Gas Chromatographic Method for the Analysis of Organic Acids in the Bio-Catalytic Conversion Process.

An analytical method for the quantification of acrylic acid (AA), 1,3-propanediol (1,3-PD) and 3-hydroxypropionic acid (3-HP) in the bio-catalytic conversion process has been developed by gas chromatography. A simple liquid-liquid extraction (LLE) procedure was used in the sample preparation. Organic acid additives such as trifluoroacetic acid were used to improve the extraction efficiency in the LLE procedure. Under optimum analysis conditions, all analytes were satisfactorily separated with no interference. In standard calibration, all correlation coefficients (r(2)) were better than or equal to 0.994. In culture media, the intra-batch precision (% relative standard deviation) and recovery (%) as the average value of the quality control samples were 2.3 and 102.4%, respectively. In addition, the inter-batch precision and recovery as the average value of the quality control samples were 5.0 and 104.0%, respectively. In phosphate buffer, the intra-batch precision and recovery as the average value of the quality control samples were 2.7 and 101.6%, respectively. In addition, the inter-batch precision and recovery as the average value of the quality control samples were 2.9 and 101.7%, respectively. The limit of detection (S/N ratio: 3) and limit of quantification (S/N ratio: 10) were 1.0 and 3.5 µg/mL, 3.0 and 10.0 µg/mL, and 9.0 and 30.0 µg/mL, respectively, for AA, 1,3-PD and 3-HP. Consequently, this method was demonstrated to be acceptable for the quantitative analysis of AA, 1,3-PD and 3-HP in culture media and phosphate buffer.

Gas chromatographic method for the determination of residual monomers, 2-(acryloyloxy)ethyl isocyanate and 2-(methacryloyloxy)ethyl isocyanate, as curing agents in an ultraviolet curable adhesive.

A gas chromatographic method is described for the determination of residual 2-(acryloyloxy)ethyl isocyanate (AOI) and 2-(methacryloyloxy)ethyl isocyanate (MOI) as curing agents in an ultraviolet curable adhesive. Pre-column derivatization was employed in the determination of AOI and MOI as a means of enhancing the response of the flame ionization detector. Urethane derivatives of AOI and MOI were derived using methanol for 30 min at room temperature. The accuracies (n = 5, three concentration levels) were in the range of 113.4 to 126.7%, and precisions (n = 5, three concentration levels) were in the range of 0.8 to 4.3% for AOI-OMe. Furthermore, the accuracies were in the range of 79.5 to 108.6% and the precisions were in the range of 1.0 to 2.4% for MOI-OMe. The correlation coefficients of six calibration standards were all greater than 0.9999 for AOI-OMe and greater than 0.9998 for MOI-OMe over the range from 10 to 100 µg/mL.

High-performance liquid chromatographic method of photoactive compounds based on diazonaphthoquinone for positive photoresists.

An analytical method for the compositional and quantitative analysis of photoactive compounds (PACs) in positive photoresist (Posi PR) has been developed by high-performance liquid chromatography (HPLC). Under optimum HPLC conditions, various types of PACs consisting of a mixture of isomers were satisfactorily separated with no interference. This method was applied to the quantitative analysis of PACs in Posi PR. All correlation coefficients were better than or equal to 0.998. The precision and accuracy showed no significant deviation and were measured with acceptable values. The intra-batch precision and accuracy (%) of quality control samples ranged from 0.80 to 1.46% and from 101.7 to 102.8%. Consequently, the method was demonstrated to be acceptable for the analysis of PACs in Posi PR. We believe that the HPLC method developed in this work can be used for the compositional and quantitative analysis of PACs in Posi PR.

Chiral recognition of N-phthaloyl, N-tetrachlorophthaloyl, and N-naphthaloyl α-amino acids and their esters on polysaccharide-derived chiral stationary phases.

