical components, the Q-marker of SFT was predicted. Results: Compared with the model group, the foot swelling of rats in SFT high and low dose groups and positive group was significantly reduced (P < 0.01), and the serum levels of IL-1β and TNF-α were significantly decreased (P < 0.01). There were 89 disease targets of RA. The pathogenesis of RA was related to abnormal cytokine-receptor pathway and RA pathway. A total of 31 components in SFT were screened and its 119 target proteins were predicted, 12 of them belong to disease targets were involved in 1 112 biological processes, such as regulation of stimulation response, regulation of cell proliferation, regulation of cell metabolism, regulation of intracellular signal transduction, and regulation of 113 signaling pathways, such as RA pathway and TNF pathway, which ultimately play a role in the treatment of RA. At the same time, 11 components were predicted to be Q-markers of SFT, including apigenin, resveratrol, bergenin, nitidine, osthol, linalool, ammidin, ethoxychelerythrine, coptisine, hesperidin, and sesamin. Conclusion: SFT can significantly reduce acute inflammation in RA rats. SFT may act on PTGS2-based targets through resveratrol and other components, and participate in regulation of RA pathway, TNF pathway and other inflammatory and immune pathways. Apigenin, resveratrol and bergenin, nitidine, osthol, linalool, ammidin, ethoxychelerythrine, coptisine, hesperidin, sesamin can be used as Q-markers for SFT quality control.

8.
Article in Chinese | WPRIM | ID: wpr-850667

ABSTRACT

Objective: To study on changes of active components in Lianzhifan solution (LZF) under different storage conditions, which may provide a reference for the application and quality improvement of the LZF. Methods: HPLC analysis was performed on Eclipse Agilent C18 column (250 mm × 4.6 mm, 5 μm). The gradient elution was performed by the mobile phase consisting of acetonitrile and 0.1% formic acid aqueous with the flow rate of 1.0 mL/min, the detection wavelength was set at 238 nm, and the column temperature was 25 ℃. A fingerprint analysis method for the chemical composition of LZF was established to analyze the change of main active components with time under sealed and unsealed storage conditions. Results: The precision, stability, repeatability and sample recovery of the HPLC fingerprint of LZF were all in accordance with the requirements, and can be used for the analysis of the chemical constituents of LZF. The similarity values of sample fingerprints and control fingerprints at different time points under both storage conditions were greater than 0.95; The nine chemical components in LZF were identified by using the external labeling method and spectral characteristic analysis. The content of alkaloids (columbamine, epiberberine, jatrorrhizine chloride, coptisine chloride, palmatine chloride, and berberine) did not change significantly under the two storage conditions. However, the content of the iridoids, genipin-1-β-D-gentiobioside and geniposide was significantly reduced, and the content of genipin was increased in the first 3 months but decreased after 3 months. It is speculated that genipin-1-β-D-gentiobioside and geniposide can be converted to genipin by the enzyme in solution, and genipin may be degraded by other enzymes produced by microorganisms. Conclusion: During the six-month storage period, LZF still has a convert tendency, which may be caused by the action of enzymes produced by microbes. In order to improve the quality consistency of LZF product, it is recommended to adopt more scientific method to judge the endpoint and it is better to inactivate enzyme and sterilize after the end of the fermentation processing.

9.
Chongqing Medicine ; (36): 2323-2326, 2018.
Article in Chinese | WPRIM | ID: wpr-692102

ABSTRACT

Objective To establish a method for determining five isoquinoline alkaloids (including berberine,jatrorrhizine,jatrorrhizind,coptisine and palmatine) in Erhuang Xiaoyan tablets simultaneously.Methods Determination by reversed phase high performance liquid chromatography (RP-HPLC).Column:YMC-Pack ODS-AM (4.6 mm× 250.0 mm,5 μm);mobile phase:elution with 0.1% (v/v) formic acid aqueous solution-acetonitrile mobile phase gradient;flow rate:0.4 mL/min;injection volume:10 μL;detection wavelength:345 nm;column temperature:27 ℃.Results Five of isoquinoline alkaloids in Erhuang Xiaoyan tablets were separated perfectly.The good linear relationships were obtained in the following ranges,1.526 4-5.342 4μg (r=0.999 9) for berberine,0.403 2-1.411 2 μg (r=0.999 6) for palmatine,0.309 7-1.083 9 μg (r=0.999 8) for coptisine,0.111 2-0.389 2 μg (r=0.999 8) for jatrorrhizine and 0.162 2-0.567 7 μg (r=0.999 6) for epiberberine.The recoveries were 98.69%,98.66%,98.67%,98.67% and 98.67%,respectively.Conclusion The method possesses high feasibility,strong specificity,high accuracy and good reproducibility,which can be used for the quality control of Erhuang Xiaoyan tablets.

