Preparing Very Thin,Uniform Cellulose Thin-Layer Plates

Abstract

A procedure is described for the preparation of very thin, uniform cellulose thin-layer plates. The cellulose is sprayed onto the plates in a fine mist from a standard chromatograph sprayer.     

Ion Exchange Application of Overpressured Thin-Layer Chromatography

Abstract

The application of overpressured thin-layer chromatography introduced into the field of ion exchange chromatography. The basic differences between overpressured thin-layer chromatography and classical thin-layer chromatography are discussed including the distinction between the separations performed on thin-layer plates containing silica gel and a mixture of ion exchanger material and silica gel. The basic increase of flow velocity of solvent front with the aid of a pressurized ultra-micro chamber and the effect of flow velocity on the height equivalent of the theoretical plates are also presented. For basic amino acids, the flow velocity vs plate height curves show optima at a moderately high rate of development.     

Structure and TLC Mobility Relationships for Tebuthiuron and Related Thiadiazoles

Abstract

Tebuthiuron and structurally-related thiadiazoles are separated by thin-layer chromatography on silica gel plates in two developing solvent systems. Relationships between chemical constitution and chromatographic mobility are discussed.     

Correlation of TLC,HPTLIC and PMD Results and Their Transfer to HPLC Systems

Abstract

The correlation of the results of five thin-layer chromatographic techniques and their transfer to high performance liquid chromatography are reported. The data obtained indicate that high performance thin-layer chromatographic results are faithfully reproduced by high performance liquid chromatography. Although plates were not activated, and mixed solvents were used, no major differences were observed between thin layer and high performance liquid chromatography.     

4-Acetylpyridine 2-Benzothiazolylhydrazone as a Chromogenic Reagent for Common Epoxides

Abstract

A simple and sensitive color reaction of common epoxides as well as other some alkylating agents with 4-acetylpyrldine 2-benzothiazolylhydrazone (ABH) is described. These compounds gave a purpllsh red color after reaction with ABH for 2 hrs at 110°C in methyl cellosolve or for 20 min at 120°C followed by treatment with plperazlne on silica gel thin-layer plates. Four aliphatic terminal epoxides could be determined spectrophotometrically in the range from 20 to 125 nmol/ml. The limits of detection of these epoxides on silica gel thin-layer plates were 10 pmol/spot order by the visual inspection. The reaction mechanism is also discussed based on the chemical structure of the isolated chromophore.     

A Combined Thin-Layer Chromatography/Micro Infrared Method

Abstract

A method is described for the automatic elution of chromatographed compounds on thin-layer chromatography plates and their subsequent identification by micro infrared spectroscopy.

The method is simple, easy to perform in a few minutes, and requires 5 μg of material, 3 mg of KBr powder, and 150 μl of solvent.     

The Role of Spacers in Displacement Thin-Layer Chromatography

Abstract

Displacement chromatography generates highly concentrated bands which migrate closely to each other along the stationary phase bed.

Spacer-displacement thin-layer chromatography is a planar method improving the observable resolution by inserting odd compounds (spacers) among the members of displacement train to be separated.

Substances were chromatographed by displacement mode of development using silica plates, chloroform carrier and triethanolamine displacer.

Resolution formula valid for elution chromatography has been adapted to the displacement type of developments. Explanation for the numerical value of required separation is given for various cases of displacement thin-layer chromatography.

Equations are suggested in order to calculate yield, loss and efficiency of displacement chromatography.     

The Application of Cross-Scanning to Conventional Thin-Layer Chromatography

Abstract

The technique of cross-scanning, the scanning of thin-layer chromatographic (TLC) plates perpendicular to the direction of development, is applicable to conventional quantitative TLC where significant advantages over normal scanning are realized by its use. Several factors are discussed which influence the ability to cross-scan, such as application of the sample, solvent development and instrumentation.     

