Determination of Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin in Danqi Xinmaikang boiled powders and pieces by quantitative analysis of multi-components by single maker / 国际中医中药杂志

Objective:To establish a quality evaluation method for the simultaneous determination of Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin in Danqi Xinmaikang boiled powders and pieces.Methods:Quantitative analysis of multi-components was performed to determine contents of Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin with Calycosin-7-O-β-D-Glucopyranoside as the reference substance by single-maker (QAMS). The chromatogram conditions were established, with C18 column as solid phase, acetonitrile-water as flowing phase, 268 nm as detecting wavelength, 1.0 ml/min as flowing rate, 30 ℃ as column temperature, and 10 μl as injection volume.Results:The relative correction factor between Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin was 1.14. There was no significant difference of measured values between the external standard method and QAMS ( P>0.05). With Calycosin-7-O-β-D-Glucopyranoside retention time of 1.00, the relative retention time of Lobetyolin was 1.51 and RSD was less than 5%. Conclusion:It is feasible and accurate to evaluate the quality of Danqi Xinmaikang boiled powders and pieces by QAMS.

Lignans with inhibitory effect on 5α-reductase from Urtica cannabina / 中国中药杂志

The lignans in Urtica cannabina were isolated by preparative HPLC, silica, and ODS column chromatographies, and identified by NMR and HR-MS. The inhibitory activities on 5α-reductase were evaluated in vitro. As a result, ten secolignans,(2R,4S)-2,4-bis(3-methoxyl-4-hydroxyphenyl)-3-butoxypropanol(1), 3,4-trans-3-hydroxymethyl-4-[bis(3,4-dimethoxyphenyl)methyl] butyrolactone(2), 3,4-trans-3-hydroxymethyl-4-[(3,4-dimethoxyphenyl)(3-methoxyl-4-hydroxyphenyl)methyl] butyrolactone(3), 3,4-trans-3-hydroxymethyl-4-[bis(3-methoxyl-4-hydroxyphenyl)methyl] butyrolactone(trans urticol, 4), 3,4-trans-3-hydroxymethyl-4-[bis(3,4-dimethoxyphenyl)methyl] butyrolactone-3-O-β-D-glucopyranoside(5), 3,4-trans-3-hydroxymethyl-4-[(3,4-dimethoxyphenyl)(3-methoxyl-4-hydroxyphenyl)methyl]butyrolactone-3-O-β-D-glucopyranoside(6), 3,4-trans-3-hydroxymethyl-4-[bis(3-methoxyl-4-hydroxyphenyl)methyl]butyrolactone-3-O-β-D-glucopyranoside(trans-urticol-7-O-β-D-glucopyranoside, 7), cycloolivil-4-O-β-D-glucopyranoside(8), isolariciresinol-4'-O-β-D-glucopyranoside(9), and olivil-4'-O-β-D-glucopyranoside(10), together with a polyphenol [α-viniferin(11)], were isolated from U. cannabina for the first time. Compound 1 was a new lignan. Compound 7 was potent in inhibiting 5α-reductase.

Distribution and dynamic changes of chemical constituents in Callicarpa nudiflora / 中草药

Objective: To provide the theoretical basis for determining the best harvesting plant organ and harvesting period, and investigate the content of chemical constituents of Callicarpa nudiflora in different plant organs and different growth periods. Methods: The contents of total flavonoids, total phenolic acid and total saponins were determined by ultraviolet spectrophotometry, and the seven components were determined by HPLC. The ANOVA and PCA methods were used to analyze the content of each constituent. Results: The dry extract rate, the contents of total flavonoids, total phenolic acid, total saponins, forsythiaside B, acteoside, isoacteoside, and apigenin-7-O-β-D-glucopyranoside in functional leaves were the highest, while the contents of caffeic acid, galuteolin and luteolin in tender leaves were the highest, and all of them were significantly different from the young shoots (P 0.05). The contents of total phenolic acid, total saponins, forsythiaside B, and acteoside were the highest in the FB period, and there was no significant difference with the EFS period (P > 0.05). The contents of galuteolin and apigenin-7-O-β-D-glucopyranoside were the highest in the earlier fruit maturation (EFM) period and the later fruit maturation (LFM) period, respectively, and there was no significant difference with the EFS period (P > 0.05). The contents of each chemical component were reduced to the minimum at the fruit-drop (FD) period, and it was significantly different from that at the EFS period and the FB period (P < 0.05). According to the comprehensive evaluation model constructed by PCA, the comprehensive score of the EFS period was the highest (F = 3.252), followed by the FB period (F = 3.011). Conclusion: Main chemical constituents of C. nudiflora were significantly different in harvesting parts and growth periods. The contents of main chemical constituents were higher in functional leaves and tender leaves, and the contents of main chemical constituents were higher from FB period to EFS period.

