Article
To read the full-text of this research, you can request a copy directly from the authors.

Abstract

In several medico legal cases bone analysis may provide the only source of toxicological information. The present study reports the development of an UHPLC-MS/MS method for the detection and quantification of 27 drugs and pharmaceuticals in human bones. The target compounds comprise pharmaceuticals (antipsychotics and antidepressants) and some of the most important groups of drugs of abuse: opiates, cocaine, cannabinoids, amphetamines and benzodiazepines. Sample pretreatment was studied and the best results were obtained after extraction with methanol, stirring and ultra-sonication. The extract, after filtration, evaporation and reconstitution was analysed on a reversed-phase column (C18) in gradient elution over 17min. The method was found to be selective, and sensitive offering limits of detection (LOD) from 0.03 to 1.35ng/g of bone. Validation included evaluation of limit of quantification (LOQ), recovery, carry-over, matrix effect, accuracy and precision (RSD%) of the method. The method performed satisfactory in relation to established bioanalytical criteria and was therefore applied to the analysis of bone and bone marrow obtained post-mortem from chronic drug abusers, offering unambiguous identification and quantitative determination of drugs in bones from legal cases where the analysis of blood was not feasible.

No full-text available

Request Full-text Paper PDF

To read the full-text of this research,
you can request a copy directly from the authors.

... Forensic toxicology consists in the search for exogenous compounds in different matrices and tissues in order to provide additional information in a medicolegal setting [1][2][3]. Conventional specimens used in forensic toxicology analysis include blood and urine [4][5][6][7]. However, when a human body is extremely decomposed, or even skeletonized, these specimens may not be available due to their degradation during decomposition or their contamination [4][5][6]. ...
... Conventional specimens used in forensic toxicology analysis include blood and urine [4][5][6][7]. However, when a human body is extremely decomposed, or even skeletonized, these specimens may not be available due to their degradation during decomposition or their contamination [4][5][6]. Thus, alternative matrices are necessary to perform toxicological analysis [4][5][6][7]. To be suitable for toxicological analyses, these alternative matrices must remain well-preserved for a long time after death to allow the detection of drugs [5,8,9]. ...
... However, when a human body is extremely decomposed, or even skeletonized, these specimens may not be available due to their degradation during decomposition or their contamination [4][5][6]. Thus, alternative matrices are necessary to perform toxicological analysis [4][5][6][7]. To be suitable for toxicological analyses, these alternative matrices must remain well-preserved for a long time after death to allow the detection of drugs [5,8,9]. ...
Article
Full-text available
In decomposed or skeletonized bodies, conventional matrices used in forensic toxicology may no longer be available for analysis. The aim of this paper was to test the survival and detection of toxicological substances in dry bone samples with over 23 years of post-mortem interval. In this perspective, bone samples from the cranium, ribs, and vertebrae of seven skeletons from the CAL Milano Cemetery Skeletal Collection, buried for over 23 years, fully decomposed and altered by taphonomic factors were selected based on their ante-mortem data, which included verified or suspected drug addictions or overdose. Qualitative and quantitative analyses were performed with Dionex™ ASE™ 350 Accelerated Solvent Extractor and Q-Exactive Orbitrap–mass spectrometry with a HPLC system. Positive results were obtained in six of the seven cases, and different psychoactive drugs (and in some cases their active metabolites) were detected, including analgesic (two opioids: methadone and buprenorphine) and anxiolytic drugs (benzodiazepines, in particular delorazepam, diazepam, nordiazepam, and lorazepam), a cannabinoid metabolite (THCCOOH) as well as metabolites of stimulants (benzoylecgonine and MDA). Consequently, this research shows that toxicological substances may be found in bone tissue after over 23 years of post-mortem interval.
... In our study, a simple extraction procedure was applied on full bones using methanol. 10,21 Drug levels in different bones from various anatomical body sites of chronic-dosed rats were compared and the accumulation of drugs and their metabolites were assessed in these sites. ...
... Observed skeletal tissue concentrations are comparable with those obtained from similar studies previously published and exceed those seen in blood. 10,26,27 Citalopram is found in substantially higher concentration than clomipramine. When drug concentrations of the different bone types are compared to each other within a rat, a big variability (>36%) is seen The relative drug−metabolite ratio for citalopram/desmethylcitalopram for each bone type for all drugs and metabolites. ...
... This explanation is not likely since bone concentrations of similar molecules showed to be well above our LOD, for example diazepam (5.84 ng/g). 10 Another explanation could be found in the potential model for drugs incorporation from Rubin et al. 8 When looking at the pKa values of clomipramine (9.2) and citalopram (9.78), it is safe to assume that they are mostly ionized at physiological pH. Following the proposed model, there is a higher chance that these drugs are incorporated in bone tissue. ...
Article
Full-text available
In recent years, the usage of skeletal tissue as alternative matrix in forensic toxicology has grown new interest. In cases where extreme decomposition has taken place, analysis of skeletal tissue is often the only option left. In this paper, a fully validated method is presented and the distribution of clomipramine, citalopram, midazolam and metabolites after chronically administration is examined within skeletal tissue. Rats were chronically dosed with respectively clomipramine, citalopram or midazolam. Extracts were quantitatively analyzed using LC‐ESI‐MS/MS. Clomipramine, citalopram and metabolites, respectively desmethylclomipramine and desmethylcitalopram are shown to be detectable in all bone types sampled. Midazolam and its metabolite α‐OH‐midazolam could not be detected. The absence of midazolam in extracts gives an indication that drugs with pKa values under physiological pH are badly or not incorporated in bone tissue. Bone and post‐mortem blood concentrations were compared. A range of different bone types were compared and showed that the concentration is strongly dependent on the bone type. In concordance with previous publications, the humerus shows the highest drug levels. Found skeletal tissue concentrations ranged from 1.1 – 587.8 ng/g. Comparison of the same bone type between the different rats showed high variances. However, the ratio of drugs/metabolite proved to have lower variances (<20%). Moreover, the drugs/metabolite ratio in the sampled bones is in close concordance to the ratios seen in blood within a rat. From this, we can assume that the drugs/metabolite ratio in skeletal tissue may prove to be more useful than absolute found concentration.
... Cone voltage and collision energy were optimized for each transition. Data acquisition and analysis was performed by MassLynx® software whereas quantitation was performed by TargetLynx application [11]. ...
... For extraction of the rib, it was used the validated method of Orfanidis et al. (2018) [11]. In brief, each bone was cleaned until any remains of muscles or other connective tissues were removed meticulously using a scalpel. ...
... The supernatant was evaporated to dryness under a steam of nitrogen. The residue was reconstituted in 300 μL (H2O: MeOH − 80:20 v/v), filtrated (0.22 μm) again and thereafter transferred into autosampler vial for analysis [11]. For brain and blood extractions, it was used the method of Goldberger et al. [12] for cocaine, opioids and alkaloids compound. ...
... among those examined, the femoral mid-diaphysis is the most frequently sampled, followed by vertebral and pelvic bone fragments and the clavicle [8,[13][14][15][16][17][18][19][20][21][22][23][24][25][26]. ...
