Morphine In Urine Research Articles

Sensitive and simple determination of zwitterionic morphine in human urine based on liquid-liquid micro-extraction coupled with surface-enhanced Raman spectroscopy

Morphine, a kind of illicit drugs, is also one of the main heroin metabolites. In consideration of a noninvasive way to monitor and identify drug abuse during forensic cases, the urine samples are usually detected. Here, colloidal gold nanorods (Au NRs) were introduced to act as active substrate, because of the strong optical extinction and spectral tunability of the longitudinal surface plasmon resonance (SPR). Thus, well surface-enhanced Raman spectra of morphine even at low concentrations could be obtained by portable Raman spectrometer. For the complex matrix environment of urine, liquid-liquid micro-extraction (LLME), a simple and inexpensive pretreatment, was employed to avoid the interferences. And then, the coupled surface-enhanced Raman spectroscopy (SERS) can give full play to the advantages of high sensitivity and unique spectroscopic fingerprint. According to the zwitterionic structure and physicochemical parameters of morphine molecules, the pH value of urine sample was adjusted to about 9 by buffer solution (KOH/NaB4O7) and the mixture of chloroform and isopropyl alcohol (V/V=9:1) was chosen as extractant. Moreover, such pretreatment was proved to be appropriate for separation and concentration of morphine from urine. The developed LLME-SERS method could provide a detection limit less than 1 ppm in the human urine environment and the whole process of detection just needed take 5–6 min. What's more, the results of urine samples from heroin users exhibited application value of the proposed technique. The excellent performance makes it promising to become a rapid, reliable, and on-spot analyzer, especially for public safety and healthcare.

Superiority of Buprenorphine over Suboxone in Preventing Addiction Relapse in Opioid Addicts under Maintenance Therapy: A Double-Blind Clinical Trial

Background: In maintenance therapy for opioid addiction, to reduce the risk of buprenorphine (BUP) abuse, the combination of BUP and naloxone (NX) has been developed and is commercially available as suboxone (BUP/NX). This study was designed to compare addiction relapse frequency in patients receiving BUP and BUP/NX as maintenance therapy. Methods: In this double-blind clinical trial with cross over design, 100 opioid abusers were randomly assigned to two treatment groups to receive either BUP or BUP/NX. After three months, without a time-out period, subjects undertook treatment with the other drug. The subjects were screened weekly for urinary morphine. Results: In each of the study arms, when the patients were given BUP/NX, the number of relapses was significantly higher compared to when they received BUP (0.13±0.24 vs. 0.04±0.09, P = 0.001). If participants� age was taken into account, the number of relapses was significantly higher when BUP/NX was given in age groups of 31 to 40 years and over 50 years (P < 0.05). The length of addiction had also a significant impact on the number of relapses, i.e., patients with over 10-year history of addiction had higher number of relapses if they were given BUP/NX compared with BUP (P < 0.05). Conclusion:BUP seems to be more effective than BUP/NX in preventing addiction relapse in opioid abusers under maintenance treatment.

Family Related Factors and Concurrent Heroin Use in Methadone Maintenance Treatment in China

ABSTRACTBackground: The use of heroin during Methadone Maintenance Treatment (MMT) is a challenging problem that contributes to poor treatment outcomes. Families may play an important role in addressing concurrent heroin use during MMT, especially in collectivist societies such as China. Objectives: In this study, we explored the relationship between family-related factors and concurrent heroin use during MMT in China. Methods: This study was conducted at 68 MMT clinics in five provinces of China. There were 2,446 MMT clients in the analysis. Demographic information, MMT dosage, family members' heroin use status, family support of MMT, family problem, and self-reported heroin use were collected in a cross-sectional survey. The most recent urinalysis of opiate use was obtained from clinical records. Results: Of the 2,446 participants, 533 (21.79%) self-reported heroin use in the previous seven days or had a positive urine morphine test result in the clinic record. Participants whose family member[s] used heroin were 1.59 times (95% CI: 1.17, 2.15) more likely to use concurrently during treatment. Those with family members who totally support them on the MMT were less likely to use (AOR: 0.75, 95% CI: 0.60, 0.94). Having more family problems was positively associated with concurrent heroin use (AOR: 2.01, 95% CI: 1.03, 3.93). Conclusions: The results highlight the importance of the family's role in concurrent heroin use during MMT programs. The study's findings may have implications for family-based interventions that address concurrent heroin use.

