ABSTRACT
@#To establish a high performance liquid chromatography(HPLC)method to determine the content of bacteriostats in the ocular extractives eye drops, Diamonsil C18(4. 6 mm×250 mm, 5 μm)column was used, with gradient elusion by 1% triethylamine solution(pH 3. 0)(mobile phase A)and methanol(mobile phase B). The detection wavelength was 256 nm; the column temperature was 40 °C; and the flow rate was 1. 0 mL/min. Under these conditions, the three bacteriostats of methylparaben, ethylparoben and chlorhexidine acetate showed good resolution. The bacteriostats exhibited good linear relationship between the peak area and the concentration in the concentration range of 0. 1- 80 μg/mL(r> 0. 999 1). The recoveries were from 97. 2% to 104. 1%, and the RSD was 0. 8% to 1. 2%. The content of bacteriostats in all the five batches of ocular extractives eye drops was less than 10% of the prescription amount. It was found that the activated carbon used in the production process had strong adsorption effect on the bacteriostat, and that the lower the temperature and the higher the concentration of activated carbon, the stronger the adsorption of bacteriostatic agent. The adsorption capacity of activated carbon for different bacteriostats is: chlorhexidine acetate > ethylparoben > methylparaben. The results showed that the established HPLC method was easy to operate with high sensitivity and good repeatability. It can be used to determine the content of bacteriostat in ocular extractives eye drops quickly and accurately. In addition, this study reveals for the first time the effect of impurity removal process on bacteriostat in the production of ocular extractives eye drops. It is not suitable to use activated carbon to remove impurities before adding parabens and chlorhexidine acetate bacteriostats. The current work provides a new guiding basis for the monitoring and improvement of the quality of ocular extractives eye drops.
ABSTRACT
Objective A high performance liquid chromatographic (HPLC) method was established for the determination of glycerol in propofol medium and long chain fat emulsion injection.MethodsThe chromatographic conditions were as follows: Kromasil 100-5-NH2 column(4.6×250mm,5μm) with the column temperature was 40℃,acetonitrile-water(8515)as mobile phase with flow rate of 1.0mL/min.Glycerol was detected by refractive index (RI) detector at 40℃.ResultsThe linear range of glycerol was 455.3916-2276.9580μg/mL(r=0.9999,n=7),the average recovery rate was 99.5%,RSD was 0.6%(n=9),the limit of detection(LOD) was 121ng and the limit of quantification(LOQ)was 364ng.ConclusionThe method was simple, rapid, strong specifity and accurate with good reproducibility, which is suitable for the content determination of glycerol in propofol medium and long chain fat emulsion injection.
ABSTRACT
Objective To compare magnetic beads kit,agrose gel recovery kit and heparinase I three methods to purify the micro DNA from crude heparin, then use q-PCR to identify the species origins and select the best method.Methods Using magnetic beads kit,agrose gel recovery kit and heparinase I to purify micro DNA from crude heparin and combined the porcine,bovine and ovine identification kits to identify the species origins and conformed the minimum detection limit of different percentage of ovine crude heparin in porcine crude heparin.Results Three pretreatment methods all can solve the pretreatment difficulties and we found that the haparinase was the best method; the minimum detection limit was 0.01%of ovine crude heparin in porcine crude heparin.Conclusion The heparinase method is the best pretreatment method and can successfully solve the pretreatment difficulties.Heparinase combine the porcine, bovine and ovine identification kits can identify the species origins from crude heparin.