... Little data concerning detectability and incorporation of drugs in bone are available. Previous studies have investigated typical basic drugs of abuse such as methamphetamine,amphetamine and opiates, and also psychopharmaceuticals such as benzodiazepines, antidepressants, and antipsychotics[9][10][11][12][13][18][19][20][21][22][23][24][25][26].However, in the above-mentioned studies, some test conditions may not be representative of forensic scenarios: in fact, in most cases, the samples were taken immediately during the autopsy and analyzed or only fresh cadavers were sampled[1,6,[12][13][14][15][19][20][21][22][23][41][42], ...
... Little data concerning detectability and incorporation of drugs in bone are available. Previous studies have investigated typical basic drugs of abuse such as methamphetamine,amphetamine and opiates, and also psychopharmaceuticals such as benzodiazepines, antidepressants, and antipsychotics[9][10][11][12][13][18][19][20][21][22][23][24][25][26].However, in the above-mentioned studies, some test conditions may not be representative of forensic scenarios: in fact, in most cases, the samples were taken immediately during the autopsy and analyzed or only fresh cadavers were sampled[1,6,[12][13][14][15][19][20][21][22][23][41][42], ...
Article
In forensic contexts of advanced decomposition, when conventional matrices are no longer available for toxicological analyses, finding alternative matrices is necessary. The skeleton, which is fundamental for anthropologists and geneticists, could be useful also for toxicological purposes. The present study aims to examine what kind of information toxicological analysis performed on bones (the cranium and the ribs) in different states of preservation could provide to the forensic practitioner. Thirty cadavers with known pharmacological history, subjected to forensic autopsy at the Institute of Legal Medicine of Milan, were selected. Rib and cranium samples were collected from each body and separated into two parts in order to create two different states of preservation: One was cleaned from soft tissues and analyzed as a well‐preserved bone sample; the other was submitted to a long maceration process, simulating complete skeletonization. All specimens were then processed with accelerated solvent extraction and the eluates analyzed using Q‐Exactive™ Orbitrap™ Mass Spectrometer. The analysis of blood and skeletal matrices showed positive results for the tested substances in 63% of cases, mainly benzodiazepines, antidepressants, and drug abuse. Significant Pearson correlations were observed between non‐macerated vs. macerated bone samples: r = 0.79 for rib samples, r = 0.61 for cranium samples, and r = 0.69 for all bone samples. As a consequence, the positive results confirm the potential of the bone tissue as an alternative matrix in forensic toxicology, even in cases of extremely decomposed bodies. This study also highlighted important elements for reconstructing the biological profile in cases of forensic anthropological concern.
... For instance, the investigation of particular bone manifestations of cocaine abuse trough CT scans [18]; discussing how homeostasis can change due to alcohol and drug use, affecting the ability to accurately assess estimation of age-at-death [19,20]; or experimental approaches with human analogues on opioids [21]. The presence of drugs in bones has been studied mainly in skeletal toxicology, where the substance is detected analytically [22][23][24][25][26][27], but very little has been done macroscopically with imaging or by direct examination of the bones. ...
... Code: Y = yes. Trazodone Cranium, rib 19 [190] Venlafaxine Cranium, rib 19 [190] Rib 7 [276] Antihistamine drugs Diphenhydramine Iliac crest, vertebra 39 [22] Antihypertensive drugs Atenolol Rib 2 [277] Bisoprolol Rib 2 [277] Antipsychotics Chlorpromazine Femur 36 [275] Clozapine Femur 36 [275] Haloperidol Cranium, rib 19 [190] Mesoridazine Femur 36 [275] Promazine Cranium, rib 19 [190] Quetiapine Cranium 19 [190] Rib 3 [189] Thioridazine Femur 36 [275] Benzodiazepines Alprazolam Cranium, rib 19 [190] Bromazepam Femur 6 [23] Delorazepam Vertebra, rib 7 [27] Cranium, rib 19 [190] Diazepam Cranium vertebra, rib 7 [27] Cranium, rib 19 [190] Iliac crest, vertebra 39 [22] Femur 36 [275] Femur 6 [23] Flurazepam Cranium, rib 19 [190] Cranium, rib 19 [190] Lormetazepam Cranium, rib 19 [190] Nordiazepam Vertebra 7 [27] Cranium, rib 19 [190] Iliac crest, vertebra 39 [ Cranium, rib 19 [190] Rib 6 [278] Iliac crest, vertebra 39 [22] Femur 6 [23] Although further research is required, results have shown that these drugs can be detected years after death. ...
... Code: Y = yes. Trazodone Cranium, rib 19 [190] Venlafaxine Cranium, rib 19 [190] Rib 7 [276] Antihistamine drugs Diphenhydramine Iliac crest, vertebra 39 [22] Antihypertensive drugs Atenolol Rib 2 [277] Bisoprolol Rib 2 [277] Antipsychotics Chlorpromazine Femur 36 [275] Clozapine Femur 36 [275] Haloperidol Cranium, rib 19 [190] Mesoridazine Femur 36 [275] Promazine Cranium, rib 19 [190] Quetiapine Cranium 19 [190] Rib 3 [189] Thioridazine Femur 36 [275] Benzodiazepines Alprazolam Cranium, rib 19 [190] Bromazepam Femur 6 [23] Delorazepam Vertebra, rib 7 [27] Cranium, rib 19 [190] Diazepam Cranium vertebra, rib 7 [27] Cranium, rib 19 [190] Iliac crest, vertebra 39 [22] Femur 36 [275] Femur 6 [23] Flurazepam Cranium, rib 19 [190] Cranium, rib 19 [190] Lormetazepam Cranium, rib 19 [190] Nordiazepam Vertebra 7 [27] Cranium, rib 19 [190] Iliac crest, vertebra 39 [ Cranium, rib 19 [190] Rib 6 [278] Iliac crest, vertebra 39 [22] Femur 6 [23] Although further research is required, results have shown that these drugs can be detected years after death. ...
Article
Full-text available
Forensic anthropologists rely on a number of parameters when analyzing human skeletal remains to assist in the identification of the deceased, predominantly age-at-death, sex, stature, ancestry or population affinity, and any unique identifying features. During the examination of human remains, it is important to be aware that the skeletal features considered when applying anthropological methods may be influenced and modified by a number of factors, and particular to this article, prescription drugs (including medical and non-medical use) and other commonly used drugs. In view of this, this paper aims to review the medical, clinical and pharmacological literature to enable an assessment of those drug groups that as side effects have the potential to have an adverse effect on the skeleton, and explore whether or not they can influence the estimation of age-at-death, sex and other indicators of the biological profile. Moreover, it may be that the observation of certain alterations or inconsistencies in the skeleton may relate to the use of drugs or medication, and this in turn may help narrow down the list of missing persons to which a set of human remains could belong. The information gathered from the clinical and medical literature has been extracted with a forensic anthropological perspective and provides an awareness on how several drugs, such as opioids, cocaine, corticosteroids, non-steroidal anti-inflammatory drugs, alcohol, tobacco and others have notable effects on bone. Through different mechanisms, drugs can alter bone mineral density, causing osteopenia, osteoporosis, increase the risk of fractures, osteonecrosis, and oral changes. Not much has been written on the influence of drugs on the skeleton from the forensic anthropological practitioner perspective; and this review, in spite of its limitations and the requirement of further research, aims to investigate the current knowledge of the possible effects of both prescription and recreational drugs on bones, contributing to providing a better awareness in forensic anthropological practice and assisting in the identification process of the deceased.