Simultaneous determination of 10 active ingredients in Yiganning Granules by UPLC / 中草药

Objective: To establish a UPLC method to simultaneously determine 10 active ingredients in Yiganning Granules (YG) and provide scientific basis for the quality control, evaluation and standard revision of YG preparations. Methods: A UPLC method was used with an Acquity UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm). The mobile phase was acetonitrile-mehanol-0.15% phosphoric acid solution with gradient elution. The flow rate was 0.3 mL/min. The column temperature was 45 ℃. The injection volume was 2 μL. Results: Ten active ingredients (chlorogenic acid, atractylenolide I, paeoniflorin, calycosin 7-O-β-D- glucopyranoside, stilbene glycoside, caffeic acid, toosendanin, kaempferol, paeonol and tanshinone ⅡA) in YG were simultaneously determined. The linearity was good (r ≥ 0.999 0), the limit of detection and quantification were 0.006-0.017 μg/mL and 0.017-0.510 μg/mL. The average recoveries were 98.8%-102.5% with RSDs of 1.13%-5.37%. Through the determination of 16 batches of samples, the average content of the above 10 ingredients was in turn (5.724 ± 0.017), (0.273 ± 0.003), (0.854 ± 0.005), (1.228 ± 0.004), (0.496 ± 0.003), (1.287 ± 0.004), (0.137 ± 0.004), (3.624 ± 0.014), (7.366 ± 0.032) and (1.754 ± 0.005) mg/g, respectively. Conclusion The established UPLC method is simple, specific, sensitive, stable, precise, accurate, and reproducible, which can be used for quality control and evaluation of YG.

Establishment of HPLC fingerprints of Qiwei Tangmaishu tablet and simultaneous determination of six constituents / 国际药学研究杂志

Objective: To establish HPLC fingerprints of Qiwei Tangmaishu tablet and simultaneously determine the contents of the six constituents, acteoside, martynoside, calycosin 7-O-β- D-glucopyranoside, schisandrin, formononetin and deoxyschizan- drin. Methods: The analysis of 50% methanol extract of this drug was performed on a 30℃ thermostatic Waters Symmetry C18 column, with the mobile phase comprising of acetonitrile(A)-0.2% formic acid solution(B)flowing at 1.0 ml/min in a gradient elution manner. The detection wavelength was set at 330 nm for acteoside and martynoside, 254 nm for calycosin 7-O-β-D-glucopyranoside, schisandrin, formononetin and deoxyschizandrin. Results: There were thirteen common peaks in the fingerprints of ten batches of samples with the similarities more than 0.9. Acteoside, martynoside, calycosin 7-O-β-D-glucopyranoside, schisandrin, formononetin and deoxyschizandrin showed good linear relationship within the ranges of 1.47-36.75, 0.66-16.50, 0.89-22.25, 2.58-64.50, 1.91-47.75 and 0.77-19.25 μg/ml(r≥0.9993), and their average recoveries were 97.57%, 99.22%, 96.69%, 100.01%, 98.79% and 96.77%, with the RSD of 0.79%, 1.54%, 0.61%, 0.64%, 0.83% and 0.36%, respectively. Conclusion: The established method is easily operable and repeatable, which could be used for quality control of Qiwei Tangmaishu tablet.

