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Extracting Technology of Total Glycosides from Barks of Ilex rotunda

Chun WANG ; Zhimao CHAO ; Xiaoyi WU ; Wen SUN

Chinese Journal of Information on Traditional Chinese Medicine.2013;(12):61-63. doi:10.3969/j.issn.1005-5304.2013.12.023

Objective To investigate an optimum extracting technology for total glycosides from dried barks of Ilex rotunda Thunb. Methods The yields of pedunculoside and syringin by high performance liquid chromatography (HPLC) determination were taken as the indexes. Some parameters of the extraction technology were evaluated with an L9(3)4 orthogonal design. The optimum extraction parameters were used to extract total glycosides in laboratory. The feasibility was checked by determining the chemical constituents by means of HPLC method. Results The optimum extracting conditions were 12 times volumes of 50% ethanol as the solvent and refluxing 90 min each time for 3 times. An extract yield of total glycosides was 19.5% from barks of Ilex rotunda. Pedunculoside (292 mg/g), syringin (59.5 mg/g), sinapaldehyde glucoside, syringaresinol 4'-O-β-D-glucopyranoside, and syringaresinol 4',4'-di-O-β-D-glucopyranoside were detected in the extract. Conclusion Some glycosides were found in the extract with the optimum extraction technology in laboratory. The extraction technology is practicable and valid.

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Simultaneous Determination of Low-molecular-weight Carbonyl Compounds in Oil-containing Herbs by HPLC Coupled with Pro-column Derivatization

Junbo XING ; Hong CAO ; Jiong ZHANG ; Tingting SHAN ; Caihong SHUI ; Yumin CHEN ; Dan HU

Chinese Journal of Information on Traditional Chinese Medicine.2013;(12):46-48. doi:10.3969/j.issn.1005-5304.2013.12.017

Objective To establish an HPLC method for determination of low-molecular-weight carbonyl compounds in the oil-containing herbs. Methods After carbonyl compounds in the samples were extracted with water, the solution reacted with 2, 4-dinitrophenylhydrazine (DNPH) in an acidic medium to form 2, 4-dinitrophenylhydrazone derivatives, which were separated on Kromasil KR100-5 C18 column (250 mm× 4.6 mm, 5 μm). The mobile phase A was water-acetonitrile-tetrahydrofuran-isopropanol (59∶30∶10∶1), mobile phase B was water-acetonitrile (35∶65), gradient elution. The flow rate was 0.8 mL/min, detection wavelength was 365 nm, and column temperature was 30 ℃. Results Good linearities were obtained in corresponding concentration ranges, with correlation coefficient over 0.999. The limits of detection of the eight DNPH derivatives were 0.002-0.008 μg/mL, and the average recoveries were 88.49%-93.65%. Conclusion The method is simple, accurate and reliable, with good reproducibility.

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Study on Quality Standard of Tongluo Tangtai Granules

Qing HU ; Ying SONG ; Caiyan LIU ; Jing TAN

Chinese Journal of Information on Traditional Chinese Medicine.2013;(12):39-41. doi:10.3969/j.issn.1005-5304.2013.12.015

Objective To establish the quality standard for Tongluo Tangtai Granules. Methods TLC was applied to the qualitative identification of Rhizoma Coptidis, Semen Sinapis Albae and Radix Scrophulariae, and HPLC was applied to the quantitative determination of berberine in the prescription. Results Rhizoma Coptidis, Semen Sinapis Albae and Radix Scrophulariae could be detected by TLC, the spots were clear and there was no interference for the negative sample. A good linearity of berberine was showed in the range of 0.037 29-2.610 3 μg (r=1.000 0). The average recovery of berberine was 98.85%, and RSD was 0.70% (n=6). Conclusion The methods are simple, sensitive, accurate with good reproducibility. It can be used for the quality control of Tongluo Tangtai Granules.

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Anticancer Effects of Xiaotansanjie Recipe on Microsatellite Instability in MKN-45 Human Gastric Cancer Orthotopic Transplantation Tumor

Min YE ; Dazhi SUN ; Pinkang WEI

Chinese Journal of Information on Traditional Chinese Medicine.2013;(12):32-34. doi:10.3969/j.issn.1005-5304.2013.12.012

Objective To explore the mechanism of Xiaotansanjie recipe on MKN-45 human gastric cancer orthotopic transplantation tumor with microsatellite instability. Methods MKN-45 human gastric cancer cell line was used to establish subcutaneous tumor model in nude mice as an experimental model which passed three generations of tumor source. Then OB glue was used to establish human gastric cancer orthotopic xenograft model. Twenty model nude were divided into model group and Xiaotansanjie recipe group (treated with Xiaotansanjie recipe). After 6 weeks of treatment intervention, tumor weight was measured, inhibitory rate was calculated and microsatellite instability (D17S250, D2S123, D5S346, Bat-25 and Bat-26) was detected. Results The tumor weight in Xiaotansanjie recipe group was significantly lower than that in model group (P<0.01). The tumor-inhibition rate was 40.84%. Five microsatellite markers (D17S250, D2S123, D5S346, Bat-25 and Bat-26) were amplified in the tumor tissue of model group, and microsatellite markers D2S123, D5S346 appeared some prolong and reduction. Microsatellite markers (D2S123, D5S346) mutation was stabilized after 6 weeks treatment of Xiaotansanjie recipe. Conclusion Xiaotansanjie recipe can inhibit the MKN-45 human gastric cancer orthotopic transplantation tumor by down-regulating the microsatellite instability.

