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To establish an HPLC-DAD method for simultaneous determination of eight ginsenosides in Renshenshouwu capsules.Ultimate C18 column (4.6 mm x 250 mm, 5 microm) was adopted for gradient elution, with acetonitrile and water as the mobile phase. The flow rate was 1 mL x min(-1), the column temperature was set at 30 degrees C, and the detection wavelength was 203 nm.A good linearity was observed in the range of 0.242-12.1, 0.222-11.1, 0.251-25.1, 0. 245-24.5, 0.232-23.2, 0.232-23.2, 0.264-26.4, 0.244-24.4 microg for ginsenoside Rg1, Re, Rb1, Rc, Rb2, Rb3, Rd and 20(S)-ginsenoside Rg3, respectively, with the average recoveries of 102.7%, 103.2%, 101.6%, 101.2%, 102.0%, 100.7%, 101.9%, 102.0%, respectively.The method was so simple, accurate and effective that it could be used for quality control of the above eight components in Renshenshouwu capsules.

Background. The method for mouse vascularized heart transplantation have been described using suture and cuff techniques. Technical problems have limited its widespread use. Here, we describe our method of modified cervical heterotopic transplantation with the cuff technique. Methods. By using a smaller Teflon cuff (external diameter 0.6 mm, internal diameter 0.4 mm) and superfine-tip forceps, it became possible to directly pull the edge of the carotid artery and evert the proximal end of the artery over the cuff. Similarly, the external jugular vein could be easily everted over a 22-gauge cuff with this direct pulling method. Results. By these modifications, the operation time was reduced. It usually takes 20 min for the donor harvest, 15 min for preparation of the cervical vessels, and 15 min for anastomosis. All procedures from the donor harvest through skin closure of the recipient mice can be completed within 1 hr, and ischemic time is within 25-40 min. Conclusions. This method can be used to investigate cyclophosphamide-induced tolerance and mechanisms of reperfusion injury.

Background.Injection of allo-spleen cells (SC) followed by a single dose of cyclophosphamide (CP) can induce tolerance of tumor and/or skin allografts in mice. To minimize the damage caused by CP, fractionation of CP that can establish long-lasting skin graft survival, stable mixed chimerism, and in

The title complex, [Ni(C3S5)(C5H12N2)] or [Ni(dmit)(dach)] (dach is 1,4-diaza­cyclo­heptane and dmit is 2-thioxo-1,3-dithiole-4,5-dithiol­ate), crystallizes with two independent mol­ecules per asymmetric unit. In each mol­ecule, the central NiII atom is coplanar with the two S atoms of one dmit ligand and two N atoms of one dach ligand. In the crystal structure, some weak S⋯S inter­actions and N—H⋯S and C—H⋯N hydrogen bonds are present. These inter­actions generate a three-dimensional framework.

Ten compounds were isolated from cultures of the fungus Armillaria mellea by silica gel and Sephadex LH-20 column chromatographies. Their structures were identified by spectroscopic data analysis and compared their spectroscopic data with those reported in the literature as 2-hydroxy-4-methoxy-6-methylbenzoic acid (1), orsellinic acid (2), melleolide (3), ergosterol (4), genistein (5), daidzein (6), daucosterol (7), genistin (8), uracil (9) and D-mannitol (10). Compounds 1-10 were isolated from cultures of A. mellea for the first time. Among them, 14, 6 and 10 had been obtained from mycelia or rhizomorphs of A. mellea. The structure of compound 3 was determined by detailed analysis of its 1D and 2D NMR data in the solvent of DMSO for the first time.

An HPLC method was developed for simultaneous quantitation of rosmarinic acid, oridonin and chrysoplenetin in the aerial parts of Isodon rubescens. Samples were analyzed on an Ultimate C18 column (4.6 mm x 250 mm, 5 microm) with methanol and water containing 0.1% formic acid as mobile phases in a linear gradient mode. The flow rate was 1.0 mL x min(-1) and the temperature was set at 30 degrees C. The PDA detector wavelengths were set at 338 nm for rosmarinic and chrysoplenetin and at 242 nm for oridonin. The linear ranges were 0.222-2.78, 0.227-2.84 and 0.005-0.071 microg for rosmarinic acid, oridonin and chrysoplenetin, respectively. The average recoveries of the three constituents were 102.9% (RSD 1.9%), 99.6% (RSD 1.1%) and 102.5% (RSD 0.94%), respectively. This method was proved to be accurate and repeatable, and can be used for quality control of the aerial parts of I. rubescens.

Simple gas chromatography –mass spectrometry (GC–MS) and highperformance liquid chromatography (HPLC) methods were developed for quantifying eight volatile compounds and 10 sesquiterpenoids, respectively. GC–MS analysis was performed on an HP-5MS capillary column (30 m30.25 mm i.d.) coated with 0.25 mm film 5% phenyl – 95% methylpolysiloxane and selected ion monitoring was used for quantification. Four volatile and previously unquantified monoterpenoids were determined. HPLC analysis was performed on a C18 column with water and acetonitrile as mobile phase. The proposed method, determined 10 non-polar and polar sesquiterpenoids simultaneously, which covered a wider polar range of analytes and had a more perfect resolution. Among them, five sesquiterpenoids were not determined before and some specific components, (4S,5S)germacrone-4,5-epoxide, curcumenone and dehydrocurdione were completely separated for the first time. Both methods were validated for linearity, limit of detection and quantification, precision, accuracy, recovery and system suitability. The methods were simple, effective, reliable and successfully applied to global detection and analysis of volatile and non-volatile components of steamed and non-steamed rhizomes of Curcuma wenyujin (Wen-E-Zhu (WEZ) and Pian-JiangHuang (PJH)). Multivariate statistical analysis was employed to distinguish PJH and WEZ and seven chemical components, including (4S,5S)-germacrone-4,5-epoxide, curcumenone, b-elemene, curzerene, borneol, isoborneol and camphor, were screened as chemical markers. The present study provided a promising method for accurate discrimination of the herbal medicines with the same origin.

