Semi-Preparative High-Performance Liquid Chromatographic Separation of Potato Glycoalkaloids

Abstract

A semi-preparative high-performance liquid chromatographic method has been developed to separate crude mixtures of the potato glycoalkaloids α-chaconine, α-solanine, commersonine and demissine. Milligram quantities of each substance can be obtained within an 8 hour period. A Zorbax semi-preparative NH₂ column and a solvent system of tetrahydrofuran-water-acetonitrile (55:20:25) were employed for the separation. The flow rate was 1.0 ml/min. Glycoalkaloid separations were monitored using both refractive index and ultraviolet detection (215 nm). Further analyses of these glycoalkaloids were done using analytical high-performance liquid chromatography (HPLC) and thin-layer chromatography (TLC) to check compound purity and identity.     

Separation of Bufotalin and Cinobufotalin by Preparative Liquid Chromatography

Abstract

Complete resolution of bufadienolides by traditional column chromatography, or by thin-layer chromatography (TLC), is quite difficult. Separation of various bufadienolide conjugates by high performance liquid chromatography (HPLC) has been described, but not for resolution of the corresponding genins. A preparative HPLC procedure has been developed for resolving the difficultly separable bufotalin (1) and cinobufotalin (2). A Lobar B column packed with LiChromprep Si-60 was used to separate these bufadienolides employing a recycling procedure.     

A Direct Fractionation Procedure for the Isolation of Synthetic Products Utilizing Normal and Reversed Phase Liquid Chromatography

Abstract

A simple, one-step fractionation scheme was developed by utilizing liquid chromatography in order to provide new and improved isolation techniques for synthetic reactions, allowing elimination of tedious, multi-step processes presently in use. Thin-layer chromatography (TLC) data, used for monitoring and optimizing synthetic reactions, were directly extrapolated to high performance liquid chromatography (HPLC) systems. Inorganic reagents and highly polar side products were removed by a silica pre-column. A reversed phase column was used for clean-up process. The desired products were rapidly fractionated from the crude reaction mixture. General applicability of this procedure was demonstrated in the syntheses of various steroid derivatives.     

Use of High Pressure Liquid Chromatography and Thin Layer Chromatography for the Separation and Detection of Testosterone and its Metabolites from in vitro Incubation Mixtures

Abstract

Testosterone and its 6β-, 7α-, and 16α-hydroxylated metabolites were resolved by high pressure liquid chromatography and thin layer chromatography. Separation by HPLC was achieved in less than 45 min on a microparticulate silica gel column using isocratic elution with isopro-panol:tetrahydrofuran:hexane (5:15:80) as the mobile phase. TLC systems utilizing silica gel on glass and plastic plates, and polysilicic acid on glass fiber sheets are presented. The monohydroxylated metabolites of testosterone formed during incubation of (¹⁴C)-testosterone with liver postmitochondrial preparations from adult male rats pre-treated with phenobarbital or Aroclor 1254 were separated and quantitated by both HPLC and TLC. The results using both techniques are compared with those obtained by paper chromatography.     

Evaluation of the Plasma Chromatograph as a Qualitative Detector for Liquid Chromatography

Abstract

The applicability of the plasma chromatograph as a sensitive, qualitative detector for liquid chromatography (LC) is demonstrated. Only a fraction of an LC column effluent directly introduced is needed to produce a qualitative mobility spectrum. Most LC carrier fluids either exhibit no response in plasma chromatographic spectra or do not interfere with LC peak components. Using an indirect technique, components separated by TLC can be identified by plasma chromatography.     

Overpressured Thin-Layer Chromatography and Its Applications

Abstract

In recent years, a rapid progress can be observed both in column and planar liquid chromatographic techniques. In the field of liquid column chromatography the most spectaular achievement was the development of high-performance liquid chromatographic/HPLC/ systems by means of several special instruments and sorbents/1, 2/. As regards planar techniques, the most significant break-through is the development of highperformance thin-layer chromatography/HPTLC//3/ based on the application of fine-particle sorbents. Both techniques proved to be very useful in many fields of chemical analysis, although the use of the latter is more restricted, mainly to micro chromatographic studies.     

Determination of Pentachlorophenol Residues in Tallow by Quantitative TLC

Abstract

A TLC method is described for screening and quantification of PCP in tallow. PCP is extracted and cleaned up by alumina column chromatography, and is determined by preadsorbent silica gel TLC with silver nitrate detection and densitometric scanning. The detection limit of the analysis was 0.50 ppm in tallow. Recoveries varied from 97.9 to 103% over the fortification range of 1 to 20 ppm. The CV of the analysis was 6.2% .     

