Anti-allergic actions of a Chinese patent medicine, huoxiangzhengqi oral liquid, in RBL-2H3 cells and in mice.

CONTEXT: Huoxiangzhengqi oral liquid (HXZQ-OL), a traditional Chinese medicine formula, has antibacterial, anti-inflammation and gastrointestinal motility regulation effects. OBJECTIVE: The study investigates the anti-allergic activity and underlying mechanism of HXZQ-OL. MATERIALS AND METHODS: IgE/Ag-mediated RBL-2H3 cells were used to evaluate the anti-allergic activity of HXZQ-OL (43.97, 439.7 and 4397 µg/mL) in vitro. The release of cytokines and eicosanoids were quantified using ELISA. RT-qPCR was used to measure the gene expression of cytokines. The level of intracellular Ca2+ was measured with Fluo 3/AM. Immunoblotting analysis was performed to investigate the mechanism of HXZQ-OL. In the passive cutaneous anaphylaxis (PCA), BALB/c mice (5 mice/group) were orally administrated with HXZQ-OL (263.8, 527.6 and 1055 mg/kg/d) or dexamethasone (5 mg/kg/d, positive control) for seven consecutive days. RESULTS: HXZQ-OL not only inhibited degranulation of mast cells (IC50, 123 µg/mL), but also inhibited the generation and secretion of IL-4 (IC50, 171.4 µg/mL), TNF-α (IC50, 88.4 µg/mL), LTC4 (IC50, 52.9 µg/mL) and PGD2 (IC50, 195.8 µg/mL). Moreover, HXZQ-OL suppressed the expression of IL-4 and TNF-α mRNA, as well as the phosphorylation of Fyn, Lyn and multiple downstream signalling proteins including MAPK and PI3K/NF-κB pathways. In addition, HXZQ-OL (527.5 mg/kg) attenuated the IgE-mediated PCA with 55% suppression of Evans blue exudation in mice. CONCLUSIONS: HXZQ-OL attenuated the activation of mast cell and PCA. Therefore, HXZQ-OL might be used as an alternative treatment for allergic diseases.

Hypouricemic Effects of Extracts from Urtica hyperborea Jacq. ex Wedd. in Hyperuricemia Mice through XOD, URAT1, and OAT1.

Urtica L. has been long used for gout in traditional Tibetan medicine and is closely related to the effect of reducing uric acid. This study aimed to investigate the effect of Urtica hyperborea Jacq. ex Wedd. (UW) on lowering uric acid and its mechanism by using HK2 cells and hyperuricemia mouse model. Petroleum ether extract (UWP), ethyl acetate extract (UWE), n-butanol extract (UWB), and alcohol-soluble extract (UWA) from UW were prepared, and HK2 cells were treated with various parts extracts to observe the expression of uric acid transporter at 25, 50, and 100 µg/mL for 24 h. Moreover, hyperuricemia mice were administered orally various parts extracts at 0.78 and 2.34 g/kg (crude drug dose converted by extraction rate) to observe the change of hepatic XOD, serum ADA, renal function, and uric acid transporter. In vitro experiments showed that UWA can remarkably elevate OAT1 expression and decrease URAT1 expression in HK2 cells. In vivo experiments showed that UWP, UWE, UWB, and UWA showed remarkable activity in reducing uric acid, rendering a substantial decline in the SUA level in hyperuricemia mice. Compared with the hyperuricemia and allopurinol groups, UWB and UWA had significant protective effects on renal injury. At the same time, UWA can significantly reduce the activity of XOD and ADA, reduce the expression of URAT1, and increase the expression of OAT1. These results indicated that UWA had an outstanding uric acid lowering effect and did not affect renal function. This may be related to increased uric acid excretion and decreased uric acid production, mediated by renal OAT1, URAT1, liver XOD, and serum ADA. UWA may be a potential drug against hyperuricemia.

[Analysis of classical prescription Jinshui Liujun Jian based on ancient literature].