Sargentodoxae Caulis was prepared from the stems of Sargentodoxa cuneata. Twenty compounds from the the stems of S. cuneata collected in Huangshan Mountain, Anhui province, were isolated and purified by column chromatography on macroporous resin (HPD100), silica gel, Sephadex LH-20 and semi-preparative HPLC. Their structures were elucidated on the basis of physico-chemical properties and spectral data analyses as (7R,8S)-3,3 '-5-trimethoxy-4,9-dihydroxy-4',7-expoxy-5',8-lignan-7'-en-9'-oic acid 4-O-beta-D-glucopyranoside(1), 1-O-(vanillic acid) -6-O-vanilloyl-beta-D-glucopyranoside(2), 4-hydroxyphenylethyl-6-O-coumaroyl-beta-D-glucopyranoside(3), citrusin B(4), cinnamoside(5), (-) -isolariciresinol 4'-O-beta-D-glucopyranoside (6), (-) -isolariciresinol 4-O-beta-D-glucopyranoside (7), 1-O-(vanillic acid) -6-(3", 5"-dimethoxy-galloyl) -beta-D-glucopyranoside (8), 4-hydroxyphenyl-ethyl-6-O-(E) -caffeoyl-beta-D-glucopyranoside (9), (-)-syringaresinol 4'-O-beta-D-glucopyranoside (10), (-)-syringaresinol di-O-beta-D-glucopyranoside (11), aegineoside (12), calceolarioside B (13), 4-hydroxy-3-methoxy-acetophenone-4-O-alpha-L-rhamnopyranosyl-(1 --6)-beta-D-glucopyranoside (14), 4-hydroxy-3-methoxy-acetophenone-4-O-beta-D-apiofuranosyl-(1 --6) -beta-D-glucopyranoside (15), (-) -epicatechin (16), salidroside (17), 3,4-dihydroxy-phenyl ethyl-beta-D-glucopyranoside (18), chlorogenic acid (19) and protocatechuic acid (20). Compound 1 was a new compound and compounds 2-7 were isolated from this plant for the first time.

Eleven compounds were isolated from the leaves of Epimedium pubescens by means of various chromatographic techniques such as silica gel, MCI, Sephadex LH-20 and preparative HPLC. Their structures were identified as anhydroicaritin (1), icariside II (2), 2'''-O-rhamonosyl-icariside II (3), desmethylanhydroicaritin (4), baohuosaide II (5), epimedokoreanin B (6), acuminatin (7), tricin(8), kaempferol (9), daidzein (10) and 4-hydroxy ethyl benzoate (11) on the basis of physicochemical properties and spectroscopic data analysis. Among them, compound 11 was isolated from Epimedium species for the first time, and other compounds were obtained from this plant for the first time.

The traditional after-harvesting drying method of C. morifolium cv. Hang-ju (HJ) is sun drying, but recently sulfur fumigation is increasingly used as a cheap and convenient method. However, the effects of sulfur fumigation on chemical constituents and potential activities of HJ were unknown. A comprehensively comparison of the chemical profiles between non-fumigated HJ (NHJ) and sulfur-fumigated HJ (SHJ) was conducted by HPLC fingerprints analysis and the discrepant peaks were identified or tentatively assigned by HPLC-ESI/MS(n). Dramatic chemical changes were found that the contents of 4 flavonoid aglycones remarkably increased while those of 7 glycosides significantly reduced which suggested that sulfur-fumigation induced flavonoid glycosides transformed into aglycons by hydrolysis reaction. A significant loss of hydroxycinnamoylquinic acids showed the sulfur fumigation was a destructive effect on HJ. Principal component analysis (PCA) was employed to rapidly discriminate NHJ and SHJ samples. By ICP-OES analysis, it was found that the residue of sulfur of SHJ were three times higher than NHJ (p

A fraction named GFC-1 with high antioxidant activities in vitro was isolated from the enzymatic hydrolysates of roasted pills of Asini Corii Colla, and the peptides in this fraction were identified. The enzymatic hydrolysates were isolated and purified with anion exchange chromatography and Sephadex G-25 filtration chromatography successively. GFC-1, a fraction isolated from the hydrolysates, exhibited the highest DPPH and ABTS scavenging capacity (DPPH 47. 95% at 2.0 g x L(-1) and ABTS 97.20% at 0.40 g x L(-1). Nine peptides from GFC-1 were identified by LC-ESI-MS/MS coupled with TurboSEQUEST search software and Swiss-Prot data base, and a high repetition core sequence GPAGPP*GPP* was also found.

To investigate the effect of different chromatographic conditions on QAMS relative correlation factors (RCF) and relative retention values (RT(R)).C18 columns were used with methyl alcohol-0.4% phosphoric acid water (85: 15) as the mobile phase. The detection wavelength was 254 nm, the column temperature was 30 degrees C, and the flow rate was 1.0 mL x min(-1). The five anthraquinones in Rhei Radix et Rhizoma were selected to be the objects of study. The RCF and RT(R) among aloe-emodin, rhein, chrysophanol, physcion and emodin were determined under different chromatographic conditions.Their RCFs showed no significant difference.The RCFs among anthraquinones established by QAMS can be used as a constant in content determination of traditional Chinese medicines/patent traditional Chinese medicines.

