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Aim To establish a method for determining the antithrombotic biological activity of Honghua injection, which can be used to evaluate and control its quality. Methods Collagen-adrenalin was used to induce mouse acute cerebral thrombosis model and hemiplegic protection rate of Honghua injection in mice as an index to investigate the antithrombotic activity of Honghua injection. The experimental conditions of the administration dosage, inducer dosage, and different strains of mice were investigated, and the experimental conditions were optimized by orthogonal design. Results Honghua injection which was made by 5. 0 g crude drug · kg-1and continuously ip for 3 d, had obvious protective effect on collagen-adrenalin-induced acute cerebral thrombosis in mice. The established biological activity limit method showed a good repeatability, intermediate precision, and reproducibility. Each sample showed different degrees of differences in biological activity. Conclusions The thrombus test in mice can be used as a method for measuring the biological activity of Honghua injection. It is recommended to use 5. 0 g crude drug · kg-1dose as the limit for bioactive process optimization and product quality controlment of Honghua injection.
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Quality control of biotech drugs has attracted increasing attentions in recent years. Deamidation reaction is one of the major concerns in quality control of biotech drugs, due to the generation of isoaspartic acid(isoAsp) . This paper describes the deamidation of asparagine(Asn) residues and its effects on the biological drugs. The detection methods currently used in China and overeas for this reaction, including pretreatment protocols and instrumental analysis were described. The identification and determination of isoaspartyl sites were also described in detail, along with the positive impact on the development of biotech drugs in China by the studies on deamidation reactions.
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OBJECTIVE: To introduce the application of the residual solvents determination methods in general chapter 0861 of Chinese pharmacopoeia 2015 edition. METHODS: Determination of the residual solvents of cefathiamidine was chosen as an example to indicate that how to use the residual solvents determination methods in general chapter 0861 of Chinese pharmacopoeia 2015 edition when the methods for the determination of residual solvent in monograph of cefathiamidine did not work. RESULTS: The residual solvents in cefathiamidine from different manufactures were determined accurately using methods in general chapter 0861 of Chinese pharmacopoeia 2015 edition. CONCLUSION: The residual solvents determination methods in general chapter 0861 of Chinese pharmacopoeia 2015 can be used to screening and confirm the unknown peaks by the two opposite polar column systems, which can help to establish the residual solvents method.
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Objective:To establish an HPLC method for the determination of compound proglumide tablets. Methods:A Diamon-sil C18(250 mm ×4.6 mm,5 μm)column was used. The mobile phase was 2% ammonium acetate-methanol (40∶60), the detection wavelength was 225nm, and an external standard method was employed. Results:The linear range of proglumide was 2. 60-208. 10 μg (r=0. 999 9), and the average recovery was 99. 8%(n=6,RSD=0. 6%). Conclusion: The HPLC determination method for com-pound proglumide tablets is accurate, simple and reproducible.
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Oleaginous microbial strains were cultivated to identify the best oil-producing strain amongst Yarrowia lipolytica (CGMCC 2.1398), Lipomyces starkeyi (CGMCC 2.1608), Rhodosporidium toruloides (CGMCC 2.1389), Mortierella isabellina (CGMCC 3.3410), Cunninghamella blakeleana (CGMCC 3.970), and Mycobacterium QJ311. A method for rapid determination of oil content and fatty acid composition was established to identify the optimum oil-producing strains. This method had a relative standard deviation of 4.09%, an average recovery ratio of 97.09% and a detection limit of 0.1–1.0 g. Mortierella isabellina CGMCC 3.3410 was identified as the best oil-producing strain amongst the six strains tested, with a total biomass of 75 g/10 L and a lipid content of 35%. A rapid screening method of oleaginous microorganisms is discussed for the first time.
