• Analytical Science and Technology
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• v.36 no.6
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• pp.332-339
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• 2023

This study aimed to verify the hypothesis that urea denatures hemoglobin in the blood, thereby exposing active sites of enzymes and enhancing the chemiluminescence of the blood-luminol reaction. When blood was pretreated with urea, higher concentrations of pretreatment urea or longer pretreatment times resulted in enhanced chemiluminescence in the blood-luminol reaction, supporting the above hypothesis. However, the chemiluminescence was enhanced when blood was treated with luminol mixed with an 8 M urea solution, although the fact that the time for urea to denature hemoglobin was shorter compared to when blood was pretreated with urea and followed by luminol. In addition, the chemiluminescence was enhanced when a transition metal without hemoglobin was reacted with urea-containing luminol. Based on these results, it is anticipated that urea not only denatures hemoglobin but also plays a role in the luminol-hydrogen peroxide reaction.

• Analytical Science and Technology
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• v.35 no.6
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• pp.242-248
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• 2022

The effect of ethylenediamine (EDA) on the reaction of luminol or Bluestar with blood and bleaches was studied. For this purpose, blood, chlorine bleach, and oxygen bleach were applied to filter paper, treated with EDA-containing luminol or Bluestar, and the changes in chemiluminescence intensity were observed. It was found that the chemiluminescence intensity of the luminol (or Bluestar)-blood reaction did not change with the increasing concentration of EDA. However, the chemiluminescence intensity of the luminol (or Bluestar)-chlorine bleach reaction decreased and the chemiluminescence intensity of the luminol (or Bluestar)-oxygen bleach reaction increased, with increasing EDA concentration. Thus, it was found that when EDA was added to luminol (or Bluestar), which is a blood-sensitive reagent, EDA suppressed the false-positive reaction induced by chlorine bleach and induced a false-positive reaction with oxygen bleach. Consequently, the addition of EDA is not recommended for enhancing bloodstains with luminol or Bluestar.

• Analytical Science and Technology
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• v.31 no.1
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• pp.47-56
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• 2018

Finding the blood left at a crime scene is very important to reconstruct or solve a criminal case. Although numerous reagents have been developed for use at crime scenes, luminol is the most representative. Bluestar Forensic has been used in recent years, but is expensive and cannot be stored after preparation. This study aims to develop a new luminol reagent that can be stored for a long period of time while maintaining the chemiluminescence intensity at the level of Bluestar Forensic. Because luminol dissolves well in aqueous alkaline solutions, the use of sodium hydroxide in the preparation of luminol reagents can promote the decomposition of hydrogen peroxide. Magnesium sulfate, sodium silicate, and potassium triphosphate have been used as hydrogen peroxide stabilizers. The effects of the addition of these substances on the chemiluminescence emission intensity and the storage period of the luminol reagents were confirmed. The addition of a hydrogen peroxide stabilizer was shown to have no significant affect on the chemiluminescence emissions intensity or stabilized pH of the luminol reagent during storage. It also greatly increases the shelf life of the reagents. The use of magnesium sulfate as a hydrogen peroxide stabilizer is the most appropriate. When sodium perborate is used instead of hydrogen peroxide as an oxidizing agent, there is no significant change in the sensitivity and chemiluminescence emissions intensity, but the storage period is shortened. However, after the reaction with blood, the pH of the mixed solution does not increase significantly, and is judged to be more suitable than a reagent made of hydrogen peroxide.

• ALGAE
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• v.28 no.3
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• pp.289-296
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• 2013

For the quantitative detection of algal toxin, microcystin, a chemiluminescence immunochromatographic assay method was developed. The developed system consists of four parts, chemiluminescence assay strip (nitrocellulose membrane), horse radish peroxidase labeled microcystin monoclonal antibodies, chemiluminescence substrate (luminol and hydrogen peroxide), and luminometer. The performance of the chemiluminescence immunochromatographic assay system was compared with high performance liquid chromatography (HPLC) detection. The detection limit of chemiluminescence immunochromatographic assay system is several orders of magnitude lower than with HPLC. The chemiluminescence immunochromatography and HPLC results correlated very well with the correlation coefficient ($r^2$) of 0.979.

• Bulletin of the Korean Chemical Society
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• v.28 no.12
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• pp.2315-2318
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• 2007

Epinephrine was determined using a lab-made chemiluminescence (CL) system with air pump. Luminolsodium IO4? chemiluminescence system was employed to produce the luminescence of epinephrine. In the reaction, epinephrine was oxidized to produce superoxide or singlet oxygen by periodate in alkaline solution, which enhanced CL of luminol. For optimization, various buffers, such as phosphate, borate, and tris, were studied in this experiment. Compared to NaOH, the phosphate and borate buffer showed better reproducibility with similar sensitivity. Small amount of sample, 22 μL, was required for a measurement. The limit of quantification for epinephrine was obtained to be ~10?9 g/mL after optimization.