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Influence of Yifeiqinghua Granules on Expression of Vascular Endothelial Growth Factors and Its Receptor KDR in Lewis Lung Cancer Mice

Feifei LI ; Hao WU ; Lu CHEN ; Bin ZHOU

Chinese Journal of Information on Traditional Chinese Medicine.2013;(12):29-31. doi:10.3969/j.issn.1005-5304.2013.12.011

Objective To clarify the mechanism of Yifeiqinghua granules on tumor angiogenesis and tumor inhibition by observing its influence on the expression of vascular endothelial growth factor (VEGF) and its receptor KDR in Lewis lung cancer mice. Methods Eighty C57BL/6 inbred mice subcutaneously planted with Lewis tumor cell suspension 0.2 mL were randomly divided into 8 groups:blank group, control group, low-, medium- and high-dose of Yifeiqinghua granules groups, Gefitinib group, Gefitinib plus medium-dose of Yifeiqinghua granules group and CTX group, 10 mice in each group. Drugs were administrated from the second day. CTX group was administered on third day and seventh day, other groups were administered for 14 days. The mice were killed to take the tumor on the 15th days. VEGF and KDR expression in tumor tissue was measured by Western Blot. Results Compared with the blank group, the expression of VEGF was significantly decreased in the low- and medium-dose of Yifeiqinghua granules groups (P<0.01). Gefitinib group’s VEGF value was lower than the blank group (P<0.05). Compared with the blank group, the expression of VEGF was significantly decreased in the high-dose of Yifeiqinghua granular group, Gefitinib plus medium-dose of Yifeiqinghua granules group and CTX group (P<0.001). Compared with the blank group, the expression of KDR was significantly decreased in the low-, medium- and high-dose of Yifeiqinghua granules groups, Gefitinib group and CTX group (P<0.05). Conclusions The possible mechanism of Yifeiqinghua granules in treating non-small cell lung cancer is reducing the expression of VEGF and KDR, thus play an inhibitory effect.

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Determination of Camphor, Borneol and Bornyl Acetate in Shenling Baizhu Powder by Gas Chromatography

Mengmeng WANG ; Jinghui WANG ; Xintong FU ; Yougen CHEN ; Bin WU

Chinese Journal of Information on Traditional Chinese Medicine.2013;(12):56-58. doi:10.3969/j.issn.1005-5304.2013.12.021

Objective To establish a GC method for simultaneous determine of camphor, borneol and bornyl acetate in Shenling Baizhu Powder. Methods Capillary column was used with 100% methyl polysiloxane as stationary phase. The temperature-programmed was as follows:starting temperature of 60 ℃, 5 ℃/min, rose to 130 ℃, kept for 5 minutes, followed by 20 ℃/min up to 230 ℃, and kept for 5 minutes. Results The average recovery rate of camphor was 99.87%, RSD was 1.3%, and good linear relationship was showed in the range of 0.003 77-0.150 8 μg (r=0.999 9). The average recovery of borneol was 100.71%, RSD was 2.1%, and good linear relationship was showed in the range of 0.002 411-0.096 4 μg (r=0.999 9). The average recovery rate of bornyl acetate was 101.95%, RSD was 1.3%, and good linear relationship was showed in the range of 0.003 02-0.121 1 μg (r=0.999 9). Conclusion The method is simple, reliable, accurate, and can effectively control the quality of Amomi Fructus in Shenling Baizhu Powder.

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Determination of Specnuezhenide in Shengxinfa Capsules by HPLC

Libin FENG

Chinese Journal of Information on Traditional Chinese Medicine.2013;(12):54-55. doi:10.3969/j.issn.1005-5304.2013.12.020

Objective To set up a method for determining Specnuezhenide in Shengxinfa Capsules. Methods With HPLC method, Agilent TC-C18 column (4.6 mm×250 mm, 5 μm) was used. Mobile phase was methanol-water (35∶65), the flow rate was 1.0 mL/min, and the detecting wavelength was at 224 nm. Results The linearity was in the range of 0.15-0.90 μg. The average recovery was 100.35%, and RSD was 1.43%. Conclusion The method is accurate, reliable, and can be used for the determination of Specnuezhenide in Shengxinfa Capsules for its quality control.

