Identification and differentiation of major components in three different "Sheng-ma" crude drug species by UPLC/Q-TOF-MS

Cimicifugae Rhizoma (Sheng ma) is a Ranunculaceae herb belonging to a composite family and well known in China. has been widely used in traditional Chinese medicine. Thecontains three varieties ((Turcz.),L. andKom.) which have been used clinically as "Sheng-ma". However, the chemical constituents of three components of "Sheng-ma" have never been documented. In this study, a rapid method for the analysis of the main components of "Sheng-ma" was developed using ultra-high performance liquid chromatography with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS). The present study reveals the major common and distinct chemical constituents of,andand also reports principal component and statistical analyses of these results. The components were identified by comparing the retention time, accurate mass, mass spectrometric fragmentation characteristic ions and matching empirical molecular formula with that of the published compounds. A total of 32 common components and 8 markers for different "Sheng-ma" components were identified. These findings provide an important basis for the further study and clinical utilities of the three "Sheng-ma" varieties.

Effect of Zingiber offiicinale and Aconitum cainichaeli before and after compatibility on contents of four gingerols / 中国中药杂志

<p><b>OBJECTIVE</b>To discuss the synergistic mechanism of compatible use of two medicinal herbs, Zingiber offiicinale and Aconitum cainichaeli, by determining single decoction of Z. offiicinale and four gingerols (6-gingerol, 8-gingerol, 6-shogaol, 10-gingerol) contained in compound decoction of Z. offiicinale and A. cainichaeli of different compatibility ratio using HPLC.</p><p><b>METHOD</b>Kromasil-C18 column (4.6 mm x 250 mm, 5 microm) was adopted. The mobile phase was acetonitrile (B) and 0.1% aqueous acetic acid (A) for gradient elution (0-30 min, 40%-90% B; 30-35 min, 90%-40% B). The flow rate was 1.0 mL x min(-1). The detection wavelength was set at 275 nm. The column temperature was 30 degrees C.</p><p><b>RESULT</b>The four gingerols were in baseline separation, with a good linearity (r > 0.999), an average recovery of 100.9% -103.5% and RSD < 3.0%. Compared with the single decoction of Z. offiicinale, the content of gingerols in the compound decoction of Z. offiicinale and A. cainichaeli was on the rise and in direct proportion with the increase in the volume of A. cainichaeli.</p><p><b>CONCLUSION</b>The synergistic mechanism of the compatibility of Z. offiicinale and A. cainichaeli can be proved with the increased release of gingerols from Z. offiicinale.</p>

Simultaneous determination of four components in sini tang by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To develop an HPLC method for determining four components in Sini Tan, benzoylmesaconine, liquiritin, glycyrrhizic acid and 6-gingerol.</p><p><b>METHOD</b>The Hypersil BDS column was adopted with gradient elution program at a flow rate of 1.0 mL x min(-1) and the detection wavelength of 235 nm.</p><p><b>RESULT</b>Benzoylmesaconine, liquiritin, glycyrrhizic acid and 6-gingerol showed good separation, with the linear range of 0.006-0.12, 0.021-0.42, 0.012-0.24 and 0.018-0.36 g x L(-1), respectively. Their average recoveries were 99.3%, 96.9%, 100% and 100%, respectively; and RSD of the above four components were 1.5%, 0.6%, 1.3% and 2.1%, respectively.</p><p><b>CONCLUSION</b>The method is proved to be so easy and accurate and practical that it can be used to determine the four components in Sini Tang.</p>

Variation regularity of chemical constituents contained in Euphorbia ebracteolata before and after roasting with vinegar / 中国中药杂志

<p><b>OBJECTIVE</b>To study the variation regularity of chemical constituents contained in Euphorbia ebracteolata after vinegar processing.</p><p><b>METHOD</b>The colorimetric method was adopted for determining the variation of total lactone content in toxic constituents contained in E. ebracteolata decoction, with Kedde as the coloring reagent. The HPLC method was used for detecting the content variation of jolkinolide B and jolkinolide C, both were active constituents contained in E. ebracteolata decoction, before and after roasting with vinegar, in which Kromasil-ODS column (4.6 mm x 250 mm, 5 microm) was adopted, with the detection wavelength of 290 nm, column temperature at 25 degrees C , gradient elution with acetonitrile and water and the flow rate of 1.0 mL x min(-1).</p><p><b>RESULT</b>After roasting with vinegar, the total lactone content in E. ebracteolata was reduced from 0.60 to 0.45 mg x g(-1) , with active constituents, jolkinolide B and jolkinolide C, increased to varying degrees. The established chromatographic fingerprint contained risk information and could reflect the overall variation regularity of chemical constituents after roasting with vinegar.</p><p><b>CONCLUSION</b>The chemical constituents of E. ebracteolata show significantly changes, especially a reduced toxicity, after roasting with vinegar. The increase in its efficacy may be related to the variation of these constituents.</p>