10.
Article in Chinese | WPRIM | ID: wpr-852006

ABSTRACT

Objective To investigate the interaction between Coptidis Rhizoma extracts and intestinal bacteria. Methods The metabolites of Coptidis Rhizoma extracts by human gut microbiota were identified by UPLC-Q-TOF/MS. Furthermore, the effects of Coptidis Rhizoma extracts on the growth of Bifiobactderia, Lactobacilli, Enterobacter, and Enterococcus were evaluated by the colorimetric method. Results Alkaloids in Coptidis Rhizoma were mainly degraded by reduction and demethoxy, among which berberine and coptisine were converted to reductive products and palmatine was metabolized to a demethoxyl product. Additionally, pathogens such as Enterobacter and Enterococcus were obviously inhibited and probiotics such as Lactobacilli and Bifiobactderia were notably promoted by Coptidis Rhizoma extracts. Conclusion Gut microflora can metabolize alkaloids from Coptidis Rhizoma by multiple ways, and Coptidis Rhizoma extracts can improve the intestinal microecology.

11.
Article in Chinese | WPRIM | ID: wpr-851764

ABSTRACT

Objective To establish an effective UHPLC-ESI-MS/MS method for the simultaneous determination of 19 active components (ginsenoside Rg1, Rb1, Rd, berberine, epiberberine, jatrorrhizine, palmatine, columbamine, coptisine, evodiamine, rutaecarpine, dehydroevodiamine, limonin, hyperin, curcumin, demethoxycurcumin, bisdemethoxycurcumin, echinacoside, and verbascoside) of different types in Xintiantai I (XI), and provide a comprehensive and efficient quality control method for traditional Chinese medicine (TCM). Methods The analysis was performed on an Agilent 1290 system with a Agilent Zorbax Eclipse Plus C18 column (150 mm × 4.6 mm, 3.5 μm) at a flow rate of 0.4 mL/min using acetonitrile and 0.1% formic acid aqueous solution as mobile phase. Mass spectrometric detection was performed on multiple reaction monitoring (MRM) in positive and negative ionization mode. The contents of 19 active components in XI were determined by monitoring the specific ions of each component. Results The 19 active components were accurately determined in 15 min and had the good linearity (r2 > 0.999) within the linear ranges. The average recovery rates of ginsenoside Rg1, Rb1, Rd, berberine, epiberberine, jatrorrhizine, palmatine, columbamine, coptisine, evodiamine, rutaecarpine, dehydroevodiamine, limonin, hyperin, curcumin, demethoxycurcumin, bisdemethoxycurcumin, echinacoside, and verbascoside were 94.80%, 96.78%, 95.59%, 96.88%, 97.74%, 100.08%, 96.27%, 100.25%, 98.32%, 97.16%, 95.60%, 95.28%, 96.81%, 95.22%, 96.85%, 95.31%, 93.86%, 94.79%, and 95.20%, respectively; The contents of three batches XI of the 19 components were in the ranges of 2.28-2.49, 0.82-0.90, 1.22-1.32, 14.44-15.50, 3.71-3.99, 3.26-3.49, 3.09-3.33, 4.39-4.72, 4.56-4.92, 0.52-0.57, 0.30-0.33, 4.46-4.76, 3.02-3.24, 2.59-2.76, 6.03-6.38, 1.47-1.58, 1.90-2.08, 3.40-3.88, and 1.53-1.74 mg/g, respectively. Conclusion The developed UHPLC-ESI-MS/MS-MRM method is fast, sensitive, and reproducible for TCM quality control. It can be used for the quality control of XI, which also provides reference for TCM quality research.

