Detection Of Ten Antipsychotic Drugs Based On UHPLC-Q-TOF-MS/MS And Its Preliminary Application In Human Hair And Nails

Background: In recent years,the use of hair to detect heavy metals and drug abuse has been increaseding significantly in the field of forensic toxicology and toxicological analysis.Compared with traditional matrices(e.g.,blood,urine)such as hematuria,hair and fingers(toes),keratinizing matrices including hair and fingers(toes)have unique advantages such as wide detection window,convenient material acquisition,non-invasive,convenient storage and transportation,and long detection time window.Given that patients with mental disorders often require long-term use of antipsychotics,such as risperidone,olanzapine,and clozapine,their metabolic and distribution characteristics of the drugs in these patients are similar to those of some with drug s of abuse.A review of the literature on using matrix to detect antipsychotics since 2010 shows that this trialthis field is still in its preliminary stage: there are relatively only few studies and applications,most of which involved only one or a small number of drugs.In addition,most existing studies used standard substances to establish detection methods and then apply applied them to a single or multiple actual samples for verification,lacking dose and other relevant clinical information(e.g.,dosage)and systematic researchinvestigation.In this study,we validated a method based on detection method for drug abuse to detect antipsychotic drugs in patients with mental disorders using hair and nail samples,in order to provide objective evidence for judging whether patients with mental disorders had taken antipsychotic drugs before prior to the occurrence of violent attack(e.g.,hurting,killing)wounding,killing or detention.And toand inferring the type,dose and duration of the identified person taking antipsychotic drugs,as well as the type and severity of the mental disorders according to the sample test results.Methods: In this study,we explored and optimized the method of detection of commonly used antipsychotic drugs(olanzapine,chlorpromazine,clozapine,perphenazine,sulpiride,haloperidol,amisulpride,quetiapine,risperidone,aripiprazole)using human hair,nail and blood simultaneously via application of Ultra High Performance Liquid Chromatography-Quadrupole-Time of Flight-Tandem Mass Spectrometry(UHPLC-Q-TOF-MS/MS)to simultaneously detected the pathway system of commonly used antipsychotic drugs(olanzapine,chlorpromazine,clozapine,perphenazine,sulpiride,haloperidol,amisulpride,quetiapine,risperidone,aripiprazole)using human hair,nail and blood.And apply applied the method to the blood,hair and nail samples from 54 patients who received fixed doses of antipsychotic drugs in clinical settings,and 2 cases subjects of who were under the psychiatric evaluation of mentalafter homicide in our forensic setting.In this study,blood samples were treated by acetonitrile protein precipitation method,hair and fingernail samples were washed with methanol,water and methanol in turn,then smashed with liquid nitrogen lapping machine and placed in methanol solution at 38℃ for 18 h.The protein precipitate and extract were filtered by 0.22 um membrane and analyzed in UHPLC-Q-TOF-MS/MS system.Chromatographic analysis system using Agilent Poroshell 120 EC-C18 2.7um(3.0 * 150 mm)column;The mobile phase consisted of 5m M ammonium acetate / 0.1% formic acid aqueous solution and acetonitrile.Multiple Reaction Monitoring(MRM)was used to determine the positive ion mode of electrospray ion source.Results:(1)The chromatographic peak shapes of all analytes in this method were performed well and there was no endogenous or exogenous interference;The limits of quantitation were 0.01-0.005 ng/ml in blood,0.01-0.005 ng/mg in hair and 0.005ng/mg in fingernails.The accuracy in different substrates was(blood: 96.0% ~ 101.6%;Hair: 97.5% ~ 102.5%;Nails: 97.6% ~ 101.7%);The Relative Standard Deviation(RSD)of precision of each analyte in different substrates was lower than 5%.Extraction rate: blood: 78.1% ~ 107.4%,hair: 73.9% ~ 93.5%,fingernails: 75.2% ~ 90.5%;Matrix effect was-36.5%-17.1% in blood,-20.6%-8.9% in hair,and-24.3%-11.3% in nails.Stability of samples within 72 h of injector: blood: 91.4% ~ 99.8%;Hair: 88.1% ~ 105.3%;Nails: 86.1% ~ 102.6%.(2)Sample test results: In this study,blood,hair and nail samples from 54 patients who received clinical fixed-dose drug therapy were analyzed,and longer hair samples(6-20cm)provided by 10 female patients were segmented.The study found that the detection range of blood drug concentration of all drugs was within the recommended range of AGNP psychiatric drug surveillance consensus guidelines,and the concentration range of drug concentration in hair was similar to the data in the published literature,while the concentration of antipsychotic drugs in fingernails was rarely studied and corresponding detection data were lacking.The concentrations of some drugs(chlorpromazine,sulpiride,aminosulpiride,risperidone and aripiprazole)in fingernails were reported for the first time in this study.(3)By comparing the relationship between drug concentration and dose in different substrates,it was found that there was significant correlation between clozapine and quetiapine dose and biological matrix(blood,hair,fingernail),among which there was extremely significant correlation between drug dose and blood concentration(clozapine: r =0.7901,P <0.0001;Quetiapine: r=0.8325,P=0.0054);Comparison of drug concentrations between different substrates showed that there was a significant correlation between drug concentrations in blood and hair of five drugs(olanzapine: r=0.5924,P=0.0096;Clozapine: r=0.6624,P=0.004;Quetiapine: r=0.7178,P=0.0294;Risperidone: r=0.6909,P=0.0005;Aripiprazole: r=0.6227,P=0.0408).(4)Hair samples segment detection: in addition to the unstable nature of olanzapine in the hair,the hair section cannot be detected after 3 ~ 4,the rest of the five kinds of drugs(clozapine,quetiapine,sulpiride,aripiprazole and risperidone)is relatively stable in the hair for examination,and was able to detect drugs exist any hair section,especially the patient who receive aripaiprazole treatment,even if 3 ~ 6 months after drug withdrawal,the corresponding drugs can still be detected in the hair stage.(5)It was found that risperidone was found in the hair and nail samples provided by the identified persons who had taken psychotropic drugs in the actual case.The police confirmed that the detected drugs were consistent with the drugs the patient had taken before.Conclusions:(1)the optimized detection system in this study has good selectivity and stability in the detection of standard substances and actual samples,and can accurately distinguish a variety of tested antipsychotic drugs.(2)Blood concentrations of olanzapine,clozapine,aripiprazole,quetiapine and risperidone were significantly correlated with hair concentration,so hair could be used as a supplement for clinical monitoring of antipsychotic drug concentration.(3)Hair segmentation detection showed that different drugs had different distribution characteristics in hairat fixed dose: drug concentration decreased gradually from hair root to hair end or remained basically unchanged.(4)The preliminary application in the identification of actual cases shows that this method is feasible to infer whether or not psychiatric patients take antipsychotic drugs and the types of antipsychotic drugs before the occurrence of harmful behaviors or custody.