Awareness survey on drug crime scene investigation and drug detection kits among drug-related police officers.

This questionnaire-based study aimed to investigate the drug crime scene experienced by drug-related police officers and the perceptions of drug test kits by them before initiating the development of drug test kits to detect 16 types of drugs. The subjects were 57 drug-related police officers. Most of the respondents (96.5%) had <10 years of experience in drug-related work. Respondents were questioned about the drug scene investigation and perceptions of drug test kits. The questionnaire about drug test kits included the question on 'simple/rapid drug test kit' and 'electronic portable drug analyzer' regarding the disadvantages of existing kits and expecting features when a new kit is developed. First, in the on-site survey, the drug-related crime occurred at the suspect's house (47.8%), and methamphetamine (35.0%) and γ-hydroxybutyric acid (19.5%) were mainly found. In the awareness survey on drug test kits, most respondents (67.2%) had an experience of using 'simple/rapid drug test kits', whereas 17.5% for the 'electronic portable drug analyzer'. In the case of 'simple/rapid drug test kit', the false-positive rate reached 53.8% by a misinterpretation due to ambiguous color change (47.6%). The inaccuracy of the result (33.3%) was the most concern in 'electronic portable drug analyzer'. Respondents most favored pipette type for sample collector when a new kit is developed. In addition, they preferred the smaller kit with short detection times in both kit types. This survey could be applied to the development of efficient and practical kits for police officers working in drug-related fields.

Screening of new psychoactive substances in human plasma by magnetic solid phase extraction and LC-QTOF-MS.

The emergence of new psychoactive substances (NPSs) is an increasing challenge in forensic toxicology. There are many extraction methods in use to isolate NPSs in biological fluids, including protein precipitation (PPT), liquid-liquid extraction (LLE), and solid phase extraction (SPE). However, there is a need to develop an effective extraction method with a short extraction time and low consumption of solvent. To meet these requirements, magnetic solid phase extraction (m-SPE) was attempted to isolate 40 NPSs in human plasma in this study. Forty NPSs (13 synthetic cannabinoids, 13 phenethylamines, 4 tryptamines, 4 other substances, 3 aminoindanes, 2 piperazines, 1 phencyclidine-type substance) were spiked in plasma and analyzed by m-SPE using COOH-functionalized multi-walled carbon nanotubes with magnetic nanoparticles (COOH-mMWCNTs). A liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) method was used for screening and identification of 40 target compounds. Method validation including limits of detection, recovery, matrix effect, and precision was performed for all 40 -target compounds. The limits of detection (LOD) of the 40 analytes were between 0.002 and 0.084 mg/L. Extraction recovery ranged from 36.9% to 110.6% (average 87%). Matrix effects ranged from -29.0% (ion suppression) to 9.8% (ion enhancement). Both intra- and inter-day precision values were less than 27.5% (RSD%). The accurate mass of QTOF-MS enabled the identification of analytes by exact monoisotopic mass and isotopic pattern. m-SPE was applied to extract 40 NPSs, and revealed less time-consuming and laborious than conventional SPE. This method proved to be an advantageous procedure to extract NPSs from biological fluids.

Magnetic Solid-Phase Extraction of Drugs and Pesticides from Human Plasma Using COOH-mMWCNTs.

