A Newly Discovered Phenylethanoid Glycoside from Stevia rebaudiana Bertoni Affects Insulin Secretion in Rat INS-1 Islet ß Cells.

The tea-like beverage Stevia rebaudiana Bertoni (Stevia) is popular in China because it reduces blood glucose and has a sweet taste. In this work, a comprehensive quality assessment of Stevia led to the discovery of five phenylethanoid glycosides, namely steviophethanoside (1), cuchiloside (2), salidroside (3), icariside D (4), and tyrosol (5). Of them, compound 1 is a novel compound. Mass spectrometry and NMR spectroscopy were employed to confirm the absolute configuration. A hydrolytic step with 4 N TFA at 95 °C for 4 h was used to confirm the monosaccharides. In addition, Discovery Studio 4.0 was used to predict the ADME and toxicity activity of compound 1. The results suggested that compound 1 was biocompatible and had poor toxicity, which was verified by rat INS-1 islet ß cells through an MTT assay. Meanwhile, a significant stimulatory effect on INS-1 cells was observed, which indicated a hypoglycemic effect of compound 1. This is the first report that describes a natural, novel, and hypoglycemic phenylethanoid glycoside in Stevia.

[On Chinese medicine quality precision in expectation].

According to the correlative analyses on Chinese medicine essence, dosage forms and quality control level, it expounds the precise concept of Chinese medicine, and its quality advantages and characteristics in this paper, furthermore discusses how to achieve the ideal drugs and Chinese medicine quality precision in expectation. Base on the Chinese medicine essence, using the concept of nature medicine and its drug system to construct Chinese medicine effective material basis and its drugs, with the correlative analyses of whole view and reductionism, the problems of uncertainty quality of original natural medicinal resources and preparations may well be solved, and further with the macroscopic to microcosmic construction of drug system, the precision in expectations of Chinese medicine quality and higher production lever may well be achieved.

[HPLC-fingerprint-based quality evaluation on a Tibetan medicine Phyllanthus emblica and its tannin parts].

This study is to establish the fingerprint for Phyllanthus emblica and their tannin parts from different habitats by HPLC for its quality control. The determination was carried out on a Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column, with methanol-0.2% glacial acetic acid as mobile phase with gradient elution at a flow rate of 1 mL x min(-1). The temperature was maintained at 30 degrees C and the detected wavelength is 260 nm, Thirteen chromatographic peaks were extracted as the common peaks of the fingerprint of P. emblica, and eleven as the common peaks of P. emblica tannin parts, and five peaks were identified by comparing with referent samples. The fingerprints of 8 samples were compared and classified by similarity evaluation, cluster analysis and principal component analysis (PCA). The similarity degrees of eight P. emblica were between 0.763 and 0.993, while tannin parts were between 0.903 and 0.991. All the samples of P. emblica and their tannin parts were classified into 3 categories. The method was so highly reproducible, simple and reliable that it could provide basis for quality control and evaluation of P. emblica from different habitats.

Gaultheria: Phytochemical and pharmacological characteristics.

The genus Gaultheria, comprised of approximately 134 species, is mostly used in ethnic drugs to cure rheumatism and relieve pain. Phytochemical investigations of the genus Gaultheria have revealed the presence of methyl salicylate derivatives, C6-C3 constituents, organic acids, terpenoids, steroids, and other compounds. Methyl salicylate glycoside is considered as a characteristic ingredient in this genus, whose anti-rheumatic effects may have a new mechanism of action. In this review, comprehensive information on the phytochemistry, volatile components and the pharmacology of the genus Gaultheria is provided to explore its potential and advance research.

In vivo metabolites and plasma exposure of TongMai Keli analyzed by UHPLC/DAD/qTOF-MS and LC/MS/MS.