APBB2 is associated with amphetamine use and plasma beta-amyloids in patients receiving methadone maintenance treatment

APBB2, amyloid beta (A4) precursor protein-binding family B member 2, has been reported to be associated with opioid dependence. In this study, we reported the first time that the genetic variants in the APBB2 gene were associated with use of amphetamine in opioid dependent patients undergoing methadone maintenance treatment (MMT). 344 heroin-dependent patients undergoing MMT were recruited and assessed for use of amphetamine and opioids by urine toxicology, withdrawal severity, and side effects. DNAs were genome-widely genotyped for all patients. Single nucleotide polymorphisms (SNPs) in APBB2 were selected for association analyses for methadone treatment responses. Gene expression levels of APBB2 were measured by real-time polymerase chain reaction (PCR) in the EBV-transformed lymphoblastoids from patients. MMT patients who used amphetamine showed a significantly higher percentage of positive results in the urine morphine test (P=0.005), and insomnia (P=0.018). In single locus association analyses, SNPs rs3935357 and rs4861075 located at intron 6 were significantly associated with amphetamine use in both genotype and allele type (general linear model (GLM), P=0.0003, and 0.0002 for genotype, and 0.0003, and 0.002 for allele type, respectively). The major allele type carriers had twice risk of amphetamine use compared to the minor allele type carriers. Subjects with the TT genotype of rs4861075 showed significantly higher levels of APBB2 gene expression in both total (P=0.02) and long-form (P=0.037) than those with CC genotype. Detailed mechanisms underlying the association of APBB2 with amphetamine use and level of plasma amyloid beta in MMT patients require further investigation.

A gold nanoparticle-single-chain fragment variable antibody as an immunoprobe for rapid detection of morphine by dipstick.

Gold nanoparticle (AuNP)-based optical assays are of significant interest since the molecular phenomenon can be examined easily with change in the color of AuNPs. Herein, we report the development of a dipstick using a AuNP-labeled single-chain fragment variable (scFv) antibody for the detection of morphine. The scFv antibodies for morphine were developed using phage display-based antibody library. Immunoglobulin variable regions of heavy (VH)- and light (VL)-chain genes were connected via a glycine–serine linker isolated from murine immune repertoire and cloned into the expression vector pIT2. The scFv was produced in Escherichia coli HB2151, yielding a functional protein with a molecular weight of approximately 32 kDa. The morphine scFv was labeled with gold nanoparticles and used as an optical immunoprobe in a dipstick. The competitive dipstick assay characterized the ability of the scFv antibody to recognize free morphine. The detection range was 1–1000 ng mL−1 with a limit of detection (LOD) of 5 ng mL−1 under optimal conditions, and the IC50 value was 14 ng mL−1 for morphine. The developed optical dipstick kit of scFv antibody was capable of specifically binding to free morphine and its analogs in a solution in less than 5 min and could be useful for on-site screening of a real sample in blood, urine, and saliva.