Enantiomeric separations of N-phthaloyl (N-PHT), N-tetrachlorophthaloyl (N-TCPHT), and N-naphthaloyl (N-NPHT) α-amino acids and their esters were examined on several kinds of polysaccharide-derived chiral stationary phases (CSPs). Resolution capability of CSPs was greater Chiralcel OF than the others for N-PHT and N-NPHT α-amino acids and their esters. In N-TCPHT α-amino acids and their esters, good enantioselectivities showed Chiralcel OG for N-TCPHT α-amino acids, Chiralpak AD for N-TCPHT α-amino acid methyl esters, and Chiralcel OD for N-TCPHT α-amino acid ethyl esters, respectively. From the results of liquid chromatography and computational chemistry, it is concluded that l-form is preferred and more retained with electrostatic interaction in case of interaction between N-PHT α-amino acid derivatives and Chiralcel OF, N-TCPHT α-amino acid derivatives and Chiralcel OD, and N-NPHT α-amino acid derivatives and Chiracel OF. On the other hand, d-form is preferred and more retained with van der Waals interaction in case of interaction between N-TCPHT α-amino acid ester derivatives and Chiralcel OG and Chiralpak AD.

LC-MS/MS method for the quantification of myo- and chiro-inositol as the urinary biomarkers of insulin resistance in human urine.

A simple LC-MS/MS method has been developed and validated for the quantification of endogenous myo- and chiro-inositol in human urine. myo- and chiro-Inositol were completely resolved from other carbohydrates and there were no interference peaks in human urine. The correlation coefficient (n = 3) was greater than 0.9991 over the range 0.05-25.0 µg/mL with the weighted (1/C²) least square method. Precision (%RSD) and accuracy (%RE) were 0-10.0% and 0-6.0% for the intra-day assay (n = 5) and 0-14.3% and 0-10.0% for the inter-day assay (n = 5). myo- and chiro-Inositol have been shown to be stable in human urine stored at room temperature and for three freeze-thaw cycles.

Analysis of polymer additives in high-temperature liquid chromatography.

A high-temperature liquid chromatographic technique is employed for the separation of commercially available polymer additives to enhance the resolution and speed. Separation efficiencies and elution behaviors for seven phthalate plasticizers and five antioxidants are evaluated at elevated column temperatures and with a thermal gradient. Diamondbond C18 (octadecylsilica), Zirchrom PS (zirconia-based polystyrene), and Zirchrom PBD (zirconia-based polybutadiene) columns are selected for the study because of their thermal stability. The temperature programming is controlled with a column oven in conjunction with an independent mobile phase preheater and a post-column effluent cooling assembly. Van't Hoff plots show that the reverse-phase liquid chromatography mechanism is maintained over a wide range of column temperatures. A 1% increase of acetonitrile in the mobile phase is estimated to have a comparable effect as a 7-7.5 degrees C column temperature increase on the retention time changes.

Liquid chromatographic enantiomer separation of racemic amine using chiral crown ether stationary phase.

Direct enantioseparation of racemic amine, amino-thiophene-2-yl-acetonitrile (TAN), on chiral crown ether stationary phase [Crownpak CR (+)] is described in this study. The elution behavior and the effect of acid additives on resolution of racemic amine, TAN, is intensely investigated. Moreover, the chiral recognition mechanism in this specific system is proposed based on computational methods with the density functional theory. Diastereomeric complexation of the ammonium ion of racemic amine inside the cavity of chiral crown ether appears essential for the chiral discrimination. The pH of the mobile phase containing acid additives also acts as an important factor for the chiral recognition.

Enantiomer separation of a novel aminothiazolecarboxamide fungicide using polysaccharide-derived chiral stationary phases.

The direct enantioseparation of a novel aminothiazolecarboxamide fungicide, ethaboxam, on polysaccharide-derived chiral stationary phases (CSPs) is described. Good resolution is achieved with several polysaccharide-derived CSPs. Chiralcel OD (OD-H) and Chiralpak AS are excellent for direct enantiomer separation of ethaboxam. The elution behavior and the effects of eluent composition on the resolution of ethaboxam are also investigated. Furthermore, the mechanism for chiral recognition using molecular mechanics is discussed.

Liquid chromatographic determination of novel aminithiazolecarboxamide fungicide residues in soil and crops using online solid-phase extraction.

