HPLC Fingerprint Analysis of Notopterygii Rhizoma et Radix with Different Commercial Specifications / 中国实验方剂学杂志

Objective: To establish HPLC fingerprint of Notopterygii Rhizoma et Radix with different commercial specifications and to provide the basis for the division of commercial grades and the quality control of Notopterygii Rhizoma et Radix on the market. Method: The market investigation and literature research were used to understand the existing situation of Notopterygii Rhizoma et Radix goods.Notopterygii Rhizoma et Radix goods were divided into three commercial specifications according to the source and appearance,such as Canqiang,Tiaoqiang and Datouqiang.Fingerprint of Notopterygii Rhizoma et Radix with different commercial specifications was established by HPLC-PDA,the mobile phase was consisted of acetonitrile-0.3% acetic acid in a gradient elution mode.Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine(version of 2004A) was used to confirm the common peaks and evaluate the similarity.SPSS 19.0 statistical software was used to make principal component analysis(PCA) for HPLC fingerprint pattern. Result: The common mode of fingerprint for Canqiang,Tiaoqiang and Datouqiang were established separately.A total of 22 common peaks were marked in Canqiang,23 common peaks were marked in Tiaoqiang,29 common peaks were marked in Datouqiang.The result of similarity evaluation and PCA showed that the quality of Notopterygii Rhizoma et Radix with the same commercial specification was stable.There were great differences in chemical compositions and their contents among Notopterygii Rhizoma et Radix with different commercial specifications. Conclusion: The fingerprint method can well distinguish commercial specifications of Notopterygii Rhizoma et Radix,and it can provide the basis for the division of commercial grades and the quality control of Notopterygii Rhizoma et Radix.

Qualitative and quantitative analysis of major cholic acids in Suis Fellis Pulvis / 中国中药杂志

This study is to establish a qualitative method for rapid identification of bile acids in Suis Fellis Pulvis based on UHPLC-LTQ-Orbitrap-MS technology,and an HPLC-ELSD internal standard method for the quantitative determination of two glycine-conjugated BAs in Suis Fellis Pulvis.The chromatographic separation of the UHPLC-LTQ-Orbitrap-MS qualitative analysis was achieved on a Waters Acquity UPLC HSS T_3column(2.1 mm×100 mm,1.8μm),with 0.2%formic acid aqueous solution(A)-acetonitrile(B)as mobile phase ingradient elution.Electrospray ionization(ESI)source was applied and operated in negative ion mode.Quantitative analysis was performed at 30℃on a Diamonsil-C_(18)column(4.6 mm×250 mm,5μm).The mobile phase consisted of 0.2%formic acid solution and acetonitrile with gradient elution and the flow rate was 1.0 m L·min~(-1).An ELSD was used with a nitrogen flow-rate of1.4 L·min~(-1)at a drift tube temperature of 60℃and the gain was 1.A total of 14 bile acids in Suis Fellis Pulvis were characterized based on the accurate mass measurements,fragmentation patterns,chromatographic retention times,and reference materials.For the quantitative analysis method,the glycohyodeoxycholic acid and glycochenodeoxycholic acid had good linear relationship in the range of26.52-265.20 mg·L~(-1)(r=0.999 8)and 19.84-198.40 mg·L~(-1)(r=0.999 1),respectively.The average recoveries(n=6)were104.1%and 103.1%,and the RSD were 2.0%and 2.4%.The UHPLC-LTQ-Orbitrap-MS technology provides a fast and efficient qualitative analysis method for identification of bile acids in Suis Fellis Pulvis.The HPLC-ELSD internal standard method is accurate and reliable,which has reference value for the quality control of Suis Fellis Pulvis.

Metabolic fingerprint analysis of RAW264.7 inflammatory cell model by using UPLC-Q-TOF/MS / 中国中药杂志

In order to reveal the properties of polar metabolome in inflammatory cells, we selected LPS-induced RAW264.7 inflammatory cell models as the carrier for the research of metabolic fingerprint analysis. In this study, an ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS)-based metabolomics protocol was optimized for the extraction of polar metabolites from RAW264.7 cell line. Then orthogonal partial least squares discriminant analysis (OPLS-DA) was used to process the metabolic data, and finally, a total of 17 metabolites were selected and identified. The results showed that MeOH-CHCl3-H2O (8∶1∶1) was chosen as the optimal extraction solvent to achieve higher number of chromatographic peaks, with the best relative extraction efficiency and stability. Comparing with the normal cells, the inflammatory cells presented an abnormal metabolism in protein, carbohydrate, nucleotide and phospholipids. In this study, a UPLC-Q-TOF/MS-based metabolomics protocol for the polar metabolites from RAW264.7 cell line was developed, which may provide important information for the study of mechanism of inflammation and the anti-inflammatory drugs.

