Identification of chemical constituents in ethyl acetate soluble extract of Sinopodophylli Fructus based on HPLC-MS~n / 中国中药杂志

A high performance liquid chromatography with a diode array detector combined with electrospray ionization ion trap time-of-flight multistage mass spectrometry(HPLC-DAD-ESI-IT-TOF-MS~n, HPLC-MS~n) method was established for qualitative analysis of the chemical components of ethyl acetate extract from Sinopodophylli Fructus. The analysis was performed on a Kromasil 100-5 C_(18)(4.6 mm×250 mm, 5 μm) column, with a mobile phase consisted of 0.1% formic acid(A) and acetonitrile(B) for gradient at a flow rate of 1.0 mL·min~(-1). Electrospray ionization ion trap time-of-flight multistage mass spectrometry was applied for qualitative analysis under positive and negative ion modes. With use of reference substance, characteristic fragmentation and their HR-MS data, 102 components were identified, including 67 flavonoids and 35 lignans. Among them, 45 compounds were reported in Sinopodophylli Fructus for the first time and 19 compounds were identified as new compounds. PharmMapper was used to predict the bioactivity of compounds that were first reported in Sinopodophylli Fructus, and 20 compounds of them were identified to have potential anticancer activity. The results showed that there were many isomers in the ethyl acetate extract of Folium Nelumbinis, and a total of 19 groups of isomers were found. Among them, C_(21)H_(20)O_8 had the highest number of isomers(18 compounds), all of which were α-peltatin or its isomers; C_(21)H_(20)O_7 ranked second, with 10 compounds, all of which were 8-prenylquercetin-3-methyl ether or its isomers. In conclusion, an HPLC-MS~n method was established for qualitative analysis of the ethyl acetate extract(with anti-breast cancer activity) from Sinopodophylli Fructus in this study, which will provide the evidence for clarifying pharmacological active ingredients of the ethyl acetate extract from Sinopodophylli Fructus against breast cancer.

Identification of chemical constituents in Sinopodophylli Fructus by HPLC-DAD-ESI-IT-TOF-MSn / 中国中药杂志

This experiment was performed to analyze and identify the chemical constituents of Sinopodophylli Fructus by HPLC-DAD-ESI-IT-TOF-MSn. The analysis was performed on an Agilent Zorbax SB-C₁₈ (4.6 mm×250 mm, 5 μm) column.The mobile phase consisted of 0.1% formic acid was used for gradient at a flow rate of 1.0 mL·min⁻¹. Electrospray ionization ion trap time-of-flight multistage mass spectrometry was applied for qualitative analysis under positive and negative ion modes. The results indicated that 54 compounds consisted of 18 lignans and 36 flavonoids from Xiaoyelian had been detected by their HRMS data, the information of literature and reference substance. Among them, 27 compounds were reported in Sinopodophylli Fructus for the first time. In conclusion, an HPLC-DAD-ESI-IT-TOF-MSn method was established to qualitative analysis of Xiaoyelian in this study, which will provide the evidence for evaluating the quality of Xiaoyelian herbs, clarifying the mechanism, and guiding the development of pharmacological active ingredients.

Screening potential mitochondria-targeting compounds from traditional Chinese medicines using a mitochondria-based centrifugal ultrafiltration/liquid chromatography/mass spectrometry method / 药物分析学报

Mitochondria regulate numerous crucial cell processes, including energy production, apoptotic cell death, oxidative stress, calcium homeostasis and lipid metabolism. Here, we applied an efficient mitochondria-based centrifugal ultrafiltration/liquid chromatography/mass spectrometry (LC/MS) method,also known as screening method for mitochondria-targeted bioactive constituents (SM-MBC). This method allowed searching natural mitochondria-targeting compounds from traditional Chinese medicines(TCMs), including Puerariae Radix (PR) and Chuanxiong Radix (CR). A total of 23 active compounds were successfully discovered from the two TCMs extracts. Among these 23 hit compounds, 17 were identified by LC/MS, 12 of which were novel mitochondria-targeting compounds. Among these, 6 active compounds were analyzed in vitro for pharmacological tests and found able to affect mitochondrial functions. We also investigated the effects of the hit compounds on HepG2 cell proliferation and on loss of cardiomyocyte viability induced by hypoxia/reoxygenation injury. The results obtained are useful for in-depth understanding of mechanisms underlying TCMs therapeutic effects at mitochondria level and for developing novel potential drugs using TCMs as lead compounds. Finally, we showed that SM-MBC was an efficient protocol for the rapid screening of mitochondria-targeting constituents from complex samples such as PR and CR extracts.