Purification of Periploca forrestii total flavonoids and assay for the in vitro inhibitory activity on xanthine oxidase / 国际药学研究杂志

Objective: To extract and purify total flavonoids from Periploca forrestii Schltr.(P. forrestii),and test the in vitro inhibitory activity of the total flavonoids and two flvaonoidal compounds in P. forrestii,so as to provide a reference for studies on the related medicinal substances in P. forrestii. Methods: Total flavonoids were extracted from P. forrestii and then purified by the column chromatography on macroporous resin and polyamide columns. The content of total flavonoids was determined according to the Lambert-Beer’s law. The in vitro xanthine oxidase(XOD)inhibitory ac- tivity was assayed for total flavonoids and the two flavonoidal compounds by the ultraviolet spectrophotometry. Results The purified total flavonoids had a content of more than 95%. The total flavonoids and two flavonoidal compounds all showed an inhibitory effect on XOD in vitro,with the inhibitory rate enhanced with increasing concentration. The IC50 of the total flavonoids as well as the two flavonoidal compounds,quercetin-3-O-α-L-pyranoside(QP)and quercetin-7-O-β- D-glucopyranoside(QG)were 608.9,221.2 and 261.2 μg/ml,respectively. Conclusion: The total flavonoids as well as the two flvaonoidal compounds QP and QG in P. forrestii all showed the in vitro inhibitory activity on XOD.

Optimization of extraction and purification technology for Huangqi Baihe Granules based on G1-entropy method and orthogonal design / 中草药

Objective To determine the optimum extraction and purification technology of Huangqi Baihe Granules (HBG) by using orthogonal design and combination empowerment based on G1-entropy method, so as to provide a reference for the industrial production. Methods With the ethanol extraction amount and paste-forming rate of calycosin7-O-β-D-glucopyranoside, hesperidin, crude polysaccharide as evaluation index, and with the extraction times, extraction time and the water adding amount as investigate factors, then the combination empowerment method based on G1-entropy method and orthogonal design were used to optimize the extraction technology of HBG. The same combined methods were used to optimize the purification technology of HBG with retention rate and removal rate of impurity of ethanol extract of calycosin7-O-β-D-glucopyranoside, hesperidin and crude polysaccharide, as evaluation index, and with the membrane pore diameter, operating pressure and filtration temperature of multi-channel tubular ceramic ultrafiltration membrane as investigate factors. Results The test results showed that the optimum extraction technology was as follows: Extracted 2 times with 20 times the amount of water and each for 75 min. The optimum ultrafiltration technology was as follows: Multi-channel tubular inorganic ceramic membrane at 50 nm, operating pressure at 0.10 MPa, filtration temperature at 45 ℃. Under such condition, there was no significant difference between verification groups of three batches. Conclusion The optimized extraction and purification process is stable and feasible by verification, which can provide experimental basis for industrial production of HBG.

Chemical constituents from roots of Veratrilla baillonii / 中草药

Objective To study the chemical constituents from the roots of Veratrilla baillonii. Methods Compounds were purified by normal and reversed column chromatographic techniques, and isolated by high performance liquid chromatography. Their structures were identified on the basis of spectral data including MS and NMR. Results Twenty-four compounds (including 10 xanthone glycosides, 8 xanthones, and 6 iridoids) were isolated from the ethyl acetate extracts of the roots of V. baillonii. They were identified as 1-hydroxy-2,3,4-trimethoxyxanthone-7-O-β-D-glucopyranoside (1), tripteroside (2), 1-hydroxy-2,7-dimethoxyxanthone-3-O-β-D- glucopyranoside (3), 1-hydroxy-3,4-dimethoxyxanthone-7-O-β-D-glucopyranoside (4), secamonoide B (5), tetrasweroside A (6), veratriloside B (7), 2,3,4,5-tetramethoxyxanthone-1-O-β-D-glucopyranoxyl-(1→6)-β-D-glucopyranoside (8), 2,3,4,7-tetra- methoxyxanthone-1-O-β-D-xylopyranoxyl-(1→6)-β-D-glucopyranoside (9), 2,3,5-trimethoxy-xanthone-1-O-β-D-xylopyranoxyl- (1→6)-β-D-glucopyranoside (10), 1,3-dihydroxy-4,7-dimethoxyxanthone (11), 1,7-dihydroxy-2,3,4-trimethoxyxanthone (12), 1,7- dihydroxy-3,4-dimethoxyxanthone (13), 1,7-dihydroxy-3-methoxyxanthone (14), 1-hydroxy-2,3,4,5-tetramethoxyxanthone (15), 1-hydroxy-2,3,4,7-tetramethoxyxanthone (16), 1-hydroxy-2,3,5-trimethoxyxanthone (17), 1-hydroxy-2,3,7-trimethoxyxanthone (18), sweroside (19), gentiopicrin (20), swertiamarin (21), deacetylcentapicrin (22), amaronitidin (23), and amarogentin (24). Conclusion Compound 1 is a new compound named 2-methoveratriloside, and compounds 2, 3, 5, 6, 8, 10-12, 14, and 22-24 are isolated from the Veratrilla genus for the first time.

