• Title/Summary/Keyword: HPLC analysis method

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The Antimicrobial Characteristics of McSSP-31 Purified from the Hemocyte of the Hard-shelled Mussel, Mytilus coruscus (참담치(Mytilus coruscus) 혈구(hemocyte)에서 분리한 McSSP-31의 항균 특성 분석)

  • Oh, Ryunkyoung;Lee, Min Jeong;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Joo-Won;Park, Jung-Youn;Seo, Jung-Kil;Kim, Dong-Gyun
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1276-1289
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    • 2017
  • This study isolated and purified the antimicrobial peptide McSSP-31 from an acidified hemocyte extract of a Mytilus coruscus. The antimicrobial peptide was purified by using a $C_{18}$ reversed-phase high-performance liquid chromatography (HPLC). The peptide was determined to be 3330.549 Da by matrix assisted-laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS). The N-terminus of a 14 amino-acid sequence was identified as P-S-P-T-R-R-S-T-S-R-S-K-S-R by Edman degradation method. The acquired sequence showed a 93% similarity with the sperm-specific protein Phi-1, which is from M. californianus. The identified open-reading frame (ORF) of peptide was 306 bp encoding 101 amino acids, which was analyzed by rapid amplification of cDNA ends (RACE), cloning and sequencing analysis. We compared the full sequence with other known proteins that reveal the sperm-specific protein Phi-1 (93.5%) of M. californianus. Synthesized antimicrobial peptide (McSSP-31) showed antibacterial activity against gram-positive bacteria including B. subtilis, S. mutans, S. aureus and gram-negative bacteria including E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa and fungi, C. albicans. Also, synthesized peptide showed strong antibacterial activity against antibiotic-resistant strains, including S. aureus. The cytotoxicity of the peptide was determined by using the HUVEC human cell line. The peptide did not exhibit any significant cytotoxic effects on the normal human cell line, and it had very low hemolytic activity with flounder hemoglobin. The results demonstrated that peptide purified from the hemocyte of a M. coruscus exhibits antibacterial activity against various bacteria and has the potential to be an alternative antibiotic agent.

Neuroprotective effects of Extract of Broccoli, Cultivated in Desalinated Magma Seawater, on neuron-like SH-SY5Y cells (제주도 탈염 용암해수 재배로 제조한 브로콜리 추출물의 신경 세포 보호 효과)

  • Rhee, Jin Seol;Jang, Youn Bi;Choi, Ge Sun;Choung, Jai Jun;Kang, Seung Woo
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.3
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    • pp.448-462
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    • 2020
  • Dietary nutrition is a critical lifestyle factor that can reduce the risk of future cognitive impairments caused by dementia. Accumulating evidence suggests that dietary supplementation with Sulforaphane may help the prevention of cognitive impairments and dementia. Thus, Sulforaphane-enriched broccoli extract would hold promise to improve cognitive impairments of dementia patients. Here, we have used broccoli extracts, prepared from broccoli cultivated in Magma Seawater, to test if the broccoli extracts can be dietary supplement to improve cognitive impairments. Magma Seawater originated from Jeju Island, Korea is unique in terms of containing high concentrations of usable minerals (Zinc, Vanadium and Germanium etc.). Broccoli, grown in Magma Seawater, would contain Sulforaphane and the extra amount of usable minerals. The chemical compositions of the broccoli extracts were analyzed using LC-Q-orbitrap to detect Sulforaphane and Glucoraphanin. Analysis method based on HPLC was developed for measurement of sulforaphane levels in the broccoli extracts. We have tested if the broccoli extracts have anti-apoptotic and anti-inflammatory effects on neuron-like SH-SY5Y cells. In addition, we examined if the broccoli extracts are able to upregulate expression of synaptic plasticity-associated proteins (BDNF and phospho-CREB) and to inhibit acetylcholine esterase (AchE) activity. We have shown that the broccoli extracts inhibited the apoptotic pathway and inflammatory responses. Finally, we present evidence showing that AchE activity was inhibited by the broccoli extracts, but expression of BDNF and phospho-CREB was upregulated. Taken together, these findings suggest that the broccoli extracts from Magma Seawater-grown broccoli would be a good source of dietary nutrition to improve cognitive impairments in the future.