A new method on investigate chemical constituents which have anti-thrombin effect by HPLC / 中国中药杂志

An in vitro anti-thrombin bioassay was developed to investigate the chemical constituents which have anti-thrombin effect from the water soluble components of Salvia miltiorrhiza. Using Chromozym TH as a probe combined with ethyl acetate Semi-micro extraction was applied to measure p-nitroaniline by HPLC. According to the results, the inactivationrate of thrombin by sodium danshensu, salvianolic acid A and salvianolic acid B under a given set of conditions were 3.06%, 77.77% and 2.35%, respectively. In the water-soluble components, salvianolic acid A has a direct inhibition of thrombin, while sodium danshensu and salvianolic acid B have no significant effect on thrombin. The method is sensitive and low consumption. It can eliminate the interference absorbed for the sample itself which can be used for screening single or multiple direct antithrombin active ingredient of herbal extract.

DNA barcoding identification of Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix based on trnL-trnF sequences / 中国中药杂志

To optimize indices of molecular identification for authentication of Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix, four indices, including sequence similarity, specific positions, genetic distance and phylogenetic tree, were compared based on trnL-trnF sequences. Total DNA was extracted from Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix, and trL-trnF sequences were amplified and sequenced. Sequence similarity was calculated by BLAST analysis. Specific positions were compared by DNAman software. Genetic distance and phylogenetic tree were analyzed by Mega software. The results showed that the inter-specific and intra-specific similarity of P. ginseng and P. quinquefolius respectively was 100% and 99. 6%. There were four specific positions at G153A, T463A, C732G and T818C. The inter-specific genetic distance (0) of trL-trnF sequences was lower than intra-specific genetic distance (0. 004). P. ginseng can be distinguished from P. quinquefolius based on the phylogenetic tree. It is concluded that Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix can be authenticated by identification indices of sequence similarity, specific positions, genetic distance and phylogenetic tree. Index of specific positions based on trnL-trnF sequences is the most efficient index to authenticate Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix.

A new herbs traceability method based on DNA barcoding-origin-morphology analysis--an example from an adulterant of 'Heiguogouqi' / 中国中药杂志

The fruit of Lycium ruthenicum is a common folk medicine in China. Now it is popular for its antioxidative effect and other medical functions. The adulterants of the herb confuse consumers. In order to identify a new adulterant of L. ruthenicum, a research was performed based on NCBI Nucleotide Database ITS Sequence, combined analysis of the origin and morphology of the adulterant to traceable varieties. Total genomic DNA was isolated from the materials, and nuclear DNA ITS sequences were amplified and sequenced; DNA fragments were collated and matched by using ContingExpress. Similarity identification of BLAST analysis was performed. Besides, the distribution of plant origin and morphology were considered to further identification and verification. Families and genera were identified by molecular identification method. The adulterant was identified as plant belonging to Berberis. Origin analysis narrowed the range of sample identification. Seven different kinds of plants in Berberis were potential sources of the sample. Adulterants variety was traced by morphological analysis. The united molecular identification-origin-morphology research proves to be a preceding way to medical herbs traceability with time-saving and economic advantages and the results showed the new adulterant of L. ruthenicum was B. kaschgarica. The main differences between B. kaschgarica and L. ruthenicum are as follows: in terms of the traits, the surface of B. kaschgarica is smooth and crispy, and that of L. ruthenicum is shrinkage, solid and hard. In microscopic characteristics, epicarp cells of B. aschgarica thickening like a string of beads, stone cells as the rectangle, and the stone cell walls of L. ruthenicum is wavy, obvious grain layer. In molecular sequences, the length of ITS sequence of B. kaschgarica is 606 bp, L. ruthenicum is 654 bp, the similarity of the two sequences is 53.32%.

Study on index components and fingerprints of crude and processed Siegesbeckia Herbs / 中国中药杂志

The change of kirenol, darutigenol and darutoside in Siegesbeckia and its first to ninth processed products were studied, and the ten fingerprints were compared, which provided the experimental basis for the study of Siegesbeckia processing tech- nology. The samples were analysed by HPLC on a SunFire-C18 column (4.6 mm x 150 mm, 5 μm) with gradient elution of acetonitrile (0.1% formic acid)-water (0.1% formic acid) at a flow rate of 1.0 mL x min(-1). Column temperaturewas 30 °C and the detected wavelength was 215, 320 nm. The calibration curves of kirenol, darutigenol and darutoside were linear in the range of 2.180-26.16, 2.900-34.80, and 1.012-6.072 mg x L(-1), respectively, and the average recoveries were 96.4%, 97.2% and 96.3% wit RSD 2.2%, 1.7% and 2.4%. This method was simple, the result was stable and had good repeatability, recovery and precision. The re- sult was the basis of the chemical contents variation in the processing of Siegesbeckia Herbs and further clarifying the effect of the changing.