Flavonoids isolated from Sinopodophylli Fructus and their bioactivities against human breast cancer cells / 中国天然药物

Four prenylated flavonoids compounds 1-4, named sinopodophyllines A-D, and a flavonoid glycoside (compound 13), sinopodophylliside A, together with 19 known compounds (compounds 5-12 and 14-24) were isolated from the fruits of Sinopodophyllum hexandrum. The structures of new compounds were elucidated by extensive spectroscopic analysis, including HRESIMS, 1D and 2D NMR. Compounds 1-6, 9-11, and 14-17 were tested for their cytotoxicity against human breast-cancer T47D, MCF-7 and MDA-MB-231 cells in vitro, and compounds 2, 5, 6, 10 and 11 showed significant cytotoxicity (IC values < 10 μmol·L) against T47D cells.

Lignans and diterpenes isolated from Tirpitzia ovoidea and their biological activities / 中国天然药物

A new lignan, tirpitzin A (17) together with 20 known compounds (1-16, and 18-21) were isolated from the ethyl acetate soluble fraction of ethanol extract of the aerial parts of Tirpitzia ovoidea. The structure of new compound was elucidated by means of spectroscopic analysis. Of the known compounds, 7-21 were isolated from Linaceae family for the first time. The pharmacological activity of the crude extracts was tested using a mouse inflammation model induced by dimethyl benzene. The results demonstrated that the ethyl acetate soluble fraction had anti-inflammatory activity. Moreover, the cytotoxic and anti-inflammatory activities of some compounds were studied. The new compound 17 showed moderate cytotoxic effect against BxPC-3 cell line (IC = 19.51μmol·L) and Compound 10 showed significant cytotoxicity against HepG2, HL-60, U87 and BxPC-3 cell lines with IC values in the range 4.2-8.3μmol·L. Additionally, Compounds 2, 10, 11, and 13 exhibited potent inhibitory effects on LPS-induced nitric oxide production in RAW 264.7 macrophages at the concentration of 50μmol·L.

Comparision and analysis on the blood concentration of common Chinese medicine and Western medicine / 药学学报

To a certain extent, the drug effect is determined by its blood concentration. It is generally accepted that the blood concentrations of constituents of Chinese medicines are very low. There is no sufficient experimental bases and references on its degree and the possibility of taking effect. In this study, 69 papers were collected and analyzed by searching the database of Scifinder, Pubmed, CNKI. The minimum effective blood concentrations of 73 common Western medicines and the maximum blood concentrations of 211 in vivo constituents of 40 Chinese medicines (single herb or compound Chinese medicine) were summarized. It was found that the maximum blood concentrations of the most in vivo constituents of Chinese medicines were much less than the minimum effective blood concentrations of the Western medicines. Specifically, the minimum effective blood concentrations of 17 Western medicines (23% of total) and the maximum blood concentrations of the 143 in vivo constituents of Chinese medicines (68% of total) were less than 100 ng ·mL-1; the minimum effective blood concentrations of 31 Western medicines (42% of total) and the maximum blood concentrations of the 20 in vivo constituents of Chinese medicines (9% of total) were more than 1 000 ng·mL-1. In this paper, a systematic summary and comparison of the blood concentrations in traditional Chinese medicines and Western medicines were conducted, which could provide a new ideas and references for the study of the pharmacodynamical material basis and its mechanism in traditional Chinese medicine.

Analysis of aristolochic acids, aristololactams and their analogues using liquid chromatography tandem mass spectrometry / 中国天然药物

