Simutaneous Determination Of Hair Steroids And Illicit Drugs With High Performance Liquid Chromatograply-tandem Mass Spectrometry

Long-term dependence of illicit drugs impairs the functional activity of the hypothalamicpituitary-adrenal(HPA)axis where cortisol and cortisone are believed to be its biomarkers.Therefore it is significant to simultaneously detect cortisol and cortisone in drug abusers' hair for monitoring the activity of HPA axis.However it is unknown that whether previous method based on high performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS)for simultaneous detection of cortisol and cortisone is affected by illicit drugs.Consequently the present study aimed to develop a sensitive method allowing for simultaneous identification of the two steroids and the five drugs and 6-monoacetylmorphine(6AM)in hair to monitor the status of drug exposure and health of drug abusers based on HPLC-MS/MS.The process of sample pretreatment is as follows: hair sample from posterior part was decontaminated with 2 mL methanol for 2 min and dried at room temperature.20 mg non-pulverized hair was cut into pieces of 1-2 mm and then 50 ?L of 20 ng/ml mixture internal standards and 950 ?L methanol were added.The resulting samples were incubated for 24 h at 30 ?.After centrifuging for 5 minutes,800 ?L supernatant was evaporated under nitrogen flux at 40 ?.Finally,the dry residue was re-dissolved in 50 ?L of mobile phase and introduced in a vial for injection into the HPLC-APCIMS/MS.The assay method showed the acceptable linearity with the square of correlation coefficients more than 0.99 in the range of 1.25-250 pg/mg for cortisol,cortisone and 3,4-methylenedioxymethamphetamine(MDMA),2.5-125 pg/mg for heroin,1.25-1250 pg/mg for ketamine,1.25-5000 pg/mg for methamphetamine(MAM)and 2.5-250 pg/mg for morphine and 6AM.The limits of detection and quantification were 0.7 and 1.6 pg/mg for cortisol,0.5 and 1.2 pg/mg for cortisone,0.7 and 1.6 pg/mg for heroin,0.4 and 1.0 pg/mg for ketamine,0.6 and 1.4 pg/mg for MAM,0.1 and 0.3 pg/mg for MDMA,0.9 and 2.1 pg/mg for morphine,and 0.5 and 1.2 pg/mg for 6AM.Extraction recoveries were achieved at 90-115 % for all analytes.Inter-day and intra-day coefficients of variation were below 10 %.Finally the method successfully detected the concentrations of the five illicit drugs,6-monoacetylmorphine and two steroids in hair among female drug addicts.Additionally we found that MAM abusers with current MAM use showed significantly higher cortisol concentrations in the first and second 1-cm hair segments than controls with drug withdrawal.What's more,the level of methamphetamine concentration was affected by washout effect,the dosage effect and the withdrawal effect.