Identification of human urine stains by HPLC analysis of 17-ketosteroid conjugates

A new method for identifying human urine stains utilizing high-performance liquid chromatographic (HPLC) analysis of five major 17-ketosteroid conjugates: dehydroepiandrosterone sulfate, etiocholanolone sulfate, etiocholanolone glucuronide, androsterone sulfate, and androsterone glucuronide was examined. Samples of urine stains were extracted with borate buffer solution (pH 9.3) and the extracts were applied onto a Sep-Pak tC18 cartridge. The analytes were eluted from the cartridge with methanol. The eluates were prelabeled with 2,4-dinitrophenylhydrazine in trichloroacetic acid-benzene solution and were separated by HPLC on a reversed-phase ODS column using a mobile phase of 80% methanol in a buffer consisting of 25 mM sodium acetate in 2% acetic acid. The eluates were monitored by a spectrophotometer at 380 nm. While all five 17-ketosteroid conjugates were clearly detected in the human urine stain samples, traces of only some of these conjugates were detected in the animal samples. Therefore, the presence of all five 17-ketosteroid conjugates indicated human specificity. In addition to the above finding, the properties of those five 17-ketosteroid conjugates were confirmed by electrospray ionization liquid chromatography-mass spectrometry (ESI-LC-MS).     

Biosafety considerations for autopsy

An autopsy may subject prosectors and others to a wide variety of infectious agents, including bloodborne and aerosolized pathogens such as human immunodeficiency virus, hepatitis B and C viruses, and Mycobacterium tuberculosis. Other hazards include toxic chemicals (e.g., formalin, cyanide, and organophosphates) and radiation from radionuclides used for patient therapy and diagnosis. These risks can be substantially mitigated through proper assessment, personal protective equipment, appropriate autopsy procedures, and facility design.     

Chromosome heteromorphisms and paternity testing

Most of the blood group systems have now been applied to paternity investigations. In a complicated case, an alleged father not excluded with blood group testing, could be excluded with only chromosome heteromorphisms. Chromosome heteromorphisms are inherited from parents to offspring as simple Mendelian laws. The authors' paternity testing is presented with the help of chromosome heteromorphism analysis.     

Estimation of age from a tooth by means of racemization of an amino acid, especially aspartic acid--comparison of enamel and dentin.

In a study of age estimation from teeth by means of measuring racemization of aspartic acid (Asp), a representative amino acid, the accuracy of data from enamel and dentin in the same tooth was compared. The correlation of D/L ratio of aspartic acid with actual age gave the following parameters: r = 0.928, sigma = +/- 5.2, k = 4.47 x 10(-4) yr-1 in enamel and r = 0.995, sigma = +/- 1.4, k = 5.75 x 10(-4) yr-1. The difference in ages between one estimated by the D/L ratio and the actual one was within +/- 3 years in dentin, while in enamel an error of from 2 to 11 years was observed. Reaction rate constants of the racemization in a dry postmortem state (15 degrees C) were calculated as k = 9.70 x 10(-8) yr-1 in enamel, and k = 1.33 x 10(-7) yr-1 in dentin. Compared to rates determined from teeth recently extracted from living subjects, the rate was higher in dentin than in enamel. These data reconfirmed that dentin is superior to enamel in making exact age estimations from teeth.     

Homicide–suicide in Hong Kong, 1989–1998

This study provides the first systematic research of homicide–suicide (HS) in a Chinese society. Data were drawn from the HK Homicide Monitoring Data-base computer file derived from investigation and death reports held by the HK Police Force and the Coroner’s Court. During the 10-year study period, 56 events involving 133 deaths were identified. The majority of offenders were males (75%) and most victims were female (64%). The mean age of offenders and victims were 41.9 and 32.3 years, respectively. Spouses and lovers comprised the majority of victims (46.4%) followed by child victims (36%). Most HS events were motivated by separation or termination of marital or sexual relations (39%), economic reasons (25%) and other domestic disputes (20%). The most frequent modes of killing were strangulation/suffocation (26%), stabbing/chopping (24%), followed by gassing/poisoning (14%) and falling from a height (14%). The commonest method of suicide was falling from a height (48%). It was followed by gassing/poisoning (22%) and strangulation/suffocation (13%). Depression (18.3%) was found to be the commonest mental disorder. Most offenders were from low-socio-economic background. Two-third were unemployed and 76.6% had 9 or less years of education. HS in HK were distinguished from those reported in the western literature in respect to the high relevance of economic factors, the absence of mercy killing between old couples, a higher percentage of pedicide–suicides and the infrequent use of firearms.     