Comparison on in vivo pharmacokinetics of brucine, total alkaloids of Strychni Semen and Strychni Semen pulveratum in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study different in vivo pharmacokinetic regularity of brucine, total alkaloids of scorched sand-prepared Strychni Semen products and Strychni Semen pulveratum in rats, and probe into mutual impact between single component and compound.</p><p><b>METHOD</b>Rats in each group were orally administered with brucine, total alkaloids of scorched sand-prepared Strychni Semen products and Strychni Semen pulveratum suspension. The in vivo plasma concentrations of brucine in rats were determined by HPLC. A compartment model was made for the blood drug concentration-time curve using 3P97 software package and the pharmacokinetic parameters of each group were calculated and compared.</p><p><b>RESULT</b>The in vivo metabolic process of brucine in rats complied with the two-compartment model, weight W = 1/C2. The results of variance analysis showed that among three existing forms of brucine with same dosage, the brucine solution group and the total alkaloids group of scorched sand-prepared Strychni Semen products showed significant differences in C(max), MRT (P < 0.05); and the brucine solution group and the Strychni Semen pulveratum suspension group showed significant differences in C(max), AUC(0-t), and AUC(0-infinity), in which the latter displayed minimum C(max), AUC(0-t) and AUC(0-infinity).</p><p><b>CONCLUSION</b>The total alkaloids group of scorched sand-prepared Strychni Semen products showed a relatively longer retention time of effective components of brucine in plasma, while the Strychni Semen pulveratum suspension group showed a lower bioavailability.</p>

Determination and comparison of plasma protein binding rate of alkaloids from seed of Strychnou nux-vomica / 中国中药杂志

<p><b>OBJECTIVE</b>To determine the plasma protein binding rates of brucine and strychnine in total alkaloids from the seed of Strychnou nux-vomica, and make comparison with the single components at the same concentration.</p><p><b>METHOD</b>Ultrafiltration was employed to determine the rat the plasma protein binding rate of the alkaloids from the seed of S. nux-vomica. The plasma concentrations were measured by RP-HPLC.</p><p><b>RESULT</b>The protein binding rates of brucine were (65.60 3.01)%, (68.20 +/- 7.80)%, (59.58 +/- 3.78)% when the plasma concentrations was 0.520, 1.300, 2.600 mg x L(-1), respectively. The protein binding rates of strychnine was (66.17 +/- 6.36)%, (67.10 +/- 2.52)%, (57.21 +/- 0.79)% when the plasma concentrations were 0.936, 2.340, 4.680 mg x L(-1) respectively. As to the total alkaloids from the seed of S. nux-vomica, The protein binding rate of brucine was (62.19 +/- 2.45)%, (69.55 +/- 5.84)%, (61.76 +/- 3.68)% when the plasma concentrations were 0.519, 1.288, 2.607 mg x L(-1), respectively. And the protein binding rates of strychnine were (54.79 +/- 3.55)%, (57.13 +/- 4.49)%, (59.31 +/- 3.65)% when the plasma concentrations were 0.940, 2.338, 4.674 mg x L(-1), respectively.</p><p><b>CONCLUSION</b>Brucine and strychnine have medium capacity in binding to plasma protein. In comparison with the single component of the same concentration, the protein binding rate of brucine in total alkaloids shows little difference, while there seems to be an obvious decrease for strychnine.</p>

Effects of different excipients on properties of Tongsaimai mixture and pellet molding / 中国中药杂志

<p><b>OBJECTIVE</b>To study preliminarily on the relationship between properties of the mixture composed of Tongsaimai extract and different excipients and pellet molding.</p><p><b>METHOD</b>The multivariate regression analysis was used to investigate the correlation of different mixture and pellet molding by measuring the cohesion, liquid-plastic limit of mixture, and the powder properties of pellets. The weighted coefficients of the powder properties were determined by analytic hierarchy process combined with criteria importance through intercriteria correlation.</p><p><b>RESULT</b>The results showed that liquid-plastic limit seemed to be a major factor, which had positive correlation with pellet molding, while cohesion had negative correlation with pellet molding in the measured range.</p><p><b>CONCLUSION</b>The physical properties of the mixture has marked influence on pellet molding.</p>

