HPLC同时测定野菊花中4种黄酮苷类的含量

目的:建立同时测定野菊花中木犀草素-7-O-β-D-葡萄糖苷、芹菜素-7-O-β-D-葡萄糖苷、香叶木素-7-O-β-D-葡萄糖苷、蒙花苷成分含量的方法。方法:采用HPLC法,色谱柱:Agilent Eclipse XDB-C18柱(250 mm×4.6 mm,5μm);流动相:乙腈-0.1%磷酸水溶液,梯度洗脱;检测波长:324 nm;流速:1.0 ml·min-1;柱温:30℃。结果:木犀草素-7-O-β-D-葡萄糖苷、芹菜素-7-O-β-D-葡萄糖苷、香叶木素-7-O-β-D-葡萄糖苷、蒙花苷的质量分别在2.296~45.920 ng、0.403~8.064 ng、0.410~8.208 ng、4.296~85.920 ng与峰面积呈良好的线性关系;平均加样回收率分别为100.52%,99.38%,98.79%,101.30%,RSD均小于2.0%(n=6)。结论:该分析方法快速、准确、重复好,适用于野菊花黄酮苷类含量测定。

野菊花提取液中总黄酮含量测定分析

目的测定野菊花提取液经聚酰胺色谱柱分离后不同洗脱层总黄酮含量。方法采用60%浓度乙醇水加热回流提取野菊花供试品,经聚酰胺色谱柱分离纯化得到不同浓度乙醇洗脱层。采用紫外可见分光光度法对不同洗脱层总黄酮含量进行测定。结果野菊花提取液50%乙醇洗脱液干膏得率为0. 975 3%,干膏得率最高。70%乙醇洗脱层黄酮类化合物含量为46. 51%,含量最高。结论聚酰胺色谱柱分离纯化法准确、灵敏、简便、重复性好,为野菊花黄酮类化合物富集纯化提供了科学基础。

应用Cephodex 微载体培养DF-1细胞增殖水貂源犬瘟热病毒的技术研究

为了建立水貂源犬瘟热病毒(CDV)的大规模悬浮培养技术,实现细胞高密度生长和病毒高效增殖,本研究应用Cephodex微载体悬浮培养鸡胚成纤维细胞系DF-1细胞,增殖弱毒株CDV3。整个过程采用摇瓶培养法,通过对病毒培养温度、病毒收获时间等关键技术条件进行优化,确立最佳培养条件。结果表明,DF-1细胞37℃培养至72 h,接种CDV3,35℃继续培养72 h收获病毒,病毒滴度每0.1 mL可达10^(5.0) TCID_(50)。CDV微载体悬浮培养技术的初步建立,为高效水貂犬瘟热疫苗的研发生产奠定了重要的基础。

Research Progress of Active Ingredients and Pharmacological Action of Honeysuckle

Honeysuckle, the dry flower bud or with open flower of Lonicerajaponica in caprifoliaceae, has been touted as a cure-all for heat-clearing and detoxifying effect. Honeysuckle contains abundant components such as volatile oil, flavonoids and organic acids, playing the role of antibiosis, antivirus, antiphlogosis, immunopotenfiation and antioxidation. The active ingredients and pharmacological action of honeysuckle are reviewed in the paper, so as to lay the foundation for further application of honeysuckle in veterinary clinic.

Determination of Berberine Hydrochloride Content in Qingwen Baidu Granule by HPLC

The paper was to establish a simple and sensitive HPLC method for determination of berbefine hydrochloride content in Qingwen Baidu granule. The HPLC system consisted of Stable Bond - C18 column （4.6 × 150 mm, 5um） and with a mobile phase of acetonitrile -0.05mol/L potassium dihydrogen phosphate solution （50：50,v：v） mixture. Berberine hydrochloride was detected at the wavelength of 345 nm, with a flow rate of 1.0 ml/min and a column temperature of 30 ℃. The excipients and solvents in the granule could be well separated from the drug under such a designated chromatogram condition and did not interfere with the assay. A good linear relationship was found between peak area and the concentration of berberine hydrochloride in the range of 16.0 -48.0 I^g/mL. The average recovery of berberine hydrochloride was 99.09%. The established approach was specific, accurate, reliable, prompt, sensitive and applicable, and could be used to control the quality of Qingwen Baidu granule.

Advance in Effect of Veterinary Drug Residues on Ecological Environment

Thousands of tons of pharmaceutically active substances are used in veterinary medicines in intensive livestock and poultry production, but most of the drugs are poorly absorbed by animals and excreted with the animal feces. The veterinary medicines can cause various kinds of harmful impact to the environment. Now the relevant research has been a hot all over the world. With the feces excreted to the environment, the veterinary medicines can be accumulated in the soil and water, and produce all sorts of reaction. Some medicines can cause toxic effect to animals, plants and microorganisms. The article summarized the residue, the environmental impact, the transformation and the development direction of the veterinary medicine in the environment to supply frame of reference for further research.