Flavonoids isolated from capitula of Coreopsis tinctoria / 中草药

Objective To study the chemical constituents of the water extract of capitula of Coreopsis tinctoria. Methods The chemical constituents from water extract were isolated and prepared by silica gel column, reversed-phase ODS, and pre-HPLC, and their structures were identified according to the physical and chemical properties and spectral data. Results A total of 13 compounds were isolated and identified as 8,3’,4’-trihydroxyflavone-7-O-β-D-glucopyranoside (1), 6-Hydroxyluteolin-7-O-β-D-glucoside (2), kaempferol (3), quercetin-7-O-β-D-glucopyranoside (4), (2S)-3’,4’,5,8-tetrahydroxyflavanone-7-O-β-D-glucoside (5), (2S)- eriodictyol-5-O-β-D-glucoside (6), butin-7-O-β-D-glucopyranoside (7), plathymenin (8), (Z)-6-O-β-D-glucopyranosyl-6,3’,4’- trihydroxyaurone (9), 5,6,3’,4’-tetrahydroxyaurone (10), 6,3’,4’-trihydroxyaurone (11), okanin-5’-O-β-D-glucopyranoside (12), and 4’-O-β-D- glucopyranosyl-3,4,2’,4’,5’-pentahydroxychalcon (13). Conclusion Thirteen compounds are isolated from C. tinctoria for the first time.

Research on HPLC Fingerprint of Jinsangqi Kangdu Dropping Pills / 中国中医药信息杂志

Objective To establish fingerprint of Jinsangqi Kangdu Dropping Pills by HPLC; To control the quality of the preparation. Methods Waters XSELECT CSH-C18 chromatographic column (4.6 mm × 150 mm, 5 μm) was used and eluted with acetonitrile - 0.1% phosphoric acid solution gradient at the flow rate of 1.0 mL/min. The detection wavelength was 260 nm with column temperature of 30 ℃. Using calycosin-7-O-β-D-glucopyranoside, rutin, liquiritin, hyperoside, quercetin and ammonium glycyrrhizinate as the object references, ten batches of Jinsangqi Kangdu Dropping Pills were tested and analyzed by similarity comparison.Results Fringerprint spectrum of Jinsangqi Kangdu Dropping Pills had 24 common peaks in total, and characteristic spectrums of Hypericum Perforatum, Mori Cortex, Astragali Radix and Glycyrrhizae Radix et Rhizoma had been found, while similarity of HPLC fingerprint was more than 0.9 among those batches of samples. Conclusion Using HPLC fingerprint can evaluate the Jinsangqi Kangdu Dropping Pills quality totality,which can provide references for improving the quality control of the preparation.

Chemical constituents from Rubus caesius and their inhibitory activities on PTP 1B / 中草药

Objective: To investigate the chemical constituents from the stem and leaves of Rubus caesius and the inhibitory activities on PTP 1B. Methods: Compounds were separated and purified by silica gel column chromatography, Sephadex LH-20 column chromatography, and preparative liquid chromatography. Their structures were identified by spectral methods. The PTP1B inhibitory activities were screened by microplate reader. Results: Five compounds were obtained from the stems of R. caesius respectively, elucidated as naringin (1), apigenin-7-O-β-D-glucopyranoside (2), isoquercitrin (3), hyperoside (4), and (-)-epicatechin 3-O-gallate (ECG) (5), and two compounds were obtained from the leaves respectively, elucidated as acteoside (6) and ellagic acid (7) respectively. Conclusion: Compounds 1-7 are isolated from this plant for the first time. Different fractions and compounds showed different PTP1B inhibitory activities and acteoside showed high PTP1B inhibitory activity with the IC50 value of (27.41 ± 0.61) μg/mL. This compound may be the main active composition of leaves ethyl acetate fraction.