Application of Thin Layer,Ion Exchange and High Performance Liquid Chromatography to Separate Pharmacologically Active Components of an African Arrow Poison of Plant Origin

Abstract

We have applied thin layer chromatography (TLC), ion exchange chromatography (IEC) and, high performance liquid chromatography (HPLC) to separate and identify the pharmacologically active components of an african arrow poison of plant origin. On the basis of Rf values obtained from TLC, the active components of the toxin are unlike d-tubocurarine, atropine and, scopolamine. Dowex 1 × 2 IEC of 630 mg of crude toxin on a 2.5″ × 33″ column with step gradient elution (NaCl, 0.1 - 1. OM and NaOH, 0.1M) led to the identification of three distinct peaks. When the components of each of the three peaks were subjected to HPLC, the results confirmed the homogeneity of each of the isolated peaks except for the third peak which was a doublet.     

Applicability of an HPLC System in the Analysis of Biodegraded Crude Oil Components

Abstract

High performance liquid chromatography has been used for the separation of biodegraded crude blend into fractions using back flushing procedure. The stationary phase was Energy analysis (3.9mm × 30cm) NH₂ column and the detection mode was UV absorptiometry. The method was compared with the liquid chromatography method.     

Separations in Thin Layer and High Performance Liquid Chromatography Using Alkyl Silica Gel Bonded Phases

Abstract

Separations in thin layer (TLC) and high performance liquid chromatography (HPLC) using alkyl bonded phases were carried out under optimum solvent conditions for each of three phases, RP-2, RP-8 and RP-18. The phases were tested for their efficiency and resolving power using three groups of compounds in three binary organic-water mobile phases. The organic solvents were acetonitrile, methanol and tetrahydrofuran, which are widely used as solvent modifiers in reversed phase liquid chromatography.

The results showed that, unlike HPLC, TLC using RP-18 plates was the most, and RP-2 plates the least, efficient. A naphthalene and biphenyl mixture which was resolved by HPLC using any of the three solvents and columns was not resolved by TLC using any plate or solvent combination, unless the plate was prewashed with an organic modifier. The addition of NaCl (1–2% wt/vol) to the solvent for TLC speeded development unless an alcohol was used, but did not greatly affect the separation.     

Modeling of chromatographic retention of the selected antiarrhythmics and structurally related compounds in the hydrophilic interactions under the TLC and HPLC conditions

Abstract

High-performance liquid chromatography (HPLC) plays an important role in testing the pharmaceutically active compounds. In despite of the advantages of HPLC, thin-layer chromatography (TLC) retains its applicability to the different experimental tasks. The experimental conditions which allow hydrophilic interactions in the chromatographic system were tested in the HPLC and TLC systems for ivabradine, its related compounds, diltiazem and verapamil. Under the TLC conditions, retention behavior of the investigated compounds was tested on silica gel modified with cyanopropyl ligands as stationary phase and acetonitrile?+?methanol containing 25% v/v formic acid. Under the HPLC conditions, we used silica gel modified with cyanopropyl ligands as a column packing and the acetonitrile + 0.25% aqueous solution of formic acid as mobile phase. Retention behavior of the investigated analytes depending on the changing volume fractions of the mobile phase modifier was characterized both for TLC and HPLC data sets by the Soczewiński–Wachtmeister equation. Linear relationships were established between the retention coefficients characterizing the retention mechanism (R_M⁰/m, logk₀/m) and molecular properties of the investigated compounds. The Quantitative Structure Retention Relationship (QSRR) modeling was performed with the use of the stepwise multiple linear regression, in order to select molecular properties which influence retention.     

A combined version of planar chromatography

A new version of thin-layer chromatography (TLC) was developed, in which a combination of conventional TLC versions, ascending, descending, and horizontal, were used on one plate. Such a combination decreases the time of analysis and increases the separation efficiency with respect to the conventional methods of planar chromatography. The method was used to analyze liquid kerosene pyrolysis products obtained using various catalytic systems.     

Determination of Selected Quinolones and Fluoroquinolones by Use of TLC

Abstract

There are many different methods of quinolones determination. The most often used method of quinolones analysis is liquid chromatography. In this work some selected quinolones (cinoxacin, pipemidic acid) and fluoroquinolones (ofloxacin, pefloxacin) were separated with thin-layer chromatography (TLC). The two different mobile phases were used as follows: buffer solution (pH = 5.5)-methanol, 40:10 (v/v) and acetonitrile-water-acetic acid, 6:40:4 (v/v/v), respectively, for quinolones and fluoroquinolones. The following chromatographic parameters were calculated for these separations: R_F, ?R_F, R_M, and R_S. The possibility of qualitative determination of cinoxacin, pipemidic acid, ofloxacin, and pefloxacin using TLC was shown.     

Application of Thin-Layer Chromatography to High-Performance Liquid Chromatographic Separation of Steroidal Hormones and Cephalosporin Antibiotics

Abstract

High-performance liquid chromatographic (HPLC) separation of steroidal hormones and cephalosporin antibiotics was investigated by adsorption chromatography and reversed-phase chromatography, respectively.

Prior to the HPLC separation of these pharmaceuticals, silica gel thin-layer adsorption chromatography of steroidal hormones and reversed-phase thin-layer partition chromatography of cephalosporin antibiotics with chemically bonded dimethylsilyl silica gel were performed in order to obtain suitable HPLC separation systems.