More than 80 aristolochic acids (AAs) and aristololactams (ALs) have been found in plants of the Aristolochiaceae family, but relatively few have been fully studied. The present study aimed at developing and validating a liquid chromatography tandem mass spectrometry (LC/MS(n)) for the analysis of these compounds. We characterized the fragmentation behaviors of 31 AAs, ALs, and their analogues via high performance liquid chromatography coupled with electrospray ionization mass spectrometry. We summarized their fragmentation rules and used these rules to identify the constituents contained in Aristolochia contorta, Ar. debilis, Ar. manshurensis, Ar. fangchi, Ar. cinnabarina, and Ar. mollissima. The AAs and ALs showed very different MS behaviors. In MS(1) of AAs, the characteristic pseudomolecular ions were [M + NH4](+), [M + H](+), and [M + H - H2O](+). However, only [M + H](+) was found in the MS(1) of ALs, which was simpler than that of AAs. Distinct MS(n)fragmentation patterns were found for AAs and ALs, showing the same skeleton among the different substituent groups. The distribution of the 31 constituents in the 6 species of Aristolochia genus was reported for the first time. 25 Analogues of AAs and ALs were detected in this genus. A hierarchical schemes and a calculating formula of the molecular formula of these nitrophenanthrene carboxylic acids and their lactams were proposed. In conclusion, this method could be applied to identification of similar unknown constituents in other plants.

Identification of chemical constituents in Scrophulariae Radix by HPLC-IT-TOF-MS / 中国中药杂志

This experiment was performed to establish a qualitative analysis on chemical constituents of Scrophulariae Radix by HPLC-ESI-IT-TOF-MS．The analysis was conducted on a C₁₈ column (Kromasil 100-5, 4.6 mm×250 mm, 5 μm) with 0.1% formic acid-acetonitrile as the mobile phase for gradient elution; ESI ion source was used for mass spectra, and data were collected innegative and positive modes. The results showed that 64 compounds from Scrophulariae Radix had been identified by analyzing negative ion mass data including element composition and by comparing with data from literature. Two new compounds (4-hydroxy-6-O-methylcatalpol and acetylangoroside C) and seventeen known compounds were detected from Scrophulariae Radix for the first time. Seventeen known compounds included twelve iridoid glycosides, three phenylpropanoid glycosides and two other kind compounds. This study will provide chemical basis for elucidation of the effective substance in the Scrophulariae Radix.

Quantitative determination of seven major absorbed volatile constituents in mice brain, liver and blood after intragastric administration of Asari Radix et Rhizoma suspension by headspace-solid phase microextraction-gas chromatography-mass spectrometry / 中国中药杂志

A headspace-solid phase microextraction-gas chromatography-mass spectrometry method（HS-SPME-GC-MS） was adopted for the quantitative study of 4-allylanisole, methyl eugenol, 2,3,5-trimethoxytoluene, 3,4,5-trimethoxytoluene, sarisan, 3,5-dimethoxytoluene and safrole in mice brain, liver tissues and blood after intragastric administration of Asari Radix et Rhizoma. A VF-WAXms (30 m×0.25 mm, 0.25 μm film thickness) capillary column and SPME fiber coated with 65 μm polydimethylsiloxane/divinylbenzene (PDMS/DVB) were used. The calibration curves of seven volatile constituents were established to validate the method's stability (RSD<15%), repeatability (RSD<9.5%), accuracy (RSD<22%), relative recovery (87.0%-108%) and extraction recovery (74.9%-102%). The validated HS-SPME-GC-MS assay was applied to determine the concentrations of seven constituents in liver, brain and blood. The detected contents were 0.22,0.14 μg•g⁻¹,0.25 mg•L⁻¹ (4-allylanisole), 1.1, 0.39 μg•g⁻¹, 0.69 mg•L⁻¹ (methyl eugenol), 0.45, 0.13 μg•g⁻¹, 0.54 mg•L⁻¹ (2,3,5-trimethoxytoluene), 0.51, 0.15 μg•g⁻¹, 0.45 mg•L⁻¹ (3,4,5-trimethoxytoluene), 0.48, 0.039 μg•g⁻¹, 0.69 mg•L ⁻¹ (sarisan), 2.2, 1.2 μg•g⁻¹, 1.5 mg•L⁻¹ (3,5-dimethoxytoluene) and 1.3, 0.67 μg•g⁻¹, 1.1 mg•L⁻¹ (safrole) respectively. This HS-SPME-GC-MS method is rapid and convenient, with a small sample size, and applicable for the analysis and determination of volatile constituents in traditional Chinese medicines, which provides scientific data for further studies on effective substances and toxic substances in Asari Radix et Rhizoma.