Simultaneous determination of 11 constituents in Shenqi Shiyiwei Granule by HPLC-DAD / 中草药

Objective: To establish HPLC-DAD method for the simultaneous determination of 23-acetate alisol B, ferulic acid, verbascoside, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, gastrodin, chrysophanol, aurantio-obtusin, calycosin 7-O-β-D-glucopyranoside, and hyperoside in Shenqi Shiyiwei Granule (SSG). Methods: The chromatographic separation was achieved on an Waters XBridge-C18 (250 mm × 4.6 mm, 5.0 μm) column with methanol-acetonitrile-water (15:80:5) and methanol-0.1% phosphoric acid (10:90) as mobile phases for gradient elution, at the flow rate of 1.0 mL/min; The column temperature was 35℃. Results: The linear ranges of 23-acetate alisol B, ferulic acid, verbascoside, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, gastrodin, chrysophanol, aurantio-obtusin, calycosin 7-O-β-D-glucopyranoside, and hyperoside were 0.4-8.0 μg/mL (r = 0.999 2), 0.2-4.0 μg/mL (r = 0.999 5), 0.2-4.0 μg/mL (r = 0.999 5), 0.1-2.0 μg/mL (r = 0.999 6), 0.1-2.0 μg/mL (r = 0.999 7), 0.1-2.0 μg/mL (r = 0.999 4), 0.5-10 μg/mL (r = 0.999 2), 0.6-12 μg/mL (r = 0.999 2), 0.4-8.0 μg/mL (r = 0.999 4), 1.0-20 μg/mL (r = 0.999 6), and 0.8-16 μg/mL (r = 0.9993). The average recoveries (n = 6) were 98.1% (RSD = 0.9%), 98.1% (RSD = 1.6%), 99.1% (RSD = 1.6%), 98.3% (RSD = 1.8%), 99.5% (RSD = 1.5%), 99.9% (RSD = 0.6%), 98.5% (RSD = 0.7%), 100.4 (RSD = 0.8%), 101.6% (RSD = 0.4%), 99.7% (RSD = 0.9%), and 101.2% (RSD = 1.1%), respectively. The contents of nine batches of 23-acetate alisol B, ferulic acid, verbascoside, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, gastrodin, chrysophanol, aurantio-obtusin, calycosin 7-O-β-D-glucopyranoside, and hyperoside were 0.081-0.089, 0.261-0.269, 0.060-0.069, 0.038-0.047, 0.030-0.037, 0.042-0.049, 0.420-0.428, 0.141-0.151, 0.178-0.189, 0.107-0.117, and 0.069-0.078 mg/g. The results showed that there was little difference among the batches. Conclusion: The method is accurate, sensitive, credible, and repeatable. It can be applied to the quality control of SSG.

Quality comparison between traditional and cultivated Astragalus membranaceus var. mongholicus grown in Shanxi Hengshan area / 中草药

Objective: To compare the chemical constituents difference between the cultivated Astragali Radix (AR) and those grown in traditional way in Hengshan area of northern Shanxi Province. Methods: 1H-NMR based metabolomics approach combined with content determination based on HPLC-UV-ELSD was used to compare the primary and secondary metabolites in AR of different growing pattern. Results: Twenty-five metabolites were identified in the NMR spectra, and the major metabolites in the aqueous methanol fraction were the primary metabolites, such as amino acids and organic acids, while the fatty acids derivatives were present in the chloroform fraction; Multivariate analysis showed that there was no significant difference between the two kinds of AR for the primary metabolites, but 8, 2'-dihydroxy-7, 4'-dimethoxyisoflavan was only detected in AR grown in traditional way. The results of content determination of six major isoflavonoids and four saponins revealed that the contents of calycosin-7-O-β-D-glucoside, ononin, and astragaloside III were higher in AR grown in the traditional way, but astragaloside I was significantly higher in the cultivated AR. Conclusion: The major differences between the cultivated AR and those grown in traditional way are in the secondary metabolites, which indicates that the growing pattern is important for the biosynthesis and accumulation of secondary metabolite in AR. The results lay the scientific foundation for the rational utilization of the AR resources in Hengshan Area.