Morphological and histopathological heart changes in autopsies of heroin abusers

Backround/Aim. Heroin is a semisynthetic opioid that may cause morphological and histopathological changes in heart: ventricular hypertrophy, myocardial fibrosis, hypertrophy of cardiomyocytes, myofibrils contraction band necrosis, loss of myocytes nuclei and cross-striation, perivascular bleeding, inflammatory cells infiltrate. The aim of the study was to show morphological and histopathological heart changes in autopsies of the long-time heroin abusers with positive toxicological analysis for 6-monoacetylmorphine (6-MAM) and morphine in blood and urine. Methods. Retrospective study was done at the Institute of Pathology and Forensic of the Medicine Military Medical Academy in Belgrade between 2010 and 2014 and included forensic autopsies of 27 examinees aged between 18 and 60. Heart ventricles thicknesses was analysed and histopathological myocard findings from processed material stained by hematoxyline-eosine (H&amp;E) and trichrome stains (Masson) were examined. 6-MAM and morphine concentration in blood and urine using high-performance liquid chromatography coupled with photodiode (HPLC-PDA) and ultraviolet (UV) detector were analysed. Results. Heart ventricles thickness was increased in all persons (27/27; 100%) left 1.74 ? 0.17 cm and right 0.6 ? 0.09 cm. Myocardial fibrosis affected 27/27 (100%) of the examined persons including perivascular one in 24/27 (88.89%) and interstitial focal fibrosis in 3/27 (11.11%); hypertrophy of cardiomyocytes was present in 22/27 (81.48%); myofibril contraction band necrosis in 22/27 (81.48%); loss of myocytes nuclei and cross-striation in 10/27 (37.04%); fresh perivascular bleeding in 23/27 (85.19%); focal inflammatory cells infiltrate in 14/27 (51.85%). In toxicological findings, in 27/27 (100%), 6-MAM and morphine were found in urine. Both 6-MAM and morphine in blood were found in 3/27 (11.11%) and only morphine in blood in 16/27 (59.26%) persons subjected to an autopsy. Conclusion. Our results indicate both morphological (left and right ventricle hypertrophy) and histopathological heart changes (myocardial fibrosis, hypertrophy of cardiomyocytes, contraction-band necrosis, loss of myocytes nuclei and cross-striation, fresh perivascular bleeding and focal inflammatory infiltrate) in long-term heroin obusers. These changes are non-specific and could be caused either by long-term heroin abuse or by other factors. Having in mind a lack of medical histories of examined we could not exclude other factors besides long-term heroin abuse as cause of heart changes.

Quality of Life and Substance Use over 1-year among Buprenorphine-treated Heroin Use Disorder Patients in Taiwan

Objective: Owing to the higher cost of Buprenorphine Maintenance Treatment (BMT) and harm reduction policy in Taiwan, the percentage of Heroin Use Disorder (HUD) patients receiving BMT is much lower than that of receiving methadone maintenance treatment in Taiwan despite the lower addictive, milder withdrawal symptoms and safer characteristics of BMT. As a consequence, the study on treatment outcomes of BMT in Taiwanese HUD patients is relatively limited. The purpose of this prospective, 1-year observational study was to investigate the effects of BMT on the quality of life, self-efficacy, substance use, and retention rate among HUD patients in Taiwan.Methods: Forty-two HUD patients aged between 20 and 60 years old without serious psychiatric illness were recruited from five medical centers and were given BMT for 12 months. Efficacy assessments using the Taiwan version of Brief Version of World Health Organization Quality of Life Instrument (WHOQoL-BREF-TW) and 8-item Drug-Taking Confidence Questionnaire (DTCQ-8), as well as urine drug test were performed at baseline and every 3 months thereafter.Results: Of the 25 subjects who comprised the Intent-To-Treat (ITT) population for efficacy analyses, significant improvements were seen in the psychological and social relation domains of the WHOQoL-BREF at 9 months. Significant improvements also observed in the DTCQ score under both pleasant and unpleasant emotions, testing self-control, and friend’s incitement circumstances throughout the study period. The 12 months retention rate was 36%. The positive rates of urine morphine maintained low during the study.Conclusion: This study indicated the long-term efficacy of BMT on the quality of life, self-efficacy, and reduction of illicit drug use in Taiwanese heroin use disorder patients who maintained in the BMT treatment.

Diagnosis of Heroin Overdose in an 8-Year-Old Boy: Reliable Contribution of Toxicological Investigations.

