Simultaneous Quantitative Determination of Amphetamines,Opiates, Ketamine,Cocaine and Metabolites in Human Hair: Application to Forensic Cases of Drug Abuse

A method using liquid chromatography–tandem mass spectrometry (LC–MS/MS) to simultaneously quantify amphetamines, opiates, ketamine, cocaine, and metabolites in human hair is described. Hair samples (50 mg) were extracted with methanol utilizing cryogenic grinding. Calibration curves for all the analytes were established in the concentration range 0.05–10 ng/mg. The recoveries were above 72%, except for AMP at the limit of quantification (LOQ), which was 48%. The accuracies were within ±20% at the LOQ (0.05 ng/mg) and between −11% and 13.3% at 0.3 and 9.5 ng/mg, respectively. The intraday and interday precisions were within 19.6% and 19.8%, respectively. A proficiency test was applied to the validated method with z-scores within ±2, demonstrating the accuracy of the method for the determination of drugs of abuse in the hair of individuals suspected of abusing drugs. The hair concentration ranges, means, and medians are summarized for abused drugs in 158 authentic cases.     

Quantification of Pubic Symphysis Metamorphosis Based on the Analysis of Clinical MDCT Scans in a Contemporary Malaysian Population

Forensic age estimation methods are biased to sex and population; in general, accuracy is reduced when applied to foreign populations. This study assessed the accuracy of the Suchey–Brooks method in contemporary Malaysian individuals and aimed to formulate population‐specific standards. Multi‐detector computed tomography scans of 355 individuals (165 male; 190 female) of 15–83 years of age were reconstructed using 3D‐volumetric rendering in RadiAnt. Pubic symphyseal phase, bias, inaccuracy, and percentage correct age classifications are examined. Transition analysis was used to develop age estimation standards. High observer agreement (κ = 0.763–0.832) and a positive relationship between age and pubic symphyseal phase (r = 0.884–0.90) were demonstrated. Mean inaccuracies were 8.62 and 8.95 years for males and females, respectively; overall correct classification was 97.8%. Transition ages between phases in males were 18.79, 23.29, 28.85, 43.64, and 61.15 years; in females, the corresponding data were 19.77, 22.53, 32.62, 41.85, and 57.39 years.     

The Applicability of the Lamendin Method to Skeletal Remains Buried for a 16-Year Period: A Cautionary Note

The Lamendin method is widely reported as one of the most reliable means of age estimation of skeletal remains, but very little is known concerning the influence of burial in soil. This study aimed at verifying the reliability of the Lamendin method on corpses buried for 16 years in a cemetery. The Lamendin and the Prince and Ubelaker methods were applied. In all age groups except the 40- to 49-year–olds, the error was higher in the buried sample. The age-at-death error ranged between 10.7 and 36.8 years for the Lamendin method (vs. the reported 7.3–18.9 years) and 9.5 and 35.7 for the Prince and Ubelaker one (vs. the original 5.2–32.6 years); in all age groups, the error is closer to that found on archeological populations. These results suggest caution in applying the Lamendin method to forensic cases of human remains buried even for a brief period under soil.     

A novel method for cyanide quantification in human whole blood using ion chromatography with amperometric detection and its application to cyanide intoxication cases

Cyanide is a highly toxic agent that has been frequently used for suicide in South Korea. It is also used in various industrial fields, such as metal plating, in which many accidental cyanide intoxications have occurred. To overcome the disadvantages of conventional cyanide analysis methods, a simple and fast method for the analysis of cyanide in whole blood using ion chromatography (IC) with amperometric detection was developed in this study. Whole blood samples were deproteinized, diluted, and analyzed using an IC–amperometric detection system. The limits of detection and quantitation were 0.1 and 0.2 mg/L, respectively. The method showed good linearity in the range of 0.2 to 50 mg/L with R² > 0.99. The intra- and inter-assay precision and accuracy values were <10%. The established method was successfully applied to analyze whole blood samples from three cyanide intoxication cases.     

A New Approach for the Carbon Monoxide (CO) Exposure Diagnosis: Measurement of Total CO in Human Blood Versus Carboxyhemoglobin (HbCO)

The aim of the study is to present the application of a headspace–gas chromatography–mass spectrometry (HS‐GC‐MS) method for the determination of the carbon monoxide (CO) blood concentration and to compare it with carboxyhemoglobin (HbCO) saturation. In postmortem cases, the HbCO measured by spectrophotometry frequently leads to inaccurate results due to inadequate samples or analyses. The true role of CO intoxication in the death of a person could be misclassified. The estimation of HbCO from HS‐GC‐MS CO measurements provides helpful information by determining the total CO levels (CO linked to hemoglobin (HbCO) and CO dissociated from hemoglobin). The CO concentrations were converted in HbCO saturation levels to define cutoff blood CO values. CO limits were defined as less than 1 μmol/mL for living persons, less than 1.5 μmol/mL for dead persons without CO exposure, and greater than 3 μmol/mL for dead persons with clear CO poisoning.     