Some Curcuma species are widely used as herbal medicines. Sesquiterpenes are their important bioactive compounds and their quantitative analysis is generally accomplished by gas chromatography (GC) or high performance liquid chromatography (HPLC), but the instability and high cost of some sesquiterpene reference standards have limited their application. It is necessary to find a practicable means to control the quality of herbal medicines. Using one stable component contained in Curcuma species to determine multiple analogues should be a practical option. In this study, a simple HPLC method for determination of sesquiterpenes using relative response factors (RRFs) has been developed. The easily available and stable active component curdione was selected as the reference compound for calculating the RRFs of the other eight sesquiterpenes, including zedoarondiol (Zedo), isozedoarondiol (Isoz), aerugidiol (Aeru), (4S,5S)-(+)-germacrone-4,5-epoxide (Epox), curcumenone (Curc), neocurdione (Neoc), germacrone (Germ) and furanodiene (Fura). Their RRFs against curdione were between 0.131-1.301, with a good reproducibility. By using the RRFs, the quantification of sesquiterpenes in Curcuma wenyujin herbal medicines and related products was carried out. The method is especially useful for the determination of (4S,5S)-(+)-germacrone-4,5-epoxide, curcumenone, germacrone and furanodiene, which often are regarded as the principle components in Curcuma species, but unstable when were purified. It is an ideal means to analyze the components for which reference standards are not readily available.

Premise of the study: Microsatellite markers were developed for Garcinia paucinervis (Clusiaceae), an endangered and endemic tree species of karst habitats, to analyze its genetic diversity and genetic structure. Methods and Results: Using shotgun sequencing on an Illumina MiSeq platform, a total of 22 microsatellite primer sets were characterized, of which 17 were identified as polymorphic. For these polymorphic loci, the total number of alleles per locus ranged from two to 12 across 54 individuals from three populations. The observed and expected heterozygosities ranged from 0.000 to 1.000 and from 0.000 to 0.850, respectively. No pair of loci showed significant linkage disequilibrium. Three loci in one population deviated significantly from Hardy–Weinberg equilibrium (P < 0.05). Seven loci (JSL3, JSL5, JSL22, JSL29, JSL32, JSL39, and JSL43) were successfully amplified in G. bracteata. Conclusions: These markers will be useful in studies on genetic diversity and population structure of G. paucinervis.

To study the optimum process of removing cadmium irons from extracts of Gentianae Radix et Rhizoma with gamma-mercaptopropyl-modified silica gel (MPS) and assess its cadmium ion-removing property.Static and dynamic adsorptions were adopted to detect the cadmium-removing rate. MPS' cadmium ion-removing property was assessed with such indicators as the cadmium-removing rate, the solid content and the HPLC fingerprint.The process parameters of the static adsorption were as follows: 0.20 g x mL(-1) of concentration of extracts, 120 minutes of adsorption time and 15:1 between raw materials and MPS. The process parameters of the dynamic adsorption were as follows: 1:3.5 times between diameter and height, 0.20 g x mL(-1) of concentration of extracts, 0.9 mL x min(-1) of flow rate of the extracts and 50:1 between raw materials and MPS. Before and after the cadmium ion-removing process, the extracts showed no notable difference in solid content and HPLC fingerprint.gamma-mercaptopropyl-modified silica gel (MPS) can effectively remove cadmium ion from the extracts of Gentianae Radix et Rhizoma with an excellent cadmium ion-removing property.

OBJECTIVE: To investigate the effect of different chromatographic conditions on QAMS relative correlation factors (RCF) and relative retention values (RT(R)). METHOD: C18 columns were used with methyl alcohol-0.4% phosphoric acid water (85: 15) as the mobile phase. The detection wavelength was 254 nm, the column temperature was 30 degrees C, and the flow rate was 1.0 mL x min(-1). The five anthraquinones in Rhei Radix et Rhizoma were selected to be the objects of study. The RCF and RT(R) among aloe-emodin, rhein, chrysophanol, physcion and emodin were determined under different chromatographic conditions. RESULT: Their RCFs showed no significant difference. CONCLUSION: The RCFs among anthraquinones established by QAMS can be used as a constant in content determination of traditional Chinese medicines/patent traditional Chinese medicines.

Objective To kinds of establish a HPLC method for determining contents of indole alkaloids and bufadienolides contained in toad medicines, and analyze two kinds of components contained in toad venom, toad skin and toad periostracum. Method As for alkaloids, Nucleosil C18 column was adopted with acetonitrile and water containing 0.5% potassium dihydrogen phosphate (6: 94, adjust pH to 3.2 with phosphate acid) as the mobile phase. The flow rate was 0.8 mL x min(-1), the detection wavelength was 275 nm, and the column temperature was 30 degrees C. As for bufadienolides, Alltima C18 column was adopted with acetonitrile and water containing 0.3% acetic acid (B) as the mobile phase. The gradient process was as follows: a linear gradient from 28% to 54% acetonitrile in the first 15 min, then kept at 54% for additional 20 min. The flow rate was 0.6 mL x min(-1), the detection wavelength was 296 nm, and the column temperature was 30 degrees C. Result The linear ranges were 0.079 6-0.796 microg for serotonin, 0.097 2-1.945 microg for N-methylserotonin, 0.074 4-0.744 microg for N,N-dimethylserotonin, 0.103-2.05 microg for N,N,N-trimethylserotonin, and 0.067 2-0.672 microg for bufothionine, respectively. The average recoveries of serotonin and N-methylserotonin were 98.6% and 91.3%, respectively. The linear ranges of gamabufotalin, bufotalin, bufalin, cinobufagin and resibufogenin were 0.004 83-0.614, 0.007 9-1.006, 0.007 95-1.016, 0.009 7-1.24 and 0.009 6-1.22 microg, respectively, and their average recoveries were 101.6%, 102.5%, 101.0%, 99.1% and 98.9%, respectively. Conclusion Toad venom has the highest contents of indole alkaloids and bufadienolides, followed by toad skin, and toad periostracum showed the lowest contents and even no detection result.