Study on HPLC digital fingerprint of Paeonia suffruticosa and determination of eight components / 中草药

Objective: To establish an HPLC digital fingerprint for flower petals of Danfeng (Paeonia ostii), and apply it in the determination of other six varieties of P. suffruticosa produced in Heze areas, while determining the contents of eight components in the flower petals. Methods: Nano-Micro C18 (250 mm × 4.6 mm, 5 μm) column was adopted and the gradient mobile phase consisted of acetonitrile (A)-0.1% formic acid (B) with a flow rate of 1.0 mL/min, the detective wavelength was 270 nm, and the column temperature was 25 ℃. Results: The chromatographic fingerprint similarity evaluation system for traditional Chinese medicines was used for analysis. The chromatographic fingerprint similarity of six batches of Danfeng samples was more than 0.993. There were altogether 14 common peaks, and eight of them were identified. A comparative analysis showed a similarity between samples of Danfeng and other six varieties produced in Heze. Meanwhile, the content of eight major chemical constituents, namely gallic acid, methyl gallate, kaempferol-3-O-β-D-glucopyranoside, apigenin-7-O-β-D-glucopyranoside, apigenin-7-O-β-D-neospheroside, dihydrokaempferol-7-O-β-D-glucopyranoside, paeoniflorin, and oxypaeoniflorin were determined in the range of 0.05-1.41, 1.01-4.41, 0.70-5.55, 4.60-18.30, 10.05-33.87, 0.09-1.76, 3.16-11.12, 0.85-2.02 mg/g. Conclusion: The combination of HPLC digital fingerprint and content determination could reflect its inherent quality in an all-round way, which provided scientific basis for the quality evaluation of P. suffruticosa.

Simultaneous determination of 10 components in Shendan Sanjie Capsule by HPLC-DAD / 中草药

Objective: To establish an HPLC-DAD method for the simultaneous determination of 10 components in Shendan Sanjie Capsule including tanshinone IIA, magnolol, honokiol, naringin, neohesperidin, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, calycosin 7-O-β-D-glucopyranoside, and atractylenolide I. Methods: The chromatographic separation was performed on a Hypersil BDS column (150 mm × 4.6 mm, 3.5 μm) with acetonitrile-merhanol-0.1% phosphate acid solution as mobile phase at the flow rate of 1.0 mL/min for gradient elution and the column temperature was 40 ℃. The detection wavelength was set at 270 nm for tanshinone IIA, 294 nm for magnolol and honokiol, 283 nm for naringin and neohesperidin, 203 nm for ginsenoside Rg1, ginsenoside Re, and ginsenoside Rb1, 260 nm for calycosin 7-O-β-D-glucopyranoside, and 220 nm for atractylenolide I. The volume of sample injection was 10 μL. Results: Ten compounds were well separated under the determined chromatographic conditions. The RSD values of precision and repeatability experiment were all less than 2% and the sample solution was stable during 10 h. All the compounds had a wide linear range and good linearity: the linear range of tanshinone IIA, magnolol, honokiol, naringin, neohesperidin, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, calycosin 7-O-β-D-glucopyranoside, and atractylenolide I were 112-560 μg/mL (r = 0.999 6), 64-320 μg/mL (r = 0.999 1), 48-240 μg/mL (r = 0.999 3), 80-400 μg/mL(r = 0.999 4), 80-400 μg/mL (r = 0.999 5), 16-80 μg/mL (r = 0.999 2), 16-80 μg/mL (r = 0.999 1), 16-80 μg/mL (r = 0.999 1), 40-200 μg/mL (r = 0.999 2), and 56-280 μg/mL (r = 0.999 3), respectively. The average recoveries were in the range of 98.43%-101.52% and the RSD values were all less than 2.0%. The content ranges of tanshinone IIA, magnolol, honokiol, naringin, neohesperidin, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, calycosin 7-O-β-D-glucopyranoside, and atractylenolide I in six batches of Shendan Sanjie Capsule were 0.829-0.840 mg/g, 0.538-0.548 mg/g, 0.360-0.369 mg/g, 0.210-0.219 mg/g, 0.111-0.118 mg/g, 0.081-0.089 mg/g, 0.070-0.078 mg/g, 0.111-0.117 mg/g, 0.072-0.080 mg/g, and 0.130-0.137 mg/g, respectively. Conclusion: The method is simple and convenient, the methodology validation shows that determination result of the method is accurate and reliable and it can be an effective approach for the quality control of Shendan Sanjie Capsule.