A simple and convenient liquid chromatographic method has been developed and applied to the analysis of the novel aminothiazolecarboxamide fungicide, ethaboxam, in soil and crops. After the isolation and concentration of analyte from soil and crops, clean-up and separation of sample solutions are performed by high-performance liquid chromatography with online solid-phase extraction. Good linearity (r2 > 0.9995), recovery [for soil, 95.3-98.4%, and crops (grape, red pepper), 92.9-95.9%], and repeatability are achieved in the calibration range of 0.1-10.3 microg/mL. The limit of detection is the 2.5 parts per billion (ppb) (40 g of soil) and the 20 ppb (25 g of crops), respectively. This assay method shows the suitability for the residual analysis of ethaboxam in soil and crops.

Determination of melagatran in rabbit plasma by high-performance liquid chromatography with automated column switching.

Melagatran is an active thrombin inhibitor showing oral and parenteral bioavailability for antithrombotic therapy. A simple and convenient liquid chromatographic method has been developed and applied to the analysis of melagatran in rabbit plasma. The clean-up and separation of the sample solutions were performed by automated on-line column switching HPLC. The method validation shows the suitability of the column switching liquid chromatographic system for the quantitation of melagatran in biological fluids.

Reversed-phase liquid chromatographic method for the analysis of aminoglycoside antibiotics using pre-column derivatization with phenylisocyanate.

A pre-column derivatization liquid chromatographic method has been developed for the analysis of aminoglycoside antibiotics using phenylisocyanate as a derivatization reagent. Derivatives including kanamycin, neomycin and gentamicin were formed by reaction of the analytes with phenylisocyanate in the presence of triethylamine. Phenylisocyanato groups were attached to corresponding amino groups of aminoglycoside and their molecular mass was confirmed by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). The experimental conditions for derivatization and separation of aminoglycoside derivatives were optimized and validated. A simple liquid chromatographic method for the determination of aminoglycoside antibiotics was demonstrated.

Enantiomeric discrimination of pyrethroic acid esters on polysaccharide derived chiral stationary phases.

Enantiomeric separation of pyrethroic acid methyl and ethyl esters was examined on cellulose-based chiral stationary phases (CSPs): chiralcel OD (cellulose tris(3,5-dimethylphenyl carbamate)) and chiralcel OF (cellulose tris(4-chlorophenyl carbamate)). The good resolution of pyrethroic acid esters was achieved on chiralcel OD and OF. Separation factors ranged from 1.19-5.12 for Chiralcel OD and 1.00-1.59 for chiralcel OF. Hexane/2-propanol (100:0.15, v/v %) was used as the eluent. The resolution capability of CSPs was greater chiralcel OD than chiralcel OF in the case of the pyrethroic acid esters. The flow rate was 0.8 ml/min and detection was set at 230 nm. The results of the chromatographic data and molecular mechanics suggest that steric effect was a major factor in the enantioseparation. Furthermore, the hydrogen bond between analytes and CSP played an important role in the chiral recognition.

Comparison of reversed-phase liquid chromatographic methods for the separation of new quinolones.

Reversed-phase high-performance liquid chromatography is widely used in the analysis of drug substances and their metabolites. The interaction of quinolones with residual silanol in a silica-based C18 stationary phase causes peak broadening and bad peak shapes and makes it hard to resolve the peak separations. This unusual interaction is studied and finally can be removed by masking the residual silanol of a silica-based C18 stationary phase, then good peak separation is achieved. We have chosen four new quinolones and ciprofloxacin and improved the peak shapes by optimizing the pH of the eluent and the quantity of the additive (N,N-dimethyloctylamine, approximately 0-40 mM) in the monomeric C18 stationary phase. The elution behavior of quinolones in the polymeric C18 stationary phase is compared with that in the monomeric C18 stationary phase under the same eluent condition. Good peak symmetry and a high plate number are achieved by this technique, which are hardly obtained with the conventional silica-based C18 stationary phase. Based on these results, we present data of the influence of the eluent composition such as pH, buffer, and additive concentration on the peak shape.