| Objective1-Bromopropane(1-BP),like Freons,possesses low boiling point(volatile)and stable chemical properties(e.g.non-flammable).In addition,compared to Freons,it has a shorter half-life in the atmosphere and is Ozone-friendly.With the increasing global emphasis on environmental protection,1-BP has gradually become one of the most widely used organic solvents and cleaning agents around the world.Our country has become one of the major producers of 1-BP in the world.As a result,the number of 1-BP occupational exposed workers and users in China has increased significantly.There are a growing number of studies on 1-BP toxicity at home and abroad,including a large number of animal experiments,which mostly used experimental biochemical indicators to determine the animal exposure levels.However,there is no report on the direct determination of 1-BP in the blood of experimental animals.Headspace gas chromatography has high sensitivity and fast speed for the analysis of trace organic volatile compounds in samples,and it is also featured the advantages of simple sample pretreatment.In particular,static headspace gas chromatography(HS-GC)is a simple,rapid and accurate detection method for volatile substances with simple pretreatment.Also this method allows avoiding the interference of non-volatile substances in the sample.This study intends to establish a rapid,simple,efficient and accurate method for detecting the concentration of trace 1-BP in animal blood samples.Our experiment adopted headspace gas chromatography to determine the concentration of 1-bromopropane in blood samples,which provided a convenient and accurate measurement method for related experiments.Method1 animal experiment:After 3 days acclimation,24 Wistar rats were randomly divided into four groups with 8 in each,i.e.1-BP low-dose group(200mg/kg),medium-dose group(400mg/kg),high-dose group(800mg/kg).Animals in each group were given in accordance with their own dosage for 12 days,then the rats were sacrificed with collecting blood。Half blood let stand for 30min,then centrifuge the blood to 3000 r/min speed for 10min.take supernatant fluid,-20℃ refrigerated processing;The other half blood-20 ℃ refrigerated processing directly.2 check test:Blood samples were added to the top 20ml bottles,use HS-20 automatic sample headspace sample of Shimadzu company。equilibrium constant temperature furnace temperature80℃,10min,balance time 1 ml sample quantity.Using GC-2010 gas chromatography,Shimadzu company RTX-5 chromatographic column,carrier gas for high purity N2,column flow rate 1.05 ml/min,linear velocity 20.0 m/SEC.3 Determination of 1-BP concentration in blood of infected rats This method was used to determine the 1-BP concentration of blood and serum,and the experimental results were compared and discussed.4 Toxicokinetics study about 1-BP the concentration of 1-bp in the serum were determined at different time points after the rats were infected,the changes in the distribution of 1-BP in the blood were analyzed.Results1 Multi-point standard addition method was used to verify the linear relationship between the concentration of 1-BP in whole blood and serum of rats measured by headspace gas chromatography,and the regression equation was obtained as follows:In whole blood:y=37636x+258912,r=0.9985;In serum:y=37728x+401557,r=0.99982 The concentrations of 1-BP in whole blood and serum of rats weremeasured by standard addition method,and the results were statistically analyzed.There was no significant difference between the two sets of data.3 After exposure,the concentration of 1-BP in the blood of the rats reached the highest value after one hour and then decreased gradually.Conclusions1 In this study,a method of direct determination of 1-BP concentration in rat blood by headspace gas chromatography was established.The method is rapid,simple,efficient and accurate.It also avoids the unnecessary introduction of organic solvents in the sample pretreatment to simplify the steps.2 Headspace gas chromatography was used to determine the concentration of 1-BP in rat whole blood and serum samples.There was no significant difference between the two groups.3 The peak concentration of 1-BP in blood was 30.8 μg/L,and the peak time was 1h. |
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