[Comparative characteristics of enzymatic hydrolysis methods for isolating toxic substances from blood and hair]. / Sravnitel'naya kharakteristika metodik fermentativnogo gidroliza dlya izolirovaniya toksichnykh veshchestv iz tsel'noi krovi i volos.

The aim of study was to develop methods for the hydrolysis with hyaluronidases for the isolation of drugs (phenobarbital, diphenhydramine hydrochloride and phenibut) from biological objects (blood, hair) and to compare their effectivenes with previously developed methods of enzymatic hydrolysis. The studies were carried out using model «blood - model drug substance (MDS)¼, a natural and artificially colored wool (hair) of laboratory animals - guinea pigs of white, red and black natural colors, which were daily given a solution of MDS and then were subjected to cosmetic effects - coloring. The isolation of MDS from the model complex «blood - MDS¼ and from wool was carried out using the developed methods of hydrolysis with proteolytic enzymes (papain, chymopsin and chymotrypsin) and hyaluronidase. Phenobarbital and diphenhydramine from hydrolysates were extracted by liquid-liquid extraction method, phenibut - by direct freezing extraction with acetonitrile. The analysis of the extracts was carried out by gas chromatography with mass selective detection. The results showed that the developed methods of enzymatic hydrolysis can be recommended for isolating substances of various properties (acidic, amphoteric and alkaline) from blood, natural and artificially colored wool (hair).

[The development and validation of the methods for enzymatic hydrolysis for the extraction of toxic compounds from the uncoloured hairs]. / Razrabotka i validatsiia metodiki fermentativnogo gidroliza dlia izolirovaniia toksicheskikh veshchestv iz neokrashennykh volos.

The objective of the present study was the development of the method for the extraction of toxic compounds from the uncoloured hairs with the use of enzymatic hydrolysis by proteolytic enzymes of the toxic compounds comprising the group of pharmaceutical substances present on the uncoloured hairs collected for the forensic medical expertise. The experiments were carried out using laboratory animals, viz. guinea pigs having hair of natural white and black colour and white rats that were given per os a phenobarbital solution and a dimedrol solution as well as daily intravenous injections of tropicamide hydrochloride administered every 28 days. The hair obtained from the experimental animals was subjected to acidic, alkaline, and enzymatic hydrolysis with chymotrypsin, trypsin, and papain. The analysis of the extracted materials was performed using the gas chromatographic technique with mass-selective detection. The results of the study give evidence that the completeness of the extraction of the toxic substances from animal hairs with the use of enzymatic hydrolysis is twice and thrice that of acidic and alkaline hydrolysis respectively. The enzymatic hydrolysis methods developed in the present study are equally efficient using the naturally coloured white and black hairs of the laboratory animals which allows to recommend them for the extraction of poisonous substances from such material. The methods were validated.

EVALUATION OF EFFICACY AND SAFETY POSTOPERATIVE PAIN MANAGEMENT BY INTRAMUSCULAR ANALGESIA AFTER DIFFERENT TYPES OF ANAESTHESIA: PILOT CLINICAL PROSPECTIVE STUDY.

MATERIAL AND METHODS: Efficacy Safety Score (ESS) with "call-out algorithm" developed in Kongsberg hospital, Norway was used for the validation. ESS consists of the mathematical sum ofscorefrom: 2 subjective (Visual Analog Scale: VAS at rest and during mobilization) and 4 vital (conscious levels, PONV circulation and respiration status) parameters and ESS > 10 is a "call-out alarm "for visit ofpatient by anaesthesiologist. Hourly registration of ESS, mobility degree and amounts of analgetics during the first 8 hours after surgery was recorded in the specially designed IPad program. According to the type ofanaesthesia all patients were allocated in 4 groups: I spinal anaesthesia (SA), II general anesthesia (GA), III peripheral blockade (PB) and IV Total intravenous anaesthesia (TIVA). RESULTS AND DISCUSSION: A total of 223 patients were included in the study. Statistically low levels of both VAS and ESS in the first 2-4 postoperative hours were found in SA and PB groups compared to GA and TIVA groups. During 8 post-operative hours, VAS> 3 was recorded in 10.5% of SA, 13.9% in GA, 12.8% in PG and 23.5% in TIVA patients. CONCLUSIONS: Intramuscular postoperative analgesia was effective in SA, GA and PG groups. More attention of anaesthesiologist must be paid to patients ofter TIVA.

[The chemical toxicological study of cyclomed].

The objective of the present study was to develop the methods for the chemical and toxicological analysis of cyclopentolate contained in the material evidence for the purpose of forensic chemical and forensic toxicological expertises. The optimal conditions for the isolation of cyclopentolate from the cyclomed preparation and biological fluids were created using a chloroform-2-ptropanol mixture at pH of the medium 10.0. The substance of interest was identified with the use of the color and flocculation reactions, UV spectroscopy, thin layer chromatography and gas chromatography with the use of a mass-selective detector. HPLC and GCh/MS were employed for the quantitative determination. The results of the two methods are in excellent agreement.

[The study of naphthyzin present in material evidence and biological fluids].

The optimal conditions for isolation of naphazoline from naphthyzin preparations and biological fluids with chloroform at pH 9.18 are described. The compound of interest was identified with the use of color and precipitation reactions, IR and UV spectroscopy, thin-layer and gas chromatography, and chemical methods including high performance liquid chromatography, chromatodensitometry, and UV spectroscopy. The results obtained by the three methods are comparable.

[The application of chromatography and spectrometry for the analysis of ketorolac and diclofenac in biological fluids].