Speed up to formulate "National processing procedures of prepared slices of Chinese crude drugs", unified the national standards of prepared slices / 中国中药杂志

The prepared slices of Chinese crude drugs are growing important in recent years, and faced with new developments and opportunities. The author analyzed the importance of formulate national processing procedures of prepared slices of Chinese crude drugs combined actual work, proposed the overall objectives and tasks for the formulation, and emphasized to need to correctly deal with several important factors during the process of formulate "National processing procedures of prepared slices of Chinese crude drugs", unified the national standards of prepared slices, solved the real problems that the prepared slices of Chinese crude drugs industry faced.

Study on apoptosis of SMMC-7721 hepatocarcinoma cells induced by Oxytropis falcata and its preliminary mechanism / 中国中药杂志

<p><b>OBJECTIVE</b>To examine apoptosis of SMMC-7721 hepatocarcinoma cells induced by total flavonoids of Oxytropis falcata (TFOF) and its preliminary mechanism.</p><p><b>METHOD</b>SMMC-7721 cells were treated for 24 h with TFOF in different concentrations. Inhibition on proliferation of SMMC-7721 cells was assessed by MTT assay. The morphology of treated SMMC-7721 cells was observed by optical microscope. Effect of TFOF on the nuclear morphology of cells was analyzed using Hoechst 33258 staining by fluorescence microscope. Annexin V-FITC/PI staining and flow cytometric measurement were used for investigating the effect of TFOF on induction of apoptosis in SMMC-7721 cells and cell cycle analysis.</p><p><b>RESULT</b>The results of MTT assay showed that TFOF could induce cytotoxicity in SMMC-7721 cells in a dose-dependent manner. Hoechst 33258 staining analysis indicated that TFOF caused typical characteristics of apoptotic programmed cell death, such as cell shrinkage, apoptotic body formation etc. Flow cytometric analysis demonstrated that TFOF caused a dose-dependent apoptosis of SMMC-7721 cells and arrested cell cycle in G1 phase.</p><p><b>CONCLUSION</b>It suggested that TFOF inhibit proliferation of SMMC-7721 cells by inducing apoptosis of the cells and arresting cell cycle in G1 phase.</p>

Pharmacokinetics of brucine in rats after intravenous administration of liposomes containing total alkaloids from seed of Strychnos nux-vomica / 中国中药杂志

<p><b>OBJECTIVE</b>To compare the pharmacokinetic characteristics of brucine following intravenous administration of liposomes, containing total alkaloids from seed of Strychnos nux-vomica, to rats with different phospholipids composition.</p><p><b>METHOD</b>Liposomes containing the total alkaloids were prepared by the method of ammonium sulfate transmembrane gradients and stealth liposome technique. The contents of total alkaloids and brucine in liposomes were determined and compared after free drug being removed. After intravenous administration of total alkaloids solution or liposomes with different composition, plasma samples were drawn at predetermined time points and the concentrations of brucine were determined by a validated method of HPLC. Pharmacokinetic analysis was performed by 3P97 program.</p><p><b>RESULT</b>The ratios of brucine to total alkaloids in liposomes hardly varied with phospholipids composition. Compared with SPC liposome, AUC of brucine was increased 13.3-fold and apparent volume of distribution was decreased to only 3.6% following intravenous administration of HSPC liposome. In addition, besides that AUC of brucine was slightly increased, most pharmacokinetic parameters were not significantly changed after administration of the novel liposome compared with those of SPC liposome.</p><p><b>CONCLUSION</b>Phospholipids composition has a significant influence on the pharmacokinetics of brucine after intravenous administration of liposomes containing total alkaloids from seed of S. nux-vomica.</p>

Effects of glycyrrhiza extract on pharmacokinetics property of daphnetin in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To research the influence of glycyrrhiza extract on the pharmacokinetics characteristic parameters of daphnetin, which was aimed to explore the rationality of concert application of drugs.</p><p><b>METHOD</b>The rats received intragastric administration of daphnetin and glycyrrhiza extract containing the same daphnetin respectively. The blood concentration of daphnetin was assayed by LC-MS. The data was processed by program DAS2.1.1.</p><p><b>RESULT</b>Glycyrrhiza extract can reduce the t(1/2), tmax and Ke of daphnetin, while increased the Ka and AUC(0-infinity).</p><p><b>CONCLUSION</b>Glycyrrhiza extract promoted the oral absorption of daphnetin, slowed down the elimination and increased the biological availability.</p>

