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Background Delayed diagnosis and improper therapy for intraocular infections usually result in poor prognosis. Due to limitations of conventional culture and polymerase chain reaction methods, most causative pathogens cannot be identified from vitreous humor (VH) or aqueous humor (AH) samples with limited volume. Methods Patients with suspected intraocular infections were enrolled from January 2019 to August 2021. Metagenomic next-generation sequencing (mNGS) was used to detected causative pathogens. Results This multicenter prospective study enrolled 488 patients, from whom VH (152) and AH (336) samples were respectively collected and analyzed using mNGS of cell-free DNA (cfDNA). Taking final comprehensive clinical diagnosis as the gold standard, there were 39 patients with indefinite final diagnoses, whereas 288 and 161 patients were diagnosed as definite infectious and noninfectious diseases, respectively. Based on clinical adjudication, the sensitivity (92.2%) and total coincidence rate (81.3%) of mNGS using VH samples were slightly higher than those of mNGS using AH samples (85.4% and 75.4%, respectively). Conclusions Using mNGS of cfDNA, an era with clinical experience for more rapid, independent, and impartial diagnosis of bacterial and other intraocular infections can be expected. [ABSTRACT FROM AUTHOR]

Excessive drinking and drunkenness are underlying factors in many fatal accidents, which make the quantitative determination of ethanol in postmortem (PM) specimens an essential part of all unnatural death investigations. The same analytical methods are used to determine ethanol in blood taken from living and deceased persons although the interpretation of the results is more complicated in medical examiner cases owing to various preanalytical factors. The biggest problem is that under anaerobic conditions ethanol can be produced naturally in decomposed bodies by microbial activity and fermentation of blood glucose. Ways are needed to differentiate antemortem ingestion of ethanol from PM synthesis. One approach involves the determination of ethanol in alternative specimens, such as bile, cerebrospinal fluid, vitreous humor and/or urine, and comparison of results with blood alcohol concentration (BAC). Another approach involves the analysis of various alcohol biomarkers, such as ethyl glucuronide, ethyl sulfate and/or phosphatidylethanol or the urinary metabolites of serotonin 5-hydroxytryptophol/5-hydroxyindoleacetic acid (5-HTOL/5-HIAA). If ethanol had been produced in the body by microbial activity, the blood samples should also contain other low-molecular volatiles, such as acetaldehyde, n -propanol and/or n -butanol. The inclusion of 1–2% w/v sodium or potassium fluoride, as an enzyme inhibitor, in all PM specimens is essential to diminish the risk of ethanol being generated after sampling, such as during shipment and storage prior to analysis. Furthermore, much might be gained if the analytical cut-off for reporting positive BAC was raised from 0.01 to 0.02 g% when PM blood is analyzed. During putrefaction low BACs are more often produced after death than high BACs. Therefore, when the cadaver is obviously decomposed, a pragmatic approach would be to subtract 0.05 g% from the mean analytical result. Any remaining BAC is expected to give a more reliable indication of whether alcohol had been consumed before death. [ABSTRACT FROM AUTHOR]

ostmortem redistribution (PMR), a well-known phenomenon in forensic toxicology, can result in substantial changes in drug concentrations after death, depending on the chemical characteristics of the drug, blood collection site, storage conditions of the body and postmortem interval (PMI). Limited PMR data are available for ∆9-tetrahydrocannabinol (THC), the primary psychoactive component in Cannabis sativa. PMR was evaluated after controlled cannabis inhalation via a smoking machine and exposure chamber in New Zealand white rabbits. Necropsies were performed on five control rabbits immediately after euthanasia, whereas 27 others were stored at room temperature (21°C) or refrigerated conditions (4°C) until necropsy at 2, 6, 16, 24 or 36 h after death. THC and its Phase I and glucuronidated Phase II metabolites were quantified in blood, vitreous humor, urine, bile and tissues by liquid chromatography–tandem mass spectrometry (LC–MS-MS). Under refrigerated temperature, heart blood THC concentrations significantly increased at PMI 2 h in rabbits, whereas peripheral blood THC concentrations showed a significant increase at PMI 16 h. Central:peripheral blood and liver:peripheral blood ratios for THC ranged from 0.13 to 4.1 and 0.28 to 8.9, respectively. Lung revealed the highest THC concentrations, while brain and liver exhibited the most stable THC concentrations over time. This report contributes much needed data to our understanding of postmortem THC behavior and can aid toxicologists in the interpretation of THC concentrations in medicolegal death investigations. [ABSTRACT FROM AUTHOR]

The intentional ingestion of sodium nitrite causes toxicity by inducing methemoglobinemia, which can lead to cyanosis, hypotension and death. The number of reported suicide cases has significantly increased in the past 10 years as sodium nitrite is readily available online. The traditional tests for nitrite and nitrate require specialized detection methods which are not typically available in a postmortem toxicology laboratory. This rise in sodium nitrite overdose cases indicates the need for a simple, quick test for suspected nitrite toxicity. In this study, a common Griess reagent color test (MQuant™ Nitrite Test Strips) was used as a presumptive method in cases where the ingestion of sodium nitrite was suspected. The test results were consistent between specimens in all cases, and vitreous humor was identified as a reliable matrix to be used in the cases of suspected sodium nitrite poisonings. Case reports of five patients who died of suicide by sodium nitrite in a 6-month span are presented. [ABSTRACT FROM AUTHOR]

There is an increasing number of people affected worldwide by mental health disorders, such as depression and anxiety. One of the main courses of treatment, along with psychotherapy, is the use of psychoactive medications, like antidepressants and benzodiazepines. Also, the unprescribed use of these substances is a concerning public health issue. Hence, the analysis of psychotropic medications is mandatory in postmortem toxicology and various biological samples can be used for this detection, among them the vitreous humor (VH) stands out. Also, there is a demand for more sustainable and more efficient extraction methodologies according to green chemistry. An example is solid phase microextraction techniques (SPME), which use a solid sorbent and small solvent amounts. Biosorbents are substances of natural origin with sorptive properties, and they have been successfully used in SPME in environmental toxicology for water analysis, mainly. This study aimed to develop a sustainable, fast, cheap and simple SPME methodology using cork sheet strips as a biosorbent, to extract antidepressants, benzodiazepines and others from VH samples by liquid chromatography coupled to tandem mass spectrometry. The extraction was conducted in a 96-well plate using 200 µL of VH and optimization of relevant parameters for extraction was performed. For solvent optimization, two simplex-centroid experiments were planned for extraction and desorption and to evaluate time and pH, a Doehlert design experiment was performed. The analytical method for the determination and quantification of 17 substances was validated. The quantification limits were 5 ng/mL for all analytes and the calibration curves were linear between 5 and 30 ng/mL. This study was able to develop an efficient, cheap, simple and fast microextraction method for 17 analytes in VH, using strips of cork sheet for extraction and a 96-well plate as a container. Furthermore, this approach system could be automated for routine toxicology laboratories. [ABSTRACT FROM AUTHOR]