On the basis of analysis and summary of current application of reference extracts of traditional Chinese medicine, reference extracts for quality assessment of traditional Chinese medicines are divided into three classes by purpose: 1. herbal or volatile oil reference extracts for TLC identification of original herbal medicines or patent traditional Chinese medicines containing original herbal medicines. 2. component reference extracts for identification of certain components or positioning of chromatographic peaks. 3. reference extracts marked with component content to be determined for quantitative analysis on samples. For the third kind of reference extracts, akebiae saponin was taken as an example for preparation thought and methods in such aspects as preparation process, component identification and content analysis.

OBJECTIVE To establish a quality evaluation method, quantitative analysis of multi-component with a single-marker (QAMS) to determine the contents of four rhubarb anthraquinones in Sanhuang tablets. METHOD Emodin was used as the internal reference substance, the relative correlation factors (RCF) of rhein, chrysophanol and physcion to emodin were calculated and evaluated. The contents of these four anthraquinones were determined by the external standard method and QAMS, respectively. Rationality, feasibility and repeatability of the QAMS method was verified by comparing the results obtained from the two different methods. RESULT RCFs of rhein, chrysophanol and physcion to emodin against Sanhuang tablets were 1.13, 1.46 and 1.01, respectively, indicating good reproducibilities. The two methods did not show significant difference in results. CONCLUSION The QAMS method can be used a new quality assessment model for quantity of anthraquinones in Sanhuang tablets.

Present study is concentrated on evaluating the climate adaptive capability of Tuzhang dwellings. The comparisons to different thermal physical properties such as thermal resistance R0, thermal inertia index data D, reduction coefficient of thermal wave transferring V0 and thermal transferring delaying time ξ0 have been performed between Tuzhang dwelling and normal brick house. Comparing results show that Tuzhang dwelling has much better thermal properties than normal brick house. Therefore, regarding to climate adaptive capability, traditional Tuzhang dwelling is vastly superior to brick house.

Objective To evaluate the inhibitory effect of total bufadienolides from toad venom against H22 tumor in mice and preliminarily analyze the structures of the metabolites in tissues. Method HPLC and LC-MS were used for analysis of the chemical composition of TBFs. High, middle and low dosages of TBFs were orally administered or intra-peritoneally injected to H22 tumor-bearing mice for thirteen days. The animals were killed and the tumors were stripped and weighed. The metabolites in the tissues such as heart, liver, spleen, lung and kidney, were analyzed by HPLC and LC-MS. Result The chemical composition of TBFs were identified by comparison of the retention times with those of reference substances, on-line UV spectra and MS data. Its main components are concerned with gamabufotalin, arenobufagin, bufotalin, resibufagin, cinobufotalin, bufalin, cinobufagin and resibufogenin. TBFs had no obvious influence on body weight of H-22 tumor-bearing mice orally administered and the inhibition rate against tumor were 14.76%, 16.38% and 10.32% for low (5 mg x kg(-1)), middle (10 mg x kg(-1)) and high dosage (20 mg x kg(-1)), respectively. The mice intra-peritoneally injected with middle and high-dose of TBFs gained body weight slower than the control mice on the 5th day and recovered on the 13th day. The inhibition rate against tumor were 17.30%, 19.80% and 40.95% for low (1.5 mg x kg(-1)), middle (3 mg x kg(-1)) and high dose (6 mg x kg(-1)), respectively. The inhibitory effect took on dose-dependent manner. Based on the HPLC analyses on heart, liver, spleen, lung and kidney, bufadienolides were found in the liver tissue and 11 compounds of them were tentatively identified by LC-DAD-MS. Conclusion TBFs by oral administration had no inhibitory effect against H22 tumor in mice, however, TBFs by intra-peritoneal injection displayed the significantly inhibitory effect, accompanying some toxicity for early duration of the study. The identification of bufadienolides in the liver provides a good basis for the further investigation of the metabolic pathways of TBFs in vivo.

OBJECTIVE To establish a new quantitative method for simultaneous determination of multi-components in Salvia miltiorrhiza by using one chemical reference substance and validate its feasibilities. METHODS The new quality evaluation method, quantitative analysis of multi-components by singer-marker (QAMS), was established and validated with S. miltiorrhiza . Five main ingredients were selected as analytes to evaluate the quatity and their relative correlation factors (RCF) were determined by HPLC-DAD. Within the linear range, the values of RCF at 280 nm of sodium danshensu to protocatechuic aldehyde, rosmarinic acid, lithospermic acid and salvianolic acid B were 7.559, 2.901, 1.825 and 1.746, respectively. The contents of danshensu in the samples of S. miltiorrhiza were determined by using the external standard method, and the contents of the four other ingredients were calculated by their RCFS. The contents of these four ingredients in all samples were also determined by the external standard method. RESULT Within a certain range, the RCFs showed good reproducibility (RSD 1.0%-2.5%). No significant differences were observed between the quantitative results of the two methods. CONCLUSION It is feasible and suitable to evaluate the quality of S. miltiorrhiza and its processed products by QAMS.