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Determination of Brucine and Strychnine in Semen Strychni and Its Extractive by HPLC

Guolan DIAO ; Guihua ZHANG ; Yunshu MA ; Wenping ZHANG ; Jine HUANG ; Hongmei LUO

Chinese Journal of Information on Traditional Chinese Medicine.2013;(12):51-53. doi:10.3969/j.issn.1005-5304.2013.12.019

Objective To develop a RP-HPLC method for the determination of Brucine and Strychnine in Semen Strychni and its extractive of total alkaloids. Methods A chromatographic column of Licrospher C18 (4.6 mm×250 mm, 5 μm) was used with the mobile phase of acetonitrile∶0.01 mol/L sodium heptane sulfonate and 0.02 mol/L potassium dihydrogen phosphate mixed with equal amount (adjusted pH to 2.8 with 10% phosphonic acid)=27∶73, detection wavelength at 260 nm, column temperature of 30 ℃ and flow rate of 1 mL/min. Results The calibration curves of Brucine and Strychnine were both in good linearity in the ranges of 0.1-1.0 μg and 0.12-1.2 μg (r=1.000) respectively. The average recovery rates of Brucine and Strychnine were 99.88% (RSD=1.06%) and 100.06% (RSD=0.78%) respectively. Conclusion The method is realiable and accurate, which can be applied to determination of Brucine and Strychnine in Semen Strychi and its extractive.

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Resources Distribution Survey of Wild and Cultivated Scutellaria baicalensis Georgi

Jing GU ; Wei HUANG ; Wensheng ZHANG

Chinese Journal of Information on Traditional Chinese Medicine.2013;(12):42-45. doi:10.3969/j.issn.1005-5304.2013.12.016

Objective To provide reference basis for protection and sustainable utility of S. baicalensis by a large scale of nationwide resources survey of Scutellaria baicalensis Georgi in China. Methods On the basis of literature research, survey was conducted on geographical distribution, resources reserves, recovery and growth of the resources status etc. of S. baicalensis. Results The wild resources of S. baicalensis was wildly distributed in the emperate zone in north of China and the distribution was discontinuous. Large differences were showed between different populations in entironment, coenotype and plant morphology. The cultivated resources of S. baicalensis were mainly distributed in Shanxi, Hebei, Gansu, Shandong provinces and Inner Mongolia autonomous region. There were diverse cultivation models and differences existed in production and quality in different regions. Conclusion Destruction of the wild resources and unstable of the resource quality were two main problems of S. baicalensis resources. It was suggested that researches on quality formation and control should be carried out, and institutional guarantee was essential to protect and sustainable use the plant resources of S. baicalensis.

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Effect of Qizhi Zhoufei Granules on Cytokines of Chronic Obstructive Pulmonary Disease Model Rats

Yi ZHANG ; Juan LI ; Shuchang WEI ; Jintian LI ; Yongqi LIU ; Kunpeng ZHAO ; Guangwei LIU

Chinese Journal of Information on Traditional Chinese Medicine.2013;(12):26-28. doi:10.3969/j.issn.1005-5304.2013.12.010

Objective To observe the effects of Qizhi Zhoufei Granules on IL-8, TNF-αand ET-1 in serum and bronchoalveolar lavage fluid (BALF) of chronic obstructive pulmonary disease (COPD) model rats, and to study its mechanism of preventing and treating COPD. Methods Wistar rats were randomly divided into blank group, model group, positive control group, Qizhi low-dose group, Qizhi medium-dose group and Qizhi high-dose group. The COPD model was established by smoking and infusing lipopolysaccharide (LPS). After 15 days, the rats were given the drugs by orally taking once a day. On the 43rd day, the change of IL-8, TNF-α and ET-1 content in serum and BALF were measured by radio-immunity assay. Results Compared with the blank group, the level of IL-8, TNF-α and ET-1 in serum and BALF were considerably raised in model group (P<0.05). Whereas compared with the model group, the level of TNF-αin serum of Qizhi high-dose group was remarkably lowered (P<0.05), the level of IL-8 in serum of Qizhi low-dose group and Qizhi high-dose group was remarkably lowered (P<0.05), the level of TNF-αin BALF of Qizhi medium-dose group was remarkably lowered (P<0.05), the level of IL-8 in BALF of Qizhi low-, medium- and high-dose group was remarkably lowered (P<0.05), and the level of ET-1 in BALF of Qizhi high-dose group and Qizhi medium-dose group was remarkably lowered (P<0.05). Conclusion Qizhi Zhoufei Granules have protective effect on lung tissue of COPD rats, and can adjust the content of IL-8, TNF-α and ET-1, which may be one of its mechanisms of intervening occurrence and development of COPD.

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