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A new inhibitive heavy metals determination method using trypsin has been developed. The enzyme was assayed using the casein- Coomassie-dye-binding method. In the absence of inhibitors, casein was hydrolysed to completion and the Coomassie-dye was unable to stain the protein and the solution became brown. In the presence of metals, the hydrolysis of casein was inhibited and the solution remained blue. The bioassay was able to detect zinc and mercury with IC50 (concentration causing 50% inhibition) values of 5.78 and 16.38 mg l-1 respectively. The limits of detection (LOD), for zinc and mercury were 0.06 mg l-1 (0.05-0.07, 95% confidence interval) and 1.06 mg l-1 (1.017-1.102, 95% confidence interval), respectively. The limits of quantitation (LOQ) for zinc and mercury were 0.61 mgl-1 (0.51-0.74 at a 95% confidence interval) and 1.35 mg l-1 (1.29-1.40 at a 95% confidence interval), respectively. The IC50 value for zinc was much higher than the IC50 values for papain and Rainbow trout, but was within the range of Daphnia magna and MicrotoxTM. The IC50 value for zinc was only lower than those for immobilized urease. Other toxic heavy metals, such as lead, silver, arsenic, copper and cadmium, did not inhibit the enzyme at 20 mg l-1. Using this assay, we managed to detect elevated zinc concentrations in several environmental samples. Pesticides, such as carbaryl, flucythrinate, metolachlor, glyphosate, diuron, diazinon, endosulfan sulphate, atrazine, coumaphos, imidacloprid, dicamba and paraquat, showed no effect on the activity of trypsin relative to control (One-way ANOVA, F12, 26 = 0.3527, p> 0.05). Of the 17 xenobiotics tested, only (sodium dodecyl sulphate) SDS gave positive interference with 150 % activity higher than that of the control at 0.25% (v/v).
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Apesar de os desarenadores serem largamente utilizados, há mais de um século, como uma das unidades que compõem o tratamento preliminar de estações de tratamento de esgoto (ETEs), pouco se sabe a respeito de suas eficiências de remoção devido à total carência de métodos analíticos padronizados para determinação da concentração de areia presente no esgoto sanitário. O presente artigo apresenta um método original para determinação da concentração de areia no esgoto, baseado no emprego de procedimentos amplamente conhecidos, como: sedimentação em cones Imhoff, oxidação com peróxido de hidrogênio, enxágües sucessivos, calcinação e pesagem em ba-lança analítica. A aplicação do método desenvolvido demonstrou que a concentração de areia no esgoto sanitário afluente da ETE Jardim das Flores (Rio Claro, SP) apresenta grande variabilidade com picos de até 200 mg/l e que a concentração média de areia encontra-se entre 20 mg/l e 73 mg/l.
Although degritters have been largely used, for more than a century, as preliminary treatment units of wastewater treatment plants (WWTP), their removal efficiency is yet poorly known due to the lack of standardized analytical methods to determine wastewater grit concentration. This paper brings up this subject and presents an original method to determine wastewater grit concentration based on use of well-known procedures, such as: Imhoff cone sedimentation, hydrogen peroxide oxidation, sequential rinse, calcination and analytical balance weighing. The application of the developed method demonstrated that grit concentration in the influent of WWTP Jardim das Flores varies significantly, with peak concentration of about 200 mg/l, and that the mean grit concentration seems to be among 20 mg/l and 73 mg/l.
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The antioxidant activity of lycopene is highest among the current carotenoids.Recently the researches on lycopene are focused on ingredient of functional foods in the world.The fermentation production of lycopene by Streptomyces rimosus was reported first time in China.The determination methods,such as spectrophotometric method and HPLC were constructed.Using a stain Fc of Streptomyces rimosus as a original strain,a high-yield lycopene producing mutant strain Fc' was selected after UV mutation.The lycopene yield of the strain is 2.5 times higher than the original strain Fc.The optimal fermentation conditions were determined by flask experiments,and the lycopene yield of strain Fc' reached 230mg/L in flask under the optimal fermentation condition,meanwhile the Streptomyces rimosus strain could produce purer lycopene without adding any blocking agents in its fermentation process.This results lay a good foundation for lycopene commercial production by fermentation using Streptomyces rimosus.
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0.05). There were significant differences among 3- or 6- or 9- or 12-hour groups and 24- or 36-hour groups in 6, 13 ℃ and 28 ℃ respectively (P