ETHNOPHARMACOLOGICAL RELEVANCE: TongMai Keli (TM) is a widely used traditional Chinese medicine preparation for the treatment of cardiovascular and cerebrovascular diseases. It is composed of Puerariae Lobatae Radix (roots of Pueraria lobata (Willd.) Ohwi), Salviae Miltiorrhizae Radix (roots of Salvia miltiorrhiza Bge.), and Chuanxiong Rhizoma (rhizomes of Ligusticum chuanxiong Hort.). The aim of this study is to identify the in vivo metabolites of TM, and to elucidate the pharmacokinetics of TM constituents and their metabolites. MATERIALS AND METHODS: For metabolites identification, TM was orally administered to rats (n=3), and the metabolites in plasma were identified by UHPLC/DAD/qTOF-MS analysis and ß-glucuronidase hydrolysis. For pharmacokinetic study, rats (n=10) were treated with TM at a clinical dose, and the plasma was analyzed by LC/MS/MS. RESULTS: A total of 25 metabolites from TM were identified in rats plasma. Glucuronide and sulfate conjugations were the major metabolic reactions, and produced 14 metabolites. The analytical method for pharmacokinetic study was fully validated with good linearity (r>0.99), wide dynamic ranges (6-6000 ng/mL), and low variations (<14.3%). The plasma concentration-time curves of puerarin and nine metabolites were profiled. CONCLUSION: Isoflavones from Puerariae Lobatae Radix were the major metabolites in rat plasma after oral administration of TM. Puerarin and other isoflavone glycosides could reach their first C(max) within 30 min, and were then rapidly eliminated, followed by their phase II metabolites.

Three new isoflavone glycosides from Tongmai granules.

Three new isoflavone glycosides, 3'-methoxydaidzein-7,4'-di-O-ß-d-glucopyranoside (1), biochanin A-8-C-ß-d-apiofuranosyl-(1 â†’ 6)-O-ß-d-glucopyranoside (2), daidzein-7-O-ß-d-glucopyranosyl-(1 â†’ 4)-O-ß-d-glucopyranoside (3), and a new natural isoflavone glycoside, daidzein-7-O-α-d-glucopyranosyl-(1 â†’ 4)-O-ß-d-glucopyranoside (4) were isolated along with 18 known isoflavones from the EtOAc and n-BuOH fractions of the aqueous extraction of Tongmai granules. All the isoflavones were obtained and determined for the first time from Tongmai granules. The structures of these compounds were elucidated by spectral methods. It was confirmed that the compounds 1-4 were originally from Puerariae Lobatae Radix based on HPLC-DAD analysis of the crude drug extract. The isoflavones isolated were tested for their antioxidative activities by measuring the capacities of scavenging the 2,2'-diphenyl-1-picrylhydrazyl radical.

[HPLC determination of 2-hydroxy-1-methoxyaporphine, pronuciferine, nuciferine and roemerine in Nelumbo nucifera and its alkaloid fraction].

OBJECTIVE: To establish an HPLC method for the determination of four alkaloids, i.e., 2-hydroxy-1-methoxyaporphine, pronuciferine, nuciferine and roemerine, in Nelumbo nucifera and its alkaloid fraction. METHOD: The determination was carried out at 35 degrees C on a Hypersil C18 column (4.6 mm x 250 mm, 5 microm), eluting with acetonitrile-water containing 0.1% triethylamine as mobile phases in gradient mode. The flow rate was 1.0 mL x min(-1) and detection at the wavelength was set at 270 nm. RESULT: The linear ranges of 2-hydroxy-1-methoxyaporphine, pronuciferine, nuciferine and roemerine were 0.110-0.658 microg (r = 0.9995), 0.0210-0.126 microg (r = 0.9995), 0.103-0.618 microg (r = 0.9998), 0.085 6-0.514 microg (r = 0.9995), with the average recoveries (n=6) were 101.5%, 99.14%, 99.21% and 98.41% for the alkaloid fraction of N. nucifera and 99.53%, 100.5%, 97.51% and 100.1% for N. nucifera respectively. CONCLUSION: The determination results of the three batches of samples showed that the method was easy and accurate which could be used to determine the contents of four components in N. nucifera and its alkaloid fraction.