Simultaneous determination of six flavonoids in Hyperici Japonici Herba by HPCE-DAD / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a high performance capillary electrophoresis method with diode array detection (HPCE-DAD) for simultaneous determination of rutin, isoquercitrin, hyperoside, quercitrin, kaempferol and quercetin in Hyperici Japonici Herba.</p><p><b>METHOD</b>Based on the mode of capillary zone electrophoresis, 40 mmol x L(-1) borax was used as buffer solution (pH 8.62), uncoated fused silica capillary (56 cm x 64.5 cm x 75 microm) was used, separation voltage was 25 kV, detection wavelength was at 206 nm, column temperature was maintained at 25 degrees C, and sample was injected at 50 mbar, 8 s.</p><p><b>RESULT</b>Six flavonoids showed good linearity (r > 0.9953) in the range of the tested concentration, the average recoveries of the method were between 98.8%-102.9%.</p><p><b>CONCLUSION</b>The method is simple, accurate and reproducible, and can be used for quality control of Hyperici Japonici Herba.</p>

Pharmaceutical properties of novel liposomes containing total alkaloids from seed of Strychnos nux-vomica / 中国中药杂志

<p><b>OBJECTIVE</b>To prepare the novel liposomes composed of hydrogenated soybean phosphatidylcholine (HSPC) and soybean phosphatidylcholine (SPC) containing the total alkaloids from seed of Strychnos nux-vomica, and to compare the pharmaceutical properties of the novel liposomes with the corresponding HSPC or SPC liposomes.</p><p><b>METHOD</b>The total alkaloids were extracted from seeds of S. nux-vomica. and further purified. Novel liposomes containing the total alkaloids were prepared by ammonium sulfate transmembrane gradients and stealth liposome technique. Pharmaceutical properties such as encapsulation efficiency (EE), size, zeta potential and drug release profile of novel liposomes and corresponding HSPC or SPC liposomes were compared intensively.</p><p><b>RESULT</b>For novel liposomes, HSPC-SPC (1:3) was the best ratio which has the highest EE. At the drug/lipid weight ratio of 1:6, the EE of novel, SPC and HSPC liposomes were (73.6 +/- 2.9)%, (62.9 +/- 1.8)% and (54.7 +/- 1.0)% (n = 3), respectively. Compared with the corresponding SPC or HSPC liposomes, the size of novel liposomes was obviously decreased but the zeta potential was not different. The results of drug release showed that the novel liposomes were more stable than the SPC liposomes in the presence of rat plasma</p><p><b>CONCLUSION</b>Taken together, high encapsulation efficiency improved stability in blood, and relative low price of phospholipids of the novel liposomes, indicate that the novel liposomes may act as promising carriers for anti-tumor traditional Chinese medicine.</p>

Study on density quality control of crude pyritum / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the quality assessment method of crude pyritum.</p><p><b>METHOD</b>The correlations of methods of modern and traditional quality appraisement with density for crude pyritum were investigated.</p><p><b>RESULT</b>Appraisement of crude pyritum using density as an evaluation index was in accordance with its traditional quality appraisement method. The density had extremely significant positive correlation with FeS2 content contained in crude pyritum (r = 0.990, P<0.001) and became smaller with the decreasing of FeS2 content. Meantime the density also had extremely significant negative correlation with total ferrum content contained in crude pyritum (r = -0.971, P<0.001) and became smaller with the increasing of total ferrum content.</p><p><b>CONCLUSION</b>It's feasible to take density as evaluation index for quality control of crude pyritum, and the testing method established is simple and convenient.</p>

HPLC determination of chikusetsusaponin IVa in Rhizoma Panacis Majoris from different producing areas / 中国中药杂志

To determinate the chikusetsusaponin IVa in Rhizoma Panacis Majoris from different producing areas. The HPLC separation was performed on a Inertsil ODS-sp column (4.6 mm x 150 mm, 5 microm). A mixture of acetonitrile and 0.2% phosphoric acid solution (35:65) as the mobile phase the column. Temperature was set in 30 degrees C. The flow rate was 1.0 mL x min(-1), and the wave length of the detector is 203 nm. The content of the chikusetsusaponin IVa in Rhizoma Panacis Majoris from Meixian, Shaanxi is the highest and the lowest is from Enshi, Hubei. There have most differerence among the content of the chikusetsusaponin IVa in Rhizoma Panacis Majoris from different producing areas.

