A highly efficient liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for etomidate and etomidate acid in urine, liver and kidney.

Etomidate (ETO) is a highly-efficient drug that can induce anesthesia with increasing doses, thus subject to strict regulation. However, an accurate and efficient method for ETO intake detection is currently lacking. Therefore, this study developed a straightforward sample preparation method using LC-MS/MS to analyze ETO and its primary metabolite, etomidate acid (ETA), in urine, liver, and kidney samples. Snap frozen pig liver and kidney samples were ground into a fine powder. Then, all the biological samples, including human urine, pig liver and kidney tissues, were deproteinized using acetonitrile and filtered for analysis. The separation was achieved in 9.01 min with gradient elution. The calibration curves ranged from 0.5 to 50 ng/mL for ETO in urine and 0.5 to 50 ng/g in liver and kidney, while the curves ranged from 1 to 100 ng/mL for ETA in urine and 1 to 100 ng/g in liver and kidney. The correlation coefficients (R2) were greater than 0.9957. The Limit of detection (LOD) and limit of quantitation (LOQ) for ETO were 0.2 and 0.5 ng/mL in urine samples and 0.2 and 0.5 ng/g in liver and kidney samples, respectively. For ETA, the LOD and LOQ were 0.5 and 1 ng/mL in urine samples and 0.5 and 1 ng/g in liver and kidney samples. This method was assessed by validation parameters, including selectivity, intra- and inter-day precision and accuracy, recovery, matrix effect, dilution integrity and stability. It was successfully applied to a practical case, revealing ETO and ETA concentrations in urine of 1.01 and 5.58 µg/mL, in liver samples of 12.30 and 1.13 µg/g, and in kidney samples of 6.95 and 4.23 µg/g. This suggests that the method is suitable for routine forensic detection of illicit ETO abuse.

[Progress in application of microbeam X-ray fluorescence spectroscopy in forensic science].

Microbeam X-ray fluorescence (micro-XRF) spectrometry has been raised as an analytical technique of microbeam during the recent years. With its advantages of high sensitivity, small sample requirement, high testing accuracy and non-destruction, the technique is widely utilized in forensic science. This review bases on recent researches at home and abroad, describes its applications including identification of gunshot residue, visualization of fingerprints, discrimination of drug source, production process, and other material evidences of analysis in crime scene. Thanks to the advances in technology, intelligent and portable micro-XRF equipment has appeared to be applied. It is believed that it may be more popular and frequent in administration of forensic science in the near future.

Detection of diatoms in water and tissues by combination of microwave digestion, vacuum filtration and scanning electron microscopy.

The detection of diatoms has been proposed to be useful in the diagnosis of drowning. In the presented paper, we describe a novel method for qualitative and quantitative analysis of diatoms in water and organs. Samples were treated by microwave digestion. The fluid obtained was vacuum-filtrated; its sediment on the membrane was coated and examined under the scanning electron microscope with the mode of automatic field scanning; and qualitative and quantitative diatom analysis was performed based on the images captured automatically and on manual identification of the suspected diatoms at a required higher magnification. The method is rapid, sensitive, labor-saving, and accurate for qualitative and quantitative diatom analysis. However, further scientific research in the field of automatic diatom identification using SEM images has to be done in order to automate the process of detection and identification of diatoms in water and tissues for the diagnosis of drowning.

[Concentration changes of potassium and hypoxanthine in vitreous humor of swine and its application to postmortem interval estimation].

OBJECTIVE: To investigate the relationship between postmortem interval (PMI) and concentration changes of components in swine vitreous humor. METHODS: Ninety-six porcine eyes from swine dying from acute massive hemorrhage, being randomly divided into 24 groups, were stored in dark situation, at temperature of (15 +/- 2) degrees C and humidity of (50 +/- 5)% for 2-96 hours separately. The vitreous humor was collected. Concentrations of K+, Na+, Cl- and hypoxanthine (Hx) were analyzed by automatic biochemical analyzer and ultra performance liquid chromatograph (UPLC). The data were statistically analyzed by SPSS software. RESULTS: Linear regression analysis showed that concentrations of vitreous K+ and Hx were positively correlated with PMI(R2=0.767 and R2 = 0.793, respectively). Binary linear regression showed a higher correlation for K+ and Hx with PMI estimation (R2 = 0.866). PMI was not significantly correlated with vitreous Na+ and Cl- concentrations. CONCLUSION: Vitreous K+ and Hx concentrations can be used as the objective markers for PMI estimation. The binary linear regression functions of vitreous K+ and Hx concentrations with PMI are more accurate for estimating the PMI.

