• Journal of Microbiology and Biotechnology
• /
• v.23 no.6
• /
• pp.826-832
• /
• 2013

We investigated the transgalactosylation reaction of chlorphenesin (CPN) using ${\beta}$-galactosidase (${\beta}$-gal)-containing Escherichia coli (E. coli) cells, in which galactose from lactose was transferred to CPN. The optimal CPN concentration for CPN galactoside (CPN-G) synthesis was observed at 40 mM under the conditions that lactose and ${\beta}$-gal (as E. coli cells) were 400 g/l and 4.8 U/ml, respectively, and the pH and temperature were 7.0 and $40^{\circ}C$, respectively. The time-course profile of CPN-G synthesis under these optimal conditions showed that CPN-G synthesis from 40 mM CPN reached a maximum of about 27 mM at 12 h. This value corresponded to an about 67% conversion of CPN to CPN-G, which was 4.47-5.36-fold higher than values in previous reports. In addition, we demonstrated by thin-layer chromatography to detect the sugar moiety that galactose was mainly transferred from lactose to CPN. Liquid chromatography-mass spectrometry revealed that CPN-G and CPN-GG (CPN galactoside, which accepted two galactose molecules) were definitively identified as the synthesized products using ${\beta}$-gal-containing E. coli cells. In particular, because we did not use purified ${\beta}$-gal, our ${\beta}$-gal-containing E. coli cells might be practical and cost-effective for enzymatically synthesizing CPN-G. It is expected that the use of ${\beta}$-gal-containing E. coli will be extended to galactose derivatization of other drugs to improve their functionality.

• Proceedings of the PSK Conference
• /
• 2003.04a
• /
• pp.305.2-306
• /
• 2003

We aimed at determining bioavailability of chlorphenesin carbamate, a muscle relaxant, and developing a simple analysis in human blood using HPLC. A rapid and sensitive HPLC method was developed and validated using reverse-phase C 18 column with retention time and limit of quantification of toferisone being 8.6 min and 0.5 ng/$m\ell$, respectively. Quantification was performed at 260 nm with tolferison as internal standard. (omitted)

• Journal of Life Science
• /
• v.25 no.10
• /
• pp.1164-1168
• /
• 2015

Previous research showed that chlorphenesin galactoside (CPN-Gal), a preservative in cosmetics, was safer than CPN against human skin cells [9]. To establish a stable and long-term process for CPN-Gal production, we investigated the repeated-batch process. In this process, β-gal-producing recombinant Escherichia coli cells were immobilized in calcium alginate beads, and CPN was converted to CPN-Gal by the transgalactosylation reaction. The process was conducted in a 300 ml flask, which contained E. coli cell-immobilized alginate beads, 33.8 mM of CPN, and 400 g/l of lactose. The pH and temperature were 7.0 and 40℃, respectively. During the repeated-batch operation, four consecutive batch operations were conducted successfully until 192 hr. The conversion yield of CPN to CPN-Gal was 64% during 192 hr, which was higher than the values in previous reports [3, 13]. Thereafter, however, the conversion yield gradually decreased until the operation was finished at 336 hr. Western blotting of immobilized E. coli cells revealed that β-gal gradually decreased after 192 hr. In addition, alginate beads were cracked when the operation was finished. It is probable that, including this loss of E. coli cells by cracks, deactivation, and product inhibition of E. coli β-gal might lead to a gradual decrease in the production of CPN-Gal after 192 hr. However, as the purification of β-gal is not necessary with β-gal-producing recombinant E. coli cells, β-gal-producing E. coli cells might be a practical and cost-effective approach for enzymatically synthesizing CPN-Gal. It is expected that this process will be extended to long-term production process of CPN-Gal for commercialization.

• Journal of the Korean Applied Science and Technology
• /
• v.34 no.4
• /
• pp.954-961
• /
• 2017

We investigated the purifications of PE-gal and CPN-gal, synthesized by transgalactosylation reaction using recombinant ${\beta}$-gal. The reaction mixture containing PE and PE-gal was first mixed with EA, and thereafter PE and PE-gal were distributed in two-phase (EA/water) system. In this system, PE and PE-gal was selectively moved into EA and water phase, respectively. Then, the water phase was collected, and silica gel chromatography was carried out using the collected water phase. Finally, we compared two purified PE-gal samples using HPLC and TLC analysis, in which the one was purified only by silica gel chromatography and the other was purified by EA extraction followed by silica gel chromatography. In the latter case, the residual PE was almost completely removed, whereas, in the former case, the residual PE was remained remarkably. Additionally, the purification yield of PE-gal was about 21% on the basis of mole. In the same purification protocol, CPN-gal was able to be purified using EA extraction followed by silica gel chromatography, in which the residual CPN was almost removed when CPN-gal was purified by EA extraction followed by silica gel chromatography.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.33 no.4
• /
• pp.263-267
• /
• 2007

Parabens are used in nearly all types of cosmetics and toiletries because they are formulated well and have broad spectrum of activity, interness, low costs and excellent chemical stability in relation to pH. 2-phenoxyethanol and chlorphenesin are common preservatives which are usually used in combination with parabens in cosmetics. Toxicity of parabens is generally low but application of parabens to damaged or broken skin has resulted in sensitization. Moreover, the possibility of their estrogenic potential, anesthetic effects and reproductive toxicity has been reported. Consequently there are some regulations in use of parabens. And the maximum permitted concentrations of chlorphenesin and 2-phenoxyethanol in cosmetic products are authorized by the same reasons. So it is important to control and estimate the amount of parabens in products. In this article, we proposed a valid method for the simultaneous determination of 8 preservatives including parabens in a short time using ultra performance liquid $chromatography^{TM}\;(UPLC^{TM})$. Separation of eight components was achieved in less than 10 min and resolutions were reasonable (USP resolution ${\geqq}\;2$). And limit of detection and quantification were evaluated. The method was suitably validated for specificity, linearity, precision (repeatability, intermediate precision) and accuracy for assay (recovery) based on International conference on harmonisation (ICH) guideline. The method was applicable to analysis of preservatives in cosmetic products.