• Applied Chemistry
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• v.15 no.1
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• pp.37-40
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• 2011

A chemiluminescent method with silver nanoparticles for determination of L-alanine has been presented. The chemilumiscence intensity was further enhanced by silver nanoparticles in the luminol system by its catalytic role. The silver nanoparticles enhanced chemiluminescent method is applicable for the determination of an amino acid such as alanine. When alanine was introduced to the luminol system with silver nanoparticles, chemiluminescence intensity was reduced with the concentration of the added alanine. The effects of pH, concentrations of luminol, hydrogen peroxide and silver nanoparticles on the chemiluminescence intensity were investigated. The calibration curve for L-alanine was linear over the range from 6.60×10^-8 M to 4.00×10^-7 M, coefficient of correlation was 0.996 and detection limit was 3.5×10^-9 M under the optimal conditions of 4.0×10^-3 M, 4.0×10^-2 M, 4.0×10^-4 M, 12.8 for the concentration of luminol, H₂O₂, silver nanoparticles and pH, respectively.

• Applied Chemistry
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• v.15 no.1
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• pp.21-24
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• 2011

A sensitive silver nanoparticles (Ag NPs) enhanced chemiluminescence (CL) method is reported for the determination of mandelic acid (MA). This method is based on the luminol-KIO₄ system catalyzed by Ag NPs to produce CL spectra. Prepared Ag NPs were characterized by UV-visible spectra and TEM image. Under optimal condition, CL spectra of the system were responded linearly with the concentration of MA in the range of 2.5×10^-9 to 2.0×10^-8 mol L^-1 (r=0.9989) with a detection limit of 1.2×10^-10 mol L^-1. The relative standard deviation of 1.0×10^-7 mol L^-1 MA was found 1.45 (n=9).

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.275-289
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• 2002

The objective of this study was to develop an in vitro assessment of sperm fertilizing capacity of bulls and investigate the factors influencing sperm function and characteristics of frozen-thawed bovine spermatozoa. in vitro fertilization (IVF), the evaluation of motility and normal morphology, HOST (hypoosmotic swelling test), Ca-ionophore induced acrosome reaction, luminol and lucigenin-dependent chemiluminescence for the measurement of reactive oxygen species (ROS), the measurement of malondialdehyde formation for the analysis of lipid peroxidation (LPO), and the evaluation of DNA fragmentation using the method of 747-mediated nick end labelling (TUNEL) by flow cytometry were performed in frozen-thawed bovine spermatozoa. Correlations between the rates of fertilization, blastocyst formation after IVF and the values of respective assays were investigated. 1. IVF rate and blastocyst formation rate averaged 64.4% and 34.3% for spermatozoa from high -fertility bull group and averaged 18.5% and 6.2% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. Sperm motility and percentage acrosome reaction averaged 79.0% and 66.2% for spermatozoa from high-fertility bull group and averaged 40.7% and 22.9% for spermatozoa from low-fertility bull group, respectivitely. There were not different between two bull groups. 2. Luminol depenent chemiluminescence, LPO and DNA fragementation averaged 6.4, 2.0 nmol and 2.6% from spermatozoa from high-fertility bull group and averaged 6.5, 3.1 nmol and 7.4% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. There was no significant difference in lucigenin dependent chemiluminescence between two bull groups. 3. Fertilization rate was positively correlated with motility and the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between fertilization rate and the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. 4. Blastocyst formation rate was positively correlated with the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between blastocyst formation rate and motility, the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. In conclusion, these data suggest that ROS significantly impact semen quality. The assays of this study may provide a basis fur improving in vitro assessment of sperm fertilizing capacity.

• Bulletin of the Korean Chemical Society
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• v.32 no.2
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• pp.639-644
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• 2011

A simple and highly sensitive chemiluminescence method to determine norfloxacin (NFLX) has been proposed by measuring the chemiluminescence (CL) intensities using a flow injection (FI) system. The CL intensity of the luminol-$H_2O_2$ system is strongly enhanced by the addition of Cu (II) in alkaline condition. The CL intensity is substantially increased after the injection of NFLX into the luminol-$H_2O_2$-Cu (II) system. The enhancement effect is attributed to a catalytic effect of Cu (II) due to the interaction with NFLX which forms a complex with the catalyst. Under the optimal conditions, the sensitizing effect of the CL intensity is proportional to the concentration of NFLX in the range of $1.5{\times}10^{-9}-5.9{\times}10^{-7}molL^{-1}$ (r = 0.9994) with a detection limit ($3{\sigma}$) of $2.98{\times}10^{-10}molL^{-1}$. The proposed method had good reproducibility with the relative standard deviation (RSD, n = 5) of 1.6% for $1{\times}10^{-7}molL^{-1}$ of NFLX. The possible reaction mechanism of the CL reaction is also discussed. This method has been successfully applied for the determination of trace amount of NFLX in pharmaceutical preparations and serum samples.

• Bulletin of the Korean Chemical Society
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• v.32 no.1
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• pp.149-152
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• 2011

A new chemiluminescence immunochromatographic analysis system with high sensitivity and high reproducibility was developed for the determination of microcystins (MCs) in water. Horse radish peroxidase (HRP) labeled microcystin monoclonal antibody was used for the sensitive chemiluminescence detection. The chemiluminescence immunochromatographic analysis system was composed of microcystin LR (MCLR)-monoclonal antibody (mAb)-Horse Radish Peroxidase (HRP) conjugate, MCLR-BSA conjugate, luminol, hydrogen peroxide mixture solution, an immunochromatographic assay strip and luminometer. To detect the concentration of microcystins in water, we utilized one spot analysis of the strip instead of flow type analysis. We could detect the microcystins in water at a concentration as low as 9.45 pg/mL with the chemiluminescence (CL) detection.