Determination of Caffeine in Small Plasma Samples by Gas-Liquid Chromatography with Thin-Layer Chromatographic Sample Clean-Up

Abstract

A sensitive method for the determination of caffeine in small plasma samples (50 μ1) is described. The samples are extracted with chloroform and the extrects are freed from interfering compounds by thin-layer chronatography. The spots of caffeine and the internal standard, 7-ethyl theophylline, are scraped off the plates and both compounds are eluted from the scrapings by means of a glass capillary collector. The caffeine in the eluate is quantitated by gas-     

Non-Coded Biosynthesis of N-acetylaspartyl Peptides in Mouse Brain Homogenates

Abstract

N-acetylaspartate and labelled glutamate, aspartate, γ-aminobutyrate and taurine were incorporated into a number of low-molecular weight peptides in mouse brain homogenates, as detected by autoradiography on thin-layer chromatography plates. Peptide biosynthesis was dependent on N-acetylaspartate, cAMP, ATP and an ATP regenerating system. The number of peptides varied from 5 to 12 after incubation for 40 min. The original labelled amino acids were determined by re-chromatography of the acid hydrolysates of the peptides, and the probable structures (N-acetylaspartyl-glutamyl-taurine, [N-acetyl] aspartyltaurine and N-acetylaspartyl [aspartyl, glutamyl]-taurine) were assumed to be present.     

Unconventional TLC systems in lipophilicity determination: A review

ABSTRACT

This review focuses on four unusual thin-layer chromatography (TLC) approaches for the determination of lipophilicity: (1) the use of medium-polar stationary phases: CN, NH₂, and DIOL instead of RP plates, together with water-based mobile phase; (2) the use of silica gel in a typical normal-phase manner and treating extrapolated retention indices as the “reversed lipophilicity”; (3) the use of oil impregnated silica gel in the reversed-phase manner; and (4) the use of salting-out mobile phases. The chromatographic indices obtained in these systems are numerously reported as well correlated with lipophilicity and they are an interesting alternative to classical RP systems approaches.     

Detection reagents used for on-plate identification of organic pesticides in biological samples with preliminary separation by TLC/HPTLC

Many specific and non-specific chromogenic spray reagents have been used to detect organic pesticides (organophosphorus, organochlorine, carbamates, and pyrethroids) on thin-layer chromatographic (TLC)/high-performance thin-layer chromatographic (HPTLC) plates. To realize high sensitivity and improved selectivity, several chromogenic reagents have been introduced. The physical properties of organic pesticides reported so far are also presented in tabular form. The colors produced on TLC plates due to reactions taking place between pesticides and spray reagents are illustrated in the form of chromatograms, and reaction mechanism is also presented.

     

Determination of Oxazepam in Pharmaceutical Formulation by HPTLC UV-Densitometric and UV-Derivative Spectrophotometry Methods

Abstract

Methods for determination of oxazepam in pharmaceutical formulation by derivative ultraviolet (UV) spectrophotometry as well as high-performance thin-layer chromatography (HPTLC) UV densitometry were described. For UV-derivative spectrophotometry, some derivatives and wavelengths may be recommended for routine quality control of the drug of interest. On the other hand, HPTLC provided good results, but only when the calibration curve was estimated using nonlinear regression analysis. The HPTLC method was developed with silica F₂₅₄ plates, a mobile phase of benzene/ethanol (5:1, v/v), and densitometric detection at 204 nm receiving R _f  = 0.47. Developed methods were validated and found to be sufficiently precise and reproducible for established conditions.     

Thin-Layer Radiochromatographic Determination of Captopril (SQ 14,225) and its Disulfide Dimer Metabolite in Blood

Abstract

A reliable thin-layer radiochromatographic (TLRC) assay has been developed for quantitation of radiolabeled captopril (CP), a new sulfhydryl-containing orally active antihypertensive agent, and its disulfide dimer metabolite (CPD) in blood. CP, which is chemically unstable in blood, was immediately converted to a stable derivative by addition of N-ethylmaleimide (NEM) to freshly collected samples. Aliquots of whole blood samples were analyzed for total radioactivity, and NEM-treated aliquots were extracted with methanol. Reconstituted residues of the extracts were applied to silica gel GF plates, developed with chloroform/ethyl acetate/glacial acetic acid (4:5:3), and analyzed for radioactivity associated with CP and CPD by zonal analysis.     