Objective To establish a new quantitative method for simultaneous determination of multi-coumarins in Fraxini Cortex by using one chemical reference substance, and validate its feasibilities. Method The new quality evaluation method, quantitative analysis of multi-components by singer-marker (QAMS), was established and validated with Fraxini Cortex. Four main coumarins were selected as analytes to evaluate the quality and their relative correlation factors (RCF) were determined by HPLC-DAD. Within the linear range, the values of RCF at 340 nm of aesculin to asculetin, fraxin and fraxetin were 1.771, 0.799, 1.409, respectively. And the contents of aesculin in samples of Fraxini Cortex were authentically determined by the external standard method, and the contents of the three other coumarins were calculated by their RCF. The contents of these four coumarins in all samples were also determined by the external standard method. Result Within a certain range, the RCF had a good reproducibility (RSD 2.5%-3.9%). Significant differences were not observed between the quantitative results of two methods. Conclusion It is feasible and suitable to evaluate the quality of Fraxini Cortex and its Yinpian by QAMS.

OBJECTIVE: To examine the feasibilities of the method, quantitative analysis of multi-components by single marker (QAMS) for determination of four alkaloids in Xianglian preparations. METHOD: A HPLC method was developed to determine the contents of four alkaloids, epiberberine, coptisine, palmatine and berberine in Xianglian preparations. Berberine was selected as the internal reference substance, and 3 relative correction factors to berberine were calculated. The contents of the four alkaloids in 16 batches of samples were determined by both external standard method and QAMS. The new method was evaluated by comparing the quantitative results with of the external standard method. RESULT: No significant differences were found between the quantitative results of four alkaloids in 16 batches of samples determined by the two methods. CONCLUSION: QAMS is feasible for determination of four alkaloids simultaneously, and the developed method can be used for quality control of Xiang Lian preparations.

OBJECTIVE To remove plumbum ion from the extract of Lonicera japonica by gamma-mercaptopropyl-modified silica gel (MPS). METHOD The reference plumbum ion solution was added into the extract of L. japonica as the tested samples. The static and dynamic adsorptions were investigated to evaluate the influent factors on the removal of plumbum ion in the extract. The solid content and HPLC profiles of the extract were determined to evaluate the process changes. RESULT MPS showed excellent adsorption charateristics to remove plumbum ion from the extract of L. japonica, with fast adsorption rate and more than 80% removal efficiency. The optimum parameters were as follows: 6 times sample volume, 10 times of height to diameter, and 6 BV x h(-1) of flow velocity, at the temperature of 25 degrees C. During the removal process, obvious changes of solid content and HPLC profiles were not observed. CONCLUSION MPS could be used to remove plumbum ion from extract of L. japonica.

OBJECTIVE To study HPLC characteristic fingerprint of Moghania philippinensis and provide a reliable method for scientific evaluation and quality control. METHOD The chromatographic separation was performed on a Kromasil C15 column (4.6 mm x 250 mm, 5 microm). The mobile phase consisted of acetonitrile and water containing 0.3% acetic acid in gradient elution. The column temperature was set at 40 degrees C and the detection wavelength was at 254 nm. RESULT The developed HPLC fingerprint method above was applied to analyze 23 batches of crude herbs of Moghania philippinensis. The total 22 peaks were selected as the characteristic peaks. And five of them were identified as genistein, genistin, 5,7,3',4'-tetrahydroxy-6,8-diprenylisoflavone, 5,7,4'-trihydroxy-6,8-diprenylisoflavone and flemiphilippinin E by matching their retention time, UV spectra data with those of the authentic chemical reference substances. CONCLUSION The method is simple, accurate and reproducible. It is suitable for the quality control of M. philippinensis.

OBJECTIVE To study the key processes of Fufang Kushen injection for technical upgrading. METHOD Total alkaloids (sum of matrine, oxymatrine, sophoridine and oxysophoridine) and macrozamin were selected as quality evaluation markers. The key processes of percolation with acetic acid and discoloration with activated carbon were optimized by orthogonal experiment design, and process of purification with alcohol was investigated by single factor method. RESULT The optimal condition of percolation process is as follows: the medicinal materials are soaked for 9 h with 4 times water containing 0.8% acetic acid, then percolation starts at flow-rate of 5 mL x min(-1) x kg(-1) and adding 2 times 0.8% acetic acid solution is added at the same velocity. Purification process is that the concentrated solution is precipitated by 60%, 80% and 90% alcohol in turn. Discoloration process is that 6 activated carbon is added into the solution which is heated at 60 degrees C for 20 minutes. CONCLUSION The optimal extraction process is not only simple, saving the industrial cycle, reducing the potential risk, but also decreasing the acetic acid amount to guarantee the acid-insoluble ash as well as the functional ingredients.