Determination of Positional Fatty Acid and Triacylglycerol Compositions of Selected Infant Formulas (영유아용 조제분유의 위치별 지방산 및 Triacylglycerols의 정성 분석)

  • Son, Jeoung-Mae;Lee, Jeung-Hee;Hong, Soon-Taek;Lee, Kyung-Su;Park, Hye-Kyung;Kwon, Kwang-Il;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.9
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    • pp.1256-1264
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    • 2011
  • Infant formula is one of the main nutritional sources for infants. In this study, the fatty acid compositions of Korean (domestic, n=8) and foreign infant formulas (n=3) were analyzed. Crude fats were extracted using the Folch method, and fatty acid compositions (total and positional) were analyzed by gas chromatography. In the fatty acid compositions of infant formulas, oleic (19.88~44.64%), palmitic (7.59~21.65%) and linoleic acids (16.72~25.87%) were the major fatty acids. Domestic infant formula products contained 35.94~56.55% total saturated fatty acid content, whereas that of foreign infant formula ranged from 34.40~42.88%. The content of monounsaturated fatty acids ranged from 20.23~44.99% in domestic products and 34.11~45.07% in foreign products. In addition, 0.17~2.57% arachidonic acid/docosahexaenoic acid and 10~13% linoleic acid/linolenic acid were detected in the analyzed products (domestic and foreign products). A small amount of trans fatty acids (0.25~1.69%) were found. In sn-2 position analysis, palmitic acids (1.84~38.74%) were detected in the analyzed formulas. Further, typical triacylglycerols in human milk, including 1,3-di-monounsaturated-2-saturated triacylglycerol, were not detected in the analyzed formulas.

Physiological Activity of Supercritical Poria cocos back Extract and Its Skin Delivery Application using Epidermal Penetrating Peptide (초임계 복령피 추출물의 생리활성 및 경피투과 펩티드를 이용한 경피 약물전달의 응용)

  • Kim, Min Gi;Park, Su In;An, Gyu Min;Heo, Soo Hyeon;Shin, Moon Sam
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.3
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    • pp.766-778
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    • 2019
  • In this study, Poria cocos bark were extracted by supercritical process, and anti-inflammatory, whitening, and antioxidant effects were measured in comparison with ethanol extract. Also, An effective percutaneous permeation method using a selected formulation of the extract and a drug delivery peptide was proposed. Pachymic acid, known as the anti-cancer and anti-inflammatory compound of the ventricle, is an indicator component and the HPLC analysis shows that the supercritical extract of the pericardium is more than twice that of the Poria cocos bark extract. In order to confirm antioxidative effect of Bombyx mori, DPPH scavenging ability and ABTS scavenging ability test showed that the ethanol extract of Poria cocos Back had lower concentration than the supercritical extract of Poria cocos back. However, RAW 264.7 Measurements of Nitric oxide (NO) production in cells showed lower NO production at the same concentration than the Poria cocos back ethanol extract. In addition, after 72 hours of processing of $20{\mu}g/mL$ of the Poria cocos back extract in B16 melanoma cells, both the intracellular and extracellular melanin extract were effective and the supercritical extract was lower melanin content. No toxicity was observed at the concentration of $800{\mu}g/mL$ in RAW 264.7 cells used in NO production experiments. However, in B16 melanoma cells, even at $50{\mu}g/mL$, both Poria cocos back ethanol extract and supercritical extract showed a survival rate of less than 60%. The liposome formulation and drug delivery peptides were shown to be useful for percutaneous permeation of Supercritical Extract of Poria cocos back using a liposome formulation and a drug delivery peptide. it is expected that there will be great potential for development as a variety of cosmetic materials for Poria cocos back.