Isolation and identification of metabolites in rat urine after ig administration of calycosin-7-O-β-D-glucopyranoside / 中草药

Objective: To study the metabolites from rats urine after ig administration of calycosin-7-O-β-D-glucopyranoside. Methods: The constituents were isolated and purified by preparative liquid chromatographic technique, and the structures were identified by spectroscopic analyses including ESI-MS, NMR, and 2D-NMR. Results: Five metabolites were isolated from rats urine after ig administration of calycosin-7-O-β-D-glucopyranoside. They were identified as calycosin (M1), 3', 4', 7-trihydroxyisoflavone (M2), daidzein (M3), calycosin-3'-O-β-D-glucuronide (M4), and calycosin-3'-O-β-D-glucuronide methyl ester (M5). Conclusion: M5 is a new compound.

Determination of 12 kinds of index components in Boerning Capsules by HPLC-DAD variable wavelength method / 中草药

Objective: To develop an HPLC-DAD wavelength switching combined with gradient elution method for the determination of the contents of calycosin 7-O-β-D-glucopyranoside, solasonine, solamargine, chonglou saponin I, chonglou saponin II, chonglou saponin VI, chonglou saponin VII, specnuezhenide, rosmarinic acid, rhein, chrysophanol, and emod in Boerning Capsules (BC) simultaneously. Methods: The chromatographic separation was achieved on an Atlantis T3 C18 (250 mm × 4.6 mm, 5 μm) with acetonitrile-methanol (A)-0.1% formic acid solution (B) as mobile phase at the flow rate of 0.8 mL/min for gradient elution; variable wavelength method; sample quantity was 20 μL. Results: The 12 active components were well separated and showed good linearity, such as calycosin 7-O-β-D-glucopyranoside, solasonine, solamargine, polyphyllin I, polyphyllin II, polyphyllin VI, polyphyllin VII, specnuezhenide, rosmarinic acid, rhein, chrysophanol, and emod 1.97—19.70 μg/mL (r = 0.999 2), 1.022—10.220 μg/mL (r = 0.999 3), 0.982—9.820 μg/mL (r = 0.999 1), 1.102—11.020 μg/mL (r = 0.999 3), 1.114—11.140 μg/mL (r = 0.999 5), 1.154—11.540 μg/mL (r = 0.999 8), 1.114—11.140 μg/mL (r = 0.999 5), 2.768—27.680 μg/mL (r = 0.999 3), 3.04—30.40 μg/mL (r = 0.999 6), 3.379—33.790 μg/mL (r = 0.999 5), 3.286—32.860 μg/mL (r = 0.999 4), and 3.507—35.070 μg/mL (r = 0.999 7). The precision and repeatability were good, and RSD values were less than 2.0%. The average recoveries and the corresponding RSD values were 100.08% (1.27%), 98.11% (1.15%), 99.68% (1.13%), 101.38% (0.87%), 101.87% (0.95%), 100.53% (0.74%), 98.52% (0.83%), 99.52% (0.88%), 97.84% (1.33%), 98.31% (0.71%), 99.66% (0.57%), and 101.73% (1.41%), respectively. The contents of 12 batches of the tevelve active components were 0.085—0.118, 0.065—0.085, 0.051—0.075, 1.822—1.888, 1.532—1.599, 1.027—1.148, 2.420—2.621, 6.428—6.937, 0.258—0.289, 0.122—0.143, 0.159—0.184, and 0.222—0.273 mg/g. Conclusion: An HPLC wavelength switching combined with gradient elution method has been successfully established for simultaneous determination of 12 components in BC. The method is simple, quick, accurate, and helpful for the quality control of BC.