To study the chemical constituents of the stems of Clematis parviloba, six compounds were isolated from a 95% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, ODS, Sephadex LH-20, and semi-preparative HPLC. Two new phenolic glycosides, 2-((E)-3-carboxybut-2-en-yl)-4-hydroxy-3-methyl-phenyl-O-beta-D-glucopyranoside (1) and 4'-hydroxy-phenol-beta-D-[6-O-(4"-hydroxy-3", 5"-dimethoxy-benzoate)] glucopyranoside (2) were isolated, together with a known phenolic glycoside, 4'-hydroxy-3'-methoxy-phenol-beta-D-[6-O-(4"-hydroxy-3", 5"-dimethoxy-benzoate)] glucopyranoside (3) as well as three known megastigmane glycosides, linarionoside A (4), linarionoside C (5), and staphylionoside K (6). Their structures were determined on the basis of spectroscopic analysis and chemical evidence. Among them, compounds 1 and 2 were named as clemaparvilosides A (1) and B (2), respectively, and compounds 3-6 were obtained from Clematis genus for the first time.

A HPLC method for limit detection of aristolochic acid A in the Chinese herbs containing aristolochic acid or suspected-containing aristolochic acid and their preparations was established. The samples were analyzed on an Alltima C18 column eluted with methanol-water-acetic acid (68:32:1.5) as the mobile phase. Flow rate was at 1.0 mL x min(-1) and the detection wavelength was at 390 nm. The calibration curve was linear over the range from 0.016 to 0.51 g (r = 0.9993) and LOD was 4 ng. The average recovery was 101.2% with RSD of 2.01%. The procedures of sample preparation were systematically investigated. The contents of aristolochic acid A in Radix et Rhizoma Asari bought from market or drugstore were fluctuated from 3.1 to 26.6 microg x g(-1) and 3 of 11 samples accorded with the quality requirement of current Chinese Pharmacopoeia. Among 15 batches samples of Chinese medicaments, only one sample was found to contain aristolochic acid A. The present investigation shows that the method is sensitive and repeatable and it could be used for the limit detection of aristolochic acid A in the Chinese herbal medicines containing trace amount of aristolochic acid A or suspected-containing aristolochic acid A and their preparations.

OBJECTIVE To develop a GC-MS method for the determination of methyl nonyl ketone in the volatile oil from the herbs of Houttuynia cordata. METHOD The sample was split in the 240 degrees C injection port, with 20:1 split ratio, and separated on a DB-5 (30 m x 0.25 mm, 0.25 microm film thickness) fused silica column with helium as the carrier gas. The temperature program was as follows: 100 degrees C for 2 min, the 5 degrees C x min(-1) to 150 degrees C, then 15 degrees C x min(-1) to 200 degrees C, and kept for 10 min. The MS transfer line temperature was set to 250 degrees C, the MS source temperature was set to 200 degrees C. The ionization mode was electron ionization (EI) and the selective ion monitor mode was used. RESULT A good linear relationship was constructed over the injection amount range of 5.5-110 ng of methyl nonyl ketone. The average recovery was 98.9%, and RSD was 2. 2%. CONCLUSION The developed method was sensitive, accurate, and can be used for the determination of methyl nonyl ketone in the volatile oil and for the quality control of H. cordata.

Objective To establish a method for quick finding the absorption ingredients of Wuzhuyu decoction in order to select the index to control its quality. Method The absorption of three concentration of Wuzhuyu decotion was investigated with the in vitro-everted intestinal sac model. The intestinal bag fluid of jejunum and ileum were collected in different time and the eight ingredients, which were evodiamine (Ev), rutaecarpine (Ru), limonin (Li), ginsenoside-Rb1, -Rg1, -Re (Rb1, Rg1, Re), isorhamnetin-3-O-beta-D-glucosyl(6''-->1'")-alpha-L-rhamnoside (Irs)and 6-gingerol (6-Gi), were detected by HPLC as the represent constituents in samples. Result Eight ingredients except Ru in samples could be detected, but Ev could not be detected in high concentration samples. The ratios between absorption ingredients were different from in Wuzhuyu decotion. Conclusion The in vitro-everted intestinal sac canc absorb the ingredients of Wuzhuyu decotion selectivity. Compare with the ileum, the jejunum can provide the more absorption information and faster, the best test time is 60-90 min.

To develop a quantitative method for determination of zizybeoside II in Ziziphus jujuba.The samples were separated at 30 degrees C on a Zorbax SB-C18 column eluted with methanol-water (20 : 80) as the mobile phase. Flow rate was set at 1.0 mL x min(-1) and the detection wavelength was set at 210 nm.The calibration curve was linear within the range from 0.046 to 0.582 microg (r = 0.999 9) and the average recovery was 97.2%. 12 batches of the crude drugs purchased from different areas were determined and the contents of zizybeoside II in Fructus Jujubae were fluctuated from 0.013% to 0.041%.The method is simple, repeatable and could be used for the quality control of Z. jujuba.

The research patterns for the pharmacodynamic chemical substances in compound prescriptions of Chinese medicinal materials in recent years were summarized, and the deficiencies of the commonly used patterns were commented on. A research pattern for the pharmacodynamic chemical substances in compound prescriptions of Chinese medicinal materials was raised, which is suitable for the characteristics of the Chinese medicine. The trend of the research work was predicted, which would provide some thinking for the pharmacodynamic chemical substances in compound prescription of Chinese medicinal materials.