12.
Chinese Pharmacological Bulletin ; (12): 552-556, 2017.
Article in Chinese | WPRIM | ID: wpr-511211

ABSTRACT

Aim To establish the pharmacokinetic-pharmacodynamic(PK-PD) modeling to characterize the antipyretic effects of coptisine, an active component in coptis chinensis on rats.Methods Nine healthy male Sprague-Dawley(SD) rats were randomly divided into three groups, each with three.The rats in the first group were injected intravenously with lipopolysaccharide(LPS,100 μg·kg-1) alone.The second and third group rats were given coptisine high-dose(3.87 mg·kg-1) and coptisine low-dose(1.93 mg·kg-1) by tail vein injection at 30 min after LPS injection, respectively.Body temperature was measured at different time points, and blood samples from tail vein were collected simultaneously.The blood concentration of coptisine was determined by ultra performance liquid chromatography.Monolix software was used to model PK-PD of coptisine mean plasma concentration and temperature effects,by population computation with non-covariates.Besides.the model with advantage was selected by the fitting goodness.Results Coptisine could inhibit body temperature of endotoxin-induced fever in rats significantly.Two-compartment linear elimination model was used to describe the final PK model.Gaussian function, an input function of body temperature changes, which was used to depict PD model, the PK and PD models were connected by the Emax model.At last, the final model was fitted better;the fitting results indicated that the EC50 of antipyretic effect of coptisine was 89.7 μg·L-1, and the Emax was 1.88℃.Conclusions Coptisine has a powerful anti-pyretic effect on endotoxin-induced pyrexia of rats with high potency, Low in vivo distribution and quick clearance.

13.
Chinese Traditional Patent Medicine ; (12): 2308-2311, 2017.
Article in Chinese | WPRIM | ID: wpr-668972

ABSTRACT

AIM To establish an HPLC method for the simultaneous content determination of six constituents in Jiaotai Pills (Coptidis Rhizoma and Cinnamomi Cortex).METHODS The analysis of 30% methanol of this drug was performed on a 30 ℃ Agilent ZORBAX SB-C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-KH2PO4flowing at 0.8 mL/min in a gradient elution manner,and the detection wavelength was set at 276 nm.RESULTS Epiberberine,jatrorrhizine hydrochloride,coptisine hydrochloride,palmatine chloride,berberine hydrochloride and cinnamaldehyde showed good linear relationships within the ranges of 0.64-41.24 μg/mL (R2 =0.999 9),0.65-43.76 μg/mL (R2 =1.000 0),0.82-52.65 μg/mL (R2 =0.999 9),0.79-50.70 μg/mL (R2 =0.999 9),3.08-197.20 μg/mL (R2=0.999 8) and 0.65-41.65 μg/mL (R2 =0.999 9),whose average recoveries were 98.06%,102.76%,99.27%,99.75%,96.74% and 101.33% with the RSDs of 0.56%,0.54%,0.39%,0.55%,0.48% and 2.14%,respectively.CONCLUSION This accurate,sensitive,stable and reproducible method can be used for the quality control of Jiaotai Pills.

14.
The Journal of Practical Medicine ; (24): 4033-4037, 2017.
Article in Chinese | WPRIM | ID: wpr-665312

ABSTRACT

Objective To explore the effect of coptisine on the growth of NCI-H1650cells and to evaluate its potential value in the treatment of human non-small cell lung cancer. Methods MTT method was used to ana-lyze cell proliferation. Protein expressions of Bax/Bcl-2 and cytochrome C in NCI-H1650 cells were detected by-Western blot.Apoptosis was analyzed using flow cytometrywithAnnexin V/PI method.ROS concentration was tested with fluorometry.Results Coptisine could significantly inhibit growth of NCI-H1650 cells in a time-and dose-de-pendent manner.Coptisine induced apoptosis in NCI-H1650 cells by inducing ROS accumulation and the following mitochondria mediated apoptosis which was identified by increased Bax expression,Bcl-2 expression was down-reg-ulated,and cytochrome C moved from mitochondria to cytoplasm.ROS inhibitor(N-acetyl cysteine)treatment dra-matically abrogated coptisine-induced growth inhibition and apoptosis.Conclusions This study suggests that copti-sine can induce ROS irritated-and mitochondria-mediated apoptosis in NCI-H1650 cells.Coptisine has a potential value in the treatment of human non-small cell lung cancer.

15.
Article in Chinese | WPRIM | ID: wpr-236057

ABSTRACT

Everted intestinal sac models were used to investigate the intestinal absorption of the 4 alkaloids(berberine, palmatine, coptisine, and epiberberine) in Fuzheng Xiaozheng Fang(FZ) at different intestine segments. The absorption parameters of each component were calculated; SPSS 20.0 software was used to analyze the data and evaluate the absorption characteristics at different intestinal segments. The results showed that all the four active ingredients conformed to zero-order absorption rate. There was significant difference in absorption rate constant (Ka) between the four ingredients at low dose and medium and high dose groups(P<0.05), but there was no significant difference in Ka between medium dose and high dose. The absorption mechanization of four ingredients presented two absorption manners: positive diffusion and passive absorption. The absorptive amount of 4 alkaloids in ileum was slightly greater than that of jejunum, but no significant differences were observed, which indicated that these four alkaloids had no specific absorption windows in intestinal segment.