Toxicological investigations are often required by clinicians in comatose patients with suspected poisoning. However, the usefulness of toxicological analyses to support a diagnosis of acute poisoning is debated among clinicians and the interpretation of laboratory tests is challenging given the wide diversity of analytical techniques available. We report the case of an 8-year-old boy who was admitted to an intensive care unit with severe respiratory depression and neurological impairment. In order to formulate appropriate hypothesizes about the diagnosis and circumstances of intoxication, clinicians consulted toxicologists for a comprehensive toxicological screening. Routine blood immunoassays were negative for common toxicants but urine tests were positive for opiates. A general unknown screening using liquid and gas chromatography combined with mass spectrometry detection confirmed the presence of morphine, codeine and related glucuronides metabolites in plasma and urine. Subsequently, morphine and codeine were quantified in plasma samples by online-SPE-LC-MS-MS. In addition, analyses performed with GC-MS and LC-MSn identified compounds used as markers when profiling illicit heroin, namely noscapine, dextromethorphan and codeine. In conjunction with the patient's history, clinical picture and circumstances of intoxication, toxicological findings strongly suggested an acute pediatric opioid overdose as a collateral damage of parental heroin abuse in the home. This case highlights the significant contribution of toxicological investigations in sensitive legal cases and the critical role of communications between clinicians and toxicologists.

The Pharmacokinetics of Morphine and Codeine in Human Plasma and Urine after Oral Administration of Qiangli Pipa Syrup.

Papaveris pericarpium, a natural source of morphine and codeine, is the principal active component in many antitussive traditional Chinese medicines. We herein report the first PK study of papaveris pericarpium in human plasma and urine following oral administration of single (15, 30, 60mL) and multiple dose (15mL) of Qiangli Pipa Syrup (MOR 0.1mg/mL, COD 0.028mg/mL) by monitoring morphine and codeine using a HPLC-MS/MS method. Their Tmax and t1/2 values are independent of dosages, while the AUC0-t linearly increased with higher dosages, indicating linear PK characteristics. AUC0-t increased obviously after multiple doses, indicating possible risk of accumulative toxicity. Urine studies suggested risks of positive opiate drug tests with a cutoff of 300ng/mL, which lasted 6-14h at different doses. These results provide important information for clinical safety, efficacy and rational drug use of Qiangli Pipa Syrup and also guide the related judicial expertise of its administration.

Codeine influences the serum and urinary profile of endogenous androgens but does not interact with the excretion rate of administered testosterone.

Today's doping tests involve longitudinal monitoring of urinary steroids including the testosterone glucuronide and epitestosterone glucuronide ratio (T/E) in an Athlete Biological Passport (ABP). The aim of this study was to investigate the possible influence of short-term use of codeine on the urinary excretion of androgen metabolites included in the steroidal module of the passport prior to and after the co-administration with testosterone. The study was designed as an open study with the subjects being their own control. Fifteen healthy male volunteers received therapeutic doses of codeine (Kodein Meda) for 6 days. On Day 3, 500 mg or 125mg of testosterone enanthate (Testoviron®-Depot) was administered. Spot urine samples were collected for 17 days, and blood samples were collected at baseline, 3, 6, and 14days after codeine intake. The circulatory concentration of total testosterone decreased significantly by 20% after 3days' use of codeine (p=0.0002) and an atypical ABP result was noted in one of the subjects. On the other hand, the concomitant use of codeine and testosterone did not affect the elevated urinary T/E ratio. In 75% of the individuals, the concentration of urinary morphine (a metabolite of codeine) was above the decision limit for morphine. One of the participants displayed a morphine/codeine ratio of 1.7 after codeine treatment, indicative of morphine abuse. In conclusion, our study shows that codeine interferes with the endogenous testosterone concentration. As a result, the urinary steroid profile may lead to atypical findings in the doping test.

Simultaneous determination of morphine-6-d-glucuronide, morphine-3-d-glucuronide and morphine in human plasma and urine by ultra-performance liquid chromatography-tandem mass spectrometry: Application to M6G injection pharmacokinetic study.

A robust ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the determination of morphine-6-d-glucuronide (M6G), morphine-3-d-glucuronide (M3G) and morphine (MOR) in human plasma and urine has been developed and validated. The analytes of interest were extracted from plasma by protein precipitation. The urine sample was prepared by dilution. Both plasma and urine samples were chromatographed on an Acquity UPLC HSS T3 column using gradient elution. Detection was performed on a Xevo TQ-S tandem mass spectrometer in multiple reaction monitoring mode using positive electrospray ionization. Matrix interferences were not observed at the retention time of the analytes and internal standard, naloxone-D5. The lower limits of quantitation of plasma and urine were 2/0.5/0.5 and 20/4/2 ng/mL for M6G/M3G/MOR, respectively. Calibration curves were linear over the concentration ranges of 2-2000/0.5-500/0.5-500 and 20-20,000/4-4000/2-2000 ng/mL for M6G/M3G/MOR in plasma and urine samples, respectively. The precision was <7.14% and the accuracy was within 85-115%. Furthermore, stability of the analytes at various conditions, dilution integrity, extraction recovery and matrix effect were assessed. Finally, this quantitative method was successfully applied to the pharmacokinetic study of M6G injection in Chinese noncancer pain patients.