Determination of Total Aflatoxins in Foods by Parallelism of ELISA and LC/MS/MS (ELISA-LC/MS/MS 병행에 의한 식품 중 aflatoxins 분석)

  • Kim, Kyeong-Yeol;Nam, Min-Ji;Nam, Bo-Ram;Ryu, Hee-Jung;Song, Jeong-Eon;Shim, Won-Bo;Lee, Soo-Hyung;Chung, Duck-Hwa
    • Journal of Environmental Health Sciences
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    • v.36 no.1
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    • pp.52-60
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    • 2010
  • High performance liquid chromatography (HPLC) and liquid chromatography mass spectrometry (LC/MS) have been widely used to quantify aflatoxins in food, but these methods are expensive, time-consuming, unsuitable for analysis of the routine screening of large sample numbers and require derivatization and high level techniques to perform. The objective of this study is to detect aflatoxins in a large number of foods by a high efficient analytical system of combined enzyme linked immunosorbent assay (ELISA) for screening and LC/MS/MS for confirmation. The samples spiked individually with aflatoxin $B_1$ (0.5 and 1.0 ng/g) and total aflatoxins (10 ng/g) were analyzed by ELISA and LC/MS/ MS, and the recoveries for ELISA and LC/MS/MS were 71.8~119.2% and 70.8~135.3%, respectively. A total of 378 samples (grains, nuts, soybean and fermented soybean foods, pepper and fermented pepper foods) were purchased from the six major cities in Korea and analyzed by ELISA-LC/MS/MS system. Twenty two (5.8%; peanut: 11, pistachio: 2, walnut: 6, almond: 1, pepper powder: 1, pepper paste: 1) out of 378 samples were screened as aflatoxin B1 positive by ELISA, but, 4 (1.1%; peanut: 2, pistachio:1, pepper powder: 1) out of the 22 samples screened were confirmed as aflatoxins positive at levels of 1.02~52.79 ng/g by LC/MS/MS. ELISA-LC/MS/MS system provides a more rapid, accurate and cost-effective method for the detection of aflatoxins in large number of samples.

Tencel Dyeing by Natural Indigo Prepared from Dyer's Knotweed (천연인디고를 이용한 텐셀직물의 염색)

  • Shin, Youn-Sook;Son, Kyung-Hee;Yoo, Dong-Il
    • Journal of the Korean Society of Clothing and Textiles
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    • v.32 no.12
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    • pp.1963-1970
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    • 2008
  • Natural indigo dye in powder form was prepared by modifying traditional Niram method, using $Ca(OH)_2$ instead of baked oyster powder for precipitating indigo dye. The prepared dye was applied to dyeing Tencel fabrics to investigate the effect of experimental conditions for the optimization of dyeing process. The indigo dye powder contained 15.2%(w/w) of indigo content and 0.757%(w/w) of indirubin content on the basis of HPLC analysis. Maximum dye uptake was obtained at $60^{\circ}C$ for 20min. Almost saturated dye uptake was obtained at 2g/L of sodium hydrosulfite concentration up to 4g/L of indigo dye and then slowly increased for further increase of sodium hydrosulfite. Whereas at higher indigo dye concentration(8g/L) more than 3g/L of reducing agent concentration was required for obtaining the maximum dye uptake. At the same indigo dye and reducing agent concentration, K/S value of the sample dyed without sodium hydroxide(pH 5.75) was 15.19, much higher than one dyed in alkaline condition(K/S 5.76). There was no difference in colorfastness ratings among samples with different color strength. However, more fading was occurred for the sample with low color strength.