We report on a case of a 35-year-old man who died suddenly and unexpectedly due to a 4-fluoroisobutyrylfentanyl (4-FIBF) mono-intoxication. Pathological, toxicological and chemical investigations were conducted at the Netherlands Forensic Institute. A full three-cavity forensic pathological examination was performed according to international guidelines. Biological samples obtained during autopsy were comprehensively investigated for the presence of toxic substances using headspace gas chromatography (GC) with flame ionization detection, liquid chromatography–time-of-flight mass spectrometry (LC–TOF-MS), GC–MS, high-performance LC with diode array detection and LC–tandem MS (LC–MS-MS). The seized crystalline substance found next to the body was investigated using a presumptive color test, GC–MS, Fourier-transform infrared spectroscopy and nuclear magnetic resonance. Pathological investigation identified minor lymphocytic infiltrates in the heart, considered irrelevant for the cause of death. Toxicological analysis of the victims' blood indicated the presence of a fluorobutyrylfentanyl (FBF) isomer, with no other compounds detected. The FBF isomer was identified in the seized crystalline substance as 4-FIBF. 4-FIBF concentrations were quantified in femoral blood (0.030 mg/L), heart blood (0.12 mg/L), vitreous humor (0.067 mg/L), brain tissue (>0.081 mg/kg), liver tissue (0.44 mg/kg) and urine (approximately 0.01 mg/L). Based on the outcomes of the pathological, toxicological and chemical investigations, the cause of death of the deceased was attributed to a fatal 4-FIBF mono-intoxication. The presented case underlines the added value of a combined bioanalytical and chemical investigative approach to identify and subsequently quantify fentanyl isomers in postmortem cases. Furthermore, it demonstrates the importance of investigating the postmortem redistribution of novel fentanyl analogs to establish reference values and to subsequently allow for correct interpretation of cause of death analysis in future casework. [ABSTRACT FROM AUTHOR]

New psychoactive substances continue to appear on the drug market, and alpha-pyrrolidinoisohexanophenone (α-PiHP) is one of the most popular cathinone derivatives. In this article, we report a case of death caused by α-PiHP. Based on the toxicological results of the studied case along with autopsy, histopathological findings and crime-scene information, fatal intoxication with α-PiHP was accepted as the final cause of death. α-PiHP and its metabolite (OH-α-PiHP) were detected and quantified in all postmortem materials (blood collected from the heart, the femoral vein and the dural venous sinuses; vitreous humor; cerebrospinal fluid; cerebral cortex; brainstem; cerebellum; bile; liver; kidney; heart; pancreas; spleen; thyroid gland; lung; adipose tissue; stomach and intestine). To date, this is the first case of determination of α-PiHP and its metabolite in postmortem specimens. In our opinion, α-PiHP and its metabolite concentration database can be helpful in the interpretation of fatal cases. [ABSTRACT FROM AUTHOR]

Benzodiazepines and z-hypnotics are detected in the majority of fatal overdose cases in Norway, often in combination with other drugs of abuse, and their concentrations in peripheral blood (PB) might be important to elucidate the cause of death. In some forensic autopsies, PB is however not available. The aim of the present study was to compare concentrations of benzodiazepines and z-hypnotics in five alternative matrices to assess whether these concentrations are comparable to concentrations in PB. A total of 109 forensic autopsy cases were included. PB, cardiac blood (CB), pericardial fluid (PF), psoas muscle (PM), lateral vastus muscle (LVM) and vitreous humor (VH) from each case were analyzed using ultra high performance liquid chromatography--tandem mass spectrometry. We were able to detect clonazepam, 7-aminoclonazepam, flunitrazepam, 7-aminoflunitrazepam, nitrazepam, 7-aminonitrazepam, diazepam, nordiazepam, oxazepam, alprazolam, midazolam, zopiclone and zolpidem in all the analyzed matrices. Concentrations measured in VH were generally much lower than those of PB for all compounds except zopiclone. 7-Amino metabolite concentrations were high compared to the parent compounds, although less so for the muscle samples. Concentrations of the parent nitrobenzodiazepines in muscles were higher than those in PB, but for the other compounds, concentrations in muscle showed good correspondence with PB. Both CB and PF were viable alternative matrices for PB, although a larger variation and a tendency for higher concentrations in PF were observed. This study shows that CB, PM, LVM and PF can give comparable concentrations to PB for benzodiazepines and z-hypnotics, while VH was less suitable. The concentrations in alternative matrices must, however, be interpreted carefully. [ABSTRACT FROM AUTHOR]

Barium is an alkaline earth metal whose toxicity is dictated by its compounded salt form: barium sulfate is insoluble and safe to ingest, but other barium salts (chloride, carbonate, sulfide, oxide and acetate) are bioavailable and therefore toxic when ingested. There have been 49 previous reports of fatal intoxications following barium consumption: 38 deemed accidental in nature, 8 suicidal, 1 homicidal and 2 of undetermined intent. In this report, we detail the first intentional fatal self-poisoning with barium chloride to be reported in the UK, along with a review of the surrounding literature. This is the first case to report quantified levels of barium in blood and vitreous humor, and by providing details of sample collection, storage and processing, this case will aid in future interpretations. [ABSTRACT FROM AUTHOR]

Headspace gas chromatography with a flame ionization detector (HS-GC–FID) is a well-established approach for determining blood alcohol concentration, including in cadaveric specimens. Although the integrity of blood specimens can be adequately guaranteed after the sampling, the quantification of ethanol in cadaveric blood can be affected by postmortem fermentative phenomena occurring between the time since death and the sampling of biofluids. The vitreous humor is less affected by putrefactive phenomena allowing compound determination and its use as an alternative biological matrix. The present work aimed to develop and validate a method using the salting-out effect and based on HS-GC–FID for the determination of ethanol in the vitreous humor. The reported analytical method is based on a simple vitreous humor pre-treatment consisting of a dilution (1:9) with a solution of 2.5 mol/L K2CO3 and 0.0012 mol/L tert -butanol (internal standard). After 1 min of incubation, part of the specimen evaporated in the headspace (2,000 µL) is injected into the chromatographic system and analyzed in isothermal mode (40°C), with a chromatographic time of 1.6 min. The method was validated in terms of selectivity, the lowest limit of detection, intraday and total imprecision, and trueness (bias). The determination of ethanol in the vitreous humor and blood was carried out in 75 cases. The correlation between the two matrices was confirmed in 61 cases. However, 14 vitreous humor specimens showed lower ethanol concentrations, and in the related blood specimens, it was possible to identify the signal of n -propanol, a typical product of postmortem microbial fermentation, that justifies the excess of ethanol in the blood specimens. [ABSTRACT FROM AUTHOR]