Carbon nanotubes (CNTs) are useful for extracting chemical compounds due to their properties, such as surface area and the potential for chemical modification. Especially the formation of CNTs with carboxylic acid functional group makes them disperse in water-based samples and have strong interaction forces with cationizable analytes. Based on these features, carboxylic acid functionalized multi-walled CNTs (COOH-MWCNTs) have been used as extraction sorbents. CNT can also be gathered using an external magnet by forming complex with iron oxide (Fe3O4) magnetic nanoparticles (MNPs). In this study, COOH-MWCNTs with MNPs were subjected to magnetic solid-phase extraction (mSPE) in order to extract the targeted substances such as diphenhydramine, doxylamine, tramadol, escitalopram, zolpidem, diphenamid, paclobutrazol, hexaconazole, cyproconazole and mepronil from human plasma samples. The following five factors were optimized: (i) the ratio of COOH-MWCNTs to MNPs as a sorbent from 1:1 to 1:4; (ii) sorbent amount starting from 12.5 to 75%; (iii) sample pH tested pH 2 to pH 10 with 1 N hydrochloride and 1 N sodium hydroxide; (iv) agitating time from 0 to 4 min and (v) elution solvent. Limit of detection of 10 targeted substances in human plasma were in the range of 0.1-0.4 mg/L. The recovery of targeted substances (except diphenamid) in human plasma was 73.06-110.28% for intra-day and 83.00-107.70% for inter-day and the precision (relative standard deviation, %) in human plasma was 0.3-13.3% for intra-day and 2.9-15.6% for inter-day. The method was applied to nine authentic biological samples from overdose patients in the emergency room of Chungnam National University Hospital. The performance of mSPE was compared with the liquid-liquid extraction method using ethyl acetate. The results showed that the newly developed method in this study can be used for screening analysis in forensic and clinical toxicology.

Comparison of legislative management for new psychoactive substances control among Taiwan, South Korea, and Japan.

For decades, the three United Nations drug conventions have served as the basis for member states' obligations and international cooperation on drug control. However, the emergence of new psychoactive substances (NPSs) poses a new risk to public health and a challenge to drug policy because of their unknown toxicological effects and easy modification of chemical structures to shun legal control. So far, there is no international consensus on legislative control of NPSs. Therefore, we compared the legislative management on NPS control among Taiwan, South Korea, and Japan. Drug-related information was obtained from the authorities of these three countries. The results indicate that despite geographic proximity and similar legal attitudes toward illegal drug use, the legislative criteria, and responses for NPS control in these three countries were quite different. Ketamine has been the major used NPS in Taiwan but seldom found in South Korea and Japan. The difference in the number of controlled NPSs in Taiwan (91) and South Korea (245) might be due to the implementation of temporary designation systems and analog controls in South Korea. The recent surge of newly controlled NPSs in Japan was because of the promulgation of designated drug regulation and subsequent control of "dangerous drugs." Although NPS use has become a potential social and medical problem among these three countries, the outcomes of NPS legislation control remain to be scrutinized. To minimize harm from NPS use, development of legislative mechanism(s) on NPS scheduling is the first step for early identification and control of NPS problems.

Analysis of hypoxanthine and lactic acid levels in vitreous humor for the estimation of post-mortem interval (PMI) using LC-MS/MS.

Hypoxanthine (Hx) is produced during terminal stages of purine catabolism in humans. The concentrations of Hx and l-lactic acid in vitreous humor highly correlate with post-mortem interval (PMI). In this study, we measured the concentrations of Hx and l-lactic acid in uncontrolled authentic vitreous humor from cadavers, and investigated the correlation between these molecules and PMI. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used for the quantitative analysis of Hx and l-lactic acid in vitreous humor. These concentrations were also corrected by four seasons such as spring, summer, autumn, and winter and temperatures on the day of death such as -10 to 5 °C, 6-15 °C, 16-25 °C, and 26-32 °C that may affect the biomarker concentrations. Vitreous humors were collected from cadavers with known time of death at the National Forensic Service (NFS), Republic of Korea. The correlation between the concentrations of Hx and l-lactic acid in vitreous humors and PMI was evaluated using 79 corpses (53 males and 26 females), with sampling time ranging from 13 to 103 h after death. The average daily ambient temperature at the time of death of each sample was investigated to calibrate the correlation with PMI. Following correction of the concentrations of Hx and l-lactic acid with temperature, the correlation of Hx and l-lactic acid with PMI increased from 0.53 to 0.59 for Hx and 0.38 to 0.42 for l-lactic acid. The highest correlation of Hx and l-lactic acid concentrations with PMI was observed in the winter season, with an R2 value of 0.80 and 0.71 for Hx and l-lactic acid, respectively. The correlation of Hx and l-lactic acid with PMI was corrected by ambient temperature for each season, resulting in an increase in the R2 value to 0.88 for Hx and 0.72 for l-lactic acid. The best correlation was observed when the temperature was corrected after dividing by season.