... 13,14 Studies utilizing LC-MS/MS are also found in the literature for the same kind of compounds. 15,16 In order to lower the detection limits and eliminate/reduce the interference effects, different extraction strategies could be used. Different microextraction procedures have been used for extraction/preconcentration of a variety of analytes in different sample matrices. ...
... The results indicated that the detection window for dental tissues was located between the detection windows for hair and body fluids. In addition to the dental tissues, bone and bone marrow were also investigated as a specimen in postmortem toxicology by Orfanidis et al. [85]. In this work, femur bone samples were collected from the chronic drug abusers, 14-20 h postmortem. ...
Article
Morphine (MO) as an opioid analgesic is used for the treatment of moderate-to-severe pains, particularly cancer-related pains. Pharmacologic studies on MO are complicated due to drugs binding to the protein or metabolization to active metabolites, and even inter-individual variability. This necessitates the selection of a reliable analytical method for monitoring MO and the concentrations of its metabolites in the biological samples for the pharmacokinetic or pharmacodynamic investigations. Therefore, this study was conducted to review all the analytical researches carried out on MO and its metabolites in the biological samples during 2007–2019 as an update to the study by Bosch et al. (2007).
... Forensic science laboratories have to be equipped to provide specific and sensitive techniques to recognize legal and illegal substances and to quantify them in biological samples such as blood and urine. There are many traditional methods for the determination of illicit drugs using chromatographic (HPLC, GC and so on) and mass spectrometric techniques [1][2][3]. Most of these techniques are based on high-cost immobile equipment, they are time-consuming, and they suffer from complex sample preparation protocols. ...
Article
Drug abuse considered a serious source of economic and social problems. The sensing of drugs of abuse is of demanding in forensic and clinical toxicology. There are many various methods for determination these materials using chromatographic and mass spectrometric techniques. Most of these techniques needs high-cost equipment, time consuming and suffer hard sample preparations. However, electrochemical methods are easy, simple and no need for complicated sample preparations cause to more interests of their use for determinations of toxics and pharmaceuticals. On the other hand, use of nanomaterials in electrochemistry found wide attentions to improve selectivity, sensitivity and limit of detections of various compounds such as pharmaceuticals, biologicals and environmental. Nanomaterials draw interests due to their low cost and unique size-dependent properties. The settling of nanomaterials into different matrices to prepare nanocomposite films founds wide interest. The unique properties of nanomaterials like mechanical, electrical, optical, catalytic and magnetic properties in addition of their significant high surface area per mass make them popular. Besides the novel properties, nanomaterials demonstrate new approaches to fabricate low cost electrodes by minimizing the materials needed and waste. The presence of nanotechnology beside modern electrochemical techniques helps to emerge of powerful, reliable electrical devices for sensing that shows benefits like increasing mass transport rate, high surface area and good control over electrode microenvironment. The aim of this review is to give an outline for the electrochemical determination based on nanomaterials of the commonly occurring illicit drugs in a various matrices such as urine, blood and saliva, which are important for determining of drugs of abuse.
... With levels of drugs detected in bone at level of ng/g, the analysis of drugs in bone is also a challenge. In the instrumentation aspect of skeletal toxicology, a validated method was developed by Orfanidis et al. [547], where samples were extracted with methanol followed by stirring and ultra-sonication. The extract, after filtration, evaporation and reconstitution was analyzed on a reversedphase column (C18) in gradient elution in the detection and quantitation protocol using UPLC-MS/MS. ...
Article
Full-text available
This review paper covers the forensic-relevant literature in toxicology from 2016 to 2019 as a part of the 19th Interpol International Forensic Science Managers Symposium. The review papers are also available at the Interpol website at: https://www.interpol.int/content/download/14458/file/Interpol%20Review%20.Papers%202019.pdf.
... When looking at previous case reports, drawing unambiguous conclusions about dosing or length of time since last drug use cannot be determined from skeletal tissue analysis. Although skeletal tissue has a number of advantages over other alternatives, it has been shown to be a depot for certain drugs [4][5][6]. Skeletal tissue also withstands putrefaction best of all specimens. Skeletal tissue consists of two major categories being bone tissue and bone marrow [7]. ...
Article
Blood analysis is the golden standard in the field of forensic toxicology. However, when extended decomposition of the remains has occurred, alternative matrices are required. Skeletal tissue may provide an appropriate sample of choice since it is very resistant to putrefaction. However, today, the absence of reference data of drug concentrations in skeletal tissue poses a problem to meaningfully and reliably conduct toxicological testing on human skeletal material. The present study investigates the viability of skeletal tissue as an alternative matrix to evaluate xenobiotic consumption in legal cases. Blood, bone tissue and bone marrow of different forensic cases were screened for 415 compounds of forensic interest. Afterwards, methadone, clomipramine, citalopram and their respectively metabolites positive samples were quantified using fully validated methods. Sample preparation was carried out by SPE (whole blood and bone marrow), methanol extraction (bone sections) or protein precipitation (whole blood). All samples were analyzed using liquid chromatography coupled to a triple quad mass spectrometer. Multiple drugs were successfully identified in all sampled matrices. In bone (marrow) not as many substances were detected as in blood but it poses a valid alternative when blood is not available. Especially bone marrow showed big potential with a concordance of 80.5% with blood. Clomipramine, citalopram and their metabolites were proven to be detectable and quantifiable in all specimens sampled. Bone marrow showed the highest concentrations followed by blood and bone tissue. When citalopram blood concentrations were correlated with the bone concentrations, a linear trend could be detected. The same was seen between blood and bone marrow for citalopram concentrations. Methadone was also proven to be detectable in all specimens sampled. However, its metabolites EMDP and EDPP were absent or below the LOD in some samples. Overall, methadone concentrations were higher in bone marrow than in bone. With exception of one case, blood concentrations were higher than bone concentrations. For methadone, a linear trend could be found between blood and bone concentration. Comparing methadone concentrations in blood and bone marrow an exponential trend could be seen. In conclusion, these findings show the potential forensic value of bone and bone marrow as an alternative matrix. Aside to that, a standard protocol for the sample collection and processing is proposed.
... Most recently, human skeletal tissue samples have been analysed for detecting presence of cannabinoids 10 and 12 by a UPLC-based method applying ESI-MS/MS, where an Acquity BEH C 18 column (150 mm × 2.1 mm; particle size: 1.7 μm) was used with a mobile phase comprising water-MeOH (both containing 0.1% HCOOH) and a run time of 17 min. 105 In another experiment, 11-OH-THC (9) along with the earlier two cannabinoids were quantified in human liver samples using a similar UPLC-ESI-MS/MS method. 38 Table 8 presents the summary of all LC-based analytical methods for the quantification of cannabinoids in various animal samples published during 2010-2019. ...
