Enantiomeric resolution of new triazole compounds by high-performance liquid chromatography

Cellulose tris(3,5-dimethylphenylcarbamate) (CDMPC) was synthesized and coated on aminopropylsilica to prepare a chiral stationary phase (CSP). HPLC methods were developed for the direct enantioseparation of 12 chiral triazole compounds on the CSP. The separations were made using normal phase methodology with a mobile phase consisting of n-hexane-alcohol (ethanol, 1-propanol, 1-butanol, 2-propanol, and t-butanol) in various portions. The column temperatures were studied for the optimization of the resolutions. The effects of structural features of the solutes on the discrimination between the enantiomers were examined. Baseline separation was easily obtained in many cases.     

Enantiomeric resolution of a series of chiral sulfoxides by high-performance liquid chromatography on polysaccharide-based columns with multimodal elution

The enantiomeric resolution of a series of 20 asymmetric sulfoxides was systematically investigated by HPLC using multimodal elution with amylose trisR(S)-1-phenylethylcarbamate], amylose tris(3,5-dimethoxyphenylcarbamate) and amylose and cellulose tris(3,5-dimethylphenylcarbamate) phases. The sulfoxide series was composed of aromatic, olefinic and ketosulfoxides, sulfinyl acids and esters. This work has shown that enantioselectivity and enantioresolution of the polysaccharide-based columns can be achieved by changing the type and composition of the mobile phase, widening the applicability of these chiral phases.     

Optical resolution of asymmetric triacylglycerols by chiral-phase high-performance liquid chromatography

A simple method for direct optical resolution of some asymmetric triacylglycerols (TGs) has been established. The method employs chiral-phase high-performance liquid chromatography (HPLC). An enantiomeric pair of TGs comprising 1-eicosapentaenoyl-2,3-dicapryroyl-sn-glycerol (ECC) and 1,2-dicapryroyl-3-eicosapentaenoyl-sn-glycerol (CCE) was resolved on a CHIRALCEL OF or on a CHIRALCEL OD column. The separation of another pair of asymmetric TGs, 1-docosahexaenoyl-2,3-dicapryroyl-sn-glycerol (DCC) and 1,2-dicapryroyl-3-docosahexaenoyl-sn-glycerol (CCD), was achieved with the CHIRALCEL OD column. The chiral-phase HPLC method in combination with silver-ion HPLC and high-temperature gas chromatography was used for monitoring two interesterification reactions, whose products were chiral TGs. Interesterification of tricapryloylglycerol with ethyleicosapentaenoate or with ethyldocosahexaenoate was performed using Rhizomucor miehei lipase as the catalyst. The products targeted were the asymmetric pair of TGs, ECC and CCE or DCC and CCD. The amounts of sn-1-substituted products (ECC or DCC) were greater than their sn-3-substituted counterparts (CCE or CCD) throughout the reaction period, suggesting that R. miehei lipase had a stereopreference towards the sn-1 position over the sn-3 position.     

Enantiomeric separation of asymmetric triacylglycerol by recycle high-performance liquid chromatography with chiral column

In our previous studies, we employed recycle HPLC for the separation of triacylglycerol (TAG)-positional isomers (PIs). In this study, a recycle HPLC system equipped with a polysaccharide-based chiral column was applied to the enantiomeric separation of some asymmetric TAGs having straight-chain C16-C18 acyl residues. As a result, 1,2-dipalmitoyl-3-oleoyl-rac-glycerol (rac-PPO), 1,2-dioleoyl-3-palmitoyl-rac-glycerol (rac-OOP), and 1,2-dipalmitoyl-3-linoleoyl-rac-glycerol (rac-PPL) were resolved into their respective enantiomers. However, neither 1,2-dioleoyl-3-linoleoyl-rac-glycerol (rac-OOL), consisting of only unsaturated fatty acids, nor 1,2-dipalmitoyl-3-stearoyl-rac-glycerol (rac-PPS), consisting of only saturated fatty acids, was resolved. These results suggest that the asymmetric TAGs, used in this study, having both a palmitic acid moiety and an oleic acid (or a linoleic acid) moiety at the sn-1 or sn-3 positions are resolved by the chiral column. This new chiral separation method can be used in combination with atmospheric pressure chemical ionization mass spectrometry to determine the sn-OOP/sn-POO ratio in palm oil. This method is applicable for the chiral separation of asymmetric TAGs in palm oil.     

Indirect detection in high-performance liquid chromatography

Indirect detection is a technically simple method to follow and quantify compounds without inherent detector response in high-performance liquid chromatography. The underlying principle can be expressed in simple equations. These equations indicate clearly how the detection sensitivity can be optimized and how disturbance effects can be avoided when the mobile phase gives detector response.     

Enantiomeric resolution of bifonazole by supercritical fluid chromatography

The enantiomeric separation of bifonazole by supercritical fluid chromatography on Chiralpak AD has been studied. The effect of different modifiers (methanol, ethanol, 2-propanol, and acetonitrile) was examined. Enantioseparation was possible with all of them, but the best results were provided by the alcohol-type ones. The resolution was higher than 5 in all cases. The isoelution temperatures Tiso were calculated from the study of the temperature effect for the different organic modifiers. The value of Tiso was below the working temperature range on using methanol, but above it with ethanol or 2-propanol.     

