• Journal of Applied Biological Chemistry
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• v.64 no.1
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• pp.13-17
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• 2021

Qantitative analysis of six compounds: (+)-catechin, benzoic acid, gallic acid methyl ester, paeonol, paeoniflorin, and albiflorin from Paeonia lactiflora extracts was performed using high-performance liquid chromatography and an ultraviolet (UV) detector, following different extraction methods. A reverse-phase column was used in a gradient elution system, and UV detection was performed at 280 nm. The results showed that the quantity of paeoniflorin was the highest in ethanol and water extracts (73.89 and 57.87 mg/g, respectively) among the six compounds. This study contributes a good analysis method for the contents of P. lactiflora and would be propitious for developing medicines and functional foods.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.244-249
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• 2023

Artemisia keiskeana and A. stolonifera are plants of the genus Artemisia, distributed in various regions, especially China and Korea. They are renowned as medicinal plants with biological and pharmacological activities. Fraxidin, isofraxidin, and daphnoretin are coumarins present in Artemisia spp.; however, research on them is limited. Therefore, this study was carried out to quantify the content of these compounds in the aerial parts of A. keiskeana and A. stolonifera in different regions in Korea. High-performance liquid chromatography was performed with a photodiode array detector and a reverse-phase INNO column. A. stolonifera only contained fraxidin with the highest amount found in Yongmun commune. A. keiskeana cultivation in Soyang commune gave the highest fraxidin and daphnoretin content. However, isofraxidin was not present in all samples. The findings suggest that the concentrations of the three compounds may differ depending on the growth site and provide a foundation for future studies.

• Korean Journal of Veterinary Service
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• v.18 no.3
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• pp.13-28
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• 1995

This study was carried out to explore the most sensitive and useful method for simultaneous determination of five sulfa drugs(sulfamethazine, sulfamerazine, sulfamonomethoxine, sulfadimethoxine, sulfaquinoxaline) in livestock productions(pork muscle, bovine muscle, chicken muscle, milk ) by HPLC with UV detector and reverse phase column. The results obtained were as follows：1. For mobile phase acetonitrile-0.01M ammonium acetate (23：77) showed more applicable sensitivity and retention times than acetonitrile-1% acetic acid(23：77). Thus acetonitrile-0.01M ammonium acetate(23：77) selected and applied to the modification test, from which it was found pH 6.75 was the most adequate. 2. Optimal wavelength of UV for SMT(sulfamethazine), SMR(sulfamerazine), SMM(sulfamonomethoxine), SD(sulfadimethoxine), and SQ(sulfaquinoxaline) were 266, 266, 265, 269 and 250nm, respectively, and that for simultaneous application it was 263nm. 3. The average recovery rate by extractant(chloroform, dichloromethane, chlorform+dich-loromethane) in the classic method was not significantly different(p>0.05) but that by chloroform higher than the others. Thus chloroform was found to be adequate as extractant in this classic method. 4. The average recovery rate was 86.5% by the MSPD(matrix solid phase disperse) method, which was significantly higher than that by the classic method(p<0.05). Also the recovery rates by method were significantly different(p<0.05) in accordance with sample and type of drug. The MSPD method was much superior to classic method on clean-up.

• YAKHAK HOEJI
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• v.41 no.6
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• pp.692-697
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• 1997

Creatine and 3,4-Dimethylhippuric acid (3,4-DMHA), a glycine conjugate of 1,2,4-trimethyl-benzene (1,2,4-TMB) were determined in the urine of workers exposed to 1,2,4-TMB vapor. The best condition for the simultaneous determination of 3,4-DMHA and creatine by high performance liquid chromatography was obtained by reverse phase $C_{18}$ column (4.6${\times}$150mm, 5${\mu}m$) as stationary phase and 20% acetonitrile in 20mM phosphate buffer (pH 3.0) containing 4mM sodium octylsulfate(SOS)as mobile phase. The recovery of 3,4-DMHA spiked to blank urine in the range of 1~5${\mu}g$/ml was about 96%. The concentration of urinary 3,4-DMHA of workers had a positive correlation with the environmental level of 1,2,4-TMB (r=0.866). The data suggest that urinary 3,4-DMHA concentration is a useful biological index for 1,2,4-TMB exposure.