"Efficacy Theory" may help to explain characteristic advantages of traditional Chinese medicines / 中国中药杂志

This article proposes the "Efficacy Theory" hypothesis of the traditional Chinese medicines (TCMs): TCMs take effects and weaken toxicities through the additive effects of numerous effective forms (including their constituents or/and metabolites) on a same target, the synergistic effects based on the overall action of the additive effects on individual targets and their toxicities scattering effects. A TCM may include approximately 1000 constituents and each constituent may produce about 100 metabolites in vivo after oral administration. Numerous effective forms of incalculable constituents and their metabolites could work like a "army group" together. When the quantity of a specific target molecule is larger than the pharmaceutical molecules, the molecules of different kinds of effective forms could combine with the target molecules successively, to exert the additive effects. When the target molecules are mostly occupied ("target most spaces occupied"), this TCM begins to work. The additive effects maybe exert not only in concentration but also in a time order way, which gives a sustained efficacy of TCM. The additive effects and the toxicities scattering effects are resulted from the same effective groups and not identical toxic groups among different effective form molecules. The "toxicities scattering effect" can be used to explain the non-toxic TCMs, but not fit for toxic TCMs. The efficacy theory showed that the variety of constituents and metabolites may participate in the process of pharmacodynamic actions, including the additive effects, synergy effects and toxicities scattering effects, which may be useful for explaining and developing the characteristic advantage of the TCMs. The questions we need to study or confirm are as follows: What are the TCMs' pharmacodynamic substance basis and mechanism made up of Why are toxicities of most TCMs' smaller How is the TCMs' "Efficacy Theory" which reflects characteristic advantage of TCMs applied in the research and development of new drugs.

Determination of markers from characteristic HPLC chromatogram of phenols in three official origins of Ephedrae Herba and quantitative analysis of four phenols / 中国中药杂志

This study is to establish the characteristic HPLC chromatogram of phenols in Ephedrae Herba, from which to pick out the marker peaks, followed by the analysis of the regularity of their distribution and content in the herbaceous stems of Ephedra sinica, E. intermedia and E. equisetina. The HPLC-DAD method for the characteristic chromatogram as well as quantitative analysis was established. The separation was carried out on a YMC-Pack ODS-A column (4.6 mm x 250 mm, 5 µm), eluted with the mobile phases as 0.01% formic acid aqueous solution (A) and acetonitrile (B) in a linear gradient (0-10 min, 17% B; 10-25 min, 17%-19% B; 25- 33 min, 19%-48% B; 33-35 min, 48%-51% B; 35-44 min, 51% B). The flow rate was kept at 1.0 mL · min⁻¹. The column tem- perature was 40 °C, and the detection wavelength was set at 350 nm (0-16 min) and 330 nm (16-44 min). Forty-six batches of collected samples from three official origins of Ephedrae Herba were detected, whose liquid chromatograms proven to be helpful to the differentiation of different origins. With principal component analysis and the analysis of distribution of peak area, twelve key peaks from the chromatogram were discussed in details on their contributions to the characteristics and differences of three official origins of the herb: peak area of peak 10, 11, 12 were found out to be significantly higher in E. equisetina than in other two origins, whose sum (higher than 146 mAU in E. equisetina) was useful for the discrimination between E. equisetina and the other two origins; peak area of 1 and 4 were respectively higher in E. sinica and E. intermedia than in other official origins, indicating their important effect on the differen- tiation of corresponding origins; peak 8 and 9 were picked out as two characteristic common peaks in three official origins of the herb, whose peak area showed little difference among different origins; further, peak area of other key peaks in the chromatogram also showed some difference among three origins, which make contributions to the differentiation of origins as well. Then, four phenols as 2"-O-α- L-rhamnosyl-isovitexin (1), vitexin (2), pollenitin B (5) and herbacetin-7-O-β-D-glucoside (6) were quantitative analyzed with the above-mentioned method, with good linear relationship and accuracy (recoveries in a range of 97.8%-102.5%). The content of the four phenols were firstly reported in Ephedrae Herba from official origins, which were respectively trace-1.55 (1), trace-0.160 (2), trace-0.284 (5) and trace-0.620 (6) mg · g⁻¹ in all of the tested samples. In addition, the content of these phenols showed differences in three official origins, especially 1, whose content in E. sinica [(0.670 ± 0.88) mg ± g⁻¹] were significantly higher than in other two origins (lower than 0.16 mg ± g⁻¹ besides sample Ei-060630-2-2), and 6, whose average content in E. equisetina [(0.260 ± 0.039 2) mg · g⁻¹] were twice as high as in E. sinica [(0.120 ± 0.270) mg · g⁻¹] and E. intermedia [(0.136 ± 0.485) mg g⁻¹], indicating the important effects of the two constituents on the differentiation among three official origins of the herb. The method established for the characteristic HPLC chromatogram and quantitative analysis of phenols was simple and accurate, and the marker constituents selected may provide new guides for the discrimination of official origins as well as the improvement of quality criteria of EphedraeHerba.