16.
Article in Chinese | WPRIM | ID: wpr-853767

ABSTRACT

Objective: To establish a quantitative analysis of multi-components by single marker (QAMS) for the simultaneous determination of six alkaloids in crude and processed Coptidis Rhizoma. Methods: An HPLC method was established to determine the six alkaloids (jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, palmatine hydrochloride, and berberine hydrochloride) by the external standard method (ESM). With this HPLC method, the berberine hydrochloride was used as the internal standard (IS) to determine five relative correction factors (RCFs) of the five other alkaloids, and their contents in all samples were calculated by their RCFs at the same time. Compared with the content results determined by the ESM and QAMS, the feasibility and accuracy of QAMS method were verified. Results: Within a certain range, the RCFs of jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, and palmatine hydrochloride to berberine hydrochloride were 1.131, 0.999, 1.011, 1.076, and 1.025, respectively, with the good repeatability in different experimental conditions. There was no significant difference between the QAMS method and ESM method. Conclusion: The QAMS method is feasible and accurate for the simultaneous determination of the six alkaloids in crude and processed Coptidis Rhizoma.

17.
Article in Chinese | WPRIM | ID: wpr-853740

ABSTRACT

Objective: To establish an HPLC method for simultaneous assay of seven main active constituents (protopine, coptisine, palmatine, dehydrocorydaline, D-tetrahydro jatrorrhizine, tetrahydropalmatine, and corydaline) in Corydalis Rhizoma, and on this basis, establishing a methodology of quantitative analysis on multi-components by single marker (QAMS) to validate the feasibility of method and technical adaptability of quality control applications for Corydalis Rhizoma. Methods: Taking seven components in Corydalis Rhizoma as indicators, two correction methods were used to establish the relative correction factor (fk/s) between each component and tetrahydropalmatine. Then the correction factor was used to calculate the amount of each component in Corydalis Rhizoma and finally to achieve this method. In the meantime, the external standard method was used to measure the above seven components to compare the difference between the calculated and measured value of the two fk/s, and to validate the correctness and adaptability of QAMS. Results: The methodology of QAMS which was used to evaluate the seven kinds of alkaloids in Corydalis Rhizoma was established; There was no significant difference between the data calculated by QAMS in different columns and instruments and the values measured by the external standard method. Conclusion: The QAMS method for measuring the components of protopine, coptisine, palmatine, dehydrocorydaline, D-tetrahydro jatrorrhizine, tetrahydropalmatine, and corydaline in Corydalis Rhizoma is reliable and accurate, it could be used to control the quality of crude drugs and herbal pieces of Corydalis Rhizoma.

18.
Article in Chinese | WPRIM | ID: wpr-853520

ABSTRACT

Objective: To study the dissolution changes of four alkaloid contents in the comptability of Coptidis Rhizoma and Rhei Radix et Rhizoma based on compatability ratio and pH valueenvironment. Methods: To determine the contents of berberine, palmatine, coptisine, and epiberberine by HPLC based on the quantitative analysis of multi-components by single marker (QAMS) method. Results: Contents of four kinds of alkaloids and their relative dissolution rates in the co-decoction or mixture of single decoction of Coptidis Rhizoma and Rhei Radix et Rhizoma have a similar reduction trend with the increasing of the ratio of Rhei Radix et Rhizoma.When the proportion of Coptidis Rhizoma and Rhei Radix et Rhizoma was 1∶2, the contents and their relative dissolution rates reached the maximum value, while the dispensing ratio has no effect relationship with the contents and the relative dissolution rates. When the pH value environment of the single decoction of Coptidis Rhizomain the hydrochloric acid solution was 5.20, the pH value environment had no effect relationship with the contents of four alkaloids and relative dissolution rate.Conclusion: There were influences on the contents of the four alkaloids and their relative dissolution rates in the compatability of Coptidis Rhizoma and Rhei Radix et Rhizoma. And both dispensing ratios and pH value environment in the different ratios of Coptidis Rhizoma and Rhei Radix et Rhizoma have the inconsistent influence on the alkaloids with thepH value environment by hydrochloric acid.