[Study on adscription of drug constituents in Beagle dog plasma after intragastric administration with Jiangzhining extract].

OBJECTIVE: To investigate the drug constituents in Beagle dog plasma after intragastric administration with Jiangzhining extract, which is a Chinese medicine prescription and it consisting of processed Radix Polygoni Multiflori, Fructus Crataegi, Semen Cassiae and Folium Nelumbinis. METHOD: The drug constituents and their origin in Beagle dogs plasma were assigned by comparison of high-performance liquid chromatographic profiles, retention time and ultraviolet spectra of the drug-free plasma, plasma contained drug, Jiangzhining extract and the ingredients of crude drug. RESULT: Twenty-four components were detected after intragastric administration of Jiangzhining extract, among which eight compounds were the original components existed in Jiangzhining extract and sixteen compounds were likely to metabolites of the some chemical constituents in ingredients of crude drug. CONCLUSION: Studies on the drug constituents and the metabolites in plasma after intragastric administration of Jiangzhining extract are useful to elucidate the effective substances of Jiangzhining prescription.

[A novel triterpenoid saponin from Prunella vulgaris].

To study the constituents of the Prunella vulgaris L, the constituents were isolated by various column chromatography and the structures were identified on the basis of chemical and spectral analysis. One saponin compound (I) and one flavone glycoside compound (II) were obtained from Prunella vulgaris L. Their structures were elucidated as 16-oxo-17-demethyl-3beta,24-dihydroxylolean-12-en-3-O-beta-D-glucuronoside (I), and acacetin-7-O-beta-D-glucopyranoside (II). Compound I is a novel triterpenoid saponin and named as prunelloside A. Compound II was obtained for the first time from the Prunella genus.

[A new chemical component of Pueraria lobata (Willd.) Ohwi].

Pueraria lobata (Willd.) Ohwi. was extracted for two times with 70% ethanol and the 70% ethanol-extracts was condensed. Various column chromatography with AB-8 macroreticular resin, Toyopearl HW-40, pharmadex LH-20, and silica gel were employed for the isolation and purification of the 70% ethanol-extracts from Pueraria lobata (Willd.) Ohwi. Five compounds were isolated and their structures were identified by physiochemical properties and spectral analysis (UV, IR, MS, 1H NMR, 13C NMR, HMQC, HMBC, etc.): (4R)-3-[ 2-hydroxy-4-methoxyphenyl]-4-(4-beta-D-glucopyranosyloxybenzyl) but-2-en-4-olide (1), 4', 8-dimethoxyl-7-O-beta-D-glucopyranosyl isoflavone (2), eicosanoic acid (3), hexadecanoic acid (4), tetracosanoid acid-2,3-dihydroxypropyl ester (5). Compound 1 was a new compound, and compounds 2, 3, 4 were isolated from this plant for the first time.

[HPLC fingerprint of flavonoids of Kushen Tang and its correlation to Scutellaria baicalensis and Sophora flavescens].