The objective of the present work was to study conditions for isolation of ketorolac and diclofenac from biological fluids. A method of their extraction with a mixture of organic solvents has been developed and the conditions for the identification of these compounds are proposed with the use of high performance liquid chromatography (HPLC), UV spectroscopy, and gas chromatography with electron capture detection (GC/ECD). The possibilities of using HPLC, UV spectrometry, and GC/ECD for quantitative determination of ketorolac and diclofenac are illustrated.

Radiographic lung density assessed by computed tomography is associated with extravascular lung water content.

BACKGROUND: We hypothesized that in acute lung injury (ALI), the volume of pulmonary tissue with aqueous density, as determined by spiral computed tomography (CT), is associated with extravascular lung water content. Our aim was to compare tissue volume index, as assessed by CT, before and after oleic acid-induced ALI, with extravascular lung water indexes (EVLWI), determined with single transpulmonary thermodilution (EVLWI(STD)), thermal-dye dilution (EVLWI(TDD)), and postmortem gravimetry (EVLWI(G)). METHODS: Seven instrumented sheep received an intravenous infusion of oleic acid 0.08 ml/kg (OA group) and four animals had vehicle only (Control group). The day before, and immediately after the experiment, sheep were anesthetized to undergo quantitative CT examinations during a short breath hold. Hemodynamics, oxygenation, EVLWI(STD), and EVLW(TDD) were registered. Linear regression analysis was used to assess the relationships between EVLWI(STD), EVLW(TDD), EVLWI(G), and lung tissue volume index (TVI(CT)) determined with CT. RESULTS: In the OA group, total lung volume increased compared with Controls. Poorly and non-aerated lung volumes increased a 3.6- and 4.9-fold, respectively, and TVI(CT) almost doubled. EVLWI(STD), EVLWI(TDD), and TVI(CT) were associated significantly with EVLWI(G) (r=0.85, 0.90, and 0.88, respectively; P<0.001). TVI(CT) deviated from the reference EVLWI(G) values to the greatest extent with a mean bias +/- 2SD of 4.0 +/- 6.0 ml/kg. CONCLUSIONS: In ovine oleic acid-induced ALI, lung tissue volume, as assessed by quantitative CT, is in close agreement with EVLWI, as determined by indicator dilution methods and postmortem gravimetry, but overestimates lung fluid content.

[The application of chromatographic and spectroscopic techniques for the analysis of certain antiinflammatory agents].

Optimal conditions for the separation of diclofenac and ketorolac in the presence of other pharmaceutical products and narcotic substances are described with the help of gas and high performance liquid chromatography. The detection limits for individual compounds were established and the characteristic ions were identified. The possibility of application of IR- and UV-spectroscopy for the analysis of selected narcotic and non-narcotic analgesics was evaluated.

[Thin layer chromatography for the analysis of certain anti-inflammatory medicines].

This work was devoted to the elucidation of conditions for isolation of ketorolac and diclofenac from biological fluids. A method is proposed for the extraction of these compounds from solutions with organic solvents at different pH values. Other methods permit to optimize identification of analytes by thin layer chromatography while the densitometric technique may be used for qualitative and quantitative analysis of their composition in biological fluids.

[Chemical-toxicological analysis of butorphanol].

The conditions of butorphanol isolation from biological fluids were studied. The method of its extraction with the mix of organic solvents by pH 12 was proposed. How to identify butorphanol with the methods of thin-layer chromatography, ultraviolet spectrometry, high-performance liquid chromatography, gas chromatography with a detector of electron capture, chromato-mass spectrometry was developed. Possibility of use ultraviolet spectrometry for quantitative assessment of butorphanol was shown.

[Assessment of current methods quantitating extravascular lung water and pulmonary aeration in inhomogeneous lung injury: an experimental study].

BACKGROUND: Single transpulmonary thermodilution (STTD) is a widely recognized technique for the quantification of extravascular lung water (EVLW). However, the accuracy of STTD can be substantially reduced in acute lung lesion (ALL) characterized by inhomogeneous distribution of edematous zones and major ventilation-perfusion mismatch. Quantitative computed tomography (CT) may be a helpful clinical adjunct allowing an assessment of pulmonary gas and tissue content. The purpose of the study was to compare the tissue volume index, as estimated by spiral CT (TVICT), with EVLW indices determined with STTD (EVLWISTTD), thermal-dye dilution (EVLWITDD), and postmortem gravimetry (EVLWIG) before and after oleic acid-induced ALL in sheep. MATERIALS: Eleven yearling sheep were randomly assigned to either an oleic acid (OA) group receiving an infusion of OA in a dose of 0.08 ml/kg i.v. or to a control group. The day before and immediately after the experiment, sheep underwent CT examinations. Pulmonary and systemic hemodynamics, oxygenation, EVLWISTTD and EVLWITDD were recorded. Linear regression analysis was used to assess the relationships between EVLWISTTD, EVLWITDD, EVLWIG, and TVICT (syngo PulmpCT, Siemens, Germany). RESULTS: OA caused 5- and 7-fold increments in poorly and nonaerated lung volumes, respectively, and increased total lung volume and TVICT, EVLWISTTD, EVLWITDD, and TVICT demonstrated a close agreement with EVLWIG (r = 0.86, 0.90, and 0.97, respectively; p < 0.001). TVICT overestimated reference EVLWIG values to the greatest extent. CONCLUSION: In a sheep model of OA-induced ALL, pulmonary tissue volume as estimated by quantitative CT closely correlates with EVLWI measured by dilutional methods and postmortem gravimetry.

[Chemicotoxicological evaluation of doxylamine].

The conditions of doxilamine isolation from biological fluids are studied. The method of its extraction with a mixture of organic solvents in pH 9 is proposed. Identification of doxilamine with techniques of thin layer chromatography, UV-, IR-spectroscopy, chromato-mass-spectrometry, densitometry, gas-liquid chromatography is described. UV-spectrometry, gas chromatography and densitometry can be used for quantitation of doxilamine.