Quality survey of different species of clematidis radix et rhizoma / 中国中药杂志

Quality survey of different species of Clematidis Radix et Rhizoma was made by determining the content of hederagenin and oleanolic acid from Clematidis Radix et Rhizoma. The result showed that only a few samples of Clematis chinensis met the quality standard for Clematidis Radix et Rhizoma in Chinese Pharmacopoeia 2010 Edition.

Two new sesquiterpene lactones from the pericarp of Illicium macranthum / 药学学报

Silica gel column chromatography was used for the isolation and purification of the chemical constituents of the pericarp of Illicium macranthum. From dichloromethane-EtOAc (1:1) fraction and EtOAc fraction of the methanol extracts, eleven compounds were identified on the basis of chemical and spectral data. Two new compounds were elucidated to be 6-deoxyneomajucin (1) and 2-oxo-6-deoxyneomajucin (2), along with nine known compounds 6-deoxypseudoanisatin (3), pseudoanisatin (4), anisatin (5), pseudomajucin (6), protocatecheuic acid (7), shikimic acid (8), shikimic acid methylester (9), beta-sitosterol (10) and daucosterol (11). Compounds 1 and 2 are new majucin-type sesquiterpene lactones.

Effect ofAangong Niuhuang pill and heavy metal constituents on EcoG of brain damage caused by LPS in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To probe the mechanism of EEG activation and Xingnao Kaiqiao, evaluate the actions of cinnabaris and realgar in Xingnao Kaiqiao of Angong Niuhuang pill, guess the significance of cinnabaris and realgar in specific indication treatment of Angong Niuhuang pill, and provide experimental bases for the rationality of Angong Niuhuang pill building-up.</p><p><b>METHOD</b>Seventy SD rats were divided into seven groups: the control, the model, the Angong Niuhuang pill (0.4 g x kg(-1)), the Angong Niuhuang pill without cinnabaris and realgar (0.32 g x kg(-1)) , the cinnabaris and realgar (0.08 g x kg(-1)), the realgar (0.04 g x kg(-1)), and the cinnabaris (0.04 g x kg(-1)). Rats in the control and model groups were given distilled water. After three days of administration, the brain damage model was made by Lipopolysaccharides (LPS) injection through caudal vein and the catecholamine (CA) and its metabolites levels in cerebral cortex, included noradrenaline (NE), adrenaline (E), 3-methocy-4-hydroxyphenylglycol (MHPG), 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), dopamine (DA), Homovanlic acid (HVA), 3,4-dihydroxyphenylacetic acid (DOPAC), were determined by high-performance liquid chromatography with electrochemical detector (HPLC-ECD). Influences of Angong Niuhuang pill, Angong Niuhuang pill without cinnabaris and realgar, cinnabaris and realgar on monoamine transmitters were observed in brain damage rats caused by LPS.</p><p><b>RESULT</b>LPS could raise NE, 5-HT, 5-HIAA levels and reduce E, DOPAC levels, but had no influence on HVA, DA, MHPG levels. Angong Niuhuang pill had the trend of raising E, DOPAC levels and reducing NE level, and could reduce 5-HIAA level obviously comparing with models. But Angong Niuhuang pill without cinnabaris and realgar was different, NE level was significantly higher compared to models and Angong Niuhuang pill, DA level was also significantly higher compared to all groups. Cinnabaris and realgar had the same action trends with Angong Niuhuang pill, and separate realgar could obviously reduce 5-HT.</p><p><b>CONCLUSION</b>Influence on CA and its metabolites levels in cerebral cortex may be one of the mechanisms of Angong Niuhuang pill's EEG activation, and cinnabaris and realgar have the same action on CA levels in cerebral cortex. The results of the present work allow us to put forward the hypothesis that cinnabaris and realgar are most likely one of the important material basis in Xingnao Kaiqiao of Angong Niuhuang pill.</p>

GC-MS analysis of the fatty components of pollen Typhae before and after being carbonized / 中国中药杂志

<p><b>OBJECTIVE</b>To study the changes of the fatty components of Pollen Typhae before and after being carbonized.</p><p><b>METHOD</b>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.</p><p><b>RESULT</b>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.</p><p><b>CONCLUSION</b>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.</p>