Effects of Shuanghuanglian Propolis Oral Liquid on Growth Performance,Slaughter Performance and Meat Quality of Broilers

[Objective] The paper was to investigate the effects of adding Shuanghuanglian propolis oral liquid in drinking water on growth performance,slaughter performance and meat quality of broilers. [Method]Ninety one-day-old broilers were randomly divided into three groups（ group Ⅰ,group Ⅱ and group Ⅲ） ： group Ⅰ was the control group,and groups Ⅱ and Ⅲ were added 0. 5 and 1. 0 mL / L Shuanghuanglian propolis oral liquid in drinking water,respectively.[Result] Compared with broilers in group I,the average daily feed intakes of broilers in group Ⅱ and group Ⅲ were increased by 0. 86%（ P 〉 0. 05） and 1. 10%（ P 〉 0. 05）,respectively; the average daily gains were increased by 2. 33%（ P 〈 0. 05） and 4. 43%（ P 〈 0. 05）,respectively; the feed gain ratios were decreased by 1. 05%（ P 〉0. 05） and 2. 63%（ P 〈0. 05）,respectively; the slaughter rates were increased by 0. 94%（ P 〉0. 05） and 1. 70%（ P 〈0. 05）,respectively; the eviscerated rates were increased by 0. 52%（ P 〉 0. 05） and 1. 15%（ P 〉 0. 05）,respectively; the semi-eviscerated rates were increased by 1. 51%（ P 〈 0. 05） and 2.23%（ P 〈 0. 05）,respectively; the breast muscle rates were increased by 3. 77%（ P 〈 0. 05） and 5. 51%（ P 〈 0. 05）,respectively; the leg muscle rates were increased by 1. 96%（ P 〉 0. 05） and 4. 09%（ P 〈 0. 05）,respectively; the lightness（ L＊） were decreased by 0. 96%（ P 〉 0. 05） and 1. 47%（ P 〉 0. 05）,respectively; the redness（ a＊） were increased by 1. 24%（ P 〈 0. 05） and 1. 86%（ P 〈 0. 05）,respectively; the yellowness（ b＊） were decreased by 0. 44%（ P 〉0. 05） and 1. 03%（ P 〈0. 05）,respectively; the drip losses were decreased by 1. 22%（ P 〈0. 05） and 1. 56%（ P 〈0. 05）,respectively; the cooking losses were decreased by 1. 76%（ P 〉 0. 05） and 2. 89%（ P 〈 0. 05）,respectively. [Conclusion]Adding 0. 5 and 1. 0 mL / L Shuanghuanglian propolis oral liquid in drinking water could improve growth performance,slaughter performance,meat quality of broilers,and the dose of 1. 0 mL / L is recommended in clinical application.

药物联用抑制白念珠菌生长的研究进展

白念珠菌是念珠菌属最常见的种类之一,是临床上重要的侵袭性念珠菌感染来源。由于传统抗真菌药物的广泛使用,临床已经分离出很多耐药白念珠菌,导致部分传统药物在临床治疗效果明显下降。同时,一些化合物自身的毒性限制了其临床应用。在这种背景下,联合疗法由于发挥了各种药物或手段的协同作用,有可能改进单独用药的不足,具有抑制白念珠菌生长的巨大潜力。

Simultaneous Determination of Sulfamonomethoxine Sodium and Trimethoprim in Compound Sulfamonomethoxine Sodium Propolis Solution by High Performance Liquid Chromatography(HPLC)

[Objective] The paper aimed to provide a basis for quality control of compound sulfamonomethoxine sodium propolis solution. [Method] High per- formance liquid chromatography （HPLC） method was used to determine the contents of sulfamonomethoxine sodium and trimethoprim simultaneously in compound sulfamonomethoxine sodium propolis solution. [Result] Sulfamonomethoxine sodium and trimethoprim showed a linear relationship within the concentration of 10 - 500 μg/mL under the following chromatographic conditions： Kromasil C18 column （250 mm × 4.6 mm, 5 μm）, mobile phase 0.02 mol/L phosphoric acid solution- methanol（80：20, V/V）, detection wavelength 270 nm, flow rate 1.0 mL/min, and column temperature 30℃. The average recoveries were 99.3% and 99.4%, respectively. [ Conclusion] The method is accurate and feasible, and can determine sulfamonomethoxine sodium and trimethoprim simultaneously in compound sul- famonomethoxine sodium propolis solution.