OBJECTIVE: To establish the HPLC fingerprint of the flavonoids of Kushen Tang, and to study its correlation to Scutellaria baicalensis and Sophora flavescens. METHOD: The fingerprints of the flavonoids of Kushen Tang were established by HPLC under two detective wavelengths 280 nm and 365 nm. The correlation existed in the common peaks of HPLC fingerprint of Kushen Tang and the Chinese meteria medicines which composed the prescription was explored by analyzing the HPLC fingerprints of different compatibilities. The chemical constituents which the common peaks stood for were analyzed by adding controls to the samples respectively. RESULT: Under the detective wavelength 280 nm, 19 common peaks were found, which came from S. baicalensis except that two peaks came from S. flavescens, and eight peaks were baicalin/trifolirhizin, wogonoside, luteolin, baicalein, oroxylin A, 5, 8-dihydroxy-6, 7-dimethoxyflavone, wogonin and formononetin/chrysin respectively. Under the detective wavelength 365 nm, 22 common peaks were found, which thirteen peaks came from S. baicalensis and six peaks came from S. flavescens. Among them, eleven peaks were baicalin, wogonoside, baicalein, oroxylin A, 5,8-dihydroxy-6, 7-dimethoxyflavone, wogonin, chrysin/7-methoxyflavone, 2', 4-dihydroxy-4', 6'-dimethoxychalcone, 4, 4'-dimethoxychalcone, xanthohumol and kuraridin respectively. CONCLUSION: The common peaks of HPLC fingerprint of the flavonoids of Kushen Tang, which were obtained under the two detective wavelengths 280 nm and 365 nm, came from S. baicalensis and S. flavescens. It was more scientific and comprehensive that establishing the HPLC fingerprint of the flavonoids of Kushen Tang under the detective wavelength 365 nm.

[Isolation and structure identification of chemical constituents from Saposhnikovia divaricata (Turcz.) Schischk].

Fourteen compounds were isolated from the ethanol extraction of Saposhnikovia divaricata (Turcz.) Schischk using column chromatographic methods after enrichment by macroporous adsorptive resins. They were identified as fangfengalpyrimidine (1), clemiscosin A (2), 5-hydroxy-8-methoxypsoralen (3), sec-O-glucosylhamaudol (4), hamaudol (5), nodakenetin (6), prim-O-glucosylcimifugin (7), cimifugin (8), 4'-O-beta-D-glucosyl-5-O-methylvisamminol (9), 5-O-methylvisamminol (10), marmesin (11), adenosine (12), daucosterol (13) and beta-sitosterol (14) by physico-chemical properties and spectral data. Compound 1 is a new compound. Compounds 2 and 3 were isolated from umbelliferae plants and Saposhnikovia divaricata (Turcz.) Schischk for the first time respectively.

[Relative adscriptions of components in the effective fractions of Yinqiao decoction and its composing individual herbs].

HPLC and LC-MS/MS were used to establish a comprehensive HPLC analytical method of Yinqiao decoction and identify the chemical constituents of the whole and individual herbs of Yinqiao decoction. YWG-C18 (250 mm x4. 6 mm ID, 10 microm) column was used; the mobile phase was composed of acetonitrile (A) and water ( B, with 3% acetic acid) with gradient elution; the flow rate was 1. 0 mL x min(-1) and the column temperature was set up at 25 degrees C. The detection wavelength was 280 nm. The chromatographic fingerprints of Yinqiao Decoction showed 30 main peaks. Peak 2, 14, 15, 17 were from Lonicera japonica Thunb, peak 3, 12, 13, 24 were from Fosythia suspense (Thunb) Vahl, peak 19, 25, 26, 27 were from Arctium lappa L. , peak 5, 6, 8, 9, 10, 11, 18, 28 were from Glycyrrhiza uralensis Fisch, peak 20, 21 were from Mentha haplocalyx Briq. , peak 22, 23 were from Schizonepeta tenuifolia Briq. , peak 1 presented in the chromatograms of Lonicera japonica Thunb, Fosythia suspense (Thunb) Vahl, Mentha haplocalyx Briq. , Schizonepeta tenuifolia Briq. and Platycodon grandiflorum (Jacq. ) A. DC. , peak 7 presented in the chromatograms of Fosythia suspense (Thunb) Vahl and Glycine max (L. ) Merr. , peak 16 presented in the chromatograms of Mentha haplocalyx Briq. and Schizonepeta tenuifolia Briq. , peak 29 presented in the chromatograms of the herbs except Mentha haplocalyx Briq. and Platycodon grandiflorum (Jacq. ) A. DC. , peak 30 presented in the chromatograms of the herbs except Platycodon grandiflorum (Jacq. ) A. DC. , peak 4 was not identified, maybe it was a new constituent produced during decoction. By comparison of the standards isolated and MS spectra, 14 peaks were identified as 2 ( chlorogenic acid) , 9 ( liquiritin ) , 10 ( 4'-O-[ beta-D-apiofuranosyl (1--> 2 ) -beta-D-glucopyranosyl] liquiritigenin), 12 (forsythiaside), 13 (rutin), 14 (4,5-O-dicaffeoylquiniic acid), 15 (3, 5-O-dicaffeoylquiniic acid ), 16 ( 4-0- [ beta-D-apiofuranosyl ( 1 -->2 ) -beta-D-glucopyranosyl ] isoliquiritigenin) , 17 ( 3, 4-O-dicaffeoylquiniic acid) , 18 (2'-O-[ beta-D-apiofuranosyl (1 -->2 ) -beta-D-glucopyranosyl] isoliquiritigenin) , 19 (arctiin) , 20 (linarin) , 25 (genistein) , 28 ( isoliquiritigenin) . The method could be used to identify the characteristics of Yinqiao decoction, and it could be used to evaluate the quality and quantity of Yinqiao decoction.