To provide the ancient literary evidence support for the clinical application and development of classical prescription based on systematical collection and analysis of the ancient Chinese medical literature containing Jinshui Liujun Jian, including its origin and development. Bibliometric analysis was used and information of Jinshui Liujun Jian in ancient Chinese medical literature was then collected for statistical analysis of formula compositions, main indications, dosage, preparation methods, etc. A total of 151 valid items of data were obtained from 48 ancient Chinese medicine books. Jinshui Liujun Jian was first recorded in Jingyue Quanshu written by ZHANG Jiebin. This prescription consisted of Rehmanniae Radix Praeparata, Angelicae Sinensis Radix, Pinelliae Rhizome, Citri Reticulatae Pericarpium, Poria and Glycyrrhizae Radix et Rhizome Praeparata cum Melle, and it was mainly used to treat the deficiency of lung and kidney, edema and excess production of phlegm, or Yin deficiency in the old, insufficient blood-qi, wind-cold evil, cough and disgusting, asthma and excessive phlegm. Doctors in later dynasties mostly followed the prescription compositions, dosages and indications in Jingyue Quanshu, and extended the clinical application of this prescription.

[Study on effect of extract from Tibetan medicine Urtica hyperborean on anti-prostatic hyperplasia].

In this study,mouse models of benign prostatic hyperplasia induced by subcutaneous injection of testosterone propionate was used to investigate the therapeutic effect and mechanism of Urtica hyperborean( UW) extracts on prostate hyperplasia in mice. The effects of UW extracts on prostate index,serum epidermal growth factor( EGF) and dihydrotestosterone( DHT) in model mice were observed,and the EGF and anti-apoptotic factor( Bcl-2) mRNA expression levels were detected as well as pathological changes in prostate tissue. The results showed that the ethyl acetate extraction and alcohol soluble fraction of the UW could significantly reduce the prostate index,reduce the serum DHT and EGF levels( P<0. 01),and significantly decrease the EGF and Bcl-2 mRNA expression( P<0. 01),significantly improved the morphological structure of prostate tissue. The above results confirmed that ethyl acetate extract and alcohol-soluble parts of UW have a good preventive effect on mice prostatic hyperplasia model,and its mechanism may be to reduce androgen levels by regulating polypeptide growth factors and/or inhibiting cell hyperproliferation and promoting apoptosis. This study laid the foundation for the further research on UW.

[Rapid identification of constituents of Urtica hyperborea using UPLC-ESI-Q-TOF-MS/MS method].

This paper deals with the application of ultra-performance liquid chromatography tandem quadrupole time of flight mass spectrometry(UPLC-ESI-Q-TOF-MS/MS) method to rapidly determine and analyze the chemical constituents of methanol extract of Urtica hyperborea. We employed UPLC YMC-Triart C18(2. 1 mm×100 mm,1. 9 µm) column to UPLC analysis with acetonitrile-water(containing 0. 4% formic acid) in gradient as mobile phase. The flow rate was 0. 3 m L·min-1 gradient elution and column temperature was 30℃; the injection volume was 4 µL. ESI ion source was used to ensure the data collected in anegative ion mode. The chemical components of U. hyperborea were identified through retention time,exact relative molecular mass,cleavage fragments of MS/MS and reported data.The results indicated that a total of 31 compounds were identified,including 8 flavonoids,14 phenolic compounds,8 phenylpropanoids(4 coumarins and 4 lignans),and 1 steroidal compound,13 of which were confirmed by comparison. The UPLC-ESI-Q-TOF-MS/MS method could rapid identify the chemical components of U. hyperborea. The above compounds were discovered in U. hyperborea for the first time,which could provide theoretical foundation for further research on the basis of the pharmacodynamics of U. hyperborea.

Sagittaria sagittifolia polysaccharide protects against isoniazid- and rifampicin-induced hepatic injury via activation of nuclear factor E2-related factor 2 signaling in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: The Sagittaria sagittifolia L. polysaccharide (SSP) is a purified form of a homogeneous polysaccharide isolated from the root tubers of S. sagittifolia, which has been used as a protectant against hepatotoxicity induced by coadministration of isoniazid and rifampicin. However, the protective effect of SSP against isoniazid- and rifampicin-induced liver injury has never been studied. AIM OF THE STUDY: In this study, the hepatoprotective effect of SSP and its underlying mechanism were investigated in mice with isoniazid- and rifampicin-induced liver injury. MATERIALS AND METHODS: Liver injury was induced in mice by intragastric administration of isoniazid and rifampicin, and the mice were divided into the following six groups: standard control (administration of saline by gavage), model (intragastric administration of isoniazid and rifampicin at 100 mg/kg/day each), positive control (100 mg/kg/day silymarin by gavage 4 h after isoniazid and rifampicin administration), and SSP-treated (200, 400, or 800 mg/kg/day SSP by gavage after isoniazid and rifampicin administration). Subsequently, blood and liver samples were collected from all the animals and were assessed. RESULTS: SSP significantly alleviated the liver injury, as evidenced by decreased activities of alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase in the serum and a decreased level of malondialdehyde in the liver, as well as by an increased level of glutathione and increased activities of superoxide dismutase and catalase in the liver. SSP also effectively reduced the pathological tissue damage. The gene and protein expression of cytochrome P450 (CYP) 2E1 and CYP3A4 was inhibited by SSP. The gene and protein expression of nuclear factor erythroid 2-related factor 2 (NRF2), glutamate-cysteine ligase, and heme oxygenase-1 were induced by SSP, whereas that of Kelch-like ECH-associated protein 1 was inhibited. CONCLUSIONS: SSP exerts a protective effect against isoniazid- and rifampicin-induced liver injury in mice. The underlying mechanisms may involve activation of NRF2 and its target antioxidant enzymes and inhibition of the expression of CYPs.