Correlation between insufficient methadone dosage and morphine positive urine on drop out of treatment in patients with access to methadone maintenance treatment

Objective: To estimate the incidence of drop out of treatment in patients with access to methadone maintenance treatment and explore the correlation and interaction between insufficient methadone dosage and morphine positive urine on the drop out in Guangxi Zhuang Autonomous Region. Methods: Face to face interview was conducted in 1 031 patients at 3 methadone maintenance treatment clinics in Guangxi. Results: The study included 1 031 participants, 40.6% of them (419/1 031) had stopped treatment. The drop out rates in urine morphine positive group and methadone dosage<100 mg/d group were 57.6% (99/172) and 37.4% (347/929) respectively, higher than those in urine morphine negative group and methadone dosage ≥100 mg/d group (42.3%, 363/859, and 26.5%, 27/102). Orderly logistic regression analysis results showed that after adjusted factors, such as gender, age, marital status, ethnic group, patients who received a dosage less than 100 mg/day (OR=3.05, 95%CI: 1.84-5.06) and had morphine positive urine (OR=2.25, 95%CI: 1.59-3.19) were more likely to drop out of the treatment. Interaction analysis showed that dosage less than 100 mg/d and morphine positive urine during treatment had additive interaction (RERI=256.46, AP=0.87, S= 8.05) and multiplication interaction (OR=2.45, 95%CI: 1.71-3.49). Conclusion: Insufficient dosage and morphine positive urine were significantly correlated with drop out of treatment in patients with access to methadone maintenance treatment.

Positive percentages of urine morphine tests among methadone maintenance treatment clients with HIV/AIDS: a 12-month follow-up study in Guangdong Province, China

ObjectiveWe aimed to assess the positive percentages of urine morphine tests and correlates among methadone maintenance treatment (MMT) clients with HIV/AIDS in Guangdong, China.SettingFourteen MMT clinics located in nine cities...

Pharmacogenomics study on cadherin 2 network with regard to HIV infection and methadone treatment outcome.

Heroin dependent patients have a high incidence of HIV infection. In contrast to the gene expression method, we developed a systemic correlation analysis method built upon the results of pharmacogenomics study in a methadone maintenance treatment (MMT) cohort consisting of 344 Taiwanese heroin dependent patients. We identified genetic variants and their encoding proteins that may be involved with HIV infection and MMT treatment outcome. Cadherin 2 (CDH2) genetic determinants were identified through the genome-wide pharmacogenomic study. We found significant correlations among HIV infection status, plasma levels of CDH2, cytokine IL-7, ADAM10, and the treatment responses to methadone. Two single nucleotide polymorphisms located within CDH2 gene showed associations with blood pressure and plasma CDH2 concentration. Plasma concentration of CDH2 showed correlations with the level of cytokine IL-7, status of HIV infection, and urine morphine test result. Plasma level of IL-7 was correlated with corrected QT interval (QTc) and gooseflesh skin withdrawal symptom score, while level of ADAM10 was correlated with plasma concentrations of vitamin D metabolite, nicotine metabolite, and R-methadone. The results suggest a novel network involving HIV infection and methadone treatment outcome.