[Study of quality standards on Qianshan Huoxue plaster].

OBJECTIVE: To establish the methods of identification and assay in Qianshan Huoxue Gao. METHOD: Using TLC to identify Sanchi, Dragon's Blood and using HPLC to determine the content of ginsenoside Rg1. RESULT: The linear range of ginsenoside Rg1 was from 0.153 9 to 1.026 microg. The average recovery was 97.4%, RSD was 2.1%. CONCLUSION: The methods are simple and have good reproducibility.

[Studies on chemical constituents in spikes of Schizonepeta tenuifolia].

OBJECTIVE: To study the chemical constituents in the spikes of Schizonepeta tenuifolia. METHOD: Compounds were isolated and purified with silica gel, ODS and Sephadex LH-20 gel column chromatography, and their structures were determined by using spectroscopic analysis including MS and NMR. RESULT: Nine compounds were isolated and identified as 5, 7-dihydroxy-6, 4'-dimethoxyflavone (1), 5, 7-dihydroxy-6, 3', 4'-trimethoxyflavone (2), ursolic acid (3), 3-hydroxy-4(8)-ene-p-menthane-3(9)-lactone (4), 5, 7, 4'-trihydroxyflavone (5), 5, 4'-dihydroxy-7-methoxyflavone (6), hesperidin (7), luteolin (8) and daucesterol (9). CONCLUSION: Compounds 1, 2, 6 were first obtained from the spikes of S. tenuifolia.

[Constituents in the alkaloid fraction of Kushen decoction].

OBJECTIVE: To study the chemical constituents of the alkaloid fraction of Kushen decoction. METHOD: Constituents were isolated by different kinds of column chromatography and their structures were elucidated with spectral methods. RESULT: Eight compounds were isolated and identified as matrine (I), sophoridine (II), sophocarpine (III), sophoramine (IV), oxymatrine (V), oxysophocarpine (VI), aloperine (VII) and sparteine (VIII). CONCLUSION: All these compounds were isolated from Kushen decoction for the first time. Aloperine was found firstly in Sophora flavescens, or Scutellaria baicalensis, or Rehmannia glutinosa which constituted Kushen decoction.

[GC-MS analysis of the fatty components of pollen Typhae before and after being carbonized].