Article
Introduction: Organic molecules that bind to cannabinoid receptors are called cannabinoids, and they have similar pharmacological properties like the plant, Cannabis sativa L. Hyphenated liquid chromatography (LC), incorporating high-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography (UPLC, also known as ultrahigh-performance liquid chromatography, UHPLC), usually coupled to an ultraviolet (UV), UV-photodiode array (PDA) or mass spectrometry (MS) detector, has become a popular analytical tool for the analysis of naturally occurring cannabinoids in various matrices. Objective: To review literature on the use of various LC-based analytical methods for the analysis of naturally occurring cannabinoids published since 2010. Methodology: A comprehensive literature search was performed utilising several databases, like Web of Knowledge, PubMed and Google Scholar, and other relevant published materials including published books. The keywords used, in various combinations, with cannabinoids being present in all combinations, in the search were Cannabis, hemp, cannabinoids, Cannabis sativa, marijuana, analysis, HPLC, UHPLC, UPLC, quantitative, qualitative and quality control. Results: Since 2010, several LC methods for the analysis of naturally occurring cannabinoids have been reported. While simple HPLC-UV or HPLC-UV-PDA-based methods were common in cannabinoids analysis, HPLC-MS, HPLC-MS/MS, UPLC (or UHPLC)-UV-PDA, UPLC (or UHPLC)-MS and UPLC (or UHPLC)-MS/MS, were also used frequently. Applications of mathematical and computational models for optimisation of different protocols were observed, and pre-analyses included various environmentally friendly extraction protocols. Conclusions: LC-based analysis of naturally occurring cannabinoids has dominated the cannabinoids analysis during the last 10 years, and UPLC and UHPLC methods have been shown to be superior to conventional HPLC methods.
... During the last decade, the interest in the usage of skeletal tissue as an alternative specimen in forensic toxicology has seen a revival [12][13][14]. It is shown that the bone tissue acts as a depot for certain drugs [12]. ...
Article
Full-text available
In the past decades, matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) has been applied to a broad range of biological samples, e.g., forensics and preclinical samples. The use of MALDI-MSI for the analysis of bone tissue has been limited due to the insulating properties of the material but more importantly the absence of a proper sample preparation protocol for undecalcified bone tissue. Undecalcified sections are preferred to retain sample integrity as much as possible or to study the tissue-bone bio interface in particular. Here, we optimized the sample preparation protocol of undecalcified bone samples, aimed at both targeted and untargeted applications for forensic and preclinical applications, respectively. Different concentrations of gelatin and carboxymethyl cellulose (CMC) were tested as embedding materials. The composition of 20% gelatin and 7.5% CMC showed to support the tissue best while sectioning. Bone tissue has to be sectioned with a tungsten carbide knife in a longitudinal fashion, while the sections need to be supported with double-sided tapes to maintain the morphology of the tissue. The developed sectioning method was shown to be applicable on rat and mouse as well as human bone samples. Targeted (methadone and EDDP) as well as untargeted (unknown lipids) detection was demonstrated. DHB proved to be the most suitable matrix for the detection of methadone and EDDP in positive ion mode. The limit of detection (LOD) is estimated to approximately 50 pg/spot on bone tissue. The protocol was successfully applied to detect the presence of methadone and EDDP in a dosed rat femur and a dosed human clavicle. The best matrices for the untargeted detection of unknown lipids in mouse hind legs in positive ion mode were CHCA and DHB based on the number of tissue-specific peaks and signal-to-noise ratios. The developed and optimized sample preparation method, applicable on animal and human bones, opens the door for future forensic and (pre)clinical investigations.
... In the last decade, there have been a number of reports describing extraction methods for the analysis of drugs in bone. Several studies have shown the possibility of using bone or bone marrow for post-mortem detection of xenobiotics[1- 32], even though the extraction of drugs from bones tissue is much more difficult compared to conventional extraction and recovery of analyte(s) from biological fluids or other types of tissue[12]. In fact, bone tissue provides a natural physical barrier that can prevent exposure to exogenous contaminating factors such as substances contained in the soil or products of soft tissue putrefaction, as well as protect against the destructive postmortem activity of fungi, ...
... It is listed in Table 1 of the 1988 Convention and restricted compounds in many countries [1,4]. Several techniques are commonly used for acetic anhydride detection, such as gas chromatography (GC), liquid chromatography (LC), mass spectrometry (MS), GC or LC coupled with MS (GC-MS and LC-MS), and capillary electrophoresis [5][6][7][8]. They are highly sensitive and selective instruments but expensive and not conveniently available; moreover, specialized skills are required to operate them [9]. ...
Article
Acetic anhydride is one of the most often used reagents in the production of heroin, and the detection of this compound in the air is a methodological challenge. This work aims to devise cellulose acetate (CA) nanofibers overlaid with chitosan (Cs) on a quartz crystal microbalance (QCM) for the detection of acetic anhydride. A base layer of CA nanofibers was coated on QCM via electrospinning and then immersed in Cs solution to fabricate CA nanofibers overlaid with chitosan (CA/Cs nanofibers). On scanning electron microscopy (SEM) images, both CA and CA/Cs nanofibers appeared very smooth and homogenous. The increased nanofiber diameter following chitosan immersion indicated that the chitosan layer covered the CA nanofibers. In addition, coverage was verified by the existence of primary amine groups of Cs in CA/Cs nanofibers on the FTIR spectra. The addition of a chitosan layer on top of CA nanofibers clearly increased the sensory response. The sensitivity of the QCM CA/ Cs nanofiber sensor exposed to 10-1000 ppm acetic anhydride vapor was up to 0.234 Hz/ppm, with a lower limit of detection (LOD) of 5 ppm. The sensor also showed a rapid response time (44 s), as well as repeatable and long-term stable behavior, during 5 months of measurements. The as-prepared sensor was also more sensitive to acetic anhydride vapor than to other vapors. A reversible nucleophilic addition interaction between acetic anhydride molecules and the primary amine groups in chitosan was believed to be responsible for the sensing mechanism. The high sensory ability implied that CA/Cs nanofiber coated QCM sensors will be highly suitable for use in the detection of acetic anhydride vapor.
... Add nanostructures such as conducting nanopolymers [16], carbon materials [17,18], metal nanoparticles [19] and redox mediators [20] to electrodes/sensors helps to the excellent performance of modified electrodes and sensors [21]. Molecularly imprinted polymers (MIPs) as synthetic polymers possessing selective molecular recognition and biomimetic recognition materials have many applications such as solid-phase extraction [22,23], electrochemical sensors [24] catalysis [25], also these structures are high-potential materials for cost-efficient the separation [26] and drugs determination [27] of various drugs and bioactive molecules [28] and somewhat complex matrices [29]. Metal organic frameworks (MOFs) are inorganic hybrid compounds via multi-dimensional networks which constructed from metal ions and organic linkers [30,31]. ...