Discuss on crispness to quality control of calcined drugs / 中国中药杂志

At present, the production and quality control of calcined drugs were lack of corresponding quantitative indicators, and only relied on traditional empirical control, thus it was susceptible to subjective factors, and the qualities of calcined drugs were affected. This paper analyzes the current status of the quality control of calcined drugs, the traditional quality requirements, modern quality controls and evaluation methods of calcined drugs were systematically summarized. Take the example for pyritum, a quality control method which reflects crispness of calcined drugs with density variety was investigated, and the feasibility of its application was discussed, therefore a new quantitative indicator for quality control of calcined drugs was supplied.

Research on X-ray diffraction fingerprints of crude and processed pyritum / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the X-ray diffraction fingerprints of crude and processed pyritum.</p><p><b>METHOD</b>X-ray diffraction fingerprint was used to analyze the crude and processed pyritum of different habitat and batch.</p><p><b>RESULT</b>The crude pyritum has 19 common peaks, while the processed ones has 11.</p><p><b>CONCLUSION</b>The X-ray diffraction can express characteristic of the crude and processed pyritum, which is specificity, accuracy and reliability and can be used for the quality control of crude and processed pyritum.</p>

MEKC-DAD fingerprint of Radix et Rhizoma Rhei / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the analytical method for the fingerprint of Radix et Rhizoma Rhei by MEKC-DAD and compare the fingerprints of Radix et Rhizoma Rhei and its processed products.</p><p><b>METHOD</b>Based on the mode of micellar electrokinetic chromatography, 25 mmol x L(-1) borax -25 mmol x L(-1) SDS-10% acetonitrile was selected for the running buffer (pH 9.2). The separation voltage was 12 kV and the detection wavelength was set at 254 nm. Rhein was used as a reference standard, the chromatographic fingerprint was determined via the data analyzed by fuzzy cluster and fingerprint similarity evaluation software to compare the similarity of samples.</p><p><b>RESULT</b>MEKC-DAD fingerprints with 11 common peaks of 10 batches of Radix et Rhizoma Rhei from the place of the genuine were established preliminarily. It was discovered that a small number of samples differed from others. Regarding to the fingerprints of Radix et Rhizoma Rhei and its processed products, there were obvious differences in the relative areas of common peaks.</p><p><b>CONCLUSION</b>The method is reliable, accurate and can be used for quality control of Radix et Rhizoma Rhei.</p>

Determination of densities of mineral drugs and their calcined products by sealing wax densimetry / 中国中药杂志

<p><b>OBJECTIVE</b>To determine densities of mineral drugs and their calcined products by using sealing wax densimetry, and to establish the testing method of crispness for calcined traditional Chinese medicine.</p><p><b>METHOD</b>Taking density as detection indicator, the accuracies of mineral drugs and their calcined products were investigated by sealing wax densimetry compared with direct densimetry, and the method of density determination was optimized.</p><p><b>RESULT</b>The density values measured by two methods investigated above were significantly different (P < 0.01), the density values of Pyritum, Haematitum, calcined Pyritum and calcined Haematitum measured by direct densimetry were increased 1.66%, 9.43%, 96.72% and 9.02%, respectively compared with that measured by sealing wax densimetry.</p><p><b>CONCLUSION</b>Sealing wax densimetry is suitable for the density determination of mineral drugs and their calcined products with its high accuracy, whereas direct densimetry'is not suitable for the density determination of mineral drugs and their calcined products.</p>

Research on variation regularity of five main flavonoids contents in Epimedium and processed Epimedium / 中国中药杂志

<p><b>OBJECTIVE</b>To research on the variation regularity of the five main flavonoids contents in Epimedium and processed Epimedium.</p><p><b>METHOD</b>The contents of Epimedin A, Epimedin B, Epimedin C, Icariin and Baohuoside I in Epimedium and processed Epimedium were determined, respectively.</p><p><b>RESULT</b>There were varieties in the contents of Epimedin A, Epimedin B, Epimedin C, Icariin and Baohuoside in Epimedium and processed Epimedium. There were different variety tendencies in different species and different batches.</p><p><b>CONCLUSION</b>The variety tendencies of the contents of five main flavonoids depend on the processing technology and the flavonoids proportion in Epimedium and processed Epimedium.</p>