Current quality control patterns are limited to industrial application, for most of the natural chemical reference substances are expensive and unavailable. Herein, a method, quantitative analysis of multi-components with single marker (QAMS), was established and validated to simultaneously determine nine ginsenosides (ginsenoside Rg1, Re, Rf, Rh1, Rb1, Rc, Rb2, Rb3, Rd) in P. ginseng and four ginsenosides (ginsenoside Rg1, Rh1, Rb1, Rd) in P. notoginseng. Using ginsenoside Rb1 as the contrast, the relative correction factors (RCF) of the other eight ginsenosides were determined by HPLC-DAD. Within the linear ranges, the values of RCF of ginsenoside Rb1 to ginsenoside Rg1, Re, Rf, Rh1, Rc, Rb2, Rb3 and Rd were 1.400, 1.215, 1.517, 1.801, 0.944, 1.012, 1.143, and 1.135, respectively. The RCF had a good reproducibility in various instruments, chromatographic columns (RSD = 0.30% - 3.9%). According to their RCF, we simultaneously determined nine ginsenosides in P. ginseng only using one marker. In addition, the RCF of ginsenosides were used to simultaneously quantitative analysis of four ginsenosides in P. notoginseng. The results of QAMS method were validated by comparing with that of external standard method, and no obvious significant difference was found.

To establish a fingerprint analysis method of Qingfengtong capsule by HPLC.The samples were extracted with 70% ethanol in an ultrasonic bath. The extracts were analyzed at 35 degrees C on a Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) with 0.1 mol x L(-1) potassium dihydrogen phosphate water-solution as mobile phase A, and methanol as mobile phase B. The analysis followed a linear gradient elution program. Initial condition of the mobile phase was 10% B for 2 minutes; then changed to 90% B in 40 minutes. The flow rate was kept at 1.0 ml x min(-1) and the detector wavelength was 262 nm.The main peaks in fingerprint chromatogram of Qingfengtong capsules were separated fairly well. The results of method validation meet the requirements for the fingerprints.The established method can be used for the quality control of Qingfengtong capsules.

Abroad review of current methods for the recovery of liquid residual accelerants in fire debris is given. A summary of current practices of sample preparation methods in U.S. laboratories is presented. Sampling of heated headspace is useful as a screening method. It is also applicable for samples that contain a high concentration of accelerant. Distillation-based methods are effective only when large amounts of accelerants are present but they permit the physical isolation of relatively pure liquids, including some water-soluble materials such as alcohols. Solvent extraction is particularly advantageous for accelerants of low volatility such as diesel fuel or highly evaporated petroleum-based products, but it is very susceptible to interferences from the matrix. Dynamic enrichment by adsorption/elution is the most popular and widely used recovery method for liquid accelerants. It offers high enrichment factors but shows weaknesses with respect to polar and low-volatility liquids. No single method is universally effective for all types of accelerants that are commonly encountered in practice. It appears that currently applied recovery methods are particularly weak for water-soluble organics such as alcohols. Sample preparation methods have to be viewed in conjunction with the method to be used to characterize and identify suspect accelerants. Gas chromatography is used almost exclusively for separation and measurement. Selective detection of components that are diagnostic of accelerants allows important trade-offs between sensitivity, chromatographic efficiency and the level of interferences that can be tolerated. Pattern recognition with comparison of chromatographic profiles is greatly simplified if interferences from the matrix can be suppressed. Application of capillary columns capable of high resolution facilitates the recognition of diagnostically important components. Petroleum-based fluids and products are ubiquitous in the human environment and it is therefore particularly important to pay attention to petroleum-based fluids that are derived from such sources. Many common inconspicuous household materials contain petroleum-based solvents and are thus a part of the natural background. The term sensitivity as it relates to the effectiveness of a recovery method must be critically viewed in this light.

To develop an HPLC method for the determination of hyodeoxycholic acid in Bile Arisaema.The methanol extracts were separated on a Diamonsil ODS column eluted at 35 degrees C with a mixture of methanol-water-glacial acetic acid (75:25:0.01) at flow-rate 1.0 mL x min(-1). The drift-tube temperature of ELSD was 85 degrees C, and the flow rate of air was 3.0 L x min(-1).Hyodeoxycholic acid in methanol extract was well separated, the relationship of logarithms of injection amount and peak area was linear (r = 0.9998) over the range of 2.4-24 microg. The average recovery was 97.1% (RSD 2.9%, n=5).The method could be used for quality control of Bile Arisaema.

To explore the utility of Principal Factor Analysis (PFA) in chromatographic data for quality control.Chromatographic fingerprints of processed root pieces of Paeonia lactiflora were determined by HPLC, the PFA was used for data processing.The quantitative differences among different growing areas and different processing batches were found with the method.The method could be used in quality control for monitoring between-batch products of traditional Chinese pharmaceutical process.

Premise of the study: Microsatellite markers were developed for Ilex kaushue (Aquifoliaceae), a medicinal plant with extremely small wild populations that exists in fragmented habitats, to assess and protect its genetic diversity. Methods and Results: Using 454 GS FLX Titanium sequencing, 16 microsatellite primer sets were isolated and characterized. Fifteen of these markers were polymorphic. The number of alleles per locus ranged from one to nine across 22 individuals from both cultivated and wild populations. The observed and expected heterozygosity in these two populations ranged from 0.000 to 1.000 and from 0.000 to 0.785, respectively. Conclusions: These markers will be useful in studies on genetic diversity of I. kaushue.

To develop a quantitative method for the determination of akebia saponin D in root of Dipacus asperoides and provide scientific basis for quality control of D. asperoides.An IPLC analytical method was established using Kromasil ODS column with acetonitrile-water (30:70) as mobile phase and the detection wavelength was 212 nm.The akebia saponin D in root extract were well separated by this method. The linear range is between 0.582 5-9.32 microg, r = 0.9999. The average recovery and RSD of repeatability are 100.3%, 2.3%, respectively.This method is simple, accurate and reliable by evaluating the method validation data and can be used for the quality control of D. asperoides and its preparations.