Nitrate and Nitrite Determination in Gunshot Residue Samples by Capillary Electrophoresis in Acidic Run Buffer,

Simultaneous determination of nitrate and nitrite in gunshot residue has been conducted by capillary electrophoresis using an acidic run buffer (pH 3.5). In previously developed capillary electrophoretic methods, alkaline pH separation buffers were used where nitrite and nitrate possess similar electrophoretic mobility. In this study, the electroosmotic flow has been reversed by using low pH running buffer without any additives. As a result of reversing the electroosmotic flow, very fast analysis has been actualized, well‐defined and separated ion peaks emerge in less than 4 min. Besides, the limit of detection was improved by employing large volume sample stacking. Limit of detection values were 6.7 and 4.3 μM for nitrate and nitrite, respectively. In traditional procedure, mechanical agitation is employed for extraction, while in this work the extraction efficiency of ultrasound mixing for 30 min was found sufficient. The proposed method was successfully applied to authentic gunshot residue samples.     

Citrate Content of Bone for Time Since Death Estimation: Results from Burials with Different Physical Characteristics and Known PMI

A recently introduced method to determine the postmortem interval (PMI) based on quantification of the citrate content in bone was applied on the temporal bones and femora of 20 individuals buried in wooden coffins (WO) and body bags (BB), respectively. Concerning known vs. calculated PMI, a significant difference between the temporal and the femur bone samples of the same individuals was observed in the BB group (p = 0.012). In contrast, differences were insignificant for the WO group (p = 0.400). Moreover, similar levels of underestimation of PMIs resulted from the analysis of the femora for both burial groups (p = 0.247). Also, there was consistently less citrate preserved in the flat temporal bones as compared to the femora, indicating that the cortical layer of the long bones should be preferentially employed for citrate‐based PMI estimations. The results call for additional research on subsurface‐buried and surface‐deposited remains to enhance the accuracy of the published PMI equation.     

Development and validation of the Cozart DDS oral fluid collection device

The testing of oral fluid for drugs of abuse has increased significantly over recent years and is now commonplace in drug rehabilitation clinics, the workplace, prisons and custody suites. The global problem of identifying drugged drivers has also led to an increase in oral fluid testing at the roadside. The main requirements for the implementation of roadside drug testing are a rapid sample collection time, collection of a known sample volume and recovery of drugs from the collection device. We report here the development of the Cozart^® DDS oral fluid collector, an oral fluid collector that combines rapid and adequate sample collection with satisfactory drug recovery. Oral fluid was collected from drug users (n = 134) and drug-free individuals (n = 137), using the Cozart^® DDS oral fluid collector. The mean time for the completion of collection (full coloration of the sample presence indicator) was 34 s for drug-free individuals and 44 s for drug users. The average volume collected was 0.34 mL (n = 271). No chemical stimulant (to induce salivation) was used to achieve the collection times observed in either the drug-free or the drug-taking sample populations. Drugs were extracted from the collector using the Cozart^® DDS buffer and drug recovery was determined by Cozart^® enzyme immunoassays. The recovery studies showed that for amphetamine, Δ⁹THC, cocaine, methadone, methamphetamine, morphine and temazepam over 90% of the drug in the sample was eluted from the collector. The Cozart^® DDS oral fluid collector provides a reliable mechanism for the collection of oral fluid at the roadside that achieves the rapid collection times required.     

CT‐Guided,Minimally Invasive,Postmortem Needle Biopsy Using the B‐Rob II Needle‐Positioning Robot

CT‐guided, minimally invasive needle biopsy techniques can be used to retrieve tissue or body fluid samples for histologic tissue diagnoses in forensic investigations. The purpose of this study was an evaluation of CT‐guided needle‐positioning robot B‐Rob II. To operate under CT guidance, we adopted the B‐Rob II robotic needle placement system and its workflow. The accuracy and speed of the procedure were tested on a gelatin phantom in a series of 21 biopsies. We achieved an average needle placement accuracy of 1.8 mm (±1.1 mm) using robotic assistance. The procedure required an average of 2 min 21 s. The needle placement accuracy for minimally invasive needle biopsies using the B‐Rob II biopsy robot is sufficiently accurate and fast for forensic postmortem examinations of focal organ changes. Further tests will be performed to test the feasibility of the robot for performing biopsies of focal organ changes in human bodies.     

