INVESTIGATION OF A MIXTURE CONTAINING ALPRAZOLAM, CODEINE AND PARACETAMOL USING THIN-LAYER AND HIGH PERFORMANCE LIQUID CHROMATOGRAPHY METHODS.

The increasing drug consumption in Lithuania and all over the world makes us think about the negative consequences - the risk of toxicity. Fast and accurate identification of material that caused the poisoning reduces the probability in death cases and makes easier to determine the main cause of death. The results have shown that the most appropriate systems of solvents for qualitative analysis by TLC method of the mixture consisting of alprazolam, codeine and paracetanol are: system "D" (trichloromethane : acetone : conc. ammonia = 55 : 40 : 5 (v/v/v)) and system "F" (trichloromethane : diethyl ether: isobutanol : conc. ammonia = 50 : 30 : 15 : 5 (v/v/v/v)). For qualitative analysis of the mixture consisting of alprazolam, codeine and paracetamol by HPLC method the chromatographic column ACE C18 (25 cm x 4.6 mm x 5 µm), gradient elution mode (mixture of 3% acetic acid and methanol and the flow rate 1 mL/min have been used. The injection volume was 10 pL. Photodiode array detector (210 - 240 nm range) has been used. UV absorption spectra of materials measured using photodiode array detector have been identical to those presented in the scientific literature.

Development and Evaluation of Two-Phase Gel Formulations for Enhanced Delivery of Active Ingredients: Sodium Diclofenac and Camphor.

The formulation of biphasic gels as potential semi-solid carriers for hydrophilic and lipophilic active substances is promising for the development of pharmaceutical preparations. The aim of this study was to design a stable bigel composition and to determine the influence of the organogel/hydrogel ratio on the gel's physical-chemical and structural-mechanical properties. The investigated compositions of organogel/hydrogel remained stable at ratios ranging from 5/95 to 40/60. After texture and microstructure analysis, bigels with 20/80 and 25/75 ratios were selected as carriers for the active ingredients, sodium diclofenac and camphor, for use as topical preparations for the treatment of muscle-joint inflammation and pain. Although other researchers have published data on the preparation and evaluation of bigels, there are no scientific results on the development of a two-phase gel with our proposed combination of APIs. Sodium diclofenac release was found to be higher when combined with camphor, which revealed the advantages of the biphasic formulation. The pseudoplastic behavior, thixotropy, and thermal stability of flow of the studied bigel samples was investigated by rheological analysis. Ongoing stability studies confirmed the minimal 6-month period.

Application of high-performance liquid chromatography for research of salicin in bark of different varieties of Salix.

Willow (Salix L.) species are widely spread in Lithuanian natural dendroflora. Willow bark contains active substances known for anti-inflammatory properties and is known as a phytotherapeutic precursor of aspirin. Bark extracts are components of analgesic and antirheumatic preparations. Therapeutic effectiveness is associated with salicin (2-(hydroxymethyl)phenyl-beta-D-glucopyranoside), which turns into salicylic acid. Increasing attention to natural preparations gives primary importance to research of plants. This study focused on 12 willow taxa and employed routine pharmacopoeia methods. High-performance liquid chromatography method was applied for the analysis of bark extractions. The investigation revealed that not all willow species accumulated a therapeutically sufficient amount of salicin. Bark samples were investigated after 1- and 2-year growth in autumn and spring. Salicin content ranged from 0.08 to 12.6%. Higher contents of active materials were determined in autumn and in 2-year-old willows. Certain willow taxa (Salix alba L., Salix mollissima L., Salix triandra L., Salix viminalis "Americana", Salix dasyclados L.) possessed extremely low salicin amounts. In the second year, analysis covered 32 willow species. Results indicated striking differences in salicin amounts (from 0.04% in Salix viminalis "Americana" to 12.06% in Salix acutifolia). Willow species, plant age, and season should be considered when collecting medicinal plant material. The amount of salicylates in 2-year-old willow bark collected in autumn exceeded by 25% that in 1-year-old willow bark collected in spring. Bark of some analyzed willow species contained the amount of salicylates too low for using as anti-inflammatory or antipyretic remedy.

[Identification of the drugs in the mixture using thin-layer chromatography in sudden poisoning cases]. / Vaistu misinio komponentu identifikavimas plonasluoksnes chromatografijos metodu uminiu apsinuodijimu atvejais.