Enantiomeric resolution of new aromatase inhibitors by liquid chromatography on cellulose chiral stationary phases

Analytical HPLC methods using derivatized cellulose chiral stationary phases were developed for the direct enantioseparation of substituted [1-(imidazo-1-yl)-1-phenylmethyl)]-benzothiazolinone and benzoxazolinone derivatives with one chiral center. Those analogues of fadrozole constitute new potent nonsteroidal inhibitors of aromatase (P450 arom). The separations were made using normal phase methodology with a mobile phase consisting of n-hexane-alcohol (ethanol, 1-propanol, or 2-propanol) in various proportions, and a silica-based cellulose tris-3,5-dimethylphenylcarbamate (Chiralcel OD-H), or tris-methylbenzoate (Chiralcel OJ). The effects of concentration of various aliphatic alcohols in the mobile phase were studied. A better separation was achieved on cellulose carbamate phase compared with the cellulose ester phase. The effects of structural features of the solutes along with the temperature of the column on the discrimination between the enantiomers were examined. Baseline separation (Rs > 1.5) was easily obtained in many cases.     

Preparation and rapid resolution of Xenopus phosvitins and phosvettes by high-performance liquid chromatography

The presence of acidic phosivitins phosvettes in Xenopus laevis yolk platelets and their purification by (NH4)2SO4 precipitation of associated lipovitellin were documented by polyacrylamide gel electrophoresis followed by staining with Stains-all. Procedures were further developed to resolve the various entities present in the crude phosvitin/phosvette fraction by size-exclusion, anion-exchange, and hydrophobic interaction chromatography, using a Pharmacia FPLC system, and their resolution was documented by both electrophoresis and two-dimensional chromatography. Four major entities (phosvitins 1 and 2; phosvettes 1 and 2) were observed, but microheterogeneity was also apparent, particularly by hydrophobic interaction chromatography. The new separation procedures require min/h rather than h days.     

Measurement of beta-carbolines by high-performance liquid chromatography with fluorescence detection

A method using high-performance liquid chromatography with fluorescence detection was developed for the determination of beta-carboline compounds norharman, harman, norharmol, and harmol in lung. Aqueous derivatization with acetic anhydride was used to facilitate the isolation and separation of the phenolic compounds and to reduce the fluorescence background of the biological samples. Harman was identified and quantitated in rat lung (1.88 +/- 0.55 ng/g) using this method and its identity confirmed by means of gas chromatography-negative-ion chemical ionization mass spectrometry.     

Determination of polyamines by high-performance liquid chromatography with chemiluminescence detection

A method was developed for the determination of polyamines (PA) by high-performance liquid chromatography with chemiluminescence detection. It is based on the unsaturated complex of PA with Cu(II) which had a strong catalytic effect on the luminol-H₂O₂ chemiluminescence reaction. The separation of PA was carried out on a reveres phase C18 column using methanol/water (25/75, v/v) as a mobile phase. The method was applied to the analysis of putrescine and the total amount of spermine and spermidine in apple leaves and strawberry fruit. The results indicated that the method is practical and useful.     

Separation of pesticides by high-performance liquid chromatography with amperometric detection

The possibility of the amperometric detection of a number of pesticides, such as benomyl, thiram, linuron, metoxuron, desmedipham, dicuron, lenacil, and fludioxonil, widely used in agrochemical practice was studied. The effect of the working electrode material (glassy carbon, nickel, and gold) and the type of the electrochemical cell on the value of the analytical signal was studied using the example of thiram. It was found that the optimum potential of the working electrode in analyzing a pesticide mixture was 1400 mV. The dependence of the analytical signal on the pesticide concentration was shown to be linear. The detection limits for the analytes were calculated. Using a 100-μL sample loop, all of the studied pesticides can be determined at the level of the maximum permissible concentration (MPC). The amperometric determination of seven pesticides at the level of MPC in real samples was shown by the examples of model mixtures dissolved in tap water and beetroot juice.     

Gunshot residue determination by high-performance liquid chromatography with electrochemical detection

A procedure for the detection of gunshot residue via the organic constituent diphenylamine is described. The method incorporates high-performance liquid chromatography with electrochemical detection.     

Computer-aided multichannel detection in high-performance liquid chromatography

Summary The development of a computer-aided rapid-scanning detector for HPLC based on the linear photodiode array UV-visible spectrophotometer is described. Chomatograms monitored at up to three wavelengths, with automatic capture of spectra for eluted peaks and post-run processing of spectral data to generate log₁₀ (A) spectra, second derivative and fourth derivative spectra, are described. Methods are reported for the analysis of forensic samples of diacetylmorphine (heroin) in the presence of the degradation products and potential contaminants caffeine, papaverine, 6-acetylcodeine, thebaine, 6-acetyl-morphine, procaine and morphine separated by HPLC. The novel use of second and higher derivative spectra for the qualitative characterisation of drugs and contaminants separated by HPLC is described.Presented at the 14th International Symposium on Chromatography London, September, 1982