The interpretation of postmortem γ-hydroxybutyric acid (GHB) concentrations is challenging due to endogenous existence and postmortem GHB production in body tissues and fluids. As an additional complication, formation of GHB was also described in stored postmortem samples. We examined cardiac blood, femoral blood, vitreous humor, cerebrospinal fluid and urine of eight different corpses (male/female 5/3, aged 33–92 years, postmortem interval 1–6 days) where no intake of GHB or one of its precursors was assumed. All samples were collected during autopsy and divided into two aliquots. To one of the aliquots, sodium fluoride (NaF, 1% w/v) was added. Both aliquots were vortexed, further divided into seven aliquots and stored at −20°C. GHB concentrations were measured immediately and subsequently 1 day, 7 days, 2 weeks, 4 weeks, 3 months and 6 months, after sample collection using trimethylsilyl derivatization and gas chromatography, coupled to single quadrupole mass spectrometry. Similar progression curves of GHB concentrations were obtained for the different matrices in the individual corpses. Femoral and cardiac blood GHB concentrations were always found to be higher than in vitreous humor, cerebrospinal fluid, and urine irrespective of stabilization and storage time. None of the obtained GHB concentrations exceed the cutoff values for postmortem matrices commonly used for the identification of an exogenous GHB intake (urine, venous blood and cerebrospinal fluid: 30 mg/L, cardiac blood and vitreous humor 50 mg/L). No significant differences were found for the GHB concentrations measured immediately and 6 months after autopsy. However, we found a significant increase for the GHB concentrations 4 weeks as well as 3 months after sample collection, which was followed by a decrease nearly to initial values. There were no significant differences between samples with and without NaF addition. The data presented are useful for the interpretation of GHB concentrations in upcoming death cases, with special attention to storage conditions and different postmortem matrices. [ABSTRACT FROM AUTHOR]

A 30 year old non-hispanic white male was found unresponsive at his workplace and admitted to the hospital in cardiac arrest. He was pronounced deceased shortly after arrival. At autopsy the pathologist noted a 176 pound, well-nourished, atraumatic, adult male with significant bilateral frothy pulmonary edema (right lung 930 g and left lung 1,130 g), cardiomegaly (430 g), dilated ventricles and slight cerebral edema. Upon completion of the systematic toxicological analysis scope for the Franklin County Coroner's Office Toxicology Laboratory, no known drugs were found. Further review of the gas chromatography--mass spectrometry (GC--MS) full-scan library summary reports showed an unknown peak in both the blood and urine solid phase extracts. An analogue of α-pyrrolidinovalerophenone (α-PVP) was identified, and a GC--MS selected ion monitoring method was developed to identify and quantitate the presence of 4-fluoro-3-methyl-α-PVP. This method quantified the drug at 26 ng/mL in gray top femoral blood, 30 ng/mL in purple top heart blood and 20 ng/mL in red top vitreous humor. Qualitative presence was also observed in the urine but was not detected in the liver. The decedent's cause of death was determined to be due to fluoro-methyl-PVP toxicity and the manner was ruled to be accidental. Investigational follow-up interviews corroborated drug use by the deceased with a preference of research chemicals and synthetic cannabinoids via the internet. No published literature is available currently, and to the author's knowledge this is the first incident of a fatal death solely attributed to this substituted cathinone. [ABSTRACT FROM AUTHOR]

Widespread access to the Internet has an increasing influence on how suicides are committed. On websites such as eBay® or Amazon.com®, highly toxic substances including cyanides are available for purchase. In the last 5 years, a few fatal intoxications associated with Internet shopping and buying "suicide kits" have been reported. The epidemiology of intoxications reported by the American Association of Poison Control Centers between 2000 and 2018 shows that about 10% of all exposures to cyanide were related to suicide attempts and intentional ingestion of this substance. In order to determine the cyanide concentration in four fatal intoxication cases associated with Internet shopping, a headspace gas chromatography with dual-column/dual-flame ionization detection method was validated and applied to casework. The method was linear in range, from 1 to 50 mg/L, with a coefficient of determination of 0.999 (R2). The limit of quantification was 1.0 mg/L; the detection limit was 0.5 mg/L. Intra- and interday validation precision and accuracy did not exceed 10% and 15%, respectively. Recovery and matrix effect values ranged from 94.8% to 103.8% and from −5.2% to 3.8%, respectively. The cyanide concentrations were determined in biological fluids (blood, urine, bile, vitreous humor and gastric content) and postmortem tissue samples (spleen, kidney, liver and brain). The headspace gas chromatographic method, which is routinely used in clinical and forensic toxicology to quantify ethanol with its congeners (methanol, acetone, isopropanol, n -propanol and n -butanol), can also be applied to determine the cyanide concentrations in intoxication cases. The global problem of a high number of suicides each year requires increasing and more restrictive control of highly toxic substances available online as well as careful monitoring of human exposure to cyanide. This old and well-known poison is being increasingly used nowadays for suicidal purposes; therefore, the determination of cyanide in biological samples is still important in terms of clinical and forensic toxicology. [ABSTRACT FROM AUTHOR]

A 60-year-old man was found dead at home by his wife. The man was known to be a regular trainer in a fitness center and was described as a long-term user of anabolic steroids. The autopsy revealed heart failure with cardiomegaly, visceral congestion, and pulmonary edema. The local prosecutor ordered toxicological analyses but no anatomic pathology test. Initial analyses by the liquid chromatography coupled to quadrupole time-of-flight mass spectrometry system (UPLC--q-TOF-MS) system indicated the presence in femoral blood of trenbolone, an anabolic steroid. Trenbolone was quantified with a liquid chromatography system coupled with a tandem mass spectrometer in various autopsy specimens, including hair. Concentrations of trenbolone were 3.9, 3.2, 231, and lower than 0.5 ng/mL in femoral blood, cardiac blood, bile, and vitreous humor, respectively. Moreover, in head hair, the concentration was 9 pg/mg. The presence of trenbolone in hair can be interpreted as repetitive exposures when compared to the limited literature data. Repetitive misuse of trenbolone can explain advanced heart failure. This paper highlights the importance of testing biological specimens for anabolic steroids in case of heart failure. [ABSTRACT FROM AUTHOR]

Ranolazine is a selective inhibitor of the late inward sodium current, approved for the treatment of chronic angina. Here, a case of a possibly suicidal death due to acute ranolazine overdosing is reported. A 41-year-old woman was found unconscious by her son and was urgently admitted to the intensive care unit. She had ingested an unknown amount of ranolazine tablets. Seventeen hours after admission, the patient died. An autopsy was performed 4 days postmortem. A routine screening analysis for drugs of abuse and medicinal drugs performed by liquid chromatography (LC)–ion trap mass spectrometry (MS) on autopsy samples of biological fluids did not detect any relevant presence of toxicologically relevant compounds, but ranolazine. A quantitative analysis was then carried out by liquid chromatography--triple quadrupole (LC–QqQ) MS in order to quantify ranolazine and its major metabolite o-desmethylranolazine in biological fluids and organs. Ranolazine concentrations in biological fluids were as follows: cardiac blood, 19.5 μg/mL; femoral blood, 12.3 μg/mL; bile, 0.87 μg/mL and vitreous humor, 15.4 μg/mL. For O-desmethylranolazine, the concentrations in cardiac blood, femoral blood, bile and vitreous humor were 10.7, 9.6, 11,103 and 11.4 μg/mL, respectively. The cause of death was attributed to ranolazine overdosing. To the best of our knowledge, this is the first report of a fatality associated with ranolazine, in which the postmortem distribution of ranolazine and its metabolite has been quantitatively assessed. The present study can therefore provide useful information for interpretation of the causes and mechanisms of death in ranolazine-associated fatalities. [ABSTRACT FROM AUTHOR]