• Food Science and Biotechnology
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• v.17 no.3
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• pp.679-682
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• 2008

A lactic acid bacterium producing an antimicrobial substance against Escherichia coli O157:H7 was isolated from raw milk and identified as Lactobacillus amylovorus ME-1. In addition to E. coli O157 :H7, the antimicrobial substance also inhibited the growth of Bacillus cereus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pyrogenes, and Yersinia enterocolitica. The antimicrobial substance was stable at pH 2-12 and $121^{\circ}C$ for 15 min and insensitive to proteinase K, protease, amylase, and catalase. Purification of the antimicrobial substance was conducted through methanol and acetonitrile/ethylacetate extraction, ultrafiltration with a 500 Da cutoff, thin layer chromatography (TLC) with silicagel 60, and high performance liquid chromatography (HPLC) with a $C_{18}$ reverse phase column. The ${\lambda}_{max}$ of the purified antimicrobial substance was determined as 192 nm by ultra violet (UV) scanning, while the molecular weight was estimated as 453 Da based on the mass spectrum. Accordingly, the current results suggest that the antimicrobial substance from the L. amylovorus ME-1 was not a bacteriocin, but rather a new non-proteinaceous substance distinct from acidophilin, acidolin, diacetyl, and reuterin.

• Journal of Applied Biological Chemistry
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• v.44 no.1
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• pp.12-15
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• 2001

The antioxidative compounds and antioxidant contents of ginger (Zingiber officinale) rhizomes were determined. Substances reextracted using ethyl acetate from crude methanol extract of fresh ginger rhizome were separated through thin layer chromatography. Ten phenolic antioxidative bands were visualized through color reactions using ferric chloride-potassium ferricyanide and 1,1-diphenyl-2-picrylbydrazyl (DPPH). The antioxidative compounds were purified through preparative TLC and high performance liquid chromatography (HPLC), among which, five antioxidants were identified as 4-, 6-, 8-. and 10-gingerols and 6-shogaol on the basis of their molecular weights determined through LC-MS. As shown in experiments using DPPH free radicals, 6-Gingerol and PT4-HP8 (unknown) were revealed to be more efficient than BHT (butylated hydroxy toluene). Contents of gingerols were determined through reverse phase HPLC. Total gingerol contents (sum of 6-,8-, and 10-gingerols) in rhizomes of different ginger varieties varied significantly. The HG55 (collected at Wanju district in Korea) and the HG52 (imported from Brazil) showed the highest gingerol contents.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.2
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• pp.118-125
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• 2011

In this study, an angiotensin I-converting enzyme (ACE) inhibitor from squid skin was purified and characterized. Squid (Todarodes pacificus) skin protein isolates were hydrolyzed using six commercial proteases: alcalase, ${\alpha}$-chymotrypsin, neutrase, papain, pepsin, and trypsin. The peptic hydrolysate had the highest ACE inhibitory activity. The ACE inhibitory peptide was purified using Sephadex G-25 column chromatography and reverse phase high-performance liquid chromatography (HPLC) with a $C_{18}$ column. The purified ACE inhibitory peptide was identified and sequenced, and found to consist of seven amino acid residues: Ser-Ala-Gly-Ser-Leu-Val-Pro (657Da). The $IC_{50}$ value of the purified ACE inhibitory peptide was 766.2 ${\mu}M$, and Lineweaver-Burk plots suggested that the purified peptide acts as a noncompetitive ACE inhibitor. These results suggest that the ACE inhibitory peptide purified from the peptic hydrolysate of squid skin may be of benefit in developing antihypertensive drugs and functional foods.