Postmortem proteomics to discover biomarkers for forensic PMI estimation.

The assessment of postmortem degradation of skeletal muscle proteins has emerged as a novel approach to estimate the time since death in the early to mid-postmortem phase (approximately 24 h postmortem (hpm) to 120 hpm). Current protein-based methods are limited to a small number of skeletal muscle proteins, shown to undergo proteolysis after death. In this study, we investigated the usability of a target-based and unbiased system-wide protein analysis to gain further insights into systemic postmortem protein alterations and to identify additional markers for postmortem interval (PMI) delimitation. We performed proteomic profiling to globally analyze postmortem alterations of the rat and mouse skeletal muscle proteome at defined time points (0, 24, 48, 72, and 96 hpm), harnessing a mass spectrometry-based quantitative proteomics approach. Hierarchical clustering analysis for a total of 579 (rat) and 896 (mouse) quantified proteins revealed differentially expressed proteins during the investigated postmortem period. We further focused on two selected proteins (eEF1A2 and GAPDH), which were shown to consistently degrade postmortem in both rat and mouse, suggesting conserved intra- and interspecies degradation behavior, and thus preserved association with the PMI and possible transferability to humans. In turn, we validated the usefulness of these new markers by classical Western blot experiments in a rat model and in human autopsy cases. Our results demonstrate the feasibility of mass spectrometry-based analysis to discover novel protein markers for PMI estimation and show that the proteins eEF1A2 and GAPDH appear to be valuable markers for PMI estimation in humans.

Urine Multi-drug Screening with GC-MS or LC-MS-MS Using SALLE-hybrid PPT/SPE.

To intoxicated patients in the emergency room, toxicological analysis can be considerably helpful for identifying the involved toxicants. In order to develop a urine multi-drug screening (UmDS) method, gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS-MS) were used to determine targeted and unknown toxicants in urine. A GC-MS method in scan mode was validated for selectivity, limit of detection (LOD) and recovery. An LC-MS-MS multiple reaction monitoring (MRM) method was validated for lower LOD, recovery and matrix effect. The results of the screening analysis were compared with patient medical records to check the reliability of the screen. Urine samples collected from an emergency room were extracted through a combination of salting-out assisted liquid-liquid extraction (SALLE) and hybrid protein precipitation/solid phase extraction (hybrid PPT/SPE) plates and examined by GC-MS and LC-MS-MS. GC-MS analysis was performed as unknown drug screen and LC-MS-MS analysis was conducted as targeted drug screen. After analysis by GC-MS, a library search was conducted using an in-house library established with the automated mass spectral deconvolution and identification system (AMDISTM). LC-MS-MS used Cliquid®2.0 software for data processing and acquisition in MRM mode. An UmDS method by GC-MS and LC-MS-MS was developed by using a SALLE-hybrid PPT/SPE and in-house library. The results of UmDS by GC-MS and LC-MS-MS showed that toxicants could be identified from 185 emergency room patient samples containing unknown toxicants. Zolpidem, acetaminophen and citalopram were the most frequently encountered drugs in emergency room patients. The UmDS analysis developed in this study can be used effectively to detect toxic substances in a short time. Hence, it could be utilized in clinical and forensic toxicology practices.

New psychoactive substances of natural origin: A brief review.