Preparation and Characterization of Phytochemical-Rich Extract from Sasa quelpaertensis Leaf (식물화합물 다량 함유 제주조릿대 잎 추출물의 제조와 특성)

  • Lee, Ju Yeop;Ko, Hee Chul;Jang, Mi Gyeong;Kim, Se Jae
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1330-1335
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    • 2016
  • Sasa species leaves have been used in traditional medicine for their anti-inflammatory, antipyretic, and diuretic properties. Sasa quelpaertensis Nakai is a small bamboo grass that grows only on Mt. Halla on Jeju Island, Republic of Korea. This small bamboo grass has recently been the focus of much attention due to its potential biomass as well as its beneficial health effects. In this study, to promote the efficient utilization of the S. quelpaertensis leaf, we established a simple preparation method for phytochemical-rich extract (PRE) by comparing phytochemical contents and biological activities according to extraction methods. high performance liquid chromatography (HPLC) analysis revealed that the contents of two major phytochemicals such as, tricin (5.35 mg/g) and p-coumaric acid (44.10 mg/g) contained in PRE were higher than those in fresh hot water extract (SQH, p-coumaric 23.39 mg/g, tricin 0.18 mg/g) and ethanol extract (SQE, p-coumaric 10.8 mg/g, tricin 0.38 mg/g). The antioxidant activities [1,1-diphenyl-2-picrylhydrazy (DPPH) radical scavenging activity, 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity, nitric oxide (NO) scavenging activity, and xanthine oxidase inhibitory activity] of PRE were higher than those of SQH and SQE. PRE effectively inhibited NO production in LSP- stimulation RAW 264.7 cells, and the growth of human promyelocytic leukemia (HL-60) cells. These results suggest that PRE has a potential as a promising antioxidant and anti-inflammatory agent.

Bioequivalence of AmbrectTM Tablet to MucopectTM Tablet (Ambroxol hydrochloride 30 mg) (뮤코펙트 정(염산암브록솔 30 mg)에 대한 암브렉트 정의 생물학적 동등성)

  • Yoo, Jeong-Yeon;Joung, Sun-Koung;Choi, Mee-Hee;Han, Sang-Beom;Lee, Kyung-Ryul;Lee, Hee-Joo
    • Journal of Pharmaceutical Investigation
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    • v.33 no.3
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    • pp.215-221
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    • 2003
  • A bioequivalence study of $Ambrect^{TM}$ tablets (Dong Wha Pharm. Ind. Co., Ltd.) to $Mucopect^{TM}$ tablets (Boehringer Ingelheim Korea, Ltd.) was conducted according to the guideline of Korea Food and Drug Administration (KFDA). Twenty four healthy male Korea volunteers received each medicine at the ambroxol hydrochloride dose of 30 mg in a $2{\times}2$ crossover study. There was a one-week wash out period between the doses. Plasma concentrations of ambroxol were monitored by a high-performance liquid chromatography for over a period of 24 hours after the administration. $AUC_t$ (the area under the plasma concentration-time curve from time zero to 24 hr) was calulated by the linear trapezoidal rule method. $C_{max}$ (maximum plasma drug concentration) and $T_{max}\;(time\;to\;reach\;C_{max})$ were compiled from the plasma concentration-time data. Analysis of variance was carried out using logarithmically transformed $AUC_t\;and\;C_{max}$. No significant sequence effect was found for all of the bioavailability parameters indicating that the crossover design was properly performed. The 90% confidence intervals of the $AUC_t$ ratio and the $C_{max}$ ratio for $Ambrect^{TM}/Mucopect^{TM}$ were 0.89-1.01 and 0.89-1.02, respectively. These values were within the acceptable bioequivalence intervals of 0.80-1.25. Thus, our study demonstrated the bioequivalence of $Ambrect^{TM}\;and\;Mucopect^{TM}$ with respect to the rate and extent of absorption.

Anti-inflammatory Effects of Wiryeong-tang Mix Soft Extract (위령탕 혼합단미연조엑스의 항염증 효과)

