• Title/Summary/Keyword: high performance liquid chromatography/mass spectrometry

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Antibacterial and Antibiofilm Activities of Diospyros malabarica Stem Extract against Streptococcus mutans (Streptococcus mutans에 대한 인도감나무 줄기 추출물의 항균활성 및 생물막 형성 억제 효과)

  • Kim, Hye Soo;Lee, Sang Woo;Sydara, Kongmany;Cho, Soo Jeong
    • Journal of Life Science
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    • v.29 no.1
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    • pp.90-96
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    • 2019
  • The objective of this study was to evaluate the potential of Diospyros malabarica stem extract, a natural materials, in oral health material. With this aim in mind, thin layer chromatography (TLC), TLC-bioautography, high-performance liquid chromatography (HPLC), electrospray ionization-mass spectrometry (ESI-MS), scanning electron microscopy (SEM), and real-time qPCR were performed. The antibacterial activity of D. malabarica stem extract against Streptococcus mutans KCTC3065 was confirmed in an n-hexane fraction with low polarity. The molecular weight of the antibacterial compound was estimated to be 188 by ESI-MS analysis. The inhibitory effects of the extract on biofilm formation and gene expression related to biofilm formation of S. mutans were determined by SEM and real-time PCR analysis. The extract inhibited the formation of S. mutans biofilms at D. malabarica stem extract concentrations of 1 mg/ml, as shown by SEM. The real-time PCR analysis showed that the expression of the gtfC gene, which is associated with biofilm formation, was significantly decreased in a dose-dependent manner. Based on the above results, it can be concluded that D. malabarica stem extracts, a natural materials, can be used in oral health products to suppress the formation of biofilms by inhibiting tooth adhesion of S. mutans, a causative agent of dental caries.

Determination of Phthalate Metabolites in Korean Children's Urine by High Performance Liquid Chromatography with Triple Quadrupole Tandem Mass Spectrometry (LC/MS/MS를 이용한 어린이 소변 중 프탈레이트 대사산물 분석)

  • Kho, Young-Lim;Jeong, Jee-Yeon;Choi, Kyung-Ho;Kim, Pan-Gyi
    • Journal of Environmental Health Sciences
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    • v.34 no.4
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    • pp.271-278
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    • 2008
  • Dialkylated phthalates have been commonly used as plasticizers and a variety of applications. Phthalate diesters have been shown to be developmental and reproductive toxicants. It is very difficult to exactly estimate the dose of dialkylated phthalates taken up by the general population because of environmental contamination. Urinary metabolites of phthalates enabled to estimate internal exposure. The objective of this study was quantitative determination of phthalate metabolites by LC/MS/MS with on-line cleanup method to analyze phthalate metabolites in Korean children's urine. We employed LC/MS/MS with on-line enrichment and column-switching techniques for this biological monitoring. Metabolites determined were 4 primary metabolites; MEHP, MnBP, MiBP, MEP and 2 secondary metabolites of DEHP; 5-OH-MEHP), 5-oxo-MEHP. We analyzed children's urine from 30 boys and 30 girls. The method detection limit of phthalate metabolites were 0.03 ng/mL for MEP, 1.05 ng/mL for MBP, 0.22 ng/mL for MEHP, 0.15 ng/mL for 5-OHMEHP and 0.16 ng/mL for 5-oxo-MEHP, respectively. Switching Column LC/MS/MS was proven to be a useful tool to determine metabolites of phthalate diesters in human urine. The correlation among phthalate metabolites was very high and statistically significant, except MEP. The children's age (months) was negatively correlated to the concentration of phthalate metabolites. The geometric mean concentration of phthalate metabolites (mg/g