Plant-based drugs of abuse are as old as recorded human history. Although traditional addictive substances, such as opium, cannabis and coca, have been controlled by the United Nations anti-drug conventions, many, if not most, natural plants with addictive or abuse liability remain elusive. Therefore, the United Nations Office on Drugs and Crime (UNODC) has warned the emerging threat from new psychoactive substances (NPS), which are mostly derived or modified from the constituents of natural origin. For example, synthetic cannabinoids and synthetic cathinones are derived from the cannabis and khat plant, respectively. In this review, we briefly discussed the chemistry, pharmacology and toxicology of five common NPS of natural origin, i.e., khat, kratom, salvia, magic mushroom and mandrake. Through the review, we hope that professionals and general public alike can pay more attention to the potential problems caused by natural NPS, and suitable control measures will be taken.

Overview of Forensic Toxicology, Yesterday, Today and in the Future.

BACKGROUND: The scope of forensic toxicology has been tremendously expanded over the past 50 years. From two general sections forensic toxicology can be further classified into 8-9 sections. METHODS: The most outstanding improvement in forensic toxicology is the changes brought by instrumental development. The field of forensic toxicology was revolutionized by the development of immunoassay and benchtop GC-MS in the 1980's and LC-MS-MS in 2000's. Detection of trace amounts of analytes has allowed the use of new specimens such as hair and oral fluids, along with blood and urine. Over a longer period of time, continuous efforts have been made to efficiently extract and separate drug and poison from biological fluids. International endeavors to develop high quality standards and guidelines for drugs and poisons in biological specimens and to promote them in order to increase reliability of laboratories are also part of the recent advancement of forensic toxicology. Interpretation of postmortem toxicology encompasses various factors including postmortem redistribution and stability. RESULTS: Considering the recent trend, the interpretation of toxicological results should account for autopsy findings, crime scene information, and related medical history. The fields of forensic toxicology will continuously develop to improve analysis of target analytes from various specimens, quality assurance program, and results interpretation. In addition, the development of analytical techniques will also contribute further advancement of forensic toxicology. CONCLUSION: The societies of forensic toxicologists, such as TIAFT, will play an important role for the advancement of forensic toxicology by collaborating and sharing ideas between toxicologists from both developed and developing countries.

Anthropogenic rare earth elements and their spatial distributions in the Han River, South Korea.

Rare earth elements (REE) consist of lanthanides (from La to Lu), together with yttrium and scandium, in which anthropogenic REE, such as gadolinium (Gd), lanthanum (La), and samarium (Sm), has emerged as micro-contaminants in natural waters in highly developed countries. Here, we collected water samples in the Han River (HR) and its tributaries flowing through Seoul Capital Area, the world's second largest metropolitan area in order to examine how and to what extent anthropogenic REE anomalies may occur. Water samples show higher light REE concentrations than heavy REE concentrations, while wastewater treatment plant (WWTP) samples display much higher heavy REE concentrations due to high Gd concentration. The PAAS-normalized REE patterns indicate that WWTP samples display the pronounced positive Gd anomalies, in which anthropogenic Gd from magnetic resonance imaging (MRI) diagnostic system occurs as a form of Gd complexation with either Cl- or SO42-. Due to the WWTP, both the HR and tributaries show also positive Gd anomalies and the anthropogenic Gd concentrations increase as a function of the distance from the Paldang dam. This result indicates a positive correlation between populaton, number of MRI instruments, and positive Gd anomaly. Similarly, positive La and Sm anomalies exist in the HR, indicating that the HR is also affected by their point sources. Based on the discharge rate and anthropogenic REE concentrations, their fluxes are estimated to be 952 ± 319 kg/yr, suggesting that this amount of fluxes could disturb REE distribution in the Yellow Sea, and pose harmful effects on aquatic ecosystems.

Prevalence of new psychoactive substances in Northeast Asia from 2007 to 2015.