Study on chemical constituents from Carpesium faberi / 中草药

Objective: To investigate the chemical constituents from the whole plant of Carpesium faberi. Methods: Compounds were isolated by various chromatographic techniques, including silica gel, ODS, sephadex LH-20, and semi-preparative HPLC, and their structures were identified by comparison of their experimental spectroscopic data with their reported data. Results: The phytochemistry investigation led to the isolation of 12 compounds, and their structures were elucidated as ent-kaurane-3β,16β,17-triol (1), 3-(hydroxy-acetyl)-1H-indole (2), 8,9,10-trihydroxythymol (3), 8-hydroxy-9,10-diisobutyryloxy-thymol (4), neryl-β-D- glucopyranoside (5), (3S)-linalyl-β-D-glucopyranoside (6), (1R,2S,4S,5R)-2,5-dihydroxy-p-menthane (7), luteolin (8), apigenin-7-O-β-D-glucopyranoside (9), medioresinol (10), pinoresinol (11), and a mixture of silybin and isosilybin (12). Conclusion: All compounds except compound 7 are not only isolated from this plant for the first time, but also from this genus for the first time.

Chemical constituents from rhizome of Smilax davidiana / 中草药

Objective: To study on the chemical constituents from the rhizome of Smilax davidiana. Methods: The compounds were separated and purified by sephadex LH-20 column chromatography and high performance liquid chromatography. Their structures were identified by spectroscopic analysis and comparison with literatures. Results: Twenty compounds were isolated and identified as aromadendrin7-O-β-D-glucopyranoside (1), dalbergin (2), 3,5,7,4′-tetrahydroxyflavanone-7-O-β-D-glucopyranoside (3), kaempferol-7-O-β-D-glucopyranoside (4), quercetin-7-O-β-D-glucopyranoside (5), quercetin-3-O-β-L-rhamnopyranoside (6), 4,6- dihydroxy-2-O-(β-D-glucopyranosyl)-acetophenone (7), 3,5-dihydroxy-4-O-(β-D-glucopyranosyl) acetophenone (8), 2,4,6- trihydroxy-acetophenone-4-O-β-D-glucopyranoside (9), 2,4,6-trihydroxylacetophenone-2,4-di-O-β-D-glucopyranoside (10), epicatechin (11), latifolin (12), 3′-O-(E-4-coumaroyl)-quinic acid (13), 5-O-caffeoylquinic acid butyl ester (14), 5,5′- dimethoxylariciresinol 4′-O-β-D-glucopyranoside (15), 1-cerotoylglycerol (16), cinchonain Ib (17), adenosine (18), resveratrol (19), and 3,4,5-trimethoxyphenyl-1-β-D-glucopyranoside (20). Conclusion: Compounds 4—9, and 12—16 are isolated from Smilax genus for the first time, and compounds 1—3, 17, and 18 are from S. davidiana for the first time.

Calycosin-7-O-β-d-glucopyranoside stimulates osteoblast differentiation through regulating the BMP/WNT signaling pathways

The isoflavone calycosin-7-O-β-d-glucopyranoside (CG) is a principal constituent of Astragalus membranaceus (AR) and has been reported to inhibit osteoclast development in vitro and bone loss in vivo. The aim of this study was to investigate the osteogenic effects of CG and its underlying mechanism in ST2 cells. The results show that exposure of cells to CG in osteogenic differentiation medium increases ALP activity, osteocalcin (Ocal) mRNA expression and the osteoblastic mineralization process. Mechanistically, CG treatment increased the expression of bone morphogenetic protein 2 (BMP-2), p-Smad 1/5/8, β-catenin and Runx2, all of which are regulators of the BMP- or wingless-type MMTV integration site family (WNT)/β-catenin-signaling pathways. Moreover, the osteogenic effects of CG were inhibited by Noggin and DKK-1 which are classical inhibitors of the BMP and WNT/β-catenin-signaling pathways, respectively. Taken together, the results indicate that CG promotes the osteoblastic differentiation of ST2 cells through regulating the BMP/WNT signaling pathways. On this basis, CG may be a useful lead compound for improving the treatment of bone-decreasing diseases and enhancing bone regeneration.