The problem of acute intoxication has become very urgent now due to a great number of various chemical preparations accumulated during the last decades in the environment. Intoxications with psychotropic drugs and their mixtures form the significant part of the intoxications; there is an increasing tendency of intoxication with several preparations at a time. Amitriptyline and codeine are the preparations, which more frequently can cause intoxication. Fluoxetine is one of the newest and often used antidepressants. Under certain circumstances, like overdose, using all preparations together, long term using or using for suicide, these preparations can be even a cause of death. In such cases amitriptyline, fluoxetine and codeine become the objects of chemical-toxicological analysis. The possibility of the separation and identification of amitriptyline, fluoxetine and codeine in the mixture using thin-layer chromatography was established. Dragendorf reagent, modified by Munje, is most suitable for the spray-distinct of the chromatographic plates for all three substances. Amitriptyline research limit, using this developer, is 0.4 microg, fluoxetine--1.6 microg, codeine--0.8 microg. Most acceptable for separation the components of the mixture are 5 mobile phases: 1. Diethyl acetate-methanol-ammonium hydroxide (concentrated solution) (85:10:5). Amitriptyline, fluoxetine and codeine R(f) medium values respectively are 0.94; 0.63; 0.51. 2. Buthylacetate-methanol-ammonium hydroxide (concentrated solution) (85:10:5). Amitriptyline, fluoxetine and codeine R(f) medium values respectively are 0.65; 0.24; 0.15. 3. Cyclohexane-diethyl acetate-diethyl amine (70:15:15). Amitriptyline, fluoxetine and codeine R(f) medium values respectively are 0.93; 0.75; 0.37. 4. Cyclohexane-buthylacetate-diethyl amine (70:15:15). Amitriptyline, fluoxetine and codeine R(f) medium values respectively are 0.92; 0.51; 0.25. 5. Acetone-1,4-dioxane-ammonium hydroxide (concentrated solution) (30:68:2). Amitriptyline, fluoxetine and codeine R(f) medium values respectively are 0.82; 0.62; 0.42. Recommended methodology for the separation and identification of amitriptyline, fluoxetine and codeine in the mixture using thin-layer chromatography is statistically reliable: when confidence level is 0.95, relative error is less than 0.05; standard deviation is from 0.007 to 0.03. Recommended methodology suits for mixture, extracted from biological liquids, components separation and identification.

[The chemical-toxicological research of psychostimulator mixture by thin-layer chromatography]. / Psichostimuliatoriu misinio cheminis-toksikologinis tyrimas plonasluoksnes chromatografijos metodu.

With increasing number of intoxications with some preparations at the same time there is a lack of literature data about chemical-toxicological research of psychostimulator mixture. The aim of the study was to analyze the possibility of identification of four preparations in the mixture. The thin-layer chromatography method was used. The most acceptable solvent systems were determined: ethylacetate-methanol-conc. ammonia hydroxide (40:4:2:25) and dioxane-benzenum-conc. ammonia hydroxide-acetone (28:14:2:25).

[UV spectrophotometric research of drug mixture in sudden poisoning cases]. / Vaistu misinio UV-spektrofotometrinis tyrimas uminiu apsinuodijimu atvejais.

UNLABELLED: The objective of this research--to develop the methodics for analysis of amitriptyline, fluoxetine and codeine in the mixture. RESULTS: The analytical method of amitriptyline, codeine and fluoxetine in the mixture identification and quantitative determination using ultraviolet spectrophotometry was established. Preparations in the mixture can't be separated, because material ultraviolet light peaks are in insufficient distance and therefore cover one another. The maximum of ultraviolet light absorption for amitriptyline is at 217-220 and 238-240 nm; fluoxetine--at 226-228 nm; codeine--at 224-248 and 284-286 nm. Using ultraviolet spectroscopy it's possible to identify amitriptyline, fluoxetine and codeine only after separating mixture components by thin-layer chromatography, the same time executing cleaning of extracts from blood and urine. Using ultraviolet spectroscopy can be identificated at least 0.5 micro g/ml amitriptyline, 1.5 micro g/ml fluoxetine and 1.0 microg/ml codeine. The intervals of the quantitative determination: 5-25 microg/ml amitriptyline; 5-30 microg/ml fluoxetine; 100-300 microg/ml codeine; relative error of the measurement, when confidence level is 95%, is from 0.66 to 1.2% for amitriptyline; from 0.66 to 1.45% for fluoxetine; from 0.33 to 0.88% for codeine. Standard deviation is from 1.15 to 2.08% for amitriptyline; from 1.15 to 2.52% for fluoxetine; from 0.57 to 1.53% for codeine. Molar absorption coefficients for all three preparations in distillated water were determinated. CONCLUSIONS: recommended methodology suits for mixture, extracted from biological liquids, components separation, identification and quantitative determination.

[Analysis of psychotropic drug mixtures using high-pressure liquid chromatography in acute poisoning cases]. / Psichotropiniu vaistu misinio tyrimas auksto slegio skysciu chromatografijos metodu uminiu apsinuodijimu atvejais.

The qualitative and quantitative method of determination of amitryptilin, codeine and fluoxetine in the mixture using high-pressure liquid chromatography is described in this paper. Chromatogram is presented which shows, that preparations are fully separated and do not interfere each others analysis. Tables with chromatographical separation characteristics are also presented. Proposed calibration curves of quantitative analysis and calculated medium relative error of quantitative ascertainment for every preparation of the mixture are shown. Final conclusion: method is applicable for qualitative and quantitative analysis of amitryptiline, fluoxetine and codeine in the mixture in hasty poisoning cases.