Article
Full-text available
In this paper, we report the Co-metal organic frameworks (MOFs) were successfully synthesized with the aid of 2,6-pyridinedicarboxylic acid ammonium as an organic ligand in polylactic acid (PLA) as a biodegradable and biocompatibility polymer substrate through the ultrasonic-assisted electrospinning method. Co-MOFs/PLA nanofiber used as a new nanocomposite for design and construction of nanosensor based on the polymer prepared of the 4-aminobenzoic acid electrolytic molecule for measuring ciprofloxacin (CIP) drug class. The molecularly imprinted polymer (MIP) and non-imprinted polymer (NIP) sensors were studied by differential pulse voltammetry (DPV) to study changes in oxidation peaks and reduction of potassium ferricyanide. The calibration curve for ciprofloxacin was in the range of 0.5–150 μM also the detection limit was 0.017 μM and the standard deviation for five consecutive measurements at 60 μM concentration was calculated1.7% respectively. For characterizing the synthesized Co-MOF/PLA nanofiber we used X-ray diffraction (XRD), scanning electron microscopy (SEM), atomic force microscopy (AFM), energy dispersive spectroscopy (EDAX), Transmission electron microscopy (TEM) and Dynamic light scattering (DLS). In this study, we present new nanosensors with Co-MOF/PLA nanofiber as a biodegradable and bioactive composite for the first time in modified carbon paste electrode using the ultrasound-assisted direct micelle (UADM) method as the determination of ciprofloxacin drug.
... It makes no sense to let the concentration of the standard of lowest concentration represent the LOD because the corresponding peak of the chromatogram would not be linked to the noise level and slope of the calibration line, which is otherwise required for the method [55]. Further, the LOD must be determined using the SD of a blank solution and the slope of the calibration line, [56] but some authors [3,13] reported an LOD that is higher than the concentration of standards of the lowest concentrations. ...
Article
Suppose that results of the scientific literature were presented in courts the same way they are presented in publications. In the present investigation, it is assumed that this is the case, and it is shown that it may potentially lead to issues with results that are being presented to the court. The determination of synthetic cannabinoids at low concentrations in blood, urine and other matrices is challenging to forensic science. Methods based upon the separation of the compounds by HPLC and detection by tandem mass spectrometry provides results at ultra-low concentrations. The uncertainty of measurements has become the key parameter of interest for decision making, and expert witnesses need to state the correct level of uncertainty. Recent developments in quality assurance indicate issues with reliability, owing to contradictory statements that originate from different methods of data management and interpretation of results that eventually may lead to compromising the truth. The levels of relative uncertainty of measurement that were close to 5 % were found to be unrealistically low when synthetic cannabinoids were analysed at ultra-low concentrations. It was proposed to introduce the principle of pooled calibrations to obtain correspondence between predicted and observed uncertainty, following standards of scientific methodology. Ten synthetic cannabinoids were analysed with pooled calibrations, and the results indicated that the uncertainty of measurement was found at levels much higher than expected, and with two out of ten synthetic cannabinoids that were impossible to quantify with relative uncertainties reaching levels over 70 %.
... In the last decade, skeletal tissues have been investigated as postmortem toxicological matrix by different authors who used a variety of methodologies for drugs extraction and detection [6][7][8][9][10][11][12][13][14][15][16][17]. Most researchers used experimental animals with the advantage of working under controlled conditions [2,4,[6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22], while studies performed using human bones [1,5,[23][24][25][26][27][28][29][30] are scarce and to date, do not use standardized protocols for sample preparation and analysis [2,29,30]. ...
Article
Full-text available
A method based on gas chromatography–mass spectrometry (GC–MS) is described for the determination of bisoprolol and atenolol in human bone. After the addition of lobivolol as internal standard, pulverized samples were incubated in acetonitrile for 1 h under ultrasounds. After adjusting the pH of the samples to 6, they were centrifuged, and the supernatants were subjected to solid phase extraction. Elution was achieved by using 3 mL of 2% ammonium hydroxide in 80:20 dichloromethane:isopropanol solution. Eluted samples were evaporated and derivatized. Chromatography was performed on a fused silica capillary column and analytes were determined in the selected-ion-monitoring (SIM) mode. The assay was validated in the range 0.1–0.3 ng/mg (depending on the drug) to 150 ng/mg, the mean absolute recoveries were 60% for bisoprolol and 106% for atenolol, the matrix effect was 69% for bisoprolol and 70% for atenolol and process efficiency was 41% for bisoprolol and 80% for atenolol. The intra- and inter-assay accuracy values were always better than 12%. The validated method was then applied to bone samples from two real forensic cases in which toxicological analysis in blood were positive for atenolol in the first case (0.65 µg/mL) and bisoprolol in the second case (0.06 µg/mL). Atenolol was found in bone samples from the corresponding case at the approximate concentration of 148 ng/mg and bisoprolol was found at 8 ng/mg.
... Multi-analyte LC-HRMS approaches to DoA quantification allow for much easier method development without the need for extensive MS optimization for each analyte and a limitation on the number of compounds investigated. This is also the case for ''pharmaco-toxics'' such as buprenorphine and ketamine, where dosing techniques are now well established [31,32]. The Q-TOF detection and quantification of cathinones, hallucinogenic phenethylamines, and piperazines after salting-out assisted LLE with acetonitrile in whole blood was presented by Pasin et al. [33]. ...
Article
Full-text available
High-resolution mass spectrometry (HRMS) is now the method of choice in sev-eral toxicology contexts. This paper reviews HRMS approaches for research and application invarious toxicology fields, focusing on drugs of abuse in clinical and forensic toxicology. Papersconcerning HRMS applications in screening, quantification and metabolism of drugs of abuse inbiological and non-biological samples were included. Specific applications for new psychoactivesubstances in contexts such as online libraries, bioinformatic tools (molecular networking) andmethods combinations were also included.
Article
In the present research, a procedure was described for the recovery of rosmarinic acid (RA) from medical extract samples using chitosan‑zinc oxide nanoparticles as a biocompatible nanocomposite (CS-ZnO-NC). The dispersive micro-solid phase extraction (D-μ-SPE) of RA from the medical extract samples was investigated by using the prepared biocompatible composite as a solid phase. The HPLC-UV method was used for measuring the extracted RA. The important variables (pH, biocompatible composite mass, contact time, and volume of eluent) associated with the extraction process were analyzed by the application of central composite design (CCD). The achieved optimum values for the mentioned variables were 7.0, 10 mg, 4 min, and 180 μL, respectively. The extraction recovery (99.68%) obtained from the predicted model was in agreement with the experimental data (98.22 ± 1.33%). In addition, under the obtained optimum conditions and over the concentration in the range of 2–3500 ng mL⁻¹, a linear calibration curve was obtained with R² > 0.993. The limit of detection (LOD) and quantification (LOQ) values were computed, and the obtained ranges were respectively from 0.060 to 0.089 ng mL⁻¹ and 0.201 to 0.297 ng mL⁻¹. In addition, the enrichment factors were obtained in the range of 93.7–110.5 with preconcentration factor of 83.3. Therefore, the D-μ-SPE-HPLC-UV method could be used for analyzing RA in the samples of the extracts obtained from the medical plants and water with the recovery values of the analyte in the range of 96.6%–105.4% and the precision with relative standard deviation <5.7%.
Chapter
In analytical toxicology, no matter how complex the equipment and careful the analysis, the results may be rendered worthless if sample collection, transport, and storage have not been performed with the analysis in mind. Biological samples may contain infective agents and must be handled with care, especially if originating from substance misusers, and must always be treated as if they are infective. Clinical samples can be divided into blood and related fluids, body fluids other than blood, excretory fluids/residues, and other clinical specimens. This chapter presents a list of some clinical sample types and provides guidelines for sample collection for analytical toxicology. It discusses the advantages and disadvantages of different sample types in analytical toxicology. The chapter provides information on the sample transport, storage, and disposal, as well as common interferences encountered in the storage of samples.
