• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1995.04a
• /
• pp.75-75
• /
• 1995

Succinic semialdehyde reductase, one of key enzyme of GABA shunt in CNS, is inactivated by o-phthalaldehyde, The inactivation followed pseudo first-order kinetics, and the second-order rate constant for the inactivation process was 28 M$\^$-1/s$\^$-1/ at pH 7.4 and 25$^{\circ}C$. The absorption spectrum(λ$\_$max/=377nm), fluorescence exitation(λ$\_$max/=340nm) and fluorescence emission spectra (λ$\_$max/=409nm) were consistent with the formation of an isoindole derivative in the catalytic site between a cysteine and a lysine residues about 3${\AA}$ apart. The substrate, succinic semialdehyde, did not protect the enzymatic activity against inactivation, whereas the coenzyme, NADPH, protected against o-phthalaldehyde induced inactivation of the enzyme. About 1 isoindole group per moi of the enzyme was formed following complete loss of the enzymatic activity. These results suggest that the amino acid residues of the enzyme participating in reaction with o-phthalaldehyde more likely residues at or near the coenzyme binding site.

• Bulletin of the Korean Chemical Society
• /
• v.31 no.7
• /
• pp.1973-1975
• /
• 2010

The electronic and adsorption structure of o-phthalaldehyde (OPA) on the H-Si(100) surface was investigated by using Near Edge X-ray Fine Structure (NEXAFS) and high resolution photoemission spectroscopy (HRPES). We confirmed that the OPA grown on the H-Si(100) surface showed good dependency with about 60 degree tilting angle using NEXAFS and a single O 1s peak by using HRPES. Hydrogen atom passivated on the Si(100) surface was found to be a seed for making one dimensional organic line that uses a chain reaction as the H-Si(100) surface was compared with the hydrogen free Si(100) surface.

• BMB Reports
• /
• v.28 no.2
• /
• pp.112-117
• /
• 1995

Succinic semialdehyde reductase was inactivated by o-phthalaldehyde. The inactivation followed pseudo-first order kinetics, and the second-order rate constant for the inactivation process was 28 $M^{-1}s^{-1}$ at pH 7.4 and $25^{\circ}C$. The absorption spectrum ($\lambda_{max}$ 337 nm) and fluorescence excitation ($\lambda_{max}$ 340 nm) and fluorescence emission spectra ($\lambda_{max}$ 409 nm) were consistent with the formation of an isoindole derivative in the catalytic site between a cysteine and a lysine residue approximately about 3 $\AA$ apart. The substrate, succinic semialdehyde, did not protect enzymatic activity against inactivation, whereas the coenzyme NADPH protected against o-phthaladehyde induced inactivation of the enzyme. About 1 isoindole group per mol of the enzyme was formed following complete loss of enzymatic activity. These results suggest that the amino acid residues of the enzyme participating in a reaction with o-phthalaldehyde are cysteinyl and lysyl residues at or near the NADPH binding site.

• Analytical Science and Technology
• /
• v.36 no.5
• /
• pp.250-257
• /
• 2023

The development of a spectrofluorimetric method for the determination of trimethoprim according to the reaction between O-phthalaldehyde (OPA) in highly alkaline media, and 2-mercaptoethanol (2ME) and this reaction gives fluorescent product measured at (458) nm when excited at (342) nm. The optimization of the analytical parameters that influence intensity was investigated. The intensity of fluorescence of the formed product was linearly related to the concentration of trimethoprim in the (100-1200) ng mL^-1 range. The limit of detection and limit of quantification were estimated to be (22.54) ng mL^-1 and (75.15) ng mL^-1 respectively. The utility of the proposed methods was successfully verified by analysis of trimethoprim in pure and real pharmaceutical preparations with high accuracy, the recovery percentages Re%, were found to be (100.5) % and (99.76) % for pure drug and pharmaceutical preparations respectively.

• Proceedings of the PSK Conference
• /
• 2001.10a
• /
• pp.287.1-287.1
• /
• 2001

• Analytical Science and Technology
• /
• v.8 no.3
• /
• pp.229-236
• /
• 1995

Multiple-step gradient method was used for the analysis of free amino acids in physiological fluids by high-performance liquid chromatography with diode array detector on the Amino Quant $C_{18}$ column under the condition of pH 7.2 of buffer solutions. Plasma samples of normal Korean people and abnormal Korean people who have schizophrenia were subjected to derivatization with o-phthalaldehyde in the presence of 3-mercaptopropionic acid. Quantitative analysis of amino acids in physiological fluids by internal standard method gave highly reproducible results within a relative standard deviation of less than 2~6%. And amino acids amounts of physiological fluids of Korean people gave some different results from those of foreigners. There was large differences in tyrosine amount between normal and abnormal man.