Simultaneous determination of seven bioactive constituents in Smilacis Glabrae Rhizoma by high-performance liquid chromatography / 中国中药杂志

This study is to develop an HPLC method for the simultaneous determination of (-)-epicatechin, 5-O-caffeoylshikimic acid, neoisoastilbin, astilbin, neoisoastilbin, isoastilbin and engeletin in Smilacis Glabrae Rhizoma. Samples of Smilacis Glabrae Rhizoma, Heterosmilacis Chinensis Rhizoma and Heterosmilacis Yunnanensis Rhizoma were separated on an Agilent Zorbax SB-C18 column with gradient elution of acetonitrile-0.05% phosphoric acid at a flow rate of 1.0 mL · min(-1). The detected wavelength was set at 230 nm and the column temperature was maintained at 35 °C. The contents of (-)-epicatechin, 5-O-caffeoylshikimic acid, neoastilbin, astilbin, neoisoastilbin, isoastilbin and engeletin in 84 Smilacis Glabrae Rhizoma samples were in the range of trace-1.381, trace-9.913, trace-3.673, 0.6706-27.08, trace-1.181, trace-4.833 and trace-2.754 mg · g(-1), respectively. The established method was used for analysis of common adulterants. The results demonstrated that the contents of (-)-epicatechin in Heterosmilacis Yunnanensis Rhizoma and Heterosmilacis Chinensis Rhizoma were 0.01163-0.06007 mg · g(-1) and 0.01583-0.08585 mg · g(-1), respectively, while the other six constituents were not detected. The method is simple and accurate, and can be used for the quality control of Smilacis Glabrae Rhizoma. The constituents of Heterosmilacis Yunnanensis Rhizoma and Heterosmilacis Chinensis Rhizoma are significantly different from Smilacis Glabrae Rhizoma, and they are not suitable for using as Smilacis Glabrae Rhizoma.

Qualitative and quantitative analysis of dodecatetraenamides A, B in Asari Radix et Rhizoma / 中国中药杂志

To develop an analytic method for qualitative and quantitative analysis of dodecatetraenamides A, B in 42 samples of two official species of Asari Radix et Rhizoma( ARR) (37 samples of Asarum heterotropoides var. mandshuricum with different collection time and 5 samples of Asarum sieboldiivar. seoulense). The HPLC-IT-TOF-MS/MS methods for the qualitative and UPLC-PDA methods for the quantitative analysis were established. Dodecatetraenamides A, B were identified by comparing the retention time, UV absorption spectrum and quasi-molecular ion peak [ M + H]+ with the reference compound using HPLC-IT-TOF-MS/MS. The content of dodecatetraenamides A and B in ARR were determined by UPLC-PDA. The separation was successfully carried out on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 µm) column eluted with mobile phases of water (A) and acetonitrile (B) in gradient program (0-3 min, 35% B; 3-5 min, 35%-36% B; 5-6 min, 36%-43% B; 6 min-11 min 43% B; 11-12 min, 43%-100% B). The column temperature was 45 °C, and the detection wavelength was set at 254 nm. The flow rate was 0.6 mL · min(-1). On one level mass spectrometry scanning, the results showed that the quasi-molecular ion [M + H] + of both dodecatetraenamides A and B were m/z 248.20. The quantitative method with UPLC-PDA has made the baseline separation of the constituents, which were reported as mixtures in the most literatures. The average recovery of dodecatetraenamides A and B were 97.90% and 99.86%, the relative standard deviation were 0.4% and 1.1%, respectively. The contents of dodecatetraenamides A, B in all ARR samples was in the range of 0.11-3.89 and 0.24-6.65 mg · g(-1). Their contents reduced with the extension of storage time. Compared with the samples of 2013, the average content of the two constituents in the samples collected in year 2002-2003 reduced 34% and 36%, respectively (P < 0.05). Compared the A. sieboldii var. seoulense and A. heterotropoides var. mandshuricum with the same collective time and production area, the average contents of the two constituents in latter were up to (1.59 ± 0.75) mg · g(-1) and (2.90 ± 1.17) mg · g(-1), respectively, significantly higher than that in A. sieboldii var. seoulense (dodecatetraenamide A were (0.78 ± 0.52) mg · g(-1), dodecatetraenamide B were (1.69 ± 0.83) mg · g(-1)) (P < 0.05). The content of the dodecatetraenamide A in overground part was in the range of 0.11-0.33 mg · g(-1), dodecatetraenamide B was 0. 24-0.60 mg · g(-1), which were much lower than that of the underground part of ARR (dodecatetraenamide A was in the range of 0.73-3.89 mg · g(-1), dodecatetraenamide B was 2.11-6.24 mg · g(-1)). The method was certified to be simple, accurate and reliable and could be used for qualitative and quantitative analysis of dodecatetraenamide A and B in different species of ARR, also can be used for the comprehensive quality control of traditional Chinese medicine, Asari Radix et Rhizoma.