The proliferation of new psychoactive substances (NPS) has been a global trend in drug abuse and its regulation has been a worldwide concern. There is no doubt that it is necessary to share information related to these emerging substances between countries and continents for the effective regulation of NPS. With efforts for the efficient regulation of NPS, many studies and information have been published for the prevalence of NPS in the United States and other countries in Europe and Oceania. However, there is lack of information available for the prevalence of NPS in Asian and African countries. Therefore, this research was focused on the investigation of legal status of certain NPS in Northeast Asian countries, including China, Japan, South Korea and Taiwan, in order to provide information on the prevalence and trend of emerging NPS in these countries. The results showed that a total of 940 NPS was reported in 4 Northeast Asian countries from 2007 to 2015. Among 940 NPS, 882 NPS are legally restricted in at least one country (94%) and 96 substances were not currently under control (6%) in these countries. The number of controlled NPS that are currently controlled in all 4 countries was only 25 (or 28%) out of 882 NPS. Each substance was categorized in 9 groups according to the classification proposed by the United Nations Office on Drugs and Crime (UNODC). In Northeast Asia, the most commonly controlled NPS were synthetic cannabinoids, synthetic cathinones, and phenethylamines. It was found that Japan is the most proactive country in terms of the NPS regulation with 41% of the total number of controlled NPS in Northeast Asia, followed by South Korea (21%), China (28%), Taiwan (10%). Comparing the number of NPS newly regulated in each country every year, NPS has been broadly scheduled in 2011 and the number of scheduled NPS has dramatically increased from 2013 to 2015. It was shown that Northeast Asia is also in danger of these emerging NPS and the effective regulation across countries is important for the prevention of NPS. Also, this study will bring attention to local law enforcement in the construction of local drug crime prevention network sharing information for these controlled substances.

Toxicology in international drug control-Prioritizing the most harmful, persistent and prevalent substances.

The nature of the global drugs market has evolved rapidly and has become more complex with the emergence of new psychoactive substances (NPS), some of which have been associated with increased abuse, hospital emergency admissions and sometimes fatalities. NPS are characterized by geographic heterogeneity, with some only transient in nature and others not satisfying the criteria for harm required for international control. Consequently, a pragmatic response of the international community is to prioritize the most harmful, persistent and prevalent substances for action - an objective, which is hampered by the paucity of data on harms. The report describes a United Nations Office on Drugs and Crime (UNODC) initiative, in collaboration with the International Association of Forensic Toxicologists (TIAFT), to collect, analyze and share toxicology data at a global level to reinforce the ability of the international community in making informed decisions using a scientific evidence-based approach, in identifying the most harmful NPS.

Patterns of drugs & poisons in southern area of South Korea in 2014.

The southern area of South Korea consists of three parts; Busan, Ulsan and Gyeongsangnam-do. Busan Institute of National Forensic Service (NFS) performed about 50,000 cases throughout the southern area in 2014, occupying over 15% of total cases covered by NFS. In this study, patterns of drugs and poisons in the southern area of South Korea were investigated. The investigation was carried out by the laboratory information management system of NFS between January and December of 2014. As results, a total of 606 autopsy cases were performed by Busan Institute of NFS in 2014. Among them, 15 cases were determined as drug intoxication or poisons as the cause of death, taking up 2.5% of total cases: 5 cases of intoxication by drugs, 5 by agricultural pesticides, 3 by illicit drugs, and 1 each by detergents and chemical substances. A total of 108 drugs in postmortem bloods were detected from the autopsy cases, and the top 5 drugs were chlorpheniramine, tramadol, diazepam, zolpidem and lidocaine. Meanwhile, a total of 1,728 cases were submitted for illicit drug testing in 2014. Among them, hair was the most common type of specimens, and the rate of positive detection of methamphetamine from the hair, urine, and seized materials in the southern area was over 50% in all cases, indicating that this is the most commonly abused drug in South Korea. A total of 12 types of novel psychoactive substances (NPSs) were detected in the southern area in 2014; 10 were identified as synthetic cannabinoids and 2 as alkyl nitrites.

