Sample preparation techniques

doi:10.1016/0379-0738(93)90266-D

Forensic Science International

Volume 63, Issues 1–3, December 1993, Pages 121-135

https://doi.org/10.1016/0379-0738(93)90266-D Get rights and content

Abstract

Evidentiary false positives are caused by passive exposure to drugs in the environment rather than by active use of drugs. The avoidance of such positives is essential for both hair and urine analysis. Hair analysis enjoys the advantage over urinalysis in having a number of approaches for making this distinction. These include: methylene blue staining of the hair specimen for selecting the appropriate wash solvent; application of hair digestion techniques for the complete release of chemically unaltered analytes; the determination of three diagnostic ratios from wash and digestion data; the measurement of metabolite: drug ratios; the use of cut-off levels setting the limits for passive endogenous drug exposure; reproducibility of results (including segmental analysis) with a newly collected hair specimen; and the reporting of results as either negative, positive, or contaminated. Our sample preparation procedures have been effectively applied to the analyses of nearly 200 000 specimens, i.e. to approximately one million drug analyses for cocaine, opiates, methamphetamine, phencyclidine or marijuana. On the basis of this experience we conclude that hair analysis is a safe and effective method for workplace drug testing.

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Measurement of gestational cocaine exposure: sensitivity of infants' hair, meconium, and urine
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Validity of hair analysis for detecting cocaine and heroin use among addicts
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Concordance of three measures of cocaine use in an arrestee population: hair, urine, and self-report
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Hair analysis for drugs of abuse

W.A. Baumgartner et al.

Hair analysis for drugs of abuse: forensic issues

Cited by (107)