In December 2019, unusual cases of acute renal failure with neurological changes were observed in the State of Minas Gerais, Brazil. Criminal investigations indicated cases of intoxication after consumption of beers contaminated with diethylene glycol (DEG). The elimination of DEG by the body is fast, but its metabolite, diglycolic acid (DA), may persist for a long time. To assess the level of intoxicated victims who consumed the contaminated beers, qualitative and quantitative methods were developed to determine DA in biological matrices by gas chromatography coupled to a mass spectrometer (GC–MS) and gas chromatography coupled to a mass spectrometer with triple-quadrupole mass filter (GC–MS-MS), respectively. The validated qualitative method presents good selectivity and limit of detection of 1 µg/mL (whole blood, urine, vitreous humor and cerebrospinal fluid) and 5 µg/g (liver and kidney), respectively. A quantitative method for whole blood presented satisfactory performance to determine DA. Twelve victims presented positive results for DA in whole blood, with concentrations ranging from 2 to 108 µg/mL. The toxicology laboratory of the Institute of Forensic Medicine of Minas Gerais was the first governmental agency to identify DA in whole blood, vitreous humor, cerebrospinal fluid, kidney and urine in victims affected by this contaminant. The results of this study legally supported the prohibition of the continued consumption of the beer and avoided further intoxications. Our results showed, for real cases of human intoxication, that DA can still be detected in alternative matrices, even when non-detectable in blood, demonstrating the importance of collecting different kinds of samples for a proper investigation. [ABSTRACT FROM AUTHOR]

Postmortem chemistry can be a useful ancillary technique that the forensic pathologist can use during a death investigation. In stark contrast, there is limited information available for use of postmortem vitreous humor analysis in animals. In order to use postmortem vitreous humor in veterinary forensic investigations, validation of a method to analyze vitreous humor is required. The goal of this study was to determine the precision, bias, TEobs and sigma (σ) of the Element DC chemistry analyzer; assess its precision using the vitreous humor collected postmortem from dogs, cats and horses and assess the stability of postmortem vitreous humor from all the three species. Analysis of quality control material (QCM) and pooled vitreous humor samples for the three species was used to test for sodium (Na), chloride (Cl), potassium (K), magnesium (Mg), creatinine (Crea) and blood/vitreous urea nitrogen. Analysis of QCM showed that the Element DC was both precise and accurate. When analyzing the pooled vitreous humors, most within-run coefficients of variance (CVs) were found to be <5% and the between-run CVs for five out of six analytes were found to be <5% for dogs, cats and horses. In all the three species, the capped samples of vitreous humor were stable out of refrigeration for up to 5 h. The results of this study show that the Element DC can successfully be used to analyze the postmortem vitreous humor from dogs, cats and horses. [ABSTRACT FROM AUTHOR]

A group of 16 volatile substances (ethyl acetate, 2-propanol, 1-propanol, methanol, acetone, ethanol, acetaldehyde, diethyl ether, methyl ethyl ketone, 1-butanol, 2-butanol, t -butanol, isobutanol, 2-methyl-1-butanol, 3-methyl-1-butanol and 1-pentanol) were qualitatively and quantitatively analyzed through a method developed for volatiles with endogenous production in putrefaction and submersion situations. The method was validated for blood, urine and vitreous humor, using a gas chromatograph (Varian 450-GC) with a flame ionization detector coupled to a headspace injector (HS-GC–FID). The vials were prepared by diluting 100 µL of the sample of interest in 1 mL of internal standard (acetonitrile 100 mg/L), using two capillary columns (VF-624ms and VF-5ms) with different polarities to ensure that all test compounds would be properly identified and undoubtedly distinguished from the rest. All volatiles were studied in a range of 50 to 2,000 mg/L in terms of selectivity/specificity, detection and quantification limits, linearity and calibration model, precision, accuracy, bias, robustness and stability according to the Scientific Working Group for Forensic Toxicology. Detection and quantification limits were between 1 to 8 mg/L and 4 to 24 mg/L, respectively, with coefficient of variation values under 10% in bias studies and in intermediate precision studies for most substances. The developed method was applied to real cases to test the method. [ABSTRACT FROM AUTHOR]

The aim of the study is to evaluate the contribution of ethanol metabolite detection in postmortem cases by showing the connection between the presence of ethanol metabolites, which are indicators of alcohol consumption, and the detection of potential postmortem ethanol formation in decomposed and diabetic cases. Determination of ethanol consumption before death is often one of the most important questions in death investigations. Postmortem ethanol formation or degradation products in the blood make it difficult to distinguish antemortem consumption or postmortem formation of ethanol and eventually may lead to misinterpretation. Decomposed bodies and diabetic cases are vulnerable to postmortem ethanol formation due to putrefaction, fermentation or other degradations. Ethyl glucuronide (EtG) and ethyl sulfate (EtS) are two metabolites of ethanol produced only in the antemortem time interval. In this study, EtG and EtS levels in urine and vitreous humor samples of 27 postmortem cases, including diabetic and degraded bodies were compared to ethanol results of their blood, urine and vitreous humor samples. EtG and EtS in urine and vitreous humor were analyzed by liquid chromatography–tandem mass spectrometry, and ethanol was assayed by routine headspace gas chromatography–flame ionization detector. These cases were devoid of other influences from forensically relevant drugs, so ethanol and/or glucose were among the only positive findings in these cases. The results of this pilot study indicate the postmortem ethanol concentrations do not correlate with the measured EtG and EtS values but are beneficial in rulings of accidental or natural deaths. This preliminary study gives additional data to help distinguish between antemortem ethanol intake and postmortem formation. EtG and EtS were well correlated positively with antemortem ethanol use instead of forming spontaneously in samples from decedents who are decomposing or have a history of diabetic hyperglycemia. [ABSTRACT FROM AUTHOR]