Study on Chemical Constituents in Polygoni Cuspidati Folium and its Preparation by UPLC-ESI-Q-TOF-MS/MS.

A rapid and credible analytical method was developed using online UPLC-ESI-Q-TOF-MS/MS to identify chemical constituents in Polygoni cuspidati folium and its preparation. By accurate mass measurements within 6.5 ppm error for [M-H]- ion in routine analysis, 26 chemical constituents, including tannin, derivatives of phenylpropionic acid, stilbene, flavonoid, anthraquinone, torachryson and its derivatives, were identified or tentatively characterized. Among them, five constituents (compounds 19-23) were firstly reported in Polygoni cuspidati folium, other 17 constituents were coexisting in both Polygoni cuspidati folium and its preparation. Fragmentation behaviors of different categories of constituents were also investigated to confirm the results. This established UPLC-ESI-Q-TOF-MS/MS method, with reliance and efficiency for the identification the major constituents, would be the basis for quality control of Polygoni cuspidati folium and its preparation.

[Resources and usage status of Tibetan medicinal plant "Bangga"].

The study is aimed to clarify the actual original plant, find out the usage status and the resource distribution of the Tibetan medicinal plant "Bangga". By using the way of the literatures survey, interview and investigation, it found out that the actual original plant of the Tibetan medicinal plant "Bangga" were the whole dried plant or the aerial part of Aconitum tanguticum or A. naviculare of Ranunculaceae, among which A. tanguticummainly distributed in Sichuan, Gansu, Qinghai, Tibet (Qamdo), and A. naviculare mainly distributed in Tibet. Sichuan, Gansu, Qinghai and other Tibetan areas mainly used the resources of A. tanguticum, Tibet (except the Qamdo area) mainly uses the A. naviculare, which resource was imminent in danger. Other species described in the literature were not used. It showed that the use of herbs related to their resources, it is recommended to strengthen the protection and guide the market.

Swertianlarin, an Herbal Agent Derived from Swertia mussotii Franch, Attenuates Liver Injury, Inflammation, and Cholestasis in Common Bile Duct-Ligated Rats.

Swertianlarin is an herbal agent abundantly distributed in Swertia mussotii Franch, a Chinese traditional herb used for treatment of jaundice. To study the therapeutic effect of swertianlarin on cholestasis, liver injury, serum proinflammatory cytokines, and bile salt concentrations were measured by comparing rats treated with swertianlarin 100 mg/kg/d or saline for 3, 7, or 14 days after bile duct ligation (BDL). Serum alanine aminotransferase (ATL) and aspartate aminotransferase (AST) levels were significantly decreased in BDL rats treated with swertianlarin for 14 days (P < 0.05). The reduced liver injury in BDL rats by swertianlarin treatment for 14 days was further confirmed by liver histopathology. Levels of serum tumor necrosis factor alpha (TNFα) were decreased by swertianlarin in BDL rats for 3 and 7 days (P < 0.05). Moreover, reductions in serum interleukins IL-1ß and IL-6 levels were also observed in BDL rats treated with swertianlarin (P < 0.05). In addition, most of serum toxic bile salt concentrations (e.g., chenodeoxycholic acid (CDCA) and deoxycholic acid (DCA)) in cholestatic rats were decreased by swertianlarin (P < 0.05). In conclusion, the data suggest that swertianlarin derived from Swertia mussotii Franch attenuates liver injury, inflammation, and cholestasis in bile duct-ligated rats.

Swertianlarin, isolated from Swertia mussotii Franch, increases detoxification enzymes and efflux transporters expression in rats.