Article
In this study quetiapine and pregabalin were analyzed in human bones. A method previously developed for the determination of antidepressants in human bone was tested for the analysis of these two substances. Bones were pulverized and subjected to the extraction protocol, and after undergoing solid-phase extraction, samples were analyzed using gas chromatography–mass spectrometry. The assay was validated in the range 0.3-500 ng/mg, mean analytical recovery was 76.9 % for quetiapine and 90.9 % for pregabalin, matrix effect was 83 % for quetiapine and 91 % for pregabalin and process efficiency was 63.8 % for quetiapine and 82.7 % for pregabalin. The intra- and inter-day precision was below 3 % in all cases and the intra- and inter-assay accuracy values were in almost all cases better than 12%. The validated method was then applied to bone samples from forensic cases. Drugs were detected in bone in 2 of the 3 blood positive cases. The approximate concentrations in bone were 40 ng/mg for pregabalin and 7 ng/mg for quetiapine. To our knowledge, this is the first time these substances were detected in bones. With this study the number of substances with a validated protocol to be used in human bones in case of necessity is expanded.
Article
A procedure based on gas chromatography-mass spectrometry was developed for the analysis of benzodiazepines (nordiazepam, oxazepam, lormetazepam, lorazepam, clonazepam, bromazepam and alprazolam) in postmortem human ribs. Powdered bone samples, including marrow remains inside, with the internal standard diazepam-d5 were subjected to enzymatic hydrolysis with 100 μL of β-glucoronidase and were incubated in sodium hydroxide for 1 h in a 70°C oven. Samples underwent liquid phase extraction and ethyl acetate was used as eluent. Chromatography was performed on a fused silica capillary column and the selected-ion-monitoring mode was used for analytes determination. The method was validated in the range 0.1-0.5 ng/mg (depending on the benzodiazepine) to 100 ng/mg with average values of recovery, matrix effect and process efficiency ranged from 83.2 to 94.3%, from 97.3 to 102.1% and from 80.5 to 91.2%, respectively. The intra- and inter-day accuracy was <15%. The procedure was tested in rib specimens obtained during routine autopsies from 20 cases where these benzodiazepines were found in blood. Benzodiazepines were detected in the combined bone and marrow samples in 60% of cases. Lorazepam was detected in bone in the range of 0.3-0.7 ng/mg, nordiazepam at 1.3-4.2 ng/mg and oxazepam at 1.1-1.2 ng/mg. To our knowledge, this protocol for the simultaneous analysis of these benzodiazepines is the first performed and validated using human ribs.
Article
A method based on gas chromatography–mass spectrometry (GC–MS) is described for the determination of venlafaxine, amitriptyline and duloxetine in human bone. Pulverized samples were incubated in methanol for 1 h under ultrasonication, after the addition of sertraline as internal standard. The samples were centrifuged, and the supernatants were evaporated. Samples were then resuspended in 0.1 M phosphate buffer pH 6 and subjected to solid phase extraction. Chromatography was performed on a fused silica capillary column and analytes were determined in the selected-ion-monitoring (SIM) mode. The assay was validated in the range 0.3–1 ng/mg (depending on the drug) to 500 ng/mg. The mean absolute recoveries ranged from 92.6% to 96.2%, the matrix effect from 76.9% to 103.3% and process efficiency from 74% to 95.9% depending on the analyte. The intra- and inter-assay accuracy values were always better than 20%. The validated method was then successfully applied to real bone samples from forensic cases in which toxicological analysis for these drugs in blood had been positive. Drugs were detected in bone in all blood positive results, the approximate concentrations being 36.4 ng/mg for amitriptyline, 19.3–3 ng/mg for duloxetine and 4.6–2 ng/mg for venlafaxine.
Article
Full-text available
Cocaine toxicity has been a subject of study because cocaine is one of the most common and potent drugs of abuse. In the current study the effect of cocaine on human liver cancer cell line (HepG2) was assessed. Cocaine toxicity (IC50) on HepG2 cells was experimentally calculated using an XTT assay at 2.428 mM. The metabolic profile of HepG2 cells was further evaluated to investigate the cytotoxic activity of cocaine at 2 mM at three different time points. Cell medium and intracellular material samples were analyzed with a validated HILIC-MS/MS method for targeted metabolomics on an ACQUITY Amide column in gradient mode with detection on a triple quadrupole mass spectrometer in multiple reaction monitoring. About 106 hydrophilic metabolites from different metabolic pathways were monitored. Multivariate analysis clearly separated the studied groups (cocaine-treated and control samples) and revealed potential biomarkers in the extracellular and intracellular samples. A predominant effect of cocaine administration on alanine, aspartate, and glutamate metabolic pathway was observed. Moreover, taurine and hypotaurine metabolism were found to be affected in cocaine-treated cells. Targeted metabolomics managed to reveal metabolic changes upon cocaine administration, however deciphering the exact cocaine cytotoxic mechanism is still challenging.
Article
Purpose Since the solely certain remnants of a performed autopsy are formalin-fixed paraffin-embedded (FFPE) samples, stored in the archives of every institute of legal medicine, we managed to extract molecules of toxicological interest from these specimens. Methods We assessed the analysis of ten fresh liver samples collected from heroin-related deaths and then histologically processed the same samples. The embedded blocks were then extracted by means of a new extracting method and the eluates were measured. We also selected five toxicological cases of heroin-related fatalities that were examined 20 years ago, collected the toxicological result documents of the analysis that were carried out at the time and then processed the corresponding FFPE liver samples that were stored in the archives. Results We managed to isolate heroine-related metabolites from 20-year-old paraffin-embedded blocks and calculated ratios to evaluate the performance of our new extraction. Conclusions According to our study, it is feasible to carry out a toxicological examination on old histological samples and, therefore, this matrix can be considered as a new alternative specimen for chemical-analytical evaluations of past cases or when fresh samples are not available anymore. The new extractive method was evaluated as efficient in treating these complex, paraffin-embedded samples. It was surprising that the target compounds could be quantitated from FFPE bocks created as long as 20 years ago.
Article
Full-text available
In several medico‐legal cases, bone samples analysis may provide the only source of toxicological information. This case study reports the analysis of a human bone specimen, belonging to a 46‐year‐old man, found 3 months after his death due to cervical–thoracic injuries in a motorcycle accident. Bone specimen was the only available material for toxicological analysis, among few skull hair and rotten skin. Analysis was performed by a newly developed and validated ultra‐high‐pressure liquid chromatography–mass spectrometry/mass spectrometry (UHPLC‐MS/MS) method, following simple and efficient sample pretreatment. The results were in accordance with the man's medical record: Alprazolam and zolpidem were found at 2.2 and 5.4 ng/g of bone, respectively. Both these drugs were prescribed to the deceased.