Etizolam is a novel psychoactive substance and novel benzodiazepine of the thienotriazolodiazepine class, which has recently seen an increasing trend in use worldwide. We report a case series of 10 decedents with etizolam and opioids in their systems. Death investigation, expanded toxicology and medical investigation information were included for contextualization of etizolam in death. Etizolam was detected and confirmed within peripheral and cardiac blood, urine, vitreous humor and, in one case, gastric fluid, by liquid chromatography–tandem mass spectrometry and liquid chromatography–quadrupole time of flight mass spectrometry methodologies. Death investigation indicated nonmedical use of most drugs. Medical investigation commonly noted pulmonary edema, cardiomegaly and cerebral swelling. The majority of the decedents appeared to be unaware of the presence of etizolam and succumbed to the mixed drug toxicity of their routine depressant and narcotic analgesic drug of abuse in combination with etizolam. Etizolam use continues to be observed and poses as a potentially lethal contribution to multiple drug toxicity, especially in the age of the opioid crisis. Assessment of analytes like etizolam requires up-to-date methodologies and vigilance in testing to better characterize the toxicology and interpret the contribution to death. [ABSTRACT FROM AUTHOR]

Identifying the source of ethanol in a decedent remained a complicated problem for forensic toxicologists because of postmortem ethanol formation. As ethanol's non-oxidative metabolites, ethyl glucuronide (EtG) and ethyl sulfate (EtS) have the potential to distinguish between antemortem ethanol consumption and postmortem ethanol formation, due to their high sensitivity and selectivity. In the current study, a simple and quick liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for the determination of EtG and EtS in human whole blood and vitreous humor (VH). A total of 20 μL of the sample was precipitated by methanol, and the analytes were detected by LC-MS/MS in a run of 6 min. This method achieved high sensitivity (limits of detection: 2 ng/mL for both EtG and EtS), with linearity in the range of 5–10,000 ng/mL in both whole blood and VH. Deviations in accuracy, inter- and intra-day precision were all lower than 15% at three quality control levels. Subsequently, this method was applied to 62 real forensic cases. Only blood samples were available in 52 cases. Paired blood and VH samples were present in 10 cases. The concentrations of EtG and EtS in blood were in the range of 0–22,264.8 ng/mL and 0–2,126.0 ng/mL, respectively. In one case with both blood and VH, the blood ethanol concentration was 1.22 mg/mL, with EtG and EtS both below limits of quantification (5 ng/mL) in VH, and no EtG and EtS found in whole blood. The results suggested that EtG and EtS were useful markers for the interpretation of ethanol resource in postmortem blood and VH. [ABSTRACT FROM AUTHOR]

In case of basic biological materials (blood, urine, vitreous humor) being unavailable, bone marrow can be used for toxicological tests. The aim of the study was to assess the concentration of various xenobiotics in the bone marrow, as well as in the blood and the vitreous humor. The analysis was performed in the biological material originating from the autopsy (n  = 120), using the LC–MS method and with liquid/liquid extraction at pH = 9. As many as 46 different xenobiotics were detected in the biological material, strong correlations between the concentration of a given xenobiotic in blood and bone marrow, as well as in the vitreous humor and bone marrow, were noted for most of them, with the exception of diazepam and 7-aminoclonazepam. The obtained results indicate the possibility of using bone marrow to determine the concentration of numerous xenobiotics in the situation of basic biological materials being unavailable, as well as using the results obtained in the future to better understand the pharmacokinetic processes and the effect of postmortem redistribution on medication and drug concentrations in the body of a deceased person. [ABSTRACT FROM AUTHOR]

Vitreous humor is a potential alternative matrix for postmortem toxicology drug screens when peripheral blood is unavailable. It is easily and reliably collected and may not suffer from the same postmortem redistribution as seen in blood. Here, we compared the concentrations of 7 acidic drugs (acetaminophen, ibuprofen, naproxen, salicylic acid, carbamazepine, phenobarbital and phenytoin) in peripheral blood and vitreous fluid collected in 89 autopsy cases. Analysis was done by high-performance liquid chromatography with diode-array detection. Overall, we found that vitreous drug concentrations were significantly lower than peripheral blood with median vitreous to peripheral blood (V/PB) ratios ranging from 0.0 to 0.6 (mean, 0.1–0.6). The correlations between the concentrations of over-the-counter analgesics in peripheral blood versus vitreous fluid were poor, with acetaminophen exhibiting the best linearity (R 2 = 0.72). The antiepileptic drugs (carbamazepine, phenytoin and phenobarbital) exhibited good correlations between peripheral blood and vitreous humor, with all exhibiting an R 2 ≥ 0.95. Overall, we have demonstrated the potential of vitreous fluid as an alternative matrix for the detection of select acidic drugs. [ABSTRACT FROM AUTHOR]

Here we report the identification and quantitation of cyclopropyl-fentanyl in a fatality casework occurred due to a poly-drug toxicity in Spain in December 2017. The cyclopropyl-fentanyl was identified in non-biological (paraphernalia) and biological samples (whole-blood, vitreous humor and urine). Conventional techniques (GC–MS) and the UV-Vis spectral allowed differencing between the two structural isomer compounds of fentanyl (cyclopropyl and crotonyl) when the CRM was not available at the laboratory. Both of the drugs have the same MS spectra but different UV-Vis spectra due to the presence of an additional chromophore group in the case of crotonyl. The cyclopropyl-fentanyl detection allowed generating an alert and contributing to the surveillance and detection of this dangerous substance found mixed in doses of clandestine sale heroin. Then, high-resolution analytical techniques (LC–HRMS-MS), showed limitations for the identification of the isobaric fentanyl compounds but they had a high potential for fentanyl metabolites identification. Two tentative metabolites were identified in urine samples: cyclopropyl-norfentanyl and N-methyl cyclopropyl-norfentanyl. Finally, the systematic routine method (LC–MS-MS) was validated and applied to the quantification of cyclopropyl-fentanyl in a blood sample. A obtained value (20.4 ng/mL) was in the range of those reported in other cases in different countries (from 8 to 30 mg/ mL), being the determined concentration of cyclopropyl-fentanyl high enough to infer that this fentanyl analog had a main role as cause of death. As far as we know, this is the first fatality reported in Spain involving cyclopropyl-fentanyl [ABSTRACT FROM AUTHOR]

In connection with medicolegal autopsies peripheral blood (e.g. from a femoral vein) is the specimen of choice for toxicological analysis, although alternative specimens are also sometimes submitted, such as bile, cerebrospinal fluid (CSF), vitreous humor (VH), bladder urine, pleural effusions and/or lung fluid. Ethanol concentrations were determined in duplicate in femoral blood and in various alternative biological specimens by headspace gas chromatography. The analysis was carried out on two different fused silica capillary columns furnishing different retention times for ethanol and both n -propanol and t -butanol were used as internal standards. The results were evaluated by linear regression using blood alcohol concentration (BAC) as dependent or outcome variable and the concentrations in an alternative specimen as independent or predictor variable. The Pearson correlation coefficients were all statistically highly significant (P  < 0.001); r  = 0.94 (bile), r  = 0.98 (CSF), r  = 0.97 (VH), r  = 0.92 (urine), r  = 0.94 (lung fluid) and r  = 0.96 (pleural cavity effusions). When the regression model was used to predict femoral BAC from the mean concentration in an alternative specimen the mean and 95% prediction intervals were 1.12 ± 0.824 g/L (bile), 1.41 ± 0.546 g/L (CSF), 1.15 ± 0.42 g/L (VH), 1.29 ± 0.780 g/L (urine), 1.25 ± 0.772 g/L (lung fluid) and 0.68 ± 0.564 g/L (pleural cavity effusions). This large uncertainty for a single new observation needs to be considered when alcohol-related deaths are evaluated and interpreted. However, the analysis of alternative specimens is recommended in medical examiner cases to provide supporting evidence with regard to the origin of ethanol, whether this reflects antemortem (AM) ingestion or postmortem (PM) synthesis. [ABSTRACT FROM AUTHOR]

