Exploring The Feasibility Of Saliva For Drug Control Based On The Correlation Of Ketamine Concentration Between The Saliva And Blood

In recent years,the number of driving accidents and accidents caused by the use of Ketamine(Ketamine,KET)has been increasing.How to conduct systematic detection has become the current difficulty in implementing drug control in China.At the same time,due to the complexity of the sampling procedure,the blood test materials limit its application in onsite real-time testing.Therefore,the subject takes the current status of road drug driving detection as a research background,and proposes to use saliva test materials instead of blood test materials for the preliminary screening work of road drug driving control,and to establish a systematic drug driving control program based on this.First,the study established a highperformance liquid chromatography tandem mass spectrometry(LC-MS / MS)detection method for saliva KET and its metabolite Norketamine(Norketamine,NKET),and used it as a confirmatory analysis method;second,through animal experiments Metabolism of KET in saliva and blood,simultaneous quantitative analysis of blood and saliva of clinical KET abusers,find out the correlation between saliva KET,NKET concentration and blood drug metabolism,and explore the feasibility of saliva as a test material Under the situation of simulating the detection of drug driving on the pavement,build and practice a systematic test plan using saliva as the test material,from initial screening to certification.The main research contents and results of the paper are as follows:1.Establish qualitative and quantitative analysis methods of KET and NKET in biological samples.By optimizing the pretreatment conditions,chromatographic conditions and mass spectrometry conditions of saliva,the established high-performance liquid chromatography tandem mass spectrometry(LC-MS / MS)detection method of KET and NKET in the test material,and from the linear range of quantification,detection limit,limit of quantification,Specificity,recovery rate,stability,clinical sample verification,etc.to verify the established method.The results showed that the detection limits of KET and NKET were both 0.01 ng / m L,and the limits of quantification were both 0.10 ng / m L.The intra-day precision of KET is 2.80% and 2.80%,the inter-day precision is 1.61% and 1.09%,the recovery rate is 97.64% ～ 100.30%,the matrix interference is-1.80% ～ 1.24%;NKET intra-day precision is respectively 4.11% and 1.40%,the day precision is 1.78% and 1.04%,the recovery rate is 98.06% ～ 100.58%,and the matrix interference is 1.70% ～ 3.54%.And 10 cases of clinical saliva samples were tested.The results show that the method has high sensitivity,short analysis time and low sample consumption,and is suitable for the inspection and analysis of large-scale samples.2.Establish a rabbit drug metabolism model of KET,collect and analyze clinical saliva and blood samples of patients with KET abuse,and analyze the correlation between saliva KET and NKET concentrations and blood drug metabolism to explore the feasibility of saliva test materials instead of blood test materials.The salivary drug concentration of KET and NKET in female rabbits had a good positive correlation with the blood concentration within 24 hours,but the ratio of the saliva drug concentration of KET and NKET to the blood concentration(S / P value)at different time points The performance is quite different;from the clinical sample research,it is found that the correlation between human saliva KET,NKET and blood drug concentration is similar to the conclusion of the animal model.First,the KET and NKET concentrations in 21 clinical saliva samples have a good positive correlation with blood drug concentration.Among them,KET’s Pearson correlation coefficient is 0.910;NKET’s Pearson correlation coefficient is 0.868;but the study found that the S / P value variation coefficient of different populations is larger.Because of the positive correlation between KET and NKET concentrations in saliva and blood drug concentration,ketamine in saliva can characterize ketamine in blood to a certain extent.Therefore,it is theoretically feasible for saliva test materials to replace blood test materials.However,because the specific quantitative relationship of S / P is difficult to determine,it is necessary to consider carefully when using saliva test materials to infer blood drug concentration.3.To study the requirements of roadside drug driving test management and control,combined with two rapid screening methods,built and practiced two saliva KET testing programs from initial screening to laboratory confirmation.One scheme is the colloidal gold method + LC-MS / MS method.This scheme is suitable for the control of road drug driving that requires the detection results on the spot.Law enforcement personnel use the colloidal gold method to send samples with positive screening results for experimental further testing and verification;another scheme is the ELISA method + LC-MS / MS method.This scheme is suitable for large-scale road sample screening based on the premise that all potential drug drivers are not missed.Law enforcement personnel send the collected saliva samples to the testing laboratory.The laboratory uses ELISA to screen the samples and further test and verify the positive and suspicious samples.Both methods can be applied to the testing of drug driving certification.Effectively make up for the gap in the detection scheme in the current drug driving control system in China.