Research on inorganic elements of Asari Radix et Rhizoma from different localities and species / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the character of inorganic elements in Asari Radix et Rhizoma (Xixin).</p><p><b>METHOD</b>The contents of 53 inorganic elements in Xixin samples from different localities and species were determined by ICP-AES and ICP-MS. The statistical data were made using SAS.</p><p><b>RESULT</b>The result demonstrated that Xixin has the high contents of Fe, Cr, Li. It has been observed that the content of Cu and Pb of the samples are much higher than the standard level. The results of hierarchical cluster analysis revealed two groups which correspond with the species of the samples. No correlations between the contents of the inorganic elements and the localities of the samples were found. Some characteristic elements were displayed in some specific areas. The difference of the contents of the 53 inorganic elements between root and rhizome of Xixin was reported for the first time. The primary form of inorganic elements in Xixin has been studied for the first time. The result demonstrated that the extraction rate between different elements varied, with the average extraction rate of (22.25 +/- 24.96)%.</p><p><b>CONCLUSION</b>The inorganic elements analysis of Xixin can provide evidence of its identification, cultivation and application.</p>

Research on chemical fingerprint chromatograms of Sinopodophyllum hexandrum / 中国中药杂志

<p><b>OBJECTIVE</b>To study the HPLC fingerprint of Sinopodophylli Fructus, compare the major chemical differences in fruit of Sinopodophyllum hexandrum as well as the roots and rhizomas of S. hexandrum, and provide scientific evidence for clinical application and quality control.</p><p><b>METHOD</b>HPLC fingerprint method was used to analyze 12 fruits of S. hexandrum. A total of 20 common peaks were confirmed, and 12 peaks in the HPLC fingerprint were identified. Furthermore, similarity evaluation method and clustering analysis method were introduced to compare HPLC chromatograms, between fruits and underground parts.</p><p><b>RESULT</b>In this paper, quercetin-3-O-beta-D-glucopyranosid, kaempferol-3-O-beta-D-glucopyranoside, 8-prenylquercetin and 8-prenylquercetin 3-methyl ether were firstly reported in Sinopodophylli Fructus. Among the existing fingerprint research, a total of 11 peaks were identified for the first time, containing 9 flavonoids and 2 lignans. The chemical constituents differed significantly in different medicinal parts of S. hexandrum. Prenylflavonoid compounds were the main constituents of Sinopodophylli Fructus. However, podophyllotoxin, flavonoids with simple substituent groups and flavonoid glycosides were the major ingredients in the roots and rhizomas of S. hexandrum.</p><p><b>CONCLUSION</b>This method can be used for the quality control of Sinopodophylli Fructus and the roots and rhizomas of S. hexandrum. It has provided a reference for the pharmacodynamic differences of the two different parts.</p>