• Analytical Science and Technology
• /
• v.9 no.1
• /
• pp.52-61
• /
• 1996

Detectable levels of several free D-amino acids were found in some food. This was accomplished by using a column switching method. The determination of total amount of D- and L-amino acids was based on an achiral separation with a $C_{18}$ column. The level of D-amino acids to L-amino acids was determined by the column switching system including the postcolumn reaction detection of the amino acids derivatized with O-phthalaldehyde. The chiral separation of the postcolumn detection system was carried out with chiral crown ether column. This system was applied for the micro-determination of D-amino acids in food such as soy sauce, fermented soy bean and beans. It turned out that the sampling process is critical for the trace analysis of D-amino acids under this achiral / chiral coupled-column system. It was found that commercial soy sauce contained 42ppm, conventional soy sauce 102ppm, fermented soy bean 8.34mg per 1g and bean 2.87mg per 1g sample for phenylalanine. D-phenylalanine was found 0.67% in commercial soy sauce, 0.34% in conventional soy sauce, less than 1.81% in fermented soy bean, and Jess than 2.82% in bean.

• Applied Biological Chemistry
• /
• v.39 no.1
• /
• pp.84-88
• /
• 1996

To determine brassinolide in rice(Oriza sativa L.) using HPLC equipped with fluoresence detector, a highly sensitive fluorescence reagent. 1-cyanoisoindole-2-m-phenylboronic acid, was synthesized from the reaction of o-phthaldehyde, m-phenylboronic acid and KCN, then was reacted with brassinolide. The formation ratio of brassinolide boronate exhibited 90% up at the ratio of $20\;:\;1({\mu}g/{\mu}g)$ of 1-cyanoisoindole-2-m-phenylboronic acid and brassinolide respectively. The detection limit of brassinolide boronate with fluoresence detector was 0.16 ng. Brassinolide was detected in heading stage(biomass : 10 g) and panicle formation stage(biomass : 100 g) of the rice(Oryza sativa L.) with quantity of $0.8\;{\mu}g\;and\;0.2\;{\mu}g$respectively. However, brassinolide was not detected in blooming and elongation stage.

• Journal of the Korean Chemical Society
• /
• v.29 no.5
• /
• pp.543-551
• /
• 1985

A new class of polyhydrazones has been synthesized by the solution polycondensation from equimolecular amounts of aromatic dialdehydes such as para, meta, ortho-phthal aldehyde, 5,5'-methylene-bis-salicyl aldehyde (PPTA, MPTA, OPTA, MBSA) and dihydrazides, 5,5'-methylene-bis-salicylic dihydrazide (MBSDH), terephthalic dihydrazide (TDH), sebacic dihydrazide (SDH) in DMF-$CH_3COOH$ solution. The solubility characteristics, spectral, and thermal properties of the synthesized polyhydrazones and their metal chelates were also studied. These polyhydrazones and their metal chelates except the polyhydrazone prepared from OPTA-MBSDH were generally insoluble in common organic solvents. The thermogravimetric analysis of polyhydrazones showed 10% weight losses at 250∼350$^{\circ}$C and residual weight at 500$^{\circ}$C were 32.5∼62.5%. The decomposition temperature of higher relatively, and the metal chelates decrease in the following orders; Zn(II)-IIa > Ni(II)-IIa > Co(II)-IIa > Cu(II)-IIa.

• Molecular & Cellular Toxicology
• /
• v.3 no.4
• /
• pp.273-281
• /
• 2007

Ceramide is involved in cell death as a lipid mediator of stress responses. In this study, we developed an improved method of ceramide quantification based on added synthetic ceramide and thin layer chromatography (TLC) separation, and applied to biological samples. Lipids were extracted from samples spiked with N-oleoyl-D-erythro-sphingosine ($C_{17}$ ceramide) as an internal standard. Ceramide was resolved by TLC, complexed with fatty-acidfree bovine serum albumin (BSA), and deacylated by ceramidase (CDase). The released sphingosine was derivatized with o-phthalaldehyde (OPA) and measured by high performance liquid chromatography (HPLC). The limit of detection for ceramide was about 1-2 pmol and the lower limit of quantification was 5 pmol. Ceramide recovery was approximately 86-93%. Ceramide concentrations were determined in biological samples including cultured cells, mouse tissues, and mouse and human plasma. TLC separation of ceramide provides HPLC chromatogram with a clean background without any interfering peaks and the enhanced solubility of ceramide by BSAceramide complex leads to the increased deacylation of ceramide. The use of an internal standard for the determination of ceramide concentration in these samples provides an accurate and reproducible analytical method, and this method can be applicable to diverse biological samples.