Monitoring of Levamisole Concentration in Serum of Traffic Participants after Cocaine Consumption

This study highlights the problem of levamisole‐adulterated cocaine in context of active traffic participation. For the purposes of levamisole concentration monitoring in human serum, an analytical method based on LC‐MS/MS and solid‐phase extraction was applied. A Luna 5 μm C18 (2) 100 A, 150 mm × 2 mm column and a mobile phase consisting of A (H₂O/methanol = 95/5, v/v) and B (H₂O/methanol = 3/97, v/v), both with 10 mM ammonium acetate and with 0.1% acetic acid (pH = 3.2), were used. The validation experiments demonstrated that the method applied was appropriate for levamisole quantification in human serum. For 23% of levamisole‐positive samples, the concentrations exceeded 20 ng/mL. Therefore, the interaction of this drug with cocaine has to be considered as important for active traffic participation. As a consequence, monitoring of levamisole concentration in human serum is recommended, as long as it is used as cocaine adulterant.     

Determination of Nitrite in Whole Blood by High‐Performance Liquid Chromatography with Electrochemical Detection and a Case of Nitrite Poisoning

Although nitrite is widely used in meat processing, it is a major toxicity hazard to children and is responsible for the blue‐baby syndrome. A simple and effective method to determine nitrite in whole blood has been devised using ion chromatography with suppressed conductivity detection. The blood sample was deproteinized by adding acetonitrile and purified with mini‐cartridges to remove hydrophobic compounds, chloride ions, and metal ions. An aliquot of the filtrate was injected onto the ion chromatography. The retention time for nitrite was 13.8 min and the detection limit of nitrite in whole blood was 0.4 μmol/L. The calibration curve was linear (r² = 0.9999) over the concentration working range. The blood nitrite concentration of a victim who attempted suicide by ingesting sodium nitrite powder was determined using the present method. The basal levels for nitrite in human blood was determined with 7.1 ± 0.9 μmol/L (n = 12).     

Less is More—Optimization of DNA Extraction from Canine Feces

Although most DNA crime laboratories may not encounter fecal samples often, they are a familiar sample type in non‐human forensic laboratories due to their prevalence in the environment. Fecal matter can be challenging due to low numbers of nucleated cells and the presence of inhibitors that impede amplification success. Sampling location (internal vs. external), sampling quantity (10–200 mg), and various extraction protocols (silica matrix, bead beating, and clean‐up column) were evaluated to maximize DNA yield. The greatest yield of intact DNA was obtained using a modified silica matrix extraction protocol (VGL‐Fecal) on 30–50 mg of fecal matter collected from the external surface of a stool that had been dried for 24 h. This optimized sampling and extraction protocol was applied to a pilot study where DNA yield and genotyping success were evaluated. By optimizing our collection, sampling, and extraction procedures, a reliable method for maximizing the yield of canine fecal DNA was developed.     

Quantitation of Synthetic Cannabinoids in Plant Materials Using High Performance Liquid Chromatography with UV Detection (Validated Method)

Plant based products laced with synthetic cannabinoids have become popular substances of abuse over the last decade. Quantitative analysis for synthetic cannabinoid content in the laced materials is necessary for health hazard assessments addressing overall exposure and toxicity when the products are smoked. A validated, broadly applicable HPLC‐UV method for the determination of synthetic cannabinoids in plant materials is presented, using acetonitrile extraction and separation on a commercial phenylhexyl stationary phase. UV detection provides excellent sensitivity with limits of quantitation (LOQs) less than 10 μg/g for many cannabinoids. The method was validated for several structural classes (dibenzopyrans, cyclohexylphenols, naphthoylindoles, benzoylindoles, phenylacetylindoles, tetramethylcyclopropylindoles) based on spike recovery experiments in multiple plant materials over a wide cannabinoid contents range (0.1–81 mg/g). Average recovery across 32 cannabinoids was 94% for marshmallow leaf, 95% for damiana leaf, and 92% for mullein leaf. The method was applied to a series of case‐related products with determined amounts ranging from 0.2 to >100 mg/g.     