Development of visual peak selection system based on multi-ISs normalization algorithm to apply to methamphetamine impurity profiling.

The aim of this study is to improve resolution of impurity peaks using a newly devised normalization algorithm for multi-internal standards (ISs) and to describe a visual peak selection system (VPSS) for efficient support of impurity profiling. Drug trafficking routes, location of manufacture, or synthetic route can be identified from impurities in seized drugs. In the analysis of impurities, different chromatogram profiles are obtained from gas chromatography and used to examine similarities between drug samples. The data processing method using relative retention time (RRT) calculated by a single internal standard is not preferred when many internal standards are used and many chromatographic peaks present because of the risk of overlapping between peaks and difficulty in classifying impurities. In this study, impurities in methamphetamine (MA) were extracted by liquid-liquid extraction (LLE) method using ethylacetate containing 4 internal standards and analyzed by gas chromatography-flame ionization detection (GC-FID). The newly developed VPSS consists of an input module, a conversion module, and a detection module. The input module imports chromatograms collected from GC and performs preprocessing, which is converted with a normalization algorithm in the conversion module, and finally the detection module detects the impurities in MA samples using a visualized zoning user interface. The normalization algorithm in the conversion module was used to convert the raw data from GC-FID. The VPSS with the built-in normalization algorithm can effectively detect different impurities in samples even in complex matrices and has high resolution keeping the time sequence of chromatographic peaks the same as that of the RRT method. The system can widen a full range of chromatograms so that the peaks of impurities were better aligned for easy separation and classification. The resolution, accuracy, and speed of impurity profiling showed remarkable improvement.

Illegal use patterns, side effects, and analytical methods of ketamine.

In Asian countries, such as China, Taiwan, and Hong Kong, ketamine (KT) is one of the most prevalent illicit use drugs. KT is regulated by various drug-related laws in many countries, such as Korea, Taiwan, China, U.S.A, Netherlands, UK, Australia, Mexico, and Canada. This review research explored pharmacology and side effects of KT, the illicit use patterns of KT, the analytical methods of KT in biological samples, and the concentrations of KT from abusers and non-abusers. Many side effects of KT have been reported mental and physical problems. Although many studies conducted various analytical methods for KT, this research focused on the urine and hair analysis and compared some parameters of samples, instruments, columns, extraction methods, internal standards, LOD/LOQ levels, metabolites, NK/K ratio, cut off values, and m/z values. Our research also compared the concentrations of KT in biological samples from abusers and non-abusers. Many rapid and precise analytical methods for illegal KT use are needed to be developed and applied to real samples. To minimize and prevent harm from KT, the authorities and appropriate agencies require a careful assessment, evaluation, early identification, and surveillance of KT users in both clinical and social settings. In addition, there is a need to construct a stricter legislative management and provide preventive education to younger individuals because illegal KT use is relatively common among the young populations.

Comparison of illegal drug use pattern in Taiwan and Korea from 2006 to 2014.