Simultaneous determination of five constituents in eight Qingyedan species derived from Swertia plants by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To develop an HPLC method for simultaneous determination of swertiamarin, gentiopicroside, sweroside, mangiferin, erythrocentaurin, and to detect these five constituents in eight Qingyedans derived from Swertia mileensis, S. cincta, S. patens, S. punicea, S. delavayi, S. nervosa, S. macrosperma and S. yunnanensis.</p><p><b>METHOD</b>The separation was carried out on a Thermo BDS Hypersil C18 (4. 6 mm x 250 mm, 5 microm) column eluted with mobile phase of water containing 0. 1% phosphoric acid and methanol (B) in gradient program (0-10 min, 18%-20% B; 10-30 min, 20%-35% B; 30-35 min, 35%-60% B). The column temperature was 32 degrees C , and the detection wavelength was set at 250, 260, 225 nm. The flow rate was 0. 7 mL . min-1 from 0 to 30 min, and be increased to 1. 0 mL . min-1 in 35 min.</p><p><b>RESULT</b>The five compounds were well separated. The linear response ranges of swertiamarin, gentiopicroside, sweroside, mangiferin, erythrocentaurin were 0. 072-13. 39, 0. 1204. 518, 0. 060-5. 050, 0. 025-1. 518, and 0. 031-0. 210 microg, respectively. The mean recoveries of five compounds were 97.03% -102. 7% (RSD 1. 8% -6.2% ). There are swertiamarin, gentiopicroside and sweroside in most samples, and mangiferin in half samples. But erythrocentaurin was only detected in a few samples. The contents of five compounds were different in different samples. The contents of swertiamarin in S. mileensis, S. patens, S. yunnanensis and S. delavayi are up to 34. 47-118.05 mg . g-1, the contents of gentiopicroside are up to 25. 91 mg . g-1 in S. cincta. In S. puncea all contents of swertiamarin, gentiopicroside, sweroside and mangiferin are higher, especially the content of sweroside. There are Xiao-Qingyedans and Da-Qingyedans called in markets, and they can be identified by the contents of swertiamarin, gentiopicroside and sweroside. S. punicea can be identified by the content of sweroside, and the ratio gentiopicroside/total content can be used for identification of S. cincta from other seven Qingyedan species.</p><p><b>CONCLUSION</b>The method was certified to be accurate and reliable and can be used for identification and quality evaluation of traditional Chinese medicine Qingyedan derived from Swertia species.</p>

Metabolite profiling of propyl gallate in rat plasma and urine by HPLC-DAD-ESI-IT-TOF-MS(n) technique / 中国中药杂志

To make a clear understanding of the in vivo metabolism of propyl gallate in rats and determine the original compounds of the metabolites of Paeoniae Radix Rubra (PRR), the urine and plasma of the rats administrated with propyl gallate were collected, and then the HPLC-DAD-ESI-IT-TOF-MS(n) technique was applied to screen and identify the metabolites of propyl gallate from these bio-samples. By comparing the collected LC-MS data, the origin of the metabolites of PRR were confirmed. Finally, 33 metabolites of propyl gallate were identified from urine sample, and 20 metabolites of propyl gallate were identified from the plasma sample. In total, 37 metabolites of propyl gallate were identified, and 17 of them were identical to the metabolites of PRR. They are mainly the phase II metabolites of propyl gallate, 3-hydroxypropyl 3,4,5-trihydroxybenzoate, gallic acid and pyrogallol. Phase I reactions (decarboxylation, hydroxylation) and phase II reactions (sulfation, glucuronidation and methylation) were observed as the main metabolic pathways of propyl gallate. In this research, the in vivo metabolism of proply gallate was reported for the first time and 37 metabolites were identified, among which 35 were new metabolites of propyl gallate, and 20 were new compounds. The results demonstrated that 17 metabolites of PRR can be formed from propyl gallate. This will enhance our understanding of the metabolism and Effective forms (the truly active structures) of propyl gallate and PRR.

Study on HPLC-fingerprint-based identification of dao-di herb and non dao-di herb of scutellariae radix / 中国中药杂志