Comparison Between Urinalysis Results and Self-Reported Heroin Use Among Patients Undergoing Methadone Maintenance Treatment in China

ABSTRACTBackground: The validity and concordance of two main measures of drug use behavior, self-report and urinalysis, has long been discussed. More understanding is needed about the underlying factors associated with discordance between these two methods. Objectives: Describe the pattern and associated factors of discordance between self-reported heroin use and the urinalysis results of opiate use among methadone maintenance therapy (MMT) patients in China. Methods: A total of 2,448 MMT patients from 68 clinics in five provinces of China participated in a survey, which collected information on demographics, drug use and MMT-related factors, depressive symptoms, and drug avoidance self-efficacy. The most recent urine morphine test result was obtained from medical records and compared with self-reported heroin use. Participants who had urinalysis within 14 days of the survey were included in the analysis. Results: Among the 1,092 participants, 70 (6.4%) self-reported heroin use and 195 (17.9%) had positive urinalysis results. The over-reporters group had significantly higher education, and the under-reporters had significantly higher level of drug-avoidance self-efficacy and lower level of depressive symptoms. Among the participants who either self-reported heroin use or had positive urinalysis results, being young, having higher education, and having lower level of depressive symptoms were associated with discordance between self-reports and urinalysis results. Conclusion: The combination of both measures in assessing drug use behavior seems necessary. The validity of self-report should be considered differently based on demographic and psychosocial characteristics.

Medication Adherence in Patients Undergoing Methadone Maintenance Treatment in Xi'an, China.

To identify medication adherence and its influencing factors among patients of 14 methadone maintenance treatment (MMT) clinics in Xi'an, China. Data were obtained from the National AIDS Information System-Community Methadone Maintenance Treatment. All patients registered in the system were not permitted to take methadone at home without professionals' supervision. Medication adherence was assessed using categorical (ie, dropout or retained) and continuous (ie, treatment time, methadone use time, and percentage of methadone use days) variables. Percentages of methadone use days of >90%, 50% to 90%, or <50% indicated good, moderate, and poor adherence, respectively. Multivariate Cox stepwise regression analysis was used to identify the influencing factors. Of the 10,398 patients, 52.2% had dropped out of MMT by December 31, 2013, whereas only 11.8% regularly visited the clinic for daily methadone (ie, >90% methadone use days) during a certain period. Protective factors were older age (>30 years); female sex; having no contact with peer drug users over the past month; no needle-sharing experience; a negative initial morphine urine test; and a higher average daily methadone dose (>20 mg) (P < 0.05). Risk factors were answering "others" for marital status; being employed; having a lack of stable income; not living with family; answering "others" for drug use type; frequently engaging in unauthorized drug use during MMT; no readmission; long travel times to the MMT clinic (>30 minutes); having no convenient MMT service time; and being dissatisfied with MMT service (P < 0.05). Based on our findings, multimodal intervention and management programs can be developed to improve poor medication adherence among the MMT patient population.

Detection of Methamphetamine and Morphine in Urine and Saliva Using Excitation–Emission Matrix Fluorescence and a Second-Order Calibration Algorithm

A new method was developed to determine the methamphetamine and morphine concentrations in urine and saliva based on excitation–emission matrix fluorescence coupled to a second-order calibration algorithm. In the case of single-drug abuse, the results showed that the average recoveries of methamphetamine and morphine were 95.3 and 96.7% in urine samples, respectively, and 98.1 and 106.2% in saliva samples, respectively. The relative errors were all below 5%. The simultaneous determination of methamphetamine and morphine in urine using two second-order algorithms was also investigated. Satisfactory results were obtained with a self-weighted alternating trilinear decomposition algorithm. The root-mean-square errors of the predictions were 0.540 and 0.0382 μg/mL for methamphetamine and morphine, respectively. The limits of detection of the proposed methods were very low and sufficient for studying methamphetamine and morphine in urine.