Swertianlarin, isolated from Swertia mussotii Franch and Enicostemma axillare, has hepatoprotective effects against cholestasis in rat models of hepatotoxicity. However, the underlying molecular mechanism is not clear. We then treated rats with swertianlarin for 7 d and then measured serum liver injury markers, lipids, and bile salts, as well as the expression of bile acid synthesis and detoxification enzymes (e.g. Cyp7a1 and Cyp3a), membrane influx and efflux transporters (e.g. Ntcp and Mrp3), nuclear receptors (e.g. Pxr and Fxr/Shp) and transcriptional factors (e.g. Nrf2 and Hnf3ß) in the liver. We found a significant induction of the expression of the basolateral efflux transporters Mrp3 and Mrp4 and canalicular transporter Mdr1 in rats treated with swertianlarin, compared with the controls (1.9-fold and 2.2-fold, P < 0.005, and 3.4-fold, P < 0.05, respectively). The expression of detoxification enzymes Cyp3a, Ugt2b, Sult2a1 and Gsta1 in rats treated with swertianlarin was significantly higher than that in controls (3.7-fold, 2.8-fold, 2.1-fold, and 1.7-fold, respectively, all P < 0.05). Expression of the synthetic enzyme, Cyp8b1, was higher in rats treated with swertianlarin than that in controls (1.8-fold at mRNA level and 3.4-flod at protein level, P < 0.05). Elevated serum levels of the conjugated bile acids, taurocholic acid and taurodeoxycholic acid, and a reduction in levels of serum ALP, unconjugated bile acid αMCA, and TG were observed (all P < 0.05). In conclusion, swertianlarin significantly up-regulates hepatic bile acid detoxification enzymes and efflux transporters in rats, which can increase the water solubility of hydrophobic bile acids and elimination of conjugated bile acids.

Discriminating Lamiophlomis rotata According to Geographical Origin by (1)H-NMR Spectroscopy and Multivariate Analysis.

INTRODUCTION: Lamiophlomis rotata (Duyiwei) is a folk herbal medicine that traditionally has been used in China as a hemostatic agent. Raw plant materials used for medicinal products from different geographical regions are often inconsistent in chemical composition. Metabolic fingerprinting provides a new approach for distinguishing the geographical origins of L. rotata. OBJECTIVE: To identify metabolites that contribute to the different geographical regions of L. rotata samples. METHODS: Lamiophlomis rotata metabolomics were performed by (1)H-nuclear magnetic resonance (NMR) spectroscopy and multivariate statistical analyses. The L. rotata metabolic profile was prepared for NMR measurements using methanol-d4 solvent. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were applied to analyse the L. rotata (1)H-NMR spectroscopy data. RESULTS: Nine iridoid glycosides, one flavonoid and three phenylpropanoid glycosides were detected in L. rotata by (1)H-NMR spectroscopy. (1)H-NMR measurements and multivariate analysis were used to successfully discriminate samples from three different locations. CONCLUSION: The NMR-based analysis of L. rotata is a more comprehensive approach than traditional chromatographic methods. Simple sample preparation, rapidity and reproducibility of are additional advantages of NMR analysis.

Metabolic discrimination of Swertia mussotii and Swertia chirayita known as "Zangyinchen" in traditional Tibetan medicine by (1)H NMR-based metabolomics.

Swertia mussotii Franch. and Swertia chirayita Buch.-Ham. have been commonly used under the same name "Zangyinchen" for the treatment of liver and gallbladder diseases in traditional Tibetan medicine. Detailed characterization and comparison of the complete set of metabolites of these two species are critical for their objective identification and quality control. In this study, a rapid, simple and comprehensive (1)H NMR-based metabolomics method was first developed to differentiate the two species. A broad range of metabolites, including iridoid glycosides, xanthones, triterpenoids, flavonoids, carbohydrates, and amino acids, were identified. Statistical analysis showed evident differences between the two species, and the major markers responsible for the differences were screened. In addition, quantitative (1)H NMR method (qHNMR) was used for the target analysis of the discriminating metabolites. The results showed that S. mussotii had significantly higher contents of gentiopicrin, isoorientin, glucose, loganic acid, and choline, whereas S. chirayita exhibited higher levels of swertiamarin, oleanolic acid, valine, and fatty acids. These findings indicate that (1)H NMR-based metabolomics is a reliable and effective method for the metabolic profiling and discrimination of the two Swertia species, and can be used to verify the genuine origin of Zangyinchen.

[Study on TLC identification of Dida commonly used in Tibetan medicine from different species].

