ABSTRACT
Long-term dependence of illicit drugs impairs the function of the hypothalamic-pituitary-adrenal (HPA) axis, which regulates the secretion of endogenous steroids, cortisol, and cortisone. Thus, the present study aimed to develop a sensitive method for simultaneous determination of the multiple illicit drugs and two steroids in hair to monitor the status of illicit drug exposure and the physiological and psychological health of drug addicts. The target analytes were extracted from hair by incubation with 1 mL methanol for 24 h at 40 °C and then determined with LC-APCI+-MS/MS. The validated method showed acceptable linearity (R 2 > 0.99) in the range of 1.25-250 pg/mg for cortisol and cortisone, 2.5-125 pg/mg for heroin, 2.5-1250 pg/mg for ketamine, 2.5-5000 pg/mg for methamphetamine (MAM), 2.5-250 pg/mg for 3, 4-methylenedioxymethamphetamine (MDMA), morphine, and 6-monoacetylmorphine (6-AM). Limits of quantification were 1.6, 1.2, 1.6, 1.0, 1.4, 0.3, 2.1, and 1.2 pg/mg for cortisol, cortisone, heroin, ketamine, MAM, MDMA, morphine, and 6-AM, respectively. Method recoveries were from 90-115% for all analytes. Inter-day and intra-day coefficients of variation were within 10%. Finally, this method was successfully applied to detect the aforementioned analytes in hair among female drug addicts who self-reported to be MAM abuser, heroin abuser, ketamine abuser, and abuser of mixture drugs of MAM and heroin. MAM abusers with current MAM use showed significantly higher concentrations of cortisol, MAM, and MDMA than controls with drug withdrawal.
Subject(s)
Chromatography, Liquid , Cortisone/analysis , Hair/chemistry , Hydrocortisone/analysis , Illicit Drugs/analysis , Tandem Mass Spectrometry , Female , Humans , Limit of DetectionABSTRACT
Nowadays, zidovudine, efavirenz, lopinavir and ritonavir are important components of the second-line antiretroviral therapeutic regimen of National Free Antiretroviral Treatment Program in China. The measurement of antiretroviral drugs in hair will facilitate for the evaluation of the long-term adherence to highly active antiretroviral therapy. Nevertheless, no study illustrated simultaneous quantitation of the four drugs in hair. The study intended to develop a simple, sensitive and selective method for simultaneous quantitation of the four drugs in hair. The detection employed high liquid chromatography tandem mass spectrometry with atmospheric pressure chemical ionization in positive mode. Hair samples were incubated in methanol at 40⯰C for 16â¯h. The method showed the limit of detection at 18, 8, 5 and 6 pg/mg for zidovudine, efavirenz, lopinavir and ritonavir, respectively. The linear range (R2â¯>â¯0.99) was 36-5000â¯pg/mg for zidovudine, 16-5000â¯pg/mg for efavirenz, 10-50,000â¯pg/mg for lopinavir and 12-12,500â¯pg/mg for ritonavir. The intra-day and inter-day coefficients of variation were <15% and the recovery varied from 88.1 to 110.5%. The population analysis revealed that patients with high adherence showed significantly higher drug concentrations in hair than those with low adherence for both AZT and EFV (psâ¯<â¯0.05). There was high association in drug contents in hair between AZT and EFV or LPV and RTV (psâ¯<â¯0.05).
Subject(s)
Anti-HIV Agents/analysis , Benzoxazines/analysis , Hair/chemistry , Lopinavir/analysis , Ritonavir/analysis , Zidovudine/analysis , Adolescent , Adult , Alkynes , Anti-HIV Agents/therapeutic use , Benzoxazines/therapeutic use , Chromatography, Liquid/methods , Cyclopropanes , Female , Humans , Limit of Detection , Linear Models , Lopinavir/therapeutic use , Male , Medication Adherence/statistics & numerical data , Middle Aged , Reproducibility of Results , Ritonavir/therapeutic use , Tandem Mass Spectrometry/methods , Young Adult , Zidovudine/therapeutic useABSTRACT
The determination of the concentrations of antiretroviral drugs in hair is believed to be an important means for the assessment of the long-term adherence to highly active antiretroviral therapy. At present, the combination of tenofovir, lamivudine and nevirapine is widely used in China. However, there was no research reporting simultaneous determination of the three drugs in hair. The present study aimed to develop a sensitive method for simultaneous determination of the three drugs in 2-mg and 10-mg natural hair (Method 1 and Method 2). Hair samples were incubated in methanol at 37⯰C for 16â¯h after being rinsed with methanol twice. The analysis was performed on high performance liquid chromatography tandem mass spectrometry with electronic spray ionization in positive mode and multiple reactions monitoring. Method 1 and Method 2 showed the limits of detection at 160 and 30â¯pg/mg for tenofovir, at 5 and 6â¯pg/mg for lamivudine and at 15 and 3â¯pg/mg for nevirapine. The two methods showed good linearity with the square of correlation coefficient >0.99 at the ranges of 416-5000 and 77-5000â¯pg/mg for tenofovir, 12-5000 and 15-5000â¯pg/mg for lamivudine and 39-50,000 and 6-50,000â¯pg/mg for nevirapine. They gave intra-day and inter-day coefficient of variation <15% and the recoveries ranging from 80.6 to 122.3% and from 83.1 to 114.4%. Method 2 showed LOD and LOQ better than Method 1 for tenofovir and nevirapine and matched Method 1 for lamivudine, but there was high consistency between them in the determination of the three drugs in hair. The population analysis with Method 2 revealed that the concentrations in hair were decreased with the distance of hair segment away from the scalp for the three antiretroviral drugs.