Characterization of organic contaminants in hair for biomonitoring purposes
2024, Environment International
Biological monitoring is one way to assess human exposure to contaminants. Blood and urine are often used as biological matrices, but hair is an innovative and effective tool for quantifying more biomarkers over a wider exposure window. In order to improve the use of hair in exposure assessment, this article identifies relevant compounds in the literature to investigate hair contamination. Statistical analysis was performed to correlate the physical-chemical properties of the relevant compounds and their concentration levels in hair. Phthalates, pyrethroids and organophosphate flame retardants were chosen for further study of the interpretation of hair measurements for exposure assessment. No significant correlation was found between the average concentration levels in the literature and the physical-chemical properties of the selected compounds. This work also explores the properties of hair and the analytical process that may impact the quantification of organic contaminants in hair. The sample preparation method (sampling, storage, washing) were also studied and adaptations were suggested to improve the existing methods.
Current trends and roles of surfactants for chromatographic and electrochemical sensing
2021, TrAC - Trends in Analytical Chemistry
Citation Excerpt :
Surfactant-based sample preparation techniques are suggested to lower the organic solvent usage in laboratories and help the miniaturization. The unique chemical properties of surfactants in extraction methods enable researchers to prepare miniaturized sample preparation techniques related to solid- or liquid-phase extraction [21,28–30]. Surfactants can be used in sample preparation steps as an emulsifier, an extraction medium.
Since surfactants reduce the interfacial tension of immiscible liquids by forming micelles through hydrophobic and hydrophilic groups in their structure, they are frequently used in different fields of analytical chemistry. The use of surfactants in analytical techniques continues to increase from past to present. They are used in chromatography to improve separation efficiency and reduce the amount of organic solvent. In electrochemical techniques, they have important functions such as increasing the selectivity and sensitivity of the sensor in the supporting electrolyte or as an electrode modifying agent. The use of surfactants in these techniques, as well as in capillary electrophoresis and sample preparation, draws attention. This review summarizes the recent applications of surfactants in these analytical techniques, along with their primary usage in chromatography, capillary electrophoresis, and electrochemical analysis. Moreover, this review also describes the structural properties of surfactants and the advantages of their use in these analytical techniques.
Temporal patterns of tramadol in hair after a single dose
2020, Forensic Science International
This controlled study aimed to measure concentrations of tramadol (TRA) and its two main metabolites, N-desmethyltramadol (NDMT) and O-desmethyltramadol (ODMT), in hair following a single dose ingestion and to investigate the distribution patterns in hair by segmental analysis of hair samples taken at several sampling time points after ingestion. An oral dose (50 or 100 mg) of TRA was administered to 17 healthy volunteers. Hair samples were collected prior to drug administration and 14, 30, 60 and 120 days after ingestion. Each sample was segmented in 5 mm segments and washed. The analytes were extracted from pulverized hair by incubation in extraction media for 18 h at 37 °C. A validated UHPLC–MS/MS method was used to quantify the analytes at a LLOQ of 0.001 ng/mg. Hair segments corresponding to the time of ingestion were positive for TRA and the metabolites of each sampling time point, although neighboring segments also showed positive results. The highest concentrations for both dosage groups were observed in the proximal segment of hair collected 14 days after ingestion for all subjects: 0.061–0.95 ng TRA/mg, 0.012–0.86 ng NDMT/mg and 0.009–0.17 ng ODMT/mg (n = 16). Generally, the TRA concentration was higher than the metabolites concentrations but depended on the CYP2D6 phenotype. The metabolite to TRA ratios were stable within a subject over the sampling time points, however it varied greatly between subjects. No significant differences in hair concentrations were found between the two dosage groups at each sampling time. Several confounding factors were identified such as hair pigmentation and internal sweat. We showed that analysis of 5 mm segments improved the determination of the exposure time after a single ingestion of TRA. In addition, in the later sampling time points the analytes were spread more between segments and the total drug amount of each later sampling time point declined up to a 100% (median: 75%) due to wash out. The presented results are important additions to the sparse literature reporting single dose of psychoactive drugs in hair.
A different insight in hair analysis: Simultaneous measurement of antipsychotic drugs and metabolites in the protein and melanin fraction of hair from criminal justice patients
2020, Forensic Science International
Previous studies have postulated that four structural compartments may be differentiated in hair: surface protein domain, water-accessible protein domain, water-inaccessible protein domain, and melanin. Drugs contained in blood, sweat, sebum, and environment would be deposited in the first two domains, with primarily drugs in blood being incorporated in the latter two domains during hair synthesis. Drugs in the first two domains would be removed by washing procedures. Use of enzymatic extraction procedures and evaluation of hair for damage from harsh cosmetic treatments might help to separately identify and quantify the drugs incorporated in the second two domains.
a) Development of an UPLC-MS/MS method for the simultaneous quantification of the following 19 antipsychotic drugs and metabolites in hair: amisulpride, aripiprazole, chlorpromazine, clotiapine, clozapine, desmethylclozapine, desmethylolanzapine, haloperidol, norchlorpromazine, 7-OH-quetiapine, 9-OH-risperidone, olanzapine, pimozine, pimpamperone, quetiapine, risperidone, sertindole, sulpride, and tiapride; b) evaluation of measurement of patient adherence to prescribed medication use, c) determination of the influence of biochemical individuality effects on hair drug content, d) evaluation of relative binding of antipsychotic drugs to protein and to melanin hair structures.