BACKGROUND: Illegal drug use has long been a global concern. Taiwan and Korea are geographically adjacent and both countries have experienced the illegal use problems of methamphetamine, a predominant prototype of New Psychoactive Substances (NPS). NPS, a term coined by the United Nations Office on Drugs and Crime (UNODC) in recent years, have not been scrutinized for their safety and may become a new threat to public health and security worldwide. To conduct evidence-based drug policy, it is imperative to estimate the trend and pattern of illegal drug use. Therefore, this study aims to analyze and compare the current status of drug-related seizures, arrests and illegal drug use, with a focus on methamphetamine and NPS, between Taiwan and Korea. METHODS: Data of illegal drug (including NPS)-related seizures and arrests were collected via anti-drug related agencies of both countries from 2006 through 2014.Since listing of NPS as controlled substances was a result of NPS abuse liability through official evaluation, the items of controlled NPS were used as an indicator of emerging use. These data obtained from Taiwan and Korea was then compared. RESULTS: The results showed that while methamphetamine remained as a predominant drug in both Taiwan and Korea for decades, different illegal drug use patterns have been observed in these two countries. In Taiwan, the major illegal drugs were methamphetamine, heroin, and ketamine, whereas in Korea those were methamphetamine and cannabis. By comparison of per capita illicit drug seizures, the illegal drug use situation in Taiwan was at a higher stake than that in Korea. In terms of NPS use, ketamine has been a major drug in Taiwan, but it was seldom found in Korea. Besides ketamine, the major type of NPS was synthetic cathinones in Taiwan whereas it was synthetic cannabinoids and phenethylamines in Korea. The difference in the numbers of controlled NPS items between Taiwan (23) and Korea (93) may be due to the implementation of temporary control on NPS in Korea since 2011. CONCLUSION: While the problem of methamphetamine still lingers, NPS have emerged as a new issue in both countries. However, the NPS pattern was different between Taiwan and Korea. Although the controlled NPS items in Taiwan were far less than those in Korea, the quantity of total NPS seizures, especially with ketamine, was much larger in Taiwan than in Korea. Different NPS pattern may also imply they were from different sources. Factors other than geographical proximity, such as drug policy and availability and accessibility to drugs, should be taken into account for the current status of illegal drug use in Korea and Taiwan.

Degradation of Kidney and Psoas Muscle Proteins as Indicators of Post-Mortem Interval in a Rat Model, with Use of Lateral Flow Technology.

We investigated potential protein markers of post-mortem interval (PMI) using rat kidney and psoas muscle. Tissue samples were taken at 12 h intervals for up to 96 h after death by suffocation. Expression levels of eight soluble proteins were analyzed by Western blotting. Degradation patterns of selected proteins were clearly divided into three groups: short-term, mid-term, and long-term PMI markers based on the half maximum intensity of intact protein expression. In kidney, glycogen synthase (GS) and glycogen synthase kinase-3ß were degraded completely within 48 h making them short-term PMI markers. AMP-activated protein kinase α, caspase 3 and GS were short-term PMI markers in psoas muscle. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was a mid-term PMI marker in both tissues. Expression levels of the typical long-term PMI markers, p53 and ß-catenin, were constant for at least 96 h post-mortem in both tissues. The degradation patterns of GS and caspase-3 were verified by immunohistochemistry in both tissues. GAPDH was chosen as a test PMI protein to perform a lateral flow assay (LFA). The presence of recombinant GAPDH was clearly detected in LFA and quantified in a concentration-dependent manner. These results suggest that LFA might be used to estimate PMI at a crime scene.

Detection of drugs in 275 alcohol-positive blood samples of Korean drivers.

Since driving under the influence of drugs (DUID) is as dangerous as drink-driving, many countries regulate DUID by law. However, laws against the use of drugs while driving are not yet established in Korea. In order to investigate the type and frequency of drugs used by drivers in Korea, we analyzed controlled and non-controlled drugs in alcohol-positive blood samples. Total 275 blood samples were taken from Korean drivers, which were positive in roadside alcohol testing. The following analyses were performed: blood alcohol concentrations by GC; screening for controlled drugs by immunoassay and confirmation for positive samples by GC-MS. For the detection of DUID related drugs in blood samples, a total of 49 drugs were selected and were examined by GC-MS. For a rapid detection of these drugs, an automated identification software called "DrugMan" was used. Concentrations of alcohol in 275 blood samples ranged from 0.011 to 0.249% (average 0.119%). Six specimens showed positive results by immunoassay: one methamphetamine and five benzodiazepines I. By GC-MS confirmation, only benzodiazepines in four cases were identified, while methamphetamine and benzodiazepine in two cases were not detected from the presumptive positive blood samples. Using DrugMan, four drugs were detected; chlorpheniramine (5)*, diazepam (4), dextromethorphan (1) and doxylamine (1). In addition, ibuprofen (1), lidocaine (1) and topiramate (1) were also detected as general drugs in blood samples ('*' indicates frequency). The frequency of drug abuse by Korean drivers was relatively low and a total 14 cases were positive in 275 blood samples with a ratio of 5%. However it is necessary to analyze more samples including alcohol negative blood, and to expand the range of drug lists to get the detailed information.

