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Objective@#To develop a micro-neutralization test for determination of neutralizing antibody against ZIKA virus (ZIKV) in human sera and to verify the acute and convalescent serum samples of 10 ZIKA virus-infected cases diagnosed by nucleic acid detection and/or virus isolation.@*Methods@#ZIKV isolated from ZIKA cases was used to determine micro-neutralization antibody. The virus solution was prepared by infecting BHK21, VERO and VERO-E6 cell lines and viral titer was tested; 100 TCID50 viral solution and 4 times diluted sera which were inactivated at 56 ℃ for 30 min were neutralized, then added the cell suspension and incubated in 5% CO2 incubator at 37 ℃ for 7 d. The CPE was observed every day.@*Results@#The sensitivity of BHK21, VERO and VERO-E6 was different after infection with ZIKA virus. VERO cell line was the most sensitive and showed typical CPE. VERO cell line was used to establish a micro-neutralization test for determination of neutralizing antibody against ZIKA virus in sera.@*Conclusions@#The neutralizing antibody test for zika virus in sera is a special and usefulmethod to diagnose human infection of ZIKV and to conduct population based epidemiological investigation.
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Objective To establish a method for the determination of peoniflorin and glycyrrhizic acid in modified Shaoyao Gancao Granules.Methods Reversed-phase high performance liquid chromatography(HPLC) was used.The chromatographic condition was as follows: Diamonsil C18 column(4.6mm ?250mm,5?m),column temperature being room temperature,mobile phase consisting of acetonitrile(A)-10g/L acetic acid(B) with adjusting pH value being 3.5,flowing rate at 1.0mL/min,detective wavelength at 250nm,and the injection volume being 20?L.The gradient elution procedure was as follows: elution with 18% A for 0～5 min,elution with 19%～29% A for 6～10 min,elution with 30%～45% A for 11～25min,and A+B=100%.Results Peoniflorin at the concentration of 1.916～122.6?g/L and glycyrrhizic acid at the concentration of 2.625～168?g/L showed a good linearity.The mean recovery was 101.12% for peoniflorin and 99.68% for glycyrrhizic acid.Conclusion This method is simple and reproducible,and can be used for the determination of peoniflorin and glycyrrhizic acid in modified Shaoyao Gancao Granules.
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【Objective】To optimize the process conditions for active compounds of Flos Datura by microwave-assisted extraction.【Methods】Uniform design was used to observe the effects of microwave power,microwave radiation time,solvent volume,soaking time and extracting time on the extraction of dry extract,total alkaloids,scopolamine of Flos Datura.The contents of total alkaloids and scopolamine of Flos Datura were determined by acid stain colorimetry and HPLC,and the obtaining rate of dry extract was measured by weight method.【Results】 The optimal process conditions for Flos Datura were as follows: soaking the medicinal material for 2 hours with 10 times of 75% alcohol,with continuous radiation for 25 min at the power of 730 W,at 65℃.【Conclusion】Microwave-assisted extraction is superior to soaking extraction method in increasing productivity and economizing the solvent.