Research Advances in Detection Techniques of High Performance Liquid Chromatography-Refractive Index Detector

As a highly sensitive and stable detector,refractive index detector is usually used for quantitative detection of substances such as polymer,sugar and organic acid. The research reviewed the application of HPLC-RID in the fields of quantitative determination of medicine and food,in order to lay a foundation for wider use of RID.

Associated Bacteriostasis Research of Coptidis Rhizoma-Folium Isatidis and Coptidis Rhizoma-Flos Poprli against Escherichia coli O2

[Objective] The paper was to explore the combined inhibitory effect of rhizoma coptidis-folium isatidis and rhizoma coptidis-flos poprli against Escherichia coli O2.[Method] Contrast test of single and associated bacteriostasis against known serotype E. coli O2 was conducted using microcheckerboard method.[Result] The MIC of rhizoma coptidis, folium isatidis and flos poprli were 1/8 extracting liquid, 1/8 extracting liquid and 1/2 extracting liquid, respectively. When combined with folium isatidis or flos poprli, the MIC of rhizoma coptidis was 1/8 extracting liq-uid or 1/16 extracting liquid compared with single use. When combined with rhizoma coptidis, the MIC of folium isatidis and flos poprli were 1/8 extracting liquid and 1/16 extracting liquid.[Conclusion] When rhizoma coptidis was combined with folium isatidis or flos poprli, the FIC values were 2 and 0.625, performing independent action and additive effect, respectively.

Advances in Research and Application of gE Gene/Protein in Prevention and Control of Swine Pseudora-bies

Research and application progresses of gE gene and its encoding gE protein in PR vaccines, diagnostic technique and epidemiological investigation are summarized, which have certain reference value for comprehensive prevention and control of PR and gradual purification of PR in different regions.

Effect of Shuanghuanglian Propolis Oral Liquid on Immune Function of Broilers

The effects on immune function of broilers was studied by adding different doses of Shuanghuanglian propolis oral liquid in drinking water, to provide a theoretical basis for application and promotion of Shuanghuanglian propolis oral liquid. Ninety one-day-old broilers were randomly divided into three groups ： group I was designed as control; groups Ⅱ and Ⅲ were added with 0.5 and 1.0 mL/L Shuanghuanglian propolis oral liquid in drinking water, respectively. Compared with group I, all tested indicators in groups Ⅱ and Ⅲ were increased at varying degrees as follows ： bursa of fabricius index increased by 5.74% （P 〈 0.05 ） and 7.66% （P 〈 0.05 ） ; thymus index increased by 1.47% （P 〉 0.05 ） and 2.06% （P 〈 0.05 ） ; spleen index increased by 5.88% （P 〉 0.05 ） and 7.84% （ P 〈 0.05 ） ; Newcastle disease virus （NDV） antibody titer increased by 11.99% （P 〈 0.05 ） and 16.94% （P 〈 0.05 ） ; IL-2 content increased by 2.71% （P 〉 0.05） and 4.52% （P 〈 0.05 ） ; IL -6 content increased by 4.04% （P 〉 0.05 ） and 7.26% （ P 〈 0.05 ）, respectively. The results showed that adding 0.5 and 1.0 mL/L Shuanghuangllan propolis oral liquid in drinking water could improve the immune function of broiler and immune effect of ND vaccine.

Comparison of Effective Components Contents of Lysimachia clethroides Duby with Three Extraction Methods

[ Objective] The differences in effective components contents of Lysimachia clethroides Duby were compared using three extraction methods. [ Method] L. clethroides was extracted by three methods, including water extraction, water extraction-alcohol precipitation and 75% ethanol extraction, to determine the effec- tive components of extracts, and contents of calcium, phosphorus and Vc were calculated. [ Result] L. clezhreides contained calcium salt, chlorine, VB1, VB2, Vc and phosphate, but did not contain citrate, sulfate, carbonate or bicarbonate, acetate, lactate, VA , VD and Vs ; the contents of calcium and phosphorus in water extract were 150 and 3.34 mg/g, respectively, greatly exceeding their contents in water extraction-alcohol precipitation and ethanol extraction, while the difference in Vc content was not significant. [ CondusionJ The paper provides a reference for practical application of L. clethroides as an additive.