Article
Full-text available
The aim of the present study was to investigate the distribution of clofazimine (CLF) in rat bone marrow cells by a validated reverse phase high performance liquid chromatography. CLF and chlorzoxazone (I.S) were extracted by liquid-liquid extraction from plasma and rat bone marrow cells. The chromatographic separation was performed in isocratic mode by the mobile phase consisting of 10mM ammonium formate (pH 3.0 with formic acid) and acetonitrile in a ratio of 50:50 (v/v). The method was accurate and precise in the linear range of 15.6-2000.0ng/mL with a correlation coefficient (r(2)) of 0.996 and 0.995 in rat plasma and bone marrow cells, respectively. After single oral dose of 20mg/kg, the maximum concentration of CLF in plasma and bone marrow cells were obtained at 12h with the concentrations of 593.2 and 915.4ng/mL, respectively. The AUC0-t and mean elimination half life (t1/2) of CLF in bone marrow cells were 54339.02ng*h/mL and 52.46h, respectively, which signified the low body clearance and high distribution of CLF in bone marrow cells. The single oral dose pharmacokinetic investigation was confirmed the CLF endure for a long period in rat due to high distribution in various tissues. The developed method was successfully applied to the estimation of the pharmacokinetic parameters of CLF in plasma and bone marrow cells after administration of single oral dose of 20mg/kg to rats.
Book
Although the specimen of choice in the US drug testing industry is urine, and serum in clinical medicine, interest has recently grown in the use of other matrices as drug testing media. Drug Testing in Alternate Biological Specimens provides researchers and forensic toxicologists with a convenient general text summarizing the state of knowledge today. The book is organized into chapters based upon specimen type, each describing the composition of the matrix, sample collection, preparation and analytical procedures, drugs detected to date, and a discussion of the interpretation of positive findings. These chapters focus specifically on the application of these practices to drugs of abuse. Comprehensive and easy to use, Drug Testing in Alternate Biological Specimens offers the means to solve difficult forensic mysteries and to better answer the critical questions of the criminal justice system.
Article
Bone marrow (BM) analysis is of forensic interest in postmortem toxicological investigation in case of limited, unavailable or unusable blood samples. However, it remains difficult to determine whether a drug BM concentration is therapeutic or represents overdose, due to the lack of studies on this alternative matrix. Given the variations in BM composition in the body, sample location was suggested to be a relevant factor in assessing BM concentration. The aim of the present study was to compare postmortem caffeine concentrations in various BM sample locations and secondly to consider the correlation between BM and blood concentrations. Six BM samples (right and left side: proximal and medial femur and 5th rib) and a blood sample were collected from 21 forensic autopsies. Gas chromatography coupled to tandem mass spectrometry was performed. Blood caffeine concentrations ranged from 60 to 7591ng/mL. Femoral and rib BM concentrations ranged from 51 to 6171ng/g and 66 to 7280ng/g, respectively. Blood concentrations were always higher than BM concentrations. As a good correlation was demonstrated between blood and rib BM and between blood and the average of the four femoral BM concentrations, blood caffeine concentrations could be correctly extrapolated from BM concentrations. BM caffeine concentration was found to depend on sample location. Rib BM caffeine concentrations appeared to be systematically greater than averaged femur values and concentrations were much more variable between the 4 femur BM samples than between the 2 ribs. From a practical point of view, for caffeine analysis, rib BM appeared more relevant than femoral BM, which requires multisampling to overcome the concentration variability problem.
Article
The effect of dose-death interval and tissue distribution on the detection of meperidine in selected skeletal tissues was examined using a rapid microwave-assisted extraction (MAE) methodology. Rats (n = 14) were dosed with 0 (n = 2) or 30 mg/kg (n = 12) meperidine (i.p.). Drug-positive rats were sacrificed with CO2 after 20, 30, 90 and 150 min (n = 3 per group). Heart blood was collected immediately after death. Tibiae were excised and frozen for further analysis. The remaining carcasses were allowed to decompose outside in secured cages to the point of complete skeletonization in a rural Northern Ontario location during the late summer months. Vertebrae and pelvi were collected for each animal.
Article
The use of microwave-assisted extraction (MAE) in screening of decomposed bone tissue for model drugs of abuse is described. Rats received 50 mg/kg (i.p.) pentobarbital (n=2), 75 mg/kg (i.p.) ketamine (n=2) or 16 mg/kg (i.p.) diazepam (n=1), or remained drug-free (control). Drug-positive animals were euthanized within 20 min of drug administration. Animal remains were allowed to decompose in a secure outdoor environment to the point of complete skeletonization. Bones (tibiae, femora, vertebrae, ribs, pelvi, humeri and scapulae) were collected and pooled (according to drug) in order to minimize effects due to inter-bone differences in drug distribution. Bones were crushed and cleaned of marrow and residual soft tissue in alkaline solution or phosphate buffer with ultrasonication. Cleaned bones were then ground and underwent MAE in phosphate buffer (pH 6), methanol or a methanol:water mixture (1:1, v/v) at atmospheric pressure in a domestic microwave oven, or passive extraction in methanol. Bone extracts (control and drug-exposed) containing methanol were evaporated to dryness before reconstitution in phosphate buffer (pH 6) and subsequent analysis by ELISA, while bone extracts containing only phosphate buffer were assayed directly by the same ELISA protocol. Measured absorbance values were expressed as the decrease in absorbance, measured as a percentage, relative to the corresponding drug-free control bone extract. The semi-quantitative nature of the ELISA assay allowed examination of the effects of extraction solvent and bone sample mass on the assay response for each drug examined, and subsequent comparison to assays of extracts obtained through passive methanolic extraction of various bone tissues. Overall, the time required for maximal extraction varied with extraction solvent and bone mass for each drug investigated, with significant extraction occurring with all solvent systems examined. MAE may represent a substantially faster extraction system than passive extraction, with significant extraction recovery observed within 1 min of exposure for all samples examined. The implications of these results in the context of the available literature on drug analysis in skeletal tissues are discussed.
Article
There is a paucity of literature detailing the disposition of drugs in bone and bone marrow. Infrequently, in deaths involving skeletonized remains, fragmentation, decomposition, and exsanguination, traditional specimens may be unavailable for toxicological testing. This study examined the utility of bone for the detection of benzodiazepines, opiates, cocaine and metabolites, and basic drugs in 39 cases. Cases were identified on the basis of a positive blood result. After specimen preparation, samples were assayed by liquid-liquid or solid phase extraction with gas chromatographic and gas chromatographic mass spectrometric detection. The majority of decedents were white males with 28% of individuals between the ages of 41 to 50 years. The cause of death was drug intoxication in 22 cases. The most prevalent drugs detected in the blood males and females were opiates and bases. Morphine, codeine, and oxycodone were detected in bone, whereas 6-acetylmorphine and hydrocodone were absent. For alkaline extractable drugs, in only 57% of blood positive specimens, the corresponding bone was also positive. These included antidepressants and antihistamines. Diazepam and nordiazepam were detected in the bone of all blood positive cases. Bone concentrations were higher than blood levels. Benzoylecgonine was the most common cocaine analyte detected in bone. These data demonstrated that drugs may be detected in bone using current technologies and that in general concentrations were higher than those observed in corresponding blood specimens. A negative result in bone, however, should be interpreted with caution because multiple factors determine the deposition of a drug in this matrix.
Article
Correlation between plasma and bone marrow tricyclic antidepressants has not been studied before. Two groups of rabbits were given 10 and 20 mg of desipramine/kg body weight, respectively. Desipramine was administered to the animals once daily by mouth for 5 days. On the fifth day the animals were sacrificed and blood and bone marrow samples were collected and analyzed using a high performance liquid chromatographic (HPLC) method. Data showed that a correlation exists between bone marrow and blood desipramine. The bone marrow desipramine concentration increased as its blood levels increased. The average ratio of bone marrow to blood desipramine +/- S.D. (standard deviation) in both dosage groups was 37.2 +/- 4.46 with a range of 30.99-44.82. This investigation is promising and shows that bone marrow could be used as an alternative tissue in the absence of a suitable blood sample.
Article
The feasibility of detecting methamphetamine and its major metabolite, amphetamine, in postmortem tissues over a 2-year period was examined. It is important to determine if the abuse and toxic effects of drugs can be proved from evidence found in decayed, submerged, or stained tissue materials. The blood, urine, liver, skeletal muscle, skin and extremity bones from rabbits given methamphetamine intravenously were kept at room temperature, under 4 different conditions: sealed in a test tube, dried in the open air, submerged in tap water and stained on gauze. Methamphetamine was present in all the samples, with slight change in concentration in case of sealed and air dried tissues. Changes varied in bones kept in water. There were considerable decreases in methamphetamine in blood and urine stains. Despite long term storage, drug abuse and/or toxicity could be determined, in all tissues examined.
Article
A 28-year-old male methamphetamine abuser, who had been buried for 5 years after being killed by strangulation, was found skeletonized. Methamphetamine and amphetamine in the significantly denatured fatty material of the bone marrow were analyzed by gas chromatography and gas chromatography-mass spectrometry. The confirmation of the chemicals was carried out by chemical ionization (CI) mass chromatography, CI mass spectrometry and CI mass fragmentography. The concentrations of methamphetamine and amphetamine determined by CI mass fragmentography were 1.0 mumol/100 g and 0.1 mumol/100 g, respectively. The method used would seem to be very useful for determination of methamphetamine and amphetamine in marked putrefied biological materials.
Article
Postmortem pentobarbital levels in rabbit heart blood and bone marrow were determined and compared. The average ratio of femur marrow/blood pentobarbital concentrations in 24 rabbits was 1.06 +/- 0.05. The average percent difference between actual plasma pentobarbital concentrations and calculated plasma pentobarbital concentrations was 5.82 +/- 1.96. Concentrations were determined by gas chromatography of extracted, derivatized pentobarbital.
Article
Methods for the quantitation of flurazepam in plasma and bone marrow were developed for the purpose of determining the relationship between flurazepam concentrations in both tissues. Albino New Zealand rabbits, given flurazepam in doses of 5, 10, or 20 mg/kg, were sacrificed either one or three hours after drug administration. Flurazepam concentrations in plasma and bone marrow were determined utilizing a gas-liquid chromatograph equipped with an electron capture detector. the average plasma/bone marrow flurazepam ratios were 0.033 +/- 0.012 and 0.024 +/- 0.012 for rabbits sacrificed one hour and three hours after dosing, respectively. This study showed that a range of plasma flurazepam levels can be estimated from known bone marrow concentrations. The overall mean plasma/bone marrow ratio for all rabbits used in this study was 0.029 +/- 0.012 with a range of 0.010 to 0.055.
Article
Analyses of the hypnotic triazolam from the remains of two human skeletons buried underground for 4 years were made for purposes of confirmation. The bone marrow and mummified muscle were digested with 2 M sodium hydroxide, efficiently extracted using a 3-step solvent extraction procedure, and selectively analyzed by gas chromatography/mass spectrometry with the negative ion chemical ionization mode. Estazolam was the internal standard used. Triazolam was detected in all the samples; the concentrations were 0.36 ng/g in the bone marrow of one victim, and 0.37 and 5.5 ng/g in the bone marrow and mummified muscle of the other victim. This method should prove useful for determination of triazolam in extensively decomposed bodies.
Article
Benzodiazepines were analyzed in different tissue samples, including hone, by ELISA. The sensitivity of detection for different benzodiazepines was consistent with the manufacturer's reports of the cross reactivities of the antibodies used, with the greatest sensitivity for midazolam and the least for diazepam; in addition the pharmacokinetics was consistent with the known duration of action of the different benzodiazepines, with midazolam cleared rapidly, and diazepam slowly. Following intramuscular injection of 300 microg of midazolam at 16 h intervals for ten days, the drug was detectable in bone tissue samples obtained from skeletonized remains buried in soil at room temperature for three weeks.
Article
Bone and bone marrow of a fatally poisoned heroin addict were analyzed by FPIA and GC-FID, immediately after death. A piece of the bone from the above case was buried for 1 year and analyzed by the same procedure. Morphine was detected in all specimens at concentrations of 195, 340 and 155 ng/g for bone marrow, bone and buried bone, respectively. A loss of 54.4% of morphine concentration was observed during 1-year burial. Such findings have potential forensic value in cases of skeletonized remains.
Article
A 40-year-old white male was found dead in bed in a group home for mentally ill adults. The decedent had been diagnosed a paranoid schizophrenic. An autopsy was performed at the Office of the Cuyahoga County Coroner in Cleveland, Ohio. Toxicological testing detected olanzapine and citalopram in post mortem specimens. Multiple fluids and tissues were assayed by liquid-liquid extraction followed by gas chromatography with nitrogen phosphorus detection, and qualitative confirmation by electron impact gas chromatography/mass spectrometry. Drug concentrations [olanzapine: citalopram; mg/L or mg/Kg] determined in this case are the highest reported to date involving these drugs- 1.38:3.35 heart blood, 1.11:1.65 femoral blood, 60.24:32.43 urine, 6.47:10:71 liver, and 38.36:49.16 lung, respectively. Drug concentrations in tissues were found to be the highest in lung for both drugs and lowest in the heart. Citalopram but not olanzapine was detected in bone. The cause of death was ruled acute intoxication by the combined effects of olanzapine and citalopram and the manner, accident.
Article
The aim of this study is to predict how long after time of death a buried body could be analyzed for opiates in soft tissues and to show the accessibility and suitability of bone marrow as a useful toxicological specimen from buried bodies. Morphine solutions were injected in nine albino rabbits. Doses ranged from 0.3 to 1.1 mg/kg with 0.1 mg/kg increments. One hour after the injections, the rabbits were sacrificed. Blood, urine and bone marrow samples were collected for analysis. After the whole bodies were buried, femur bone marrow specimens were collected on the seventh and fourteenth days. CEDIA was used to monitor morphine contents of the collected samples. All experimental cases showed that the increase in the given morphine doses correlated with the increase in blood and bone marrow morphine concentrations. High morphine concentrations were detected in urine samples, but there was no correlation between the urine and blood or urine and bone marrow morphine concentrations. Statistically meaningful increases in bone marrow morphine concentrations were found parallel to increase of blood morphine concentrations. Seventh and fourteenth day postmortem morphine concentrations also followed this correlation. Morphine concentrations in bone marrow at 7 and 14 day postmortem decreased consistently when compared with bone marrow morphine concentrations collected immediately after death. We conclude that in sudden death when other specimens are unavailable due to degradation, bone marrow can be a most useful specimen. Further experimental research in this area is required to validate bone marrow as an alternative tissue.