To provide experimental data for the quality control of processed Paeonia lactiflora, a Chinese herbal medicine.Traditional processing of P. lactiflora was simulated, content of paeoniflorin and water extracts among different preparations were assayed by HPLC; The quantitative correlations among different processing conditions were analyzed, the effects of processing parameters on the contents of paeoniflorin and water extracts were assayed a nanalysed.The controlled processing parameters were correlated with covariables which showed that processing procedures was controllable, and the heating temperature was a factor impacting the content of paeoniflorin.

To provide the experimental data for the quality control of Paeonia lactiflora a prepared Chinese medicinal herb.The contents paeoniflorin among different growing areas and different processing methods methods were assayed by HPLC.The quantitative differences of paeoniflorin content among different growing areas and different processing methods wereanalyzed quantitatively.The quantitative differences among different growing areas were not significant and the quantitative differences among different processing methods within 10%.

To study the cholelitholytic effect and safety of the preparation of Chinese medicine compound.Experiments were performed to study the efficacy of gallstones dissolution of the preparation of Chinese medicine compound in vitro and in vivo. The toxicity was studied by pathological and blood biochemical changes.The preparation of Chinese medicine compound dissolved cholesterol and mixing gallstones in 7 days in vitro and in vivo, no influence on the blood biochemistry and the other organs of the rabbits.The preparation of Chinese medicine compound is effective and safe for treatment of chololithiasis.

To develop an HPLC method for determination of fingerprints and evaluate green tangerine peel (GTP) processed with vinegar.The samples were extracted with 50% ethanol using ultrasonic bath and analyzed on a Chromasil C18 column eluted with methanol and water containing 0.2% acetic acid in a linear gradient program. The flow rate was 1.0 mL x min(-1), and detection wavelength was set at 280 nm.The RSD values of relative retention times and peak areas of main peaks were all less than 5%. The fingerprints of Geqingpi and Sihuaqingpi were different. The fingerprints of the processed and the unprocessed were almost same, and the fingerprint differences among the different batches processed with the same technology from the same unprocessed GTP were not obvious.The influence of processing with vinegar on 50% ethanol extracts were not evident, and the technological pross was stable. It was found that the fingerprints of Geqingpi and Sihuaqingpi were different, but it should be study further.

The title compound, [Zn2(C5H6O4)2(C13H14N2)]n or [Zn2(glu)2(bpp)]n, is a novel zinc polymer based on mixed flexible glutarate (glu) and 1,3-di-4-pyridylpropane (bpp) ligands. The Zn(II) center has a distorted tetrahedral geometry and the central atom of the bpp ligand is located at a special site with a C2 axis passing through it. A layer is formed by Zn-glu bonding. Such layers are pillared by bpp ligands, forming a three-dimensional framework with large channels. The inverted interpenetration of two three-dimensional frameworks completes the molecular structure.

Ly-49A is an inhibitory receptor that binds H-2Dd and H-2Dk. The downregulation of Ly-49A is thought to mediate NK self tolerance in vivo. In this study, we analyzed the regulation of Ly-49A, D and G2 on NK cells in an in vivo rejection model. After injection with 1 x 10(8) B10.D2 spleen cells (SC) into B 10 mice, we found Ly-49A downregulated within 3 h on NK cells of B10 mice, whereas expressions of Ly-49D and G2 were augmented. To investigate effects of different expression patterns of Ly-49 receptors on NK cells, Ly-49A, D or G2-depleted B10 mice were inoculated with B10.D2 SC. NK cells from SC of Ly-49A-depleted and B10.D2 SC-injected B10 mice showed enhanced cytotoxicity to Dd-positive targets in vitro. Furthermore, reduced numbers of B10.D2 SC were observed in Ly-49A or G2-depleted B10 mice, whereas increased numbers of B10.D2 SC were observed in Ly-49D-depleted B10 mice after inoculation with B10.D2 SC in vivo. These findings indicated that the downregulation of Ly-49A and the augmentation of Ly-49D expression may mediate NK cells to recognize and kill Dd antigen efficiently. In conclusion, each Ly-49 isoform may play independent roles in the regulation of activation or inhibition on NK cells.

To Establish a rapid and objective quantitative fluorogenic real-time PCR assay for early detection of human respiratory syncytial virus (hRSV).Two pairs of primers and one TaqMan Fluorogenic probe that are specific for the recognition of the most conservative N gene of hRSV for virus detection with LighCycler PCR in 93 nasopharyngeal secretion specimens collected from infants and young children. The assay was compared with virus isolation, routine PCR, nested PCR, and enzyme-linked immunosorbent assay (ELISA).This TaqMan assay had a sensitivity of 1 x 10(2) cDNA copies/microl with a dynamic range between 1 x 10(2) and 1 x 10(7) cDNA copies/microl, which was the same as that of nested PCR, but 10 times more sensitive than routine PCR. The specificity of the assay was evaluated by comparing hRSV with polivirus type 1, coxsackie virus type 2, influenza A, influenza B and adenovirus type 7. A PCR product of the expected size (195 bp) was produced and fluorescence signal detected for hRSV, but not for any of the other viruses. The results in LightCycler and Rotor-Gene instrument were consistent. Forty-four specimens (43.9%) were hRSV-positive with this assay and 4 (4/93,4.3%) were hRSV-positive with ELISA, showing rather low correlation between the two methods. No visible relation was found between the concentration of hRSV RNA and severity of the disease.This assay is rapid, sensitive, specific and quantitative, and has the potential of wide application for early diagnosis of hRSV infection and evaluation of the therapeutic effect.