Determination of glyphosate and its metabolite in emergency room in Korea.

The number of glyphosate intoxication cases has been increased after the regulation of paraquat. Unfortunately, there are no reports on the potential concentration of glyphosate for those acute intoxicated patients admitted to emergency rooms and the correlation between the concentration of glyphosate and clinical symptoms in Korea up to our knowledge. As a nonselective herbicide, analysis of glyphosate requires derivatization because of its amphoteric and strongly polar nature. In order to develop a method to determine the concentration of glyphosate and its metabolite, aminomethylphosphonic acid (AMPA) in blood samples without derivatization, liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was utilized with a hydrophilic interaction chromatography (HILIC) column. The validation of this method showed that the limits of detection (LODs) and limits of quantitation (LOQs) for glyphosate and AMPA were 50 and 100ng/mL, respectively. In addition, matrix effect, recovery rate, and accuracy and precision in intra and inter-day were evaluated during the validation study of this method. Blood samples acquired from five glyphosate intoxicated patients were analyzed to investigate the correlation between the concentration of glyphosate and clinical symptoms. These patients were previously admitted to the emergency room at a University Hospital in Korea after glyphosate was self-administered in suicide attempts or by accident. As results of blood sample study, the concentration of glyphosate and AMPA were found in the range of 1.0-171.1 and 0.2-2.6µg/mL, respectively. The concentration ratio of glyphosate to AMPA was 55-71. According to the clinical reports for those patients, they were in the age between 47 and 82 years old and administered about 50-400mL. The blood samples were collected within 2-5h after administration of glyphosate. Among the intoxicated patients, the most common clinical symptom was metabolic acidosis, identified in four patients. The comparison between the concentration of glyphosate and administered dosage did not show the correlation, which suggests further investigation on the effects of surfactants in glyphosate from different vendors.

Deaths from recreational use of propofol in Korea.

Propofol, a short-acting and sedative-hypnotic agent, induces and maintains anesthesia. Since it is known to produce mild euphoria and hallucinations, the recreational use of propofol has been a big issue in Korea. Furthermore, many deaths have occurred due to its abuse and misuse. In order to study the prevalence of abuse and deaths due to propofol, all autopsy cases conducted between 2005 and 2010 at the NFS (National Forensic Service, Korea) were monitored by checking its concentrations in the blood. Propofol was detected in 131 cases (0.88%) out of 14,673 autopsied cases within 6 years. Propofol alone was detected in 49 of 131 fatal cases, while the combination of drugs was detected with propofol in the remaining 82 cases. The concentrations of propofol from autopsied cases ranged from 0.05 to 8.83 mg/L (mean 1.66; median 0.9) and from 0.08 to 8.65 mg/L (mean 1.71; median 1.05) in the heart (n=31) and the femoral blood (n=32), respectively. The investigation of the ratio of heart to femoral blood and the difference between the concentrations in heart and femoral blood (n=15) from the same body revealed the ratio from 0.45 to 3.66 (mean 1.53; median 1.40). The autopsy resulted in accidental death after self-administration in 16 autopsied cases among 131 autopsied cases. In 16 cases, their ages ranged from 17 to 56 and 75% of them were in their 20's and 30's and 75% were female. Half of them were medical personnel including 19% of doctors and 38% of nurses. The combination of drugs was detected in 6 cases. Fluoxetine was detected in three and vecuronium was detected in two along with propofol. The cause of death in 14 cases was drug intoxication, while that in 2 cases was hanging. Due to its prevalence, Korea has become the first country that regulates propofol as a psychotropic substance.