Imprinted functionalized silica sol-gel for solid-phase extraction of triazolamin.

A triazolam-imprinted silica microsphere was prepared by combining a surface molecular-imprinting technique with the sol-gel process. The results illustrate that the triazolam-imprinted silica microspheres provided using γ-aminopropyltriethoxysilane and phenyltrimethoxysilane as monomers exhibited higher selectivity than those provided from γ-aminopropyltriethoxysilane and methyltriethoxysilane. In addition, the optimum affinity occurred when the molar ratio of γ-aminopropyltriethoxysilane, phenyltrimethoxysilane, and the template molecule was 4.2:4.7:0.6. Retention factor (k) and imprinting factor (IF) of triazolam on the imprinted and non-imprinted silica microsphere columns were characterized using high performance liquid chromatography (HPLC) with different mobile phases including methanol, acetonitrile, and water solutions. The molecular selectivity of the imprinted silica microspheres was also evaluated for triazolam and its analogue compounds in various mobile phases. The better results indicated that k and IF of triazolam on the imprinted silica microsphere column were 2.1 and 35, respectively, when using methanol/water (1/1, v/v) as the mobile phase. Finally, the imprinted silica was applied as a sorbent in solid-phase extraction (SPE), to selectively extract triazolam and its metabolite, α-hydroxytriazolam, from human urine samples. The limits of detection (LOD) for triazolam and α-hydroxytriazolam in urine samples were 30 ± 0.21 ng mL(-1) and 33 ± 0.26 ng mL(-1), respectively.

[Determination of acid-resistant silicic particles in lung by micro-XRF and its application in diagnosis of drowning].

OBJECTIVE: Using microbeam X-ray fluorescence (Micro-XRF) analyzer for determination of acid-resistant silicic particles in lung, and to explore its potential application in diagnosis of drowning. METHODS: Thirty two white rabbits were divided randomly into drowning group (n=12), post-mortem immersion group (n=10) and control group (n=10). Lungs and water sample were collected for determination of area concentration of acid-resistant silicic particles using Micro-XRF method. RESULTS: The area concentration of acid-resistant silicic particles for the drowning water sample was 4.4 mm2/mL. For the lungs of drowning group, the post-mortem immersion group and the control group, the determined average values were (25.30 +/- 10.95) mm2/g, (1.68 +/- 0.63) mm2/g and (1.65 +/- 0.85) mm2/g, respectively, with a statistically significant difference between the drowning group and the other two groups. CONCLUSION: The area concentration of acid-resistant silicic particles in lungs may be used as an indicator of drowning. The method is highly sensitive and rapid. It provides a potential application in drowning diagnosis.

Trajectory reconstruction through analysis of trace evidence in bullet-intermediate target interaction by SEM/EDX.

A young male was shot to death by a police officer with a Chinese Type 64 7.62 mm pistol when he was dealing with an aggravated assault arising from a traffic accident. By using scanning electron microscopy and energy dispersive X-ray spectroscopy (SEM/EDX), trace deposits on the discharged bullet and the intermediate target, i.e., a concrete telegraph pole at the scene, were identified to be from each other. The result demonstrated the bullet causing the death ricocheted from the concrete telegraph pole before striking the victim, thus indicating the incident was accidental. The case report illustrates the evidential value of trace materials derived, respectively, from discharged bullets and intermediate targets in bullet-intermediate target interaction for trajectory reconstruction. In addition, it indicates that the SEM/EDX method with its nondestructive nature, compared to other methods, may be more helpful in certain situations in determining the origins of trace evidentially valuable deposits on substrates.