Studies on the nasal epithelium toxicity of adjuvants and recombination hirudin (rHV2) nasal spary / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the nasal epithelium toxicity of adjuvants and rHV2 nasal spary(HVS).</p><p><b>METHOD</b>Ciliary movement were evaluated with in situ toad palate model; The histology assessment of nasal epithelium were carried out after long-lasting and repeated use of HVS.</p><p><b>RESULT AND CONCLUSION</b>Adjuvants included SDS, Brij 35, azone, lecithin, EDTA, menthol, nipagin and thiomersal were able to significantly inhibited the ciliary movement, while tween80, glycyrrhizic acid monoammonium salt, benzalkonium bromide, sodium benzoate and adhensive materials investigated had less influence on it. HVS was able to damaged the nasal epithelium, but this effect recovered soon after stopping administration. It was demonstrated that SDS, Brij 35, azone,lecithin, EDTA, menthol, nipagin and thiomersal. It had significant cilitoxity, while tween80, glycyrrhizic acid monoammonium salt, benzalkonium bromide, sodium benzoate and adhensive materials investigated had no significance; Chitosan co-administration with some adjuvants may make the cillitoxity severer; It is available that rHV2 be administered by nasal spary.</p>

Analysis of volatile constituents from Anemarrhena asphodeloides by GC-MS / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of the volatile oil from the rhizome of Anemarrhena asphodeloides.</p><p><b>METHOD</b>The volatile oil was steam distillation. Chemical constituents were separated and analyzed by GC-MS. The relative content of each component was determined by area nomalization.</p><p><b>RESULT</b>24 volatile compounds were isolated and identified for the first time, representing 70.83% of the total oil.</p><p><b>CONCLUSION</b>The main constituents of this oil were aldehydes (31.15%), terpene and their oxide (20.66%), alkyls (8.35%), Furan heterocyclic compounds (6.41%), non terpene alcohol (4.26%). There are 12 compounds with contents over 3%. Among them, borneol has the highest content (9.35%).</p>

Applications of solid-phase microextraction technique in natural product analysis / 中国中药杂志

Solid-phase microextraction is a new technique of analysis. It has many merits and expanse foreground. A Review of the principle, recent development and applications of solid-phase microextraction is given, focusing on natural product analysis, especially on Chinese traditional medicine. Twenty-nine references are cited in the paper.

The underground part growth distribution pattern of Glycyrrhiza uralensis and its effects on glycyrrhizinic acid content / 中国中药杂志

<p><b>OBJECTIVE</b>To ascertain the relationship between glycyrrhizinic acid content and the underground part growth character of Glycyrrhiza uralensis and provide the theoretical evidence for wild resources protection and artificial cultivation method of G. uralensis.</p><p><b>METHOD</b>Through the analytical investigation on the underground part of G. uralensis and analysis of glycyrrhizinic acid content in different organs, parts, ages, and diameter medicinal materials, the systematic study on the relationship between glycyrrhizinic acid content and the underground part growth character of glycyrrhiza uralensis was carried out.</p><p><b>RESULT</b>The underground part of a G. uralensis seedling consisted of seed root, random root, horizontal underground stem, vertical underground stem and assimilating root. The glycyrrhizinic acid content in horizontal underground stem with the age below two years old or in random root with the diameter below 0.5 cm was low. The difference of glycyrrhizinic acid content among horizontal underground stem, random root and vertical underground stem was obvious, but the difference between horizontal underground stem and random root was not obvious.</p><p><b>CONCLUSION</b>The horizontal underground stem was of G. uralnesis acts as a link that can connect random root, vertical underground and stem assimilating root, so that the whole underground part constructs one huge underground net system. The glycyrrhizinic acid accumulation is a effected by organ type, growth age, root diameter and grow position, and the distribution pattern of random root and vertical underground stem has influence on glycyrrhizinic acid distribution in horizontal underground stem.</p>

The changes of some chemical components in banxiaxiexintang decoction of different combinations / 中国中药杂志

<p><b>OBJECTIVE</b>The contents of bererine, palmatine and glycyrrhizin acid in Banxiaxiexintang decoction of different combinations were determined by PR-HPLC.</p><p><b>METHOD</b>A Shim-pack CLC-ODS column was used with a mobile phase of CH3CN-H2O (31:69; 0.005 moL.L-1 -pentanesulfonic acid sodium salt, H3PO4: pH 3.0) for bererine andpalmatine, which were detected at the wavelength of 275 nm. A YWG-C18 column was used with a mobile phase of CH3OH-H2O-HAc(62:37:1) for glycyrrhizin acid which was detected at the wavelength of 260 nm.</p><p><b>RESULT</b>Each herbs' combination influences the contents of the 3 components.</p><p><b>CONCLUSION</b>The experiment is an attempt to study the comical foundation of traditional Chinese prescription.</p>