On-Plate Electrochemical Detection for Thin-Layer Chromatography

Abstract

Voltammetric electrochemical detection (ECD) coupled with thin-layer chromatography (TLC) was demonstrated for the quantification of trace organic compounds directly on a TLC plate. the electrochemical detection solvent was a thin layer of aqueous potassium chloride. For undeveloped plates, detection limits for p-anisidine and p-phenetidine were 10 ng and 13 ng, respectively. Linearity was demonstrated over nearly 2 orders of magnitude. After development, detection limits increased by approximately a factor of ten. Advantages of this method over other quantitative TLC methods include sensitivity, speed, simplicity, and cost. in addition, this method is selective for electrochemically active compounds. Major sources of experimental error include spot size reproducibility, working electrode placement, and supporting electrolyte film thickness.     

Experience with Quantitative Thin-Layer Chromatography in the Analysis of Drugs in Plasma

Abstract

A survey is presented concerning the possible techniques of quantitative thin-layer chromatography. Most of the possibilities mentioned have been used successfully in our laboratory for the determination of drugs and metabolites in plasma. With respect to our own experience, the advantages and disadvantages of quantitative thin-layer chromatography are discussed.

Although gas chromatography and high-performance liquid chromatography are more popular, quantitative thin-layer chromatography has a high potential in the analysis of drugs in samples of biological origin and should not be neglected in the development of an assay.     

Derivatisation of sympathomimetics with primary amino groups on thin-layer plates by spotting the sample spots with fluorescamine

Summary A method has been developed for the quantitative determination of sympathomimetics containing a primary amino group on thin-layer plates with fluorescamine. The reaction is carried out by spotting fluorescamine solution in acetone on top of the sample spots. The fluorescamine derivatives are subsequently separated using appropriate solvent systems. Spotting a buffer before reaction and spraying with triethanolamine after development is unnecessary. The consumption of reagent is extremely low. For ten thin-layer plates with ten sample spots per plate only 0.2 cm³ reagent solution are needed. The method has been applied to the quantitative determination of some compounds with primary amino groups in pharmaceutical preparations.Dedicated to Prof. Dr. G. Zigeuner on his 60^th birthday.     

TLC–direct bioautography as a method for evaluation of antibacterial properties of Thymus vulgaris L. and Salvia officinalis L. essential oils of different origin

ABSTRACT

The dot-blot bioautography was used to evaluate the antibacterial properties of Thymus vulgaris L. and Salvia officinalis L. essential oils (EOs) produced by three different manufacturers. The whole samples were applied at three concentrations on thin-layer chromatography (TLC) plates which were then subjected to bioautography against Bacillus subtilis. The samples of the highest activity were found. Then, they were separated using TLC and once again subjected to bioautography against B. subtilis. As was proved, only the essential oils of T. vulgaris L. possessed strong antibacterial properties for which mostly thymol and carvacrol were responsible. Their contents were calculated using TLC–UV densitometry. The highest contents were found in the essential oils of the highest antibacterial activity revealed in the dot-blot test. It means that a dot-blot test can be used for simple and fast evaluation of antibacterial properties of essential oils.     

Thin-layer chromatographic separation of dicrotophos,ethion (or phorate), fensulfothion,oxydemeton-methyl,phosmet, phospholan,and trichlorfon from each other

A thin-layer chromatography method is reported for the separation of dicrotophos, ethion (or phorate), fensulfothion, oxydemeton-methyl, phosmet, phospholan, and trichlorfon. The procedure involves the use of commercially prepared silica gel 1B Baker-flex plates and developing with 2,2,4-trimethylpentane:methyl cyclohexane:n-hexyl alcohol:acetone (18:9:9:9). The pesticides are located by spraying with ammoniacal silver nitrate solution in acetone follwed by exposure to longwave UV light. The method does not separate ethion and phorate from each other. A method is also reported for the thin-layer chromatographic separation of ethion from phorate in the presence of the other six pesticides using solvent system 2,2,4-trimethylpentane:n-hexane:chloroform (18:18:12) on silica gel 1B Baker-flex plates.     

Chemische Vitamin D3-Bestimmung in Lebertran

The different aspects of Vitamin D₃ determination and the data reported in the literature are discussed. The spectrophotometric determination of Vitamin D₃ in cod-liver oil is carried out after the alkaline saponification, extraction of unsaponified parts, precipitation of accompanying sterols and the column- and thin-layer chromatographic purification and separation of vitamin D₃ on 40 cm plates from other vitamins. The dyestuff α-naphtholbenzein is suited well as standard substance for the better location and identification of vitamin D₃ zone on the thin-layer plate. The results obtained from the chemical method were checked through the simultaneous biological determination.