In the separation of steroidal hormones, the same binary mobile phase ratios of TLC did not give satisfactory results in HPLC. For the sharp separation in HPLC, solvent strength in the binary solvent mixture used for TLC had to be decreased.

The difference in solvent strength for efficient separation between TLC and HPLC might be attributed to the fact that in HPLC the solvent elution power acts in an isocratic manner while in TLC it acts in a gradient manner.

On the other hand, a correlation of mobility between TLC and HPLC separation for cephalosporin antibiotics was obtained, and the possibility of direct transfer of chromatographic systems from TLC to HPLC for separation of these antibiotics was confirmed.     

Chromatography of Trichothecene Mycotoxins

Abstract

Trichothecene mycotoxins occur in agricultural commodities and can cause problems from feed refusal to death in animals. This paper describes chromatographic methods for selective analysis for trichothecene mycotoxins. These methods include gas chromatography (GC), thin layer chromatography (TLC), and high pressure liquid chromatography (HPLC). The trichothecene analysis methods by GC and TLC are shown to have a greater sensitivity than in HPLC for the underivatized mycotoxins.     

Review of Sub-Microliter (Nanoliter) Injection Techniques in Liquid Chromatography

Abstract

In liquid chromatography with “low-dispersion methods”, there is an increasing need to reproducibly inject nanoliter sample volumes. Low-dispersion methods produce very narrow peaks because of short column length, narrow column bore, small particle packing, low particle surface area, open tubular configuration, or combinations of these parameters.

This paper reviews methods used for sub-microliter injections for use with these low-dispersion liquid chromatography methods. Some nanoliter injection methods for open-tubular electrophoresis are also described.     

Multiresidue Analysis of Some Insect Growth Regulators by Reversed-Phase High-Performance Liquid Chromatography

Abstract

A general procedure has been developed for the analysis of 8 different insect growth regulators (IGRs) by using reversed-phase high-performance liquid chromatography with gradient solvent systems. The method has been used to identify and separate 8 insect growth regulators from a mixture of the standards. The method has been evaluated with different column conditions and under different solvent systems. Best resolution was obtained by using a double column and methanol/water gradient system.     

Structural and Quantitative Analyses of 4-Chloroaniline-Derived Oligomers

Abstract

The oxidation of 4-chloroaniline by a peroxidase resulted in eight oligomeric products. A reverse-phase high performance liquid chromatography (HPLC) method was developed so that substrate disappearance and the corresponding product formations could be quantitatively monitored. The product mixture was isolated from the aqueous reaction solution with solid-phase extraction and the extract components were separated by thin-layer chromatography (TLC). The individual TLC bands were extracted for mass spectrometric and proton nuclear magnetic resonance analyses. The product mixture was found to contain dimers, trimers, and tetramers with benzoquinone monoimine, benzopuinone di-mine, diaminobenzene, and azobenzene structures. Analytical methodologies were specific for the study of the oxidative transformation of 4-chloroaniline, but they should be applicable to other anilinederived oligomers.     

Separation and identification of multicomponent mixture by thin-layer chromatography coupled with Fourier transform-infrared microscopy

A fast and convenient method based on coupled thin-layer chromatography (TLC) with Fourier transform infrared (FTIR) microscopy has been established for separation and identification of multicomponent mixtures. In this study, the method was developed and consummated with more perfect TLC spots transferral process and consistent FTIR testing conditions. A newly developed technique, solid-phase extract (SPE) was introduced for sample pre-treatment instead of using traditional column chromatography. It is a new field for SPE that has already been widely applied in many other fields. It not only overcomes the backwards (low separation efficiency, time consuming and solvent consumption) of column chromatography but also makes it much easier to choose an optimum TLC sheet and to set suitable TLC loading. With all the above-mentioned modifications and supplements, the analytical method of coupled TLC with FTIR microscopy for separation and identification of multicomponent mixtures becomes more convenient and more efficient. In addition, a very complex sample (a die-cast release agent) was used as an example to demonstrate the technique.     

Simultaneous isolation of seven compounds from Glehnia littoralis roots by off‐line overpressured layer chromatography guided by a TLC antioxidant autographic assay

A combinative method using overpressured layer chromatography (OPLC) and TLC bioautography against O₂^?? was developed to separate antioxidants from Glehnia littoralis roots. Through target‐directed isolation by the TLC bioautographic method, seven compounds including five antioxidants were rapidly isolated by OPLC and identified as 1‐linoloyl‐3‐palmitoylglycerol, facarindiol, panaxynol, isoimperatorin, β‐sitosterol, scopoletin, and umbelliferone from G. littoralis roots. This OPLC method is a very suitable separation technique for light‐sensitive polyacetylenes (panaxynol and facarindiol) with higher isolated yields compared to conventional open column chromatography. This is the first report on the separation of polyacetylenes by OPLC.