Approximately 10 mg of intact hair were decontaminated with isopropanol and phosphate buffer, and then enzymatically digested overnight with dithiothreitol. After centrifugation, the supernatant digest (protein fraction) was separated from the remaining melanin hair pellet (melanin fraction). Melanin fraction was washed with water, and the drugs were extracted with dimethyl sulfoxide with ball-mill pulverization. Both fractions were purified with solid-phase cation exchange cartridges and injected in the UHPLC-MS/MS system.
Validation of the method was carried out on the protein fraction following international guidelines. The limits of quantification ranged from 1.6–40 pg/mg. The method was applied to 59 head hair samples from prisoners from an antipsychotic compliance study in the criminal justice system in US. The patients were under chlorpromazine, haloperidol, risperidone, olanzapine, or quetiapine multiple antipsychotic treatment, during incarceration. The first head hair centimeter, closest to the scalp, was analyzed. The results were evaluated in relation to the type of hair, colour, hair damage, drug melanin affinity, and prescribed dose. In general, no good correlation between the prescribed dose/concentration in hair was obtained. A wide range of antipsychotic concentrations were observed ‘dose mg/day (d); pg/mg protein fraction (A)’: chlorpromazine (d:50-400;A:<LOQ->1600) and its metabolite norchlorpromazine (A: <LOQ->1600), haloperidol (d:4-20;A: <LOQ-2902), olanzapine (d:5-20;A: <LOQ-223) and its metabolite desmethylolanzapine (A: <LOQ -136), quetiapine (d:4-400;A: <LOQ -2754) and its metabolite 7-OH-quetiapine (A: <LOQ -1448), risperidone (d:2-20;A: <LOQ->1600) and its metabolite 9-OH-quetiapine (A:<LOQ-296;B). Melanin was found to have a 2- to 85-fold higher drug-binding affinity relative to the protein fraction
This report describes a different viewpoint method of quantifying the antipsychotics in the melanin and water-inaccessible protein fraction of hair, separately. No correlation between the prescribed dose and the concentrations found in the protein and melanin domains were observed. The results show that hair melanin would have a much higher affinity for the antipsychotic medications than do hair proteins, being influenced mainly by biochemical individuality effects and less by hair colour. Future studies would be of interest using the proposed extraction method applied to hair of different colours.
Hair testing for cortisol by UPLC–MS/MS in a family: External cross-contamination from use of cortisol cream
2019, Forensic Science International
Citation Excerpt :
The distal hair of 8–12 cm from the mother also had similar or higher concentrations than the proximal 0–4 cm (a chromatogram of cortisol in hair from the mother is shown in Fig. 2). If the measured concentration of cortisol in the last aqueous wash fraction is more than 10% of the content in the relevant hair segment, the sample is interpreted as being contaminated, according to [35]. In this study, the mother’s hair cortisol levels in the last aqueous wash fraction were 12–21% of the levels in the proximal hair segment (0–4 cm), suggesting some external contamination, and below 10% of the levels in the distal hair segment (8–12 cm).
In the present study, an ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was developed, validated, and applied for measuring cortisol in human hair. Baseline levels of cortisol in hair were taken from 12 control subjects, with concentrations for adult controls (n = 8) of 1.7 to 9.1 pg/mg and a median of 4.7 pg/mg and for child controls (n = 4) of 1.1 to 7.2 pg/mg and a median of 3.1 pg/mg. However, the concentrations in the hair of two children whose mother had been applying a cortisol-containing hand cream 2–3 times per week ranged from 30 to 390 pg/mg. No external contamination was observed with the children as judged from wash water concentrations. The mother had hair cortisol concentrations of 80–220 pg/mg. External contamination was observed in her proximal hair segments (0–4 cm) but not in distal ones (8–12 cm). In an experiment, cortisol cream (1%) was applied on the fingers of a subject, who then scratched the head hair once in a while. Hair was collected 1, 5, and 30 days after exposure to the cream. The cortisol level in the hair one day after exposure was 20–186 times higher than the pre-exposure level. High levels in the wash fraction agreed with external contamination. Cortisol concentrations in the hair at 5 and 30 days after exposure were 15–38 and 9–11 times higher, respectively, than the pre-exposure levels. However, no external contamination was suggested from the wash water concentrations in the hair collected 5 and 30 days after exposure. The results showed that the externally applied cortisol had, after some time, been incorporated into the hair matrix and was not removed by a pre-analysis washing. Therefore, the use of a standard decontamination procedure prior to analysis of hair may not be able to prevent the spread of cortisol from applied hand cream within a family.
Assay for opium alkaloids
2016, Neuropathology of Drug Addictions and Substance Misuse
The use of opioids (opium alkaloids, opium, synthetic prescription opioids, and other illicit opioids) is an ever-persistent societal problem. Their controlled and limited availability in well-defined therapeutic fields has not eliminated the risk of their misuse.
Along with various societal efforts to deal with the increasing incidence of drug abuse, there is an emerging need to develop analytical methods that are convenient for the determination of these substances in biological materials of drug addicts.
In this chapter, developments in the bioanalysis of opium alkaloids and related opioids, covering current best practices in the field, are presented. Focus is placed on the state-of-the-art techniques for sample preparation and separation of mostly (ab)used opioids and their metabolites.

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^☆: Presented at the First International Meeting on Hair Analysis as a Diagnostic Tool for Drugs of Abuse Investigation, December 10–11, 1992, Genova, Italy.

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Sample preparation techniques☆

Abstract

J. Pediatrics

Radioimmunoassay of hair for determining opiate-abuse histories

J. Nucl. Med.

Detection of phencyclidine in hair

J. Forensic Sci.

Radioimmunoassay of cocaine in hair

J. Nucl. Med.

Hair analysis for drugs of abuse

J. Forensic Sci.

Hair analysis for the detection of drug use in pretrial, probation and parole populations

Fed. Probat.

Validity of hair analysis for detecting cocaine and heroin use among addicts

Int. J. Addict.

Concordance of three measures of cocaine use in an arrestee population: hair, urine, and self-report

J. Psychoactive Drugs

Radioimmunoassay of hair: a valid technique for determining maternal cocaine abuse

Subst. Abuse

Hair analysis for drugs of abuse: some forensic and policy issues

Hair analysis for drugs of abuse

Hair analysis for drugs of abuse: forensic issues