Evaluation of Common Adulterants in Morphine Urine Drug Screening

Background: Manipulating drug screens, changing sample quality, and confounding test results are common practices seen among opium users. These practices are intended to deceive drug screening systems using a wide variety of commonly available chemical compounds such as lemon juice and fertilizer. These actions create a challenge for addiction screening reference labora- tories. Objectives: Therefore, we studied the eects of lemon juice, nitrogen-based chemical fertilizers, carbon copy paper, and urine sam- ple dilution on morphine urine screening test results. PatientsandMethods: We studied the urine samples of all individuals who were referred to the drug laboratory at Sabzevar public health center from August 2014 to August 2015. Out of 8012 urine samples tested in one year, morphine metabolites were identified in 493 (6%) of the urine samples using a rapid morphine slide test and thin-layer chromatography. Next, 180 confirmed positive samples were randomly assigned to three groups of 60 samples each, to which were added either carbon paper, lemon juice, or nitrogen-based fertilizer. Results: The test results showed that in the carbon paper and lemon juice groups, 59 samples tested positive for morphine, and one sample was distorted. In the fertilizer group, 57 samples were positive and 3 were distorted. None of the groups showed negative test results. Moreover, negative results were not seen when testing standard samples that had been adulterated in the test kit's sensitivity range and higher. However, fertilizer and lemon juice adulteration, but not carbon paper, can lead to false negative results with lower morphine concentrations (150 ng/mL). Conclusions: We found that adding nitrogen-based fertilizer, lemon juice, or carbon copy paper to urine samples does not facilitate false negative results when using a rapid slide morphine urine test. However, testing samples of various standard concentrations of morphine showed that at lower morphine concentrations, adding lemon juice or fertilizer can aect the test results.

Ultrasonic-assisted magnetic solid phase extraction of morphine in urine samples by new imprinted polymer-supported on MWCNT-Fe3O4-NPs: Central composite design optimization

Multiwalled carbon nanotubes (MWCNTs) were magnetized with Fe3O4 nanoparticles (MWCNTs-Fe3O4-NPs) and subsequently coated by vinyl end groups (Vinyltrimethoxysilane). MWCNT-Fe3O4-NPs were used as support for a new morphine (MO) molecularly imprinted polymer (MWCNT-Fe3O4-NPs@MO-MIP) by surface imprinting polymerization method. The MWCNT-Fe3O4-NPs@MO-MIP was characterized by FTIR, VSM and SEM techniques and successfully used for determination of MO. Ultrasonic-assisted magnetic solid phase extraction followed by UV–vis spectrophotometer (UAMSPE-UV–vis) was investigated for MWCNT-Fe3O4-NPs@MO-MIP and compared with non-imprinted polymer (NIP) using batch method. Central composite design under response surface methodology was used for the evaluation of the effect of variables, individually, as well as their possible interaction effects on the adsorption process. The variables such as sonication time, MWCNT-Fe3O4-NPs@MO-MIP mass, initial concentration of MO and pH were investigated in this study. At optimum experimental conditions, UAMSPE-UV–vis method was exhibited a linear range of 0.8–8.7mgL−1 of the MO concentration with a detection limit of 0.18mgL−1. The relative standard deviation for the analyte was found to be lower than 2.32%. The MWCNT-Fe3O4-NPs@MO-MIP adsorption capacity was found to be 37.01mgg−1. The enrichment and preconcentration factors were found to be 107.01 and 98.21, respectively. The developed method was finally applied successfully to the determination of MO in urine and wastewater samples with the recoveries ranged from 96.40 to 105.6%.

Homicidal Poisoning of Heroin and Estazolam: Autopsy and Pathological Findings, Toxicological Analysis

We reported an unusual homicidal case in which a 40-year-old woman was deceived into drinking a cup of milk that had 72 tablets of estazolam (2mg/tablet) dissolved in, and then being injected heroin aqueous solution on the right deltoid region by the criminal. At autopsy, pinpoint pupils and a new injection site on the right deltoid region were found. The pathologic pictures showed multiple patchy hemorrhages and considerable amounts of foreign amorphous substance with yellow appearance at the injection site. Some double refracting crystals with the forms of Maltese cross, acicular, rhomb or irregular were found by polarizing microscope, which may result from the diluent in heroin such as starch. Toxicological qualitative analysis by gas chromatography mass spectrometry (GC-MS) demonstrated the presence of benzodiazepine and morphine in blood and urine, and heroin in the injector left at scene. Quantitative analysis was also performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS), providing the data on distribution of 6-monoacetylmorphine, morphine and estazolam in the woman’s body. And the cause of death was determined to polydrug heroin-related deaths due to the combined poisoning of heroin and estazolam. It taught a lesson that the determination of other drugs, particularly central nervous system depressants in heroin poisoning were quite important in forensic expertise.