Morphoscopic Trait Expression in “Hispanic” Populations

This study evaluates population variation of eight cranial morphoscopic traits using samples of known southwest Hispanics (n = 72), Guatemalans (n = 106), American Blacks (n = 146), and American Whites (n = 218). We applied the support vector machine (SVM) method to build a prediction model based on a subsample (20%) of the data; the remainder of the data was used as a test sample. The SVM approach effectively differentiated between the four groups with correct classification rates between 72% (Guatemalan group) and 94% (American Black group). However, when the Guatemalan and southwest Hispanic samples were pooled, the same model correctly classified all groups with a higher degree of accuracy (American Black = 96%; American White = 77%; and the pooled Hispanic sample = 91%). This study also identified significant differences between the two Hispanic groups in six of the eight traits using univariate statistical tests. These results speak to the unique population histories of these samples and the current use of the term “Hispanic” within forensic anthropology. Finally, we argue that the SVM can be used as a classification model for ancestry estimation in a forensic context and as a diagnostic tool may broaden the application of morphoscopic trait data for the assessment of ancestry.     

A Comparison of the Effectiveness of Swabbing and Flossing as a Means of Recovering Spermatozoa from the Oral Cavity

This study examined whether flossing the teeth is a more effective collection method in recovering spermatozoa than conventional swabbing techniques. It was hypothesized that inclusion of flossing as a collection method would extend the recovery of spermatozoa to longer postcoital intervals (PCIs). Eighteen individuals provided 174 oral cavity samples. Successful recovery of spermatozoa was assessed with respect to the collection method and reported activity in the oral cavity during the PCI. Samples were subjected to a differential extraction procedure prior to microscopic evaluation of the extracted pellet. The results indicate that swabbing is more effective than flossing when the PCI falls within 1.5–12 h. However, spermatozoa were recovered from seven floss samples where the corresponding swabs gave negative results. When combining the results from the two collection methods, the percentage of subjects from whom spermatozoa are recovered increases for each PCI beyond the 0‐h interval.     

Prepared Disappearing Ink and Deciphering of Documents

Disappearing inks can be used for forgeries in many fields. In this study, thymolphthalein indicator solution was prepared as a disappearing ink. A total of 54 different solutions containing thymolphthalein were prepared at six different concentrations and nine different pH values. Among the prepared solutions, 0.16 g/50 mL and 0.32 g/65 mL thymolphthalein at pH 14 were the optimum solutions that were not distinguishable from regular ink when applied to promissory note. Fountain pens were filled with the abovementioned ink solutions and applied to promissory notes for experimental purposes. After 40 h, ink residues were not visible on promissory notes written with the ink with the optimum pH (pH = 14) value. To decipher the entries that disappeared, an instrumental method (Video Spectral Comparator‐8000) and a chemical method (using NH₃ vapor) were used, succeeded at 0.32 g/65 mL, but failed at 0.16 g/50 mL thymolphthalein. Therefore, NH₃ had to be sprayed for 0.16 g/50 mL with negligible harm.     

Reanalysis of the Trotter Tibia Quandary and its Continued Effect on Stature Estimation of Past‐Conflict Service Members

Forensic casework from past‐conflicts relies on the corrected historical Trotter data for stature estimation in Fordisc. For roughly 10 years’, stature estimation using this data has produced point estimates for the tibia that are on average 1.25 inches less than the other long bones. This issue was identified after applying the equations derived from Fordisc to the USS Oklahoma commingled assemblage. Reevaluation of Fordisc revealed that a correction factor of 20 mm, instead of 10 mm, was mistakenly applied to the Trotter tibia data. Historical forensic anthropology reports written at the Defense POW/MIA Accounting Agency were utilized to identify that the overcorrection is isolated to Fordisc 3 with an error rate of 5% of known antemortem statures falling outside of the prediction intervals that relied on the tibia. Further evaluation of the Oklahoma sample indicates the 10 mm correction is still producing point estimates less than the other long bones.     

Use of Postmortem Temperature Decay Response Surface Plots of Heat Transport in the Human Eye to Predict Time of Death

A finite element heat transfer model of the human eye was previously constructed and applied to experimental postmortem temperature decay curves collected in eyeballs of ten human bodies. The model was applied in the early postmortem period of 0–24 h under conditions of natural convection–radiation. Based upon this previous model, response surfaces for postmortem temperature decay were constructed based upon variable ranges of the natural convective–radiation heat transfer coefficient from 7–13 W/m² K, ambient temperatures of 10–33°C, and times of 0–24 h. Mathematical equations to describe these response surfaces have been developed. This response surface method is demonstrated for use by coroners/medical personnel to estimate time of death from recorded field temperature data collected over a 30‐min period. Sensitivity of the model to small changes in the key variable of ambient temperature is explored. The response surface model is applied to two cases of previously collected experimental eyeball temperature data. This response surface model method is only valid for constant surrounding temperatures, conditions of natural convection, no radiation effects, and postmortem times of 0–24 h.