Simultaneous observation of catecholamine, serotonin and their metabolites in incised skin wounds of guinea pig.

In order to examine the vital reaction in wounds, catecholamines, serotonin (5-HT) and their metabolites in the incised skin wounds of guinea pigs were analyzed simultaneously by high-performance liquid chromatography (HPLC) using electrochemical detection (ECD). The principal changes in the levels of these compounds in vital wounds were as follows: a considerable decrease in norepinephrine (NE) content was observed 12-24 h after injury which persisted to at least 7 days. 3,4-Dihydroxyphenylacetic acid (DOPAC) decreased slightly for up to 30 min and then showed no significant difference compared with postmortem levels. Epinephrine and dopamine were barely detected by the HPLC-ECD method employed. 5-HT concentrations which showed an increase up to 24 h showed maximum levels 800 microns from the wound edge at 10 and 30 min after injury. 5-Hydroxyindoleacetic acid (5-HIAA) was significantly higher than the postmortem level over almost the entire period of these experiments. A 5-HIAA content of at least twice the postmortem level was observed 800 microns from the wound edge of a 10-min-old vital wound. Therefore, 5-HIAA is a likely candidate as a new marker for evaluating the vital reaction in wounds. The vital characteristics of NE, DOPAC, 5-HIAA and 5-HT in 10-min-old wounds persisted for up to 12 h at room temperature after death. These results suggest that the HPLC-ECD method used here is very useful for simultaneous examination of the vital reaction in wounds from the earliest to the later stages of the wound-healing period.     

Application of immunoblotting to serum protein phenotyping with reference to alpha 2HS-glycoprotein (AHS) typing of bloodstains

Sera and bloodstain extracts were subjected to isoelectric focusing in polyacrylamide gels. The focused proteins were transferred to nitrocellulose membranes by diffusion or electrophoretically, then allowed to react with specific antiserum and, after washing, with peroxidase-labeled anti-rabbit IgG. The immune complexes formed on the membranes were detected with 4-chloro-1-naphthol and hydrogen peroxide. Serum group-specific component, alpha 2HS-glycoprotein, the sixth and the seventh component of complement, factor 13B, and plasminogen could be phenotyped with high sensitivity. Bloodstains as old as 6 months could be correctly typed for alpha 2HS-glycoprotein by the blotting technique.     

Abuse of smoking methamphetamine mixed with tobacco: I. Inhalation efficiency and pyrolysis products of methamphetamine

Experiments of smoking methamphetamine in tobacco have been investigated. Inhalation efficiencies of methamphetamine into tar were 6 to 17% according to the additive amounts, suction volume, and intervals of smoking. Major pyrolysis products of methamphetamine in tar were identified as methamphetamine, amphetamine, phenylacetone, dimethylamphetamine, N-formyl-, N-acetyl-, N-propionyl-, and N-cyanomethyl-methamphetamine by the spectral analysis of infrared spectra (IR), mass spectra (MS), and proton magnetic resonance spectra (PMR), and comparison with the samples synthesized from authentic samples by one step. The largest pyrolysis product was N-cyanomethylmethamphetamine which is a new compound and easily metabolized to methamphetamine in the body. Methamphetamine itself transferred into tar was not so large, but the total active compounds in tar which would be metabolized to methamphetamine in the body were considerably larger.     

An ELISA using avidin-biotin complex for the determination of ABH group from saliva

The ABH group in a trace amount of saliva could be determined by an enzyme-linked immunosorbent assay (ELISA) using an avidin-biotin-peroxidase complex (ABC) technique. In this method ABH blood group substances as a solid phase are adsorbed to wells of a microtiter plate made of polystyrene. The primary antibody corresponding to the blood antigen adheres onto the wells, and reacts with the biotinylated secondary antibody. The previously formed ABC reagent is then added to the above wells, and finally the absorbance produced by the interaction of the peroxidase activity with a chromogenic substance is measured at 492 nm. This method proved to be clearly more sensitive for the detection of ABH blood groups in secretor-saliva than the conventional hemagglutination inhibition test. Also the ABH group of non-secretor-saliva could be easily determined by this method.