Gamma-hydroxybutyrate (GHB) is a naturally occurring molecule present in the human body as a catabolite of the neurotransmitter gamma-aminobutyrate (GABA). In the USA, GHB has a history of being manufactured illicitly and abused, with misguided proposed benefits for the body-building community and a persistent party drug with reported GHB overdoses occurring worldwide. The interpretation of GHB in postmortem biological fluids is complicated by the endogenous nature of the molecule. Analysis often requires more than one biological matrix to detect exogenous exposure, typically in urine. The analysis is further complicated by the endogenous de novo production of GHB in postmortem specimens. This work sought to examine the prevalence of endogenous GHB concentrations in postmortem toxicology samples from Orange County, CA, and to establish suitable in-house secondary matrices to confirm or rule out exogenous GHB exposure. A total of 348 postmortem heart blood samples were randomly selected and analyzed for GHB using liquid–liquid extraction followed by gas chromatography–mass spectrometry with selective ion monitoring and GHB-d6 as an internal standard. Of the 348 cases analyzed, 39 cases resulted in positive GHB detection with the median concentration of 22.45 mg/L (10.4–62.16 mg/L). None of the positive samples had suspected GHB ingestion or usage from the case report. GHB concentrations were then examined in secondary matrices collected at autopsy from the positive cases that included (when available) peripheral blood, urine, vitreous humor, liver homogenate and brain homogenate. Within the secondary matrices, GHB levels in peripheral blood compared to that of heart blood, while liver homogenate levels were variable. Quantifiable GHB levels were not identified in vitreous humor and brain homogenate samples. Our findings reaffirm the importance of multi-matrix analysis in postmortem toxicology and further confirm the utility of vitreous humor and brain tissue to distinguish exogenous GHB exposure from endogenous production. [ABSTRACT FROM AUTHOR]

A specific, sensitive, fast and simple method for analysis of Δ9-tetrahydrocannabinol (THC), 11-hydroxy-Δ9-THC (THC-OH) and 11-nor-Δ9-THC-9-carboxylic acid (THC-COOH) in routine postmortem cases using LC–MS–MS was developed and validated. Prior to solid phase extraction, urine, stomach contents and bile were pretreated using alkaline hydrolysis, while blood and vitreous humor were pretreated with protein precipitation. The distribution of THC, THC-OH and THC-COOH were investigated in 31 postmortem cases that tested positive for cannabinoids. This revealed new information regarding the distribution of THC in stomach contents and vitreous humor. Alkaline hydrolysis was sufficient for the deglucuronidation of THC-COOH-glucuronide to its free form, THC-COOH, in urine, bile and stomach contents. However, the THC-OH concentration in bile reported in this study is considerably high compared to that of previous studies. In conclusion, including THC and its metabolites (THC-OH and THC-COOH) is crucial for any forensic toxicology detection method to most accurately determine the role of cannabinoids in deaths. [ABSTRACT FROM AUTHOR]

The analysis of biological specimens collected at autopsy for the presence of exogenous insulin(s) is of special interest in select death investigations as they may be suspected in the cause of a death. Technical challenges include the limited stability of insulin, and the forensic requirement of differentiating endogenous insulin from pharmaceutical analogs. A novel method was developed for the detection and quantification of human insulin, Glulisine, Lispro, Aspart, Glargine and Detemir in vitreous fluid. An immunoaffinity extraction procedure is performed followed by separation of the insulin α- and β-chains. Liquid chromatography tandem mass spectrometry analysis of the β-chain allows for the unequivocal identification of each insulin analog. The analytical measurement range for each insulin was 0.5–25 ng/mL. The method was evaluated for accuracy, precision, carryover, interferences and stability. Eight vitreous fluid samples collected from cases where untoward insulin use was suspected were subjected to analysis. Positive results were obtained from three samples, and a detailed case history is provided for one of these cases. Even though insulin instability in postmortem biological fluid remains a challenge, this method allows for a reliable forensic-level analysis in vitreous fluid. [ABSTRACT FROM AUTHOR]

Vitreous humor (VH) shows excellent potential as a matrix of choice for postmortem analytical toxicology due to the ease of sampling and low metabolic activity. This study demonstrates a simple and rapid analytical method to identify and quantify 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine and 3,4-methylenedioxy-ethylamphetamine in VH. Samples were collected with a simple eye puncture procedure, followed by liquid–liquid extraction and derivatization with heptafluorobutyric anhydride and analysis via gas chromatography–mass spectrometry. The accuracy of the method ranged 97–103%, intra-assay precision was between 4.54 and 9.14% relative standard deviation (RSD) and interassay precision ranged from 6.92 to 10.59% RSD. Limits of detection and quantification ranged from 1.0 to 2.5 ng/mL and 10 ng/mL, respectively. The validated method was successfully applied to detect methylenedioxyamphetamine derivatives in VH samples collected from victims of fatal car crashes. [ABSTRACT FROM AUTHOR]

Unintentional laser exposure is an increasing concern in many operational environments. Determining whether a laser exposure event caused a retinal injury currently requires medical expertise and specialized equipment that are not always readily available. The purpose of this study is to test the feasibility of using dynamic light scattering (DLS) to non-invasively detect laser retinal injuries through interrogation of the vitreous humor (VH). Three grades of retinal laser lesions were studied: mild (minimally visible lesions), moderate (Grade II), and severe (Grade III). A pre-post-treatment design was used to collect DLS measurements in vivo at various time points, using a customized instrument. VH samples were analyzed by liquid chromatography/tandem mass spectrometry (LC-MS/MS) and relative protein abundances were determined by spectral counting. DLS signal analysis revealed significant changes in particle diameter and intensity in laser-treated groups as compared with control. Differences in protein profile in the VH of the laser-treated eyes were noted when compared with control. These results suggest that laser injury to the retina induces upregulation of proteins that diffuse into the VH from the damaged tissue, which can be detected non-invasively using DLS. [ABSTRACT FROM AUTHOR]