OBJECTIVE: To establish a method of TLC identification for Dida commonly used in Tibetan medicine from different species. METHOD: With silica gel G as the stationary phase, and chloroform-methanol (40: 1) as mobile phase, oleanolic acid from different species of Dida was separated and identified. RESULT: Oleanolic acid was detected in 70 kinds of Dida derived from the Gentianaceae Swertia, Halenia, Gentianopsis, Lomatogonium, and Saxifragaceae saxifrage, except for the saxifrage, there are some differences among different genera or subjection. CONCLUSION: This TLC method can be used for identification of oleanolic acid in Dida from different species except saxifrage.

[Evaluation of medicinal reasonableness and resource utilization of Dida (Zang Yin Cheng) commonly used in Tibetan medicine].

OBJECTIVE: To evaluate the medicinal reasonableness and resource utilization of Dida from different species. METHOD: With common characteristic absorption peaks of HPLC fingerprints and SPSS cluster, the composition similarity of Dida from different species was evaluated. RESULT: The composition similarity of HPLC fingerprints of 33 Dida samples from 15 species and 1 variety originated from Swertia, Halenia, Gentianopsis, Lomatogonium was difference. The original species can be clustered into four groups by the relative area of 10 common characteristic peaks of HPLC fingerprints. The compositions of four different genera are quite different. CONCLUSION: Because of containing iridoids, xanthones, and triterpenes which have liver protection and cholagogue functions, all of species from Swertia, Halenia, Gentianopsis and Lomatogonium in Gentianaceae are classified as Dida in Tibetan medicine. According to the composition difference among different species, the HPLC fingerprints established for Dida from different source are an effective means to identify nd control the quality of Dida.

[Difference of chemical constituents contained in Tibetan herb Jiadiranguo from different habitats by HPLC fingerprint].

OBJECTIVE: To establish the HPLC fingerprint for Halenia elliptica herbs, a traditional Tibetan medicine, in order to study constituents contained in H. elliptica from different habitats and compare their differences. METHOD: HPLC analysis was made on a Welchrom-C18 (4.6 mm x 250 mm, 5 microm) with water and acetonitrile as mobile phase. The wavelength was detected as 265 nm, the flow rate was 1.0 mL x min(-1), and the column temperature was 40 degrees C. The software for chromatographic fingerprint was applied to analyze the similarity. And principal component analysis was conducted. RESULT: Twelve common chromatographic peaks were identified by fingerprint, showing a low similarity in constituent and variety. The significant difference in the proportion between xanthones and aglycones in each batch of herbs indicated no notable correlation between constituent characteristics and geographic locations of habitats. CONCLUSION: The method is so simple, exclusive, stable and highly repeatable that it can provide reference for identification and quality assessment of H. elliptica herbs.

Metabolic discrimination of rhizoma coptidis from different species using 1H NMR spectroscopy and principal component analysis.

Rhizoma coptidis, a broadly used medicinal plant, originates from the dried rhizomes of three species in Chinese pharmacopoeia, namely, Coptis chinensis Franch, Coptis deltoidea C. Y. Cheng et Hsiao, and Coptis teeta Wall. In this study, a novel approach using (1)H NMR spectroscopy combined with multivariate analysis was introduced to differentiate the three species and identify potential metabolic markers for better controlling the quality of rhizoma coptidis. A broad range of metabolites including alkaloids, sugars, organic acids, amino acids, and fatty acids present in rhizoma coptidis were detected by means of (1)H NMR spectroscopy. Principal component analysis (PCA) of the (1)H NMR data set showed a clear separation between all samples by PC1 and PC3, and some metabolites that could be responsible for the discrimination of the three species were identified. An analysis of variance (ANOVA) was performed to statistically verify the significance of differences in metabolite levels between species. By combining PCA and ANOVA, significantly higher contents of palmatine, coptisine, epiberberine, columbamine, and fatty acids together with lower contents of jateorrhizine were found in Coptis chinensis, whereas Coptis deltoidea and Coptis teetA showed the highest levels of sucrose and chlorogenic acid, respectively. This study indicates that metabolites of rhizoma coptidis vary with the species and the proposed method is suitable for metabolic fingerprinting analysis to check the genuine origin of rhizoma coptidis.

[Determination and quality assessment of 10 ingredients gentiopicroside and sweroside and so on in Tibetan medicine Jia Di (Swertia chirayita)].