OBJECTIVE: To study the changes of the fatty components of Pollen Typhae before and after being carbonized. METHOD: Pollen Typhae and Pollen Typhae carbonisatus were extracted with petroleum ether (60-90 degrees C) respectively. The two kinds of extracts were analyzed by GC-MS after saponificated and methanolized, and their constituents were searched through NIST. The contents of the constituents were determined by method of normalization. RESULT: Either in Pollen Typhae or in Pollen Typhae carbonisatus, 32 components were identified, among which 20 components were the same and 6 were different respectively. Among the same components, the relative contents of 3-methyl-2-butenoic acid-2-phenylethyl ester, hexanedioic acid-dimethyl ester, dimethyl phthalate, diethyl phthalate, diphenylamine, sebacic acid dimethyl ester, 1,2-benzenedicarboxylic acid, ethyl methyl ester, methyl-2-ethylhexyl phthalate and diisooctyl phthalate etc. increased obviously, and the relative contents of nonanedioic acid-dimethyl ester, diisobutyl phthalate and stigmastan-3,5-dien etc. decreased greatly. Among the different components, 8-hydroxy-octanoic acid-methyl ester, 9-hydroxy-nonanoic acid-methyl ester, 10-octadecenoic acid-methyl ester, m-hydroxycinnamic acid-methyl ester,3-[4-( acetyloxy)-3-methoxyphenyl]2-propenoic acid-methyl ester and 11-octadecenoic acid-methyl ester were detected in Pollen Typhae, 3-hydroxyspirost-8-en-11-one, benzenepropanoic acid-methyl ester, 2,4-dimethylhexanedioic acid; 2,4-bis (1,1-dimethylethyl)-phenol, undecanedioic acid-dimethyl ester and 9,10-dihydroxy-octadecanoic acid-methyl ester were detected in Pollen Typhae carbonistatus. CONCLUSION: The species and contents of the fatty components in Pollen Typhae changed before and after being carbonized, but their chemical types didn't change too much.

[Studies on the chemical constituents in herb of Mentha haplocalyx].

OBJECTIVE: To study the chemical constituents of Mentha haplocalyx. METHOD: The chemical constituents were isolated and purified with column chromatography and the structures were elucidated by spectral analysis. RESULT: Nine compounds were obtained and identified as emodin (I), chrysophanol (II), physcione (II), benzoic acid (IV), trans-cinnamic acid (V), beta-sitosterol (VI), aloe-emodin (VII), ursolic acid (VIII) and daucosterol (IX). CONCLUSION: Compounds I, II, III, V, VII were first isolated from M. haplocalyx.

[Determination of phillyrin and forsythoside A in Lianqiao Bingdu Qing capsule by RP-HPLC].

OBJECTIVE: To establish methods for the determination of phillyrin and forsythoside A in Lianqiao Bingdu Qing capsule by RP-HPLC. METHOD: The determination of phillyrin was carried out on YWG-C18 column (4.6 mm x 250 mm, 10 microm), using acetonitrile-water (25:75) as mobile phase at a flow rate of 0.8 mL x min(-1) and detected at the wavelength 277 nm. The determination of forsythoside A was carried out with YWG-C18 column(4.6 mm x 250 mm,10 microm), using acetonitrile-water-Acetic acid (17:83:0.4) as mobile phase at a flow rate of 1.0 mL x min(-1) and detected at the wavelength 280 nm. RESULT: The average recovery of phillyrin was 99.6%, RSD = 1.9% (n = 5). The average recovery of forsythoside A was 101.3%, RSD = 2.5% ( n = 5). CONCLUSION: The methods were simple and accurate and could be used to control the quality of the Lianqiao Bingdu Qing capsule.

[Isolation and identification of a new cucurbitacin from Picria fel-terrae].

AIM: To study compounds isolated from Picria fel-terrae. METHODS: The chemical constituents were separated and purified by column chromatography on silica gel and MCI. Their structures were identified on the basis of spectral data (IR, UV, MS, ID NMR and 2D NMR). RESULTS: A new cucurbitacin, along with a known one, were obtained from the 60% EtOH extract of the whole plant. CONCLUSION: The new compound was identified as 11, 24-dioxo-5, 21-diene-cucurbit-3alpha-O-beta-D-xylopyranosyl-16alpha-O-alpha-L-rhamnopyranoside (dehydrobryogenin glycoside). The known one, hexanorcucurbitacin F, was obtained for the first time from Picria fel-terrae.