  • Kim, Myoung-Jin;Kim, Se-Jin;Nam, Won-Hee;Leem, Hyun-Hee;Jeon, Young-Hee;Son, Su-Mi;Kim, Jung-Ok
    • Herbal Formula Science
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    • v.28 no.4
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    • pp.339-349
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    • 2020
  • Objectives : Wiryeong-tang (WRT) is a traditional herbal medicine used to treat kidney-related diseases. However, the anti-inflammatory and anti-gastritis effect of Wiryeong-tang was not well known. Therefore, we experimented to confirmed the anti-inflammatory and anti-gastritis effects of Wiryeong-tang. Methods : The RAW 264.7 cells were pre treated with Wiryeong-tang mix soft extract (WRT-mse; 50, 100 and 200 ㎍/mL) for 1 hrs, and then incubated with lipopolysaccharide (LPS; 500 ng/mL). Cell viability was measured by the MTT method, and nitric oxide (NO) was measured with griess reagent. In addition, pro-inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). For anti-gastritis effect in vivo, acute gastritis was induced using 150 mM HCl/60% ethanol used ICR mice. WRT-mse (133 mg/kg) was pre treated for 3 days and then treated with 150 mM HCl/60% ethanol 1 hrs later. Then gastritis was observed and inflammatory cytokines in the gastric tissue was measured. Results : The 8 marker components of the WRT-mse were determined by simultaneous analysis using HPLC. WRT-mse was not toxic and inhibited pro-inflammatory cytokines such as IL-1β, IL-6 and TNF-α at NO production, protein and mRNA levels. Also, it was confirmed that WRT-mse improved bleeding and edema in gastritis, and suppresses inflammatory cytokines. Conclusion : In summary, our results suggest that the treatment of the WRT-mse reduced and improved the 150 mM HCl/60% ethanol induced acute gastritis and the inflammation caused by LPS stimulation in RAW 264.7 cells. Therefore, this study may provide useful drug or clinical evidence for WRT-mse to prevent inflammation.

Development of HPLC Determination Method for Trace Levels of 1-, 2-Nitropyrenes and 2-Nitrofluoranthene in Airborne Particulates and Its Application to Samples Collected at Noto Peninsula

  • Hayakawa, Kazuichi;Tang, Ning;Sato, Kosuke;Izaki, Akihiko;Tatematsu, Michiya;Hama, Hirotaka;Li, Ying;Kameda, Takayuki;Toriba, Akira
    • Asian Journal of Atmospheric Environment
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    • v.5 no.3
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    • pp.146-151
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    • 2011
  • 1-Nitropyrene (1-NP), 2-NP and 2-nitrofluoranthene (2-NFR) are useful markers for studying the atmospheric behaviors of polycyclic aromatic hydrocarbons (PAHs) and nitropolycyclic aromatic hydrocarbons (NPAHs). However, present methods for measuring trace levels of these compounds are lesssensitive and laborious. Here we describe several improvements to a previously reported high-performance liquid chromatography-chemiluminescence detection system that allows it to determine trace levels of 1-, 2-NPs and 2-NFR. The proposed system was equipped with a reducer column packed with Pt/Rh instead of zinc whose life-time was limited. The combination of Cosmosil MS-II (monomeric ODS) and AR-II (polymeric ODS) columns was used instead of polymeric ODS columns as the separator column to improve the separation. An ethanol mixture with acetate buffer (pH 5.5) was used in place of an acetonitrile mixture with the same buffer to activate the reducer column. The same ethanol mixture was used as the mobile phase for the clean-up column. The switching time of the column switching valve was optimized to concentrate the amino-derivatives of above NPAHs quantitatively on the concentrator column. The concentrations of bis(2,4,6-trichlorophenly) oxalate and hydrogen peroxide in the chemiluminescence reagent solution were optimized to 0.4 mM and 30 mM, respectively, to increase the sensitivity. Under the above conditions, the detection limits (S/N=3) of 1-, 2-NPs and 2-NFR were 1 fmol (0.25 pg), 10 fmol (2.5 pg) and 4 fmol (1 pg), respectively. The proposed system was effectively used to determine trace levels of 1-, 2-NPs and 2-NFR in airborne particulates collected at Noto Peninsula. The atmospheric concentrations of 1-, 2-NPs and 2-NFR were not more than sub pg $m^{-3}$ levels. They were higher in winter (January) than in summer (July). In both seasons, the concentrations were in decreasing order, [2-NFR]>[1-NP]>[2-NP].