OBJECTIVE: To establish a method for determination of 10 ingredients such as gentiopicroside, sweroside, and mangiferin in India swertia, and settle the index components and their limits. METHOD: By Welch materials AQ-C18 column, determination was conducted by the gradient elution with methanol and 0.4% formic acid as mobile phase, with column temperature 30 degrees C, flow rate at 1.0 mL x min(-1), and 254 nm as the detection wavelength. RESULT: The linear relatives of 10 ingredients were good. The method showed the high precision and good reproducibility, and recovery rates were between 97% and 103%. The ingredients of market com-modities varied greatly. CONCLUSION: This method is simple, sensitive, reproducible, and applicable to the determination of the main ingredients in India Swertia. Sweroside and mango glycosides were suggested as the index components for determination in Jia Di (Swertia chirayita), and their content limits are not less than 0.1%, 0.3%, respectively.

[Dynamic accumulation of effective components and biomass of Coptis chinensis in Hongya county].

OBJECTIVE: To study the dynamic accumulation of the effective components and biomass of Coptis chinensis, so to provide the experimental date of optimal harvest time for C. chinensis in Hongya county. METHOD: The samples of three to five years were gathered from the same field and time. The biomass was analyzed by weighed. The jatrorrhizine, columbamine, epiberberine, coptisine, palmatine and berberine in C. chinensis were analyzed by HPLC. RESULT: With the increasing of years of growth, the rootstalk biomass of C. chinensis was increasing continuously. The biomass growth of four-year-old C. chinensis was the fastest in the year. From September to October was the fastest season of the growth of rootstalk. The dynamic accumulation in rootstalk C. chinensis had regularity in the certain extend. The contents of six alkaloids and all alkaloids in 4-year-old C. chinensis were more than that in 3-years-old and 5-year-old. The contents of six alkaloids were mostly highest in August. From July to December, there is no significant difference in the contents of columbamine, epiberberine, coptisine, palmatine, berberine and all alkaloids in 4-years-old C. chinensis. CONCLUSION: According to the biomass and the accumulation pattern of the effective components in the C. chinensis, the optimal harvest time is from September to October of 4-year-old C. chinensis.

[Simultaneous determination of six alkaloids in Coptis chinensis of different regions by RP-HPLC].

A reversed-phase HPLC method for simultaneous determination of gatrorrhizine, columbamine, epiberberine, coptisine, palmatine and berberine in Coptis chinensis was developed. Analysis was carried out on an Xtimate C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetonitrile-30 mmol x L(-1) ammonium bicarbonate solution (including 0.7% ammonia and 0.1% triethylamine) by gradient elution. The detective wavelength was 270 nm, the column temperature was 30 degrees C, and the flow rate was 1.0 mL x min(-1). By the above method, the linear ranges of gatrorrhizine, columbamine, epiberberine, coptisine, palmatine and berberine were 0.85-16.96 (r = 0.9997), 1.25-24.96 (r = 0.999 5), 2.05-40.96 (r = 0.999 9), 3.65-72.96 (r = 0.999 9), 2.88-57.60 (r = 0. 999 8),13.25-264.96 mg x L(-1) (r = 0.999 6), respectively. The average recoveries (n = 6) of the six alkaloids were 101.6% (RSD 1.3%),102.5% (RSD 1.5%), 100.8% (RSD 1.9%),102. 6% (RSD 1.2%), 97.80% (RSD 1.3%), 99.01% (RSD 1.5%), respectively. The determined results demonstrate that there is a significant difference in the contents of six alkaloids and total alkaloids among the tested samples. The method is accurate, reliable and repeatable for simultaneous determination of gatrorrhizine, columbamine, epiberberine, coptisine, palmatine and berberine in C. chinensis.

[Simultaneous determination of jatrorrhizine, palmatine, berberine, and obacunone in Phellodendri Amurensis Cortex by RP-HPLC].

OBJECTIVE: To develop a HPLC method to determine the content of jatrorrhizine, palmatine, berberine, and obacunone in Phellodendri Amurensis Cortex simultaneously. MEHTOD: The separations were carried out at 25 degrees C on a Phenomenex Gemini C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetonitril and water containing 0.1% phosphoric acid in gradient mode. The flow rate was 1.0 mL x min(-1), detection wavelengthes were 345 nm for jatrorrhizine , palmatine, berberine and 210 nm for obacunone. RESULT: The average recoveries of jatrorrhizine, palmatine, berberine, and obacunone were 98.94%, 101.17%, 96.22% and 98.90%, respectively. CONCLUSION: The method is simple, accurate and repeatable, and can be used in content determination of jatrorrhizine, palmatine, berberine, and obacunone in Phellodendri Amurensis Cortex.