<p><b>OBJECTIVE</b>To compare the discrepancies between chemical constituents in Dao-di herb and non Dao-di herb of Huangqin (the root of Scutellaria baicalensis), study the impact of habitat and growth pattern (including cultivated and wild Huangqin) on chemical substances of Huangqin, and then provide evidence for the identification of Dao-di herb and quality evaluation of Huangqin.</p><p><b>METHOD</b>The chemical constituents in Huangqin collected from different habitats and under different growth patterns, were analyzed using HPLC fingerprint. The fingerprints obtained were then evaluated by hierarchical clustering analysis, principal component analysis and components peak area pattern.</p><p><b>RESULT</b>The fingerprints' chemical profiles of Dao-di herb and non Dao-di Huangqin had significant disparity. The fingerprints of modem Dao-di herb Huangqin samples originated from Chengde (Hebei Province) were significantly different from those from other habitats, though the fingerprints of the non Dao-di Huangqin collected from Chifeng (Inner Mongolia) and Chengde had high similarity to each other. The chemical characteristics of Huangqin samples collected from the habitats recorded in ancient herbals, such as Qingyang (Gansu Province), Yan'an (Shaanxi Province), Linyi (Shangdong Province), Changzhi and Jinzhong (Shanxi Province) were similar. The fingerprints of modern non Dao-di samples collected from Dingxi and Longnan (Gansu Province) and Shangluo (Shaanxi Province) had high similarity. In addition, the content of acteoside in wild Huangqin was higher than that in cultivated Huangqin.</p><p><b>CONCLUSION</b>Dao-di herb and non Dao-di herb of Huangqin could be distinguished using the developed HPLC fingerprints. The results obtained may provide evidence for the quality control and pharmcodynamical research of Dao-di herb and non Dao-di Huangqin.</p>

Study on fingerprints of chemical constituents of cinnamomi ramulus and cinnamomi cortex / 中国中药杂志

<p><b>OBJECTIVE</b>To compare the difference of chemical constituents of Cinnamomi Ramulus and Cinnamomi Cortex,and study the characteristics of their chemical constituents, in order to provide basis for clinical application and quality control.</p><p><b>METHOD</b>HPLC fingerprint method was used to analyze chemical constituents of Cinnamomi Ramulus and Cinnamomi Cortex compare the difference between.</p><p><b>RESULT</b>Cinnamomi Ramulus and Cinnamomi Cortex had similar HPLC fingerprint profiles, but with difference in peak areas of their chemical constituents. Specifically, the average peak areas of cinnamyl alcohol, cinnamic acid and 2-methoxy cinnamic acid were higher than that of Cinnamomi Cortex, whereas the average peak areas of 2-hydroxyl cinnamaldehyde, coumarin, cinnamaldehyde and 2-methoxy cinnamaldehyde in Cinnamomi Cortex were higher than that of Cinnamomi Ramulus.</p><p><b>CONCLUSION</b>This method can be used for evaluating quality of Cinnamomi Ramulus and Cinnamomi Cortex. The ratio between the peak areas of cinnamaldehyd and cinnamic acid can be used to discriminate most Cinnamomi Ramulus samples (<23) and Cinnamomi Cortex samples (>23). Cinnamomi Ramulus and Cinnamomi Cortex have the similar types of chemical constituents but with difference in content. This study provides reference for the pharmacodynamic difference of Cinnamomi Ramulus and Cinnamomi Cortex.</p>

Comparative study on anti-inflammatory and antipyretic effects between Dao-di herb and non Dao-di herb of Huangqin / 中国中药杂志

<p><b>OBJECTIVE</b>To do some comparative study on anti-inflammatory and antipyretic effects between the Dao-di herb and non Dao-di herb of Huangqin (the roots of Scutellaria baicalensis) and provide thinking and evidence for study on geoherbalism and clinical usage of Huangqin.</p><p><b>METHOD</b>The anti-inflammatory action was assessed by auricular swelling induced by dimethylbenzene in mice and the antipyretic action was monitored by dried yeast-induced mice fever.</p><p><b>RESULT</b>All samples of both Dao-di and non Dao-di herbs of Huangqin showed antipyretic effect. The Dao-di Huangqin samples showed antipyretic effect between 61% to 53% , whereas the non Dao-di Huangqin samples between 53% to 43%. Six Dao-di Huangqin samples showed better antipyretic effect than four non Dao-di Huangqin samples. All samples of both Dao-di and non Dao-di Huangqin showed anti-inflammatory effect. Dao-di Huangqin showed anti-inflammatory effect between 73% to 54%, whereas non dao-di Huangqin between 53% to 34%. Six Dao-di Huangqin showed better anti-inflammatory effect than four non Dao-di Huangqin. In totality, results from analysis of geoherbalism showed that geoherbal production areas of Huangqin had better effect than that of the non geoherbal production areas in anti-inflammatory and antipyretic effects.</p><p><b>CONCLUSION</b>Both the Dao-di and non Dao-di Huangqin have effects of anti-inflammatory and antipyretic to a certain extent, but the efficacy of the Dao-di Huangqin surpass the non Dao-di Huangqin.</p>