• Proceedings of the PSK Conference
• /
• 2003.04a
• /
• pp.304.1-304.1
• /
• 2003

We aimed at determining bioavailability of tolperisone, a muscle relaxant. and developing a simple analysis in human blood using HPLC. A rapid and snsitive HPLC method was developed and validated using reverse-phase C18 column with retention time and limit of quantification of toferisone being 7.3 min and 20 ng/$m\ell$, respectively. Quantification was performed at 260 nm with chlorphenesin as internal standard. (omitted)

• Journal of Korean society of Dental Hygiene
• /
• v.8 no.4
• /
• pp.31-41
• /
• 2008

Steven-Johnson syndrome (SJS) and toxic epidermal necrolysis(TEN) are severe mucocutaneous reaction which are most frequently caused by drugs. Although the incidence of SJS and TEN is known to be relatively low, outcomes may be fatal. A systematic approach is required because morbidity rate is currently increasing and oral lesion is frequent. We investigated the clinical features and outcomes of 6 patients diagnosed as SJS and TEN and referred from the department of dermatology, Chonnam National University Hospital for oral care. Ketoconazol, Ofloxacin, Chlorphenesin, Amoxicillin, Pontal, Harnal, and Ciprofloxacin were suspected as the causative drugs. Average treatment period was 3.2 weeks, and two patients were referred to 'burn-patients' hospital. Most of oral lesion were cured be normal tissue, but scares with discoloration were observed. For intraoral management, antibiotic disinfection and steroid application were performed according to systemic treatment principles. Additionally, ingestion of zinc, antioxidants, and vitamin was recommended. The establishment of oral treatment principles is demanded because it has not been yet. Also, through investigation of drug side effect and careful prescription are required.

• Archives of Pharmacal Research
• /
• v.29 no.4
• /
• pp.339-342
• /
• 2006

A simple high performance liquid chromatographic (HPLC) method was developed for the determination of tolperisone in human plasma. Tolperisone and internal standard (chlorphenesin) were isolated from 1 mL of plasma using 8 mL of dichlormethane. The organic phase was collected and evaporated under nitrogen gas. The residue was then reconstituted with 300 mL aliquot of mobile phase and a 100 mL aliquot was injected onto the $C_{18}$ reverse-phased column. The mobile phase, $45\%$ methanol containing $1\%$ glacial acetic acid and $0.05\%$ 1-hexanesulfonic acid was run at a flow rate of 1 mL/min. The column effluent was monitored using UV detector at 260 nm. The retention times for tolperisone and the internal standard were approximately 7.1 and 8.4 min, respectively. The standard curve was linear with minimal intra-day and inter-day variability. The quantification limit of tolperisone in human plasma was 10 ng/ mL. The proposed method has been applied to the determination of pharmacokinetic profile of tolperisone in Koreans. The T max of tolperisone in Koreans $(0.94{\pm}0.42\;h)$ was not significantly differ from that reported in Europeans (0.5-1 h), but the mean half-life in Koreans $(1.14{\pm}0.27\;h)$ was shorter than that in Europeans $(2.56{\pm}0.2\;h)$. The proposed HPLC method is simple, accurate, reproducible and suitable for pharmacokinetic study of tolperisone.

• Journal of the Korea Convergence Society
• /
• v.8 no.11
• /
• pp.159-165
• /
• 2017

We were investigated the content of 7 preservatives for sheet mask samples(n=42) sold in markets of Daejeon metropolitan city in 2016. &3.3%(n=35) of all samples contained at least one of preservatives. In samples of 30.95(n=14) and 2.39%(n=1) was detected with 2 and 3 preservatives. Phenoxyethaol(PE), methylparaben(MP), chlorphenesin(CP) and benzyl alcohol(BA) showed detection rate of 76.19(n=32), 16.67(n=9), 21.43(n=7) and 2.38%(n=1), respectively. Also The content of detected preservative showed range of 0.06~0.71, 0.18~0.35, 0.06~0.71 and 0.32% and was within the maximum allowed amount established by Korean FDA. However ethylparaben(EP), propylparaben(PP) and benzylparaben(BP) in all samples was not detected. These results can be useful for sharing in emotion-fusioned information and supplying right to know of user.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.32 no.1 s.55
• /
• pp.65-68
• /
• 2006

Sensory irritation is one of the important side effects of cosmetics and it is required to develop new products that are more tolerable to the consumers. There are lots of cosmetic ingredients known to induce sensory irritation such as lactic acid, glycolic acid, ethanol, preservatives, fragrances and menthol etc. It is also known that sensory irritation increases by change of pH as well as additional occlusive conditions. The aim of this study is to know various factors affecting sensory irritation due to preservatives (methylparaben, propylparaben, phenoxyethanol and chlorophenecin). We also wanted to investigate the effect of preservatives to sensory irritation according to change of formulations. Our results showed that sensory irritation increased with the conditions of increasing absorption such as packs. We have also found that sensory irritation increased synergistically when to apply two different preservatives together. For example, phenoxyethanol and chlorphenesin themselves have weak sensory irritation potentials, but we observed phenoxyethanol combined with chlorpenesin synergistically increase of sensory irritation potentials. In conclusion, formulation and combination of different preservatives should be considered to reduce the unwanted sensory irritation of preservatives.