• Biomolecules & Therapeutics
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• v.15 no.2
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• pp.123-126
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• 2007

A sensitive, simple and highly selective liquid chromatography method of determination for extraction of pseudoephedrine hydrochloride from Allegra D tablet was developed. The chief benefit of the present method is the minimal sample preparation, as the procedure is only filtering through pore syringe filter. Two drugs (pseudoephedrine hydrochloride, fexofenadine) were separated on a C$_{18}$ column and analyzed by high performance liquid chromatography (HPLC). The method had a chromatographic run time of 8.0 min. 1 ml of pseudoephedrine hydrochloride solution (1 mg/ml) was filtered through 0.22 um pore syringe filter. 50 ul of filtering solution was injected to HPLC pump and we knew the retention time (1.85 min) of separating of pseudoephedrine hydrochloride using UV detector at 280 nm. We used C$_{18}$ column (4.6 mm${\times}$250 mm), mobile phase solution (<0.05 mol/L NaH$_2$PO$_4$, 2 ml/L H$_3$PO$_4$>/CH$_3$CN / sodium dodesyl sulfate = 60 ml / 40 ml / 1 g). We separated psedoephedrine hydrochloride at run time of 1.85 min from Allegra D tablet solution (1 mg/ml) filtered through 0.22 um pore syringe filter using UV detector at 280 nm. Flow rate was set at 1.0 ml/min and the column temperature was set at 40$^{\circ}C$. Psedoephedrine hydrochloride solution (1 mg/ml) separated from Allegra D tablet was filtered through 0.22 um pore syringe filter and injected 50 ul. We confirmed the peak of psedoephedrine hydrochloride at same retention time and the separating solution was freeze-dried. In conclusion, A simple isocratic reverse-phase HPLC method has been developed that provides excellent separation of pseudoephedrine from Allegra D tablet.

• Journal of Microbiology and Biotechnology
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• v.26 no.10
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• pp.1781-1789
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• 2016

Glargine insulin is a long-acting insulin analog that helps blood glucose maintenance in patients with diabetes. We constructed the pPT-GI vector to express prepeptide glargine insulin when transformed into Escherichia coli JM109. The transformed E. coli cells were cultured by fed-batch fermentation. The final dry cell mass was 18 g/l. The prepeptide glargine insulin was 38.52% of the total protein. It was expressed as an inclusion body and then refolded to recover the biological activity. To convert the prepeptide into glargine insulin, citraconylation and trypsin cleavage were performed. Using citraconylation, the yield of enzymatic conversion for glargine insulin increased by 3.2-fold compared with that without citraconylation. After the enzyme reaction, active glargine insulin was purified by two types of chromatography (ion-exchange chromatography and reverse-phase chromatography). We obtained recombinant human glargine insulin at 98.11% purity and verified that it is equal to the standard of human glargine insulin, based on High-performance liquid chromatography analysis and Matrix-assisted laser desorption/ionization Time-of-Flight Mass Spectrometry. We thus established a production process for high-purity recombinant human glargine insulin and a method to block Arg (B31)-insulin formation. This established process for recombinant human glargine insulin may be a model process for the production of other human insulin analogs.

• Analytical Science and Technology
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• v.21 no.6
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• pp.518-525
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• 2008

Mandipropamid is a new mandelamide-type fungicide to control foliar Oomycete pathogens in some vegetables. An analytical method was developed to determine mandipropamid residues in agricultural commodities using high-performance liquid chromatography (HPLC) and liquid chromatography/mass spectrometry (LC/MS). Mandipropamid was extracted with methanol from grape, tomato, green pepper, Chinese cabbage and potato samples. The extract was diluted with saturated sodium chloride solution and distilled water, and dichloromethane partition was followed to recover the mandipropamid from the aqueous phase. Florisil column chromatography was employed to further remove interfering co-extractives prior to HPLC analysis. Reverse-phased HPLC was successfully applied to determine mandipropamid in sample extracts with the detection at its ${\lambda}_{max}$ (223 nm). Overall recoveries of mandipropamid from fortified samples averaged $99.8{\pm}1.7$ (n=6), $89.3{\pm}5.3$ (n=6), $98.7{\pm}2.2$ (n=6), $99.7{\pm}6.8$ (n=6) and $91.1{\pm}3.1$ (n=6) for grape, tomato, green pepper, Chinese cabbage and potato, respectively. Limit of quantification of the method was 0.02~0.04 mg/kg for all samples. A LC/mass spectrometry with selected-ion monitoring was also provided to confirm the suspected residue. The proposed method was reproducible and sensitive enough to determine the terminal residue of mandipropamid in agricultural commodities.