19.
Article in Chinese | WPRIM | ID: wpr-853489

ABSTRACT

Objective: To establish and study the UPLC-MS/MS fingerprint, to ascertain the medicinal source of chromatographic peaks, and to identify the chemical constituents for providing scientific basis in effective substance foundation and quality control of Wuji Pill. Methods: The fingerprint of Wuji Pill was developed with ultra-performance liquid chromatography (UPLC), and the Acquity UPLC BEH C18 column (100 mm×2.1 mm, 1.7 μm) was used in the gradient elution with a mobile phase of acetonitrile-water (0.05% formic acid): The flow rate was 0.4 mL/min, the column temperature was 40℃. Similarity evaluation was used to evaluate the quality of herbs from different areas. Results: The fingerprint of Wuji Pill was established with good precision, reproducibility, and stability obtaining within 55 min, and 38 peaks in the fingerprint were designed. Twenty samples could be classified into two clusters. According to the reference, and qualitative and references identified a total of nine peaks were berberine, palmatine, jatrorrhizine, coptisine, epiberberine, evodiamine, rutaecarpine, evodine, and paeoniflorin inferred that the four common peaks were magnolia base magnoflorine, albiflorin, theophylline, and methyl-2-nonyl-4(1H)-quinolone. Conclusion: The establishment of UPLC fingerprint of Wuji Pill and application of chemical pattern recognition can provide a more comprehensive reference for the quality control of herbs.

20.
Article in Chinese | WPRIM | ID: wpr-231017

ABSTRACT

To establish a method for determining the contents of six alkaloids (jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, palmatine hydrochloride, berberine hydrochloride) in six types of Coptidis Rhizoma pieces (crude pieces, ginger juice stir-fried pieces, vinegar stir-fried pieces, wine steamed pieces, wine stir-fried pieces, evodiae juice stir-fried pieces) by RP-HPLC, and explore the relationship with the curative effect of traditional Chinese medicine (TCM) and pharmacodynamics results. The chromatographic column was Welch XtimateTM C₁₈ (4.6 mm×250 mm, 5 μm), with 0.1% triethylamine solution (adjust pH at 10 with ammonium bicarbonate and ammonia) as mobile phase A and acetonitrile as mobile phase B for gradient elution (0-15 min, 10%-25%B; 15-25 min, 25%-30%B; 25-40 min, 30%-45%B) at a rate of 1.0 mL•min⁻¹. The column temperature was set at 30 ℃, and the wavelength was set at 270 nm. The six alkaloids showed a good linear relationship within the range of 0.85-16.96 mg•L⁻¹ (r=0.999 7), 1.25-24.96 mg•L⁻¹ (r=0.999 9), 2.05-40.96 mg•L⁻¹ (r=0.999 9), 3.65-72.96 mg•L⁻¹ (r=0.999 9), 2.88-57.60 mg•L⁻¹ (r=0.999 8), and 13.25-264.96 mg•L⁻¹ (r=0.999 6) respectively. The average recoveries (n=9) of the six alkaloids were 102.4% (RSD 1.2%), 101.8% (RSD 1.3%), 100.3% (RSD 1.8%), 100.7%(RSD 1.8%), 101.2% (RSD 1.5%) and 97.90% (RSD 2.0%) respectively, and their average contents were 3.55, 4.49, 9.12, 19.17, 15.69, 62.56 mg•g⁻¹, respectively. This determination method was accurate and repeatable, which could be used for the content determination in six types of Coptidis Rhizoma pieces. Data analysis on contents determination and preliminary pharmacodynamics results was conducted by using principal component analysis (PCA) and hierarchical clustering analysis (HCA). The analysis results showed that three types of Coptidis Rhizoma pieces (wine steamed pieces, wine stir-fried pieces, and evodiae juice stir-fried pieces) had significant differences with crude pieces, and the wine steamed Coptidis Rhizoma pieces showed most difference with crude pieces especially, mainly related to triglyceride (TG) and fasting blood glucose levels (FBG) in serum. In addition, columbamine hydrochloride was most affected among the six alkaloids. Those three types of Coptidis Rhizoma pieces (wine steamed pieces, wine stir-fried pieces, and evodiae juice stir-fried pieces), had more advantages for "anti-diabetes" in TCM clinical application, especially in the treatment of diabetic hyperlipidemia.

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