Identification of Coptis chinensis and Forsythia sus-pensa from Qingwen Baidu Granules by TLC

[ Objective] The paper was to establish the identification method of Coptis chinensis and Forsythia suspensa in Qingwen baidu granules. [ Method ] With benzene-ethyl acetate-methanol-isopropanol-cancentrated ammonium liquid （ 12：6：3：3：1 ） as the developer, the samples were outspread in the cylinder with sat- urated ammonia steam by thin layer chromatography （TLC）, and the existence of C. chinensis and berberine hydrochloride was checked under UV lamp. With tri- chloromethane-methanol （5：1） as the developer and 10% sulfuric acid ethanol solution as the chromogenic reagent, the samples were heated at 105 ℃ for 5 min to check the existence of F. suspense under natural light. [Result] Qingwen baidu granules had the same spots with C. chinensis control and berberine hydrochloride at the same Rf value under 365 nm UV lamp; Qingweu baidu granules had the same spots with F. suspensa control at the same Rfvalue under natural light. [ Condusion] A TLC method detecting C. chinensis and F. suspensa in Qingwen baidu granules was established, which had good specificity and repeatability, suitable for rapid detection of C. chinensis and F. suspensa. It could be well applied to control the quality of Qingwen baidu granule.

Identification of Gardenoside and Sarsasapogenin from Qingwen Baidu Granules by TLC

[ Objective ] We aimed to establish the identification method of gardenoside and sarsasapogenin in Qingwen baidu granules. [ Method] With ethyl ace- tate-acetone-formic acid-water （10：7：2：0.5） as the developer and 10% sulfuric acid ethanol solution as the chromogenic reagent, the samples were heated at 100 ~C until the spots were clearly visible, and the existence of gardenoside was checked under natural light. With toluene-acetone（9：l ） as the developer and 5% vanillin sulfuric acid solution as the chromogenic reagent, the samples were heated at 105 ~C until the spots were clearly visible, and the existence of sarsasapogenin was checked under natural light. [Result] Qingwen baidu granules had the same spots with gardenoside and sarsasapogenin at the same Rfvalue under natural light. [ Conclusion] A TLC method detecting gardenoside and sarsasapogenin in Qingwen baidu granules was established, which had good specificity and repeatability, suitable for rapid detection of gardenoside and sarsasapogenin.

Preparation and Quality Evaluation of Toltrazuril Nanoemulsion

To determine the prescription and preparation method of toltrazuril nanoemulsion, we selected liquid paraffin, ethyl acetate, soybean oil, IPM and oleic acid as the oil phases, while RH-40, EL-40, OP-10, Tween-80 and Span-80 were the surfactants, and alcohol, 1,2-ropanediol, ispropanl, glycerol and PEG-400 were the co-surfactants. The best prescription was acquired through pseudo-ternary phase diagrams. The result showed that ethyl acetate, Tween-80 and 1,2-propanediol could facilitate the formation of nanoemulsion. The optimum prescription was surfactant：co-surfactant：oil phase：water phase =2：1：2：5, with the solubilizer of 10%. This prescription can prepare stable toltrazuril nanoemulsion.

Advances in Laboratory Diagnostic Methods for Duck Circovirus Infection

Duck circovirus infection is an immunosuppressive disease that is characterized by growth retardant and odd deaths. Its infection is usually combined with other pathogens, making the clinical diagnosis more difficult. With the rapid development of molecular biological and immunological technologies, the laboratory diagnostic methods for duck circovirus infection also advance greatly. The paper summarizes the research advances in various laboratory detection methods for duck cireovirus infection including PCR, nested PCR, multiple PCR, fluorescence quantitative PCR, LAMP, nucleic acid probe and ELISA, as well as their ad- vantages and shortages, aiming at providing reference for finding novel detection methods and for the diagnosis and comprehensive prevention and control.

Chicken Embryo Inhibition Test of Recombinant Freeze-drying Chicken Interferon against Newcastle Disease Virus( NDV) F_(48)E_(10)

The paper was to study the inhibitory effect of recombinant freeze-drying chicken interferon against Newcastle disease virus （NDV） F48E10. Nine-day-old chicken embryos were inoculated with recombinant freeze-drying chicken interferon via allaotoic sack, while 10-day-old chicken embryos were inoculated with NDV F48E10, and in vivo protective efficacy of interferon on chichen embryos was studied. The results showed that the recombinant freeze-drying chicken interferon at the dose of 1.28 mg/embryo reached the protection ratio of 90% on chicken embryo infected by F48E10.

Drug Sensitivity Test and Regression Verification of Escherichia coli from Rex Rabbit

[ Objectives] The paper aimed to select drugs reasonably for treatment of rex rabbit colibacillosis, and to isolate the pathogenicity of Escherichia coll. [ Methods ] Pathogen isolation, drug sensitivity test and pathogen regression test were performed with rex rabbits killed by E. coli in clinic. [ Results] The isolate was E. coli 0-23, susceptible to amikacin and cefotaxime sodium; when the challenge dose was 1.0 mL/rabbit （about one billion E. coli）, the test animal would discharge mucous feces. [ Conclusions] The results provided model support for clinical medicine selection against rex rabbit colibacillosis.