The analytical value of vitreous humor as a specimen in postmortem forensic toxicology has been known for some time. Numerous medical examiner laboratories outsource the analysis of this important specimen for electrolyte and glucose measurements. This can be both time-consuming and costly. The utility of the i-STAT®1 medical device to measure electrolytes and glucose in whole blood samples has been demonstrated for over two decades in a clinical setting through single-use disposable cartridges that introduce samples to the i-STAT®1. Different cartridge types allow for the analysis of various analytes including sodium, potassium, chloride, creatinine, urea nitrogen and glucose. With only 100 µL of sample, results are obtained in under 4 min. In this study, we utilized the i-STAT®1 using an alternative specimen matrix, postmortem vitreous humor and quantitatively determined the validity and reliability of the instrument for this purpose. Acceptable criterion was used for each test as suggested by the Scientific Working Group for Forensic Toxicology. All analytes of interest, except creatinine, demonstrated a percent error < ±10% for both accuracy and precision studies. Drug interference and stability studies were performed with many of the analytes demonstrating a percent error < ±20%. Throughout drug interference and stability studies, all analytes of interest were detectable except for potassium, which gave inconclusive results. Significant interference with commonly found drugs were shown for creatinine and chloride but must be evaluated carefully. Volume additions to ethanol spiked samples caused significant interference for all analytes and is considered a limitation for this method of analysis that requires additional studies. As vitreous humor continues to be used in forensic medicine to aid in diagnostic interpretation, the i-STAT®1 has the potential to give accurate results in a timely and cost-effective manner. [ABSTRACT FROM AUTHOR]

Alternative specimens have been occasionally considered as substitutes for whole blood for postmortem toxicology testing. We studied the applicability of vitreous humor, and evaluated whether it would be suitable to replace (or augment) whole blood for routine drug screening. Results showed that from 51 autopsy cases, we were able to identify an aggregate of 209 findings in whole blood compared with 169 in vitreous. The total number of compounds identified was 71 for whole blood and 60 for vitreous humor. Quantitative analysis showed that whole-blood concentrations of trazodone were several fold higher than vitreous humor concentrations (1.42 ± 0.57 vs. 0.15 ± 0.05 mg/L, respectively) and similar results were also obtained for diazepam (0.37 ± 0.06 vs. 0.13 ± 0.01, respectively). For other drugs such as oxycodone, hydrocodone and doxylamine, a trend suggesting higher concentrations in vitreous humor vs. whole blood was observed; however, this was not significant. Our results are consistent with the limited work of other investigators, and suggest that vitreous humor could be an appropriate matrix for drug screening in postmortem toxicology. [ABSTRACT FROM AUTHOR]

In forensic bioanalytical methods, there is a general agreement that calibrators should be prepared by fortifying analytes in matrix-based blank samples (matrix-based). However, in the case of vitreous humor (VH), the collection of blank samples for the validation and for routine analysis would require the availability of many cadavers. Besides the difficulty of obtaining enough blank VH, this procedure could also represent an ethical issue. Here, a study of matrix effect was performed taking into consideration human and bovine vitreous and saline solution (SS) (NaCl 0.9%). Tricyclic antidepressants [amitriptyline (AMI), nortriptyline (NTR), imipramine (IMI) and desipramine (DES)] were used as model analytes and were extracted from samples by means of liquid-phase microextraction and detected by gas chromatography--mass spectrometry. Samples of human and bovine VH and SS were prepared in six different concentrations of antidepressants (5, 40, 80, 120, 160 and 200 ng/mL) and were analyzed. Relative matrix effect was evaluated by applying a two-tailed homoscedastic Student's t-test, comparing the results obtained with the set of data obtained with human VH and bovine VH and SS. No significant matrix effect was found for AMI and NTR in the three evaluated matrices. However, a great variability was observed for IMI and DES for all matrices. Once compatibilities among the matrices were demonstrated, the method was fully validated for AMI and NTR in SS. The method was applied to six VH samples deriving from real cases whose femoral whole blood (FWB)was analyzed by a previously published method. An average ratio (VH/FWB) of ~0.1was found for both compounds. [ABSTRACT FROM AUTHOR]

A GC–MS method for simultaneous analysis of cocaine (COC), amphetamines (AMPs), opiates, cannabinoids and their metabolites in vitreous humor (VH) was developed and fully validated. VH samples were extracted using solid phase extraction and injected into the GC–MS, using a selected ion monitoring mode. Linearity ranged from 10 to 1000 ng/mL; the exception was anhydroecgonine methyl ester (AEME), for which linearity ranged from 10 to 750 ng/mL. Inter-assay imprecision lay from 1.2 to 10.0%, intra-assay imprecision was <10.4% for all the analytes and accuracy ranged from 95.6 to 104.0%. An limit of quantitation for all drugs was 10 ng/mL and recoveries ranged from 70.4 to 100.1% for basic and neutral compounds; the acid compounds had poor recovery—<40%. The validated method was applied to 10 VH samples taken from individuals whose blood had screened positive for drugs of abuse. All the individuals screened positive for COC in the blood (seven samples) also had positive results in VH; COC concentration ranged from 30.81 to 283.97 ng/mL (mean 186.98 ng/mL) and benzoylecgonine concentration ranged from 11.47 to 460.98 ng/mL (mean 133.91 ng/mL). It was also noticed that, in five cases, cocaethylene was detected. AEME was also quantified in one case. The use of AMP detected by blood analysis was confirmed in the VH of one individual (24.31 ng/mL). However, samples taken from three individuals whose blood tested positive for carboxy-tetrahydrocannabinol presented negative results. The results demonstrated that VH is a suitable alternative biological sample to determine COC, AMPs, opiates and their metabolites. [ABSTRACT FROM PUBLISHER]

The article presents a case study of a 30 years old man who was found in a critical state in a staircase and died despite resuscitation attempts. It mentions that eight small plastic bags containing white powder were found in the jacket of the dead male, which was tested using gas chromatography–mass spectrometry and high-pressure liquid chromatography. The analysis of blood and vitreous humor samples showed the presence of mephedrone, a type of metamphethamine used as designer drug.

The article focuses on a study related to the deaths caused due to intoxication of a drug 1,1-difluoroethane in the U.S. As per the study, 17 cases from the San Diego County Medical Examiner's Office in California showed the presence of drug in postmortem tissues. It states that ethanol screening method evaluates the concentrations of drug in peripheral blood, central blood and vitreous humor. It further concludes that many cases death occurs in minutes after intentional abuse of drug.

The article reports on a study which validates the applicability of photodiode-array deletion method in determining the presence of benzodiazepines in vitreous humor. It states that the study was carried out by chromatographic observation of thirty microliters of an extract with vitreous humor components. It asserts that high-performance liquid chromatography, in which photodiode-array deletion method is an example, is a good method for benzodiazepine analysis.

The article presents a study that investigates the applicability of vitreous humor as an alternative medium for drug screening by liquid chromatography-time-of-flight mass spectrometry (LC-TOFMS) in postmortem examinations. It mentions that findings between vitreous humor and urine in 50 autopsy cases were compared. It states that vitreous humor is applicable for qualitative screening analysis, although its representativeness was not as good as urine due to less frequent metabolite detection.

The article examines the relationship between immunoassay results and gas chromatography-mass spectrometry quantification of oxycodone in postmortem vitreous humor and blood. It was found that the vitreous humor and postmortem blood precipitate immunoassay responses in oxycodone-positive cases were correlated. Confirmation of quantification of oxycodone in vitreous humor was linear. There are few cases wherein disparities between vitreous fluid and blood oxycodone concentrations were seen.

The article presents a study regarding the use of vitreous humor for immunoassay screening. Researchers from Microgenics Cloned Enzyme Donor Immunoassay (CEDIA DAU) state that patients who undergo drug analysis are positive in opioids. In addition, they use chromatography-nitrogen-phosphorous detection (GC-NPD) screening method.

The article talks about the loss of ethanol from vitreous humor in deaths caused by drowning. As in the case of two different deaths, wherein the victims died by drowning with extended periods underwater, more or less 2 and 4 weeks, the ethanol concentrations found in their central blood were 260 and 280 based on the postmortem analysis. Moreover, this ethanol concentration was also reduced in the vitreous with 50 and 80, and 330 and 320 in urine (mg/100 mL). The serotin metabolite ratios support the explanation behind ethanol's diffusion from the vitreous fluid towards the surrounding water, instead of the ethanol's postmortem production in blood as the main cause of the differences on the blood-vitreous ethanol.

Describes the analysis of gamma-hydroxybutyric acid (GHB) and 4-methyl-GHB in postmortem matrices after long-term storage. Extraction procedure; Case selection; Method validation; Vitreous humor concentrations in documented cases of GHB positive fatalities; Postmortem endogenous range for GHB in vitreous humor.

The possibility of performing reliable post-mortem analysis of carbohydrate-deficient transferrin (CDT) concentration in vitreous humour (VH) by using a commercial assay designed for serum analysis (CDTect™) as well as the usefulness of VH-CDT as a marker of alcohol misuse and possible withdrawal-related death were evaluated in a forensic sample. Detectable VH-CDT was found in 20 of 21 alcoholic subjects and in two of seven controls. By using the detection limit of the CDTect™ method (VH-CDT = 5 U/l) as cut-off level for a positive test, the alcoholic group was significantly separated from the control group (P = 0.0024, Fisher's exact test). The sensitivity and specificity of the test was 95% and 71%, giving a positive and a negative predictive value of 91% and 83%, respectively. Time-dependent changes of VH-CDT in the dead body could not unequivocally be excluded, which must be considered when selecting cases suitable for VH-CDT analysis. We conclude that adding VH-CDT analysis to ordinary alcohol tests may become useful in forensic medicine for establishing the so-called ‘alcoholic state’, which may provide a tool in research dealing with the relation between alcohol withdrawal and various causes of death in alcoholics. [ABSTRACT FROM PUBLISHER]

Metaxalone (Skelaxin) is a prescription medication used primarily as a centrally acting skeletal muscle relaxer and is rarely implicated in drug fatalities. We present a case study involving a relatively young decedent where metaxalone is implicated as the sole agent causing death with little in the way of confounding factors. The concentration of metaxalone in hospital admission blood was determined to be 37.4 mcg/mL. In postmortem specimens the concentrations were shown to be 13.5mcg/mL (heart blood), 4.9 mcg/mL (vitreous humor), 69.4 mcg/g (liver) and 74.0 mcg/g (brain). Additionally a blood-to-plasma (b/p) ratio was estimated using antemortem blood and serum specimens taken at the same time on the second day following admission. The b/p ratio was calculated to be 1.4 implying a higher proportion of the drug to be found in whole blood versus plasma/serum samples, an important factor which should be taken into account when comparing blood concentrations to published therapeutic ranges determined in serum/plasma. [ABSTRACT FROM AUTHOR]

The article presents a study on the distribution of phencyclidine (PCP) into vitreous humor. It mentions that heart blood, subclavian blood and vitreous humor specimens from 26 PCP-positive postmortem cases were analyzed to evaluate the distribution of PCP in vitreous humor. It mentions that the specimens were analyzed by solid-phase extraction followed by gas chromatography-mass spectrometry. It declares that the results reveals that all positive blood specimens were associated with positive vitreous humor specimen, and on average, the blood PCP concentrations were greater than the vitreous humor PCP concentrations. It concludes that there were considerable variability between cases, which shows that the detection of PCP in postmortem cases' interpretative value is limited.

Investigates ethanol concentrations in postmortem blood and vitreous humor samples stored long-term. Refrigerated storage; Stability of ethanol; Reliability of vitreous humor as a reliable matrix for ethanol analysis.

Background Ocular syphilis is an infrequent presentation of Treponema pallidum infection. It is often seen in association with neurosyphilis but can occur in isolation as well. Here we describe a case series of 10 patients with ocular syphilis seen at our institution. Methods Patients with a diagnosis of ocular syphilis at the three sites of Mayo Clinic were identified from June 2006 to February 2019. Patient's baseline characteristics, clinical presentation, diagnostic testing, and treatment were abstracted from Electronic Medical Records. Results We identified 10 patients (17 eyes) during the study period, half being diagnosed in the last 5 years. The median age was 51 years (IQR 21–66), and the majority were males (7/10). Characteristics of the patients are outlined in Table 1. Common presenting symptoms included vision loss (8/10), photophobia (4/10), and irritation (5/10). Seven of 10 patients had bilateral involvement. Of the 17 eyes, 59% had pan-uveitis and 82% had disc edema (Table 2). Rapid Plasma Reagin (RPR) was positive in 9 patients and median serum RPR was 1:128 (IQR 1:2-1:1024). One patient was diagnosed with a positive TP-PA, and characteristic ocular findings. One patients had positive 16S Ribosomal RNA Gene PCR/Sanger Sequencing on vitreous humor specimen for Treponema species. The majority of patients had isolated ocular symptoms at presentation; two patients also had an associated skin rash. All patients underwent a lumbar puncture (LP). Half had pleocytosis (TNC > 5) and elevated protein elevated (mean 63.6 mg/dL), although not significantly high. 3/10 patients had VDRL positive in the CSF. Protein levels were much higher in those with positive VDRL. Each patient was treated with intravenous (IV) penicillin for at least 2 weeks. Nine patients received ocular steroids as well. At last available follow-up, 8 had complete and 1 had partial resolution. One had worsened eye findings with eventual scarring. Conclusion Our findings are congruent with increasing incidence of ocular syphilis nationwide. Bilateral eye involvement and pan-uveitis were the most common ocular findings. Although only 3 patients had a positive VDRL in the CSF, pleocytosis and elevated protein were found in a large number of patients. Majority of the patients had clinical resolution with appropriate treatment. Disclosures All authors: No reported disclosures. [ABSTRACT FROM AUTHOR]

Presents the abstract of the study "Interpreting Postmortem Tricyclic Antidepressant Levels in Vitreous Humor," by Uttam Garg and colleagues.

Presents the abstract of the study "Stability of Ethanol in Postmortem Blood and Vitreous Humor in Long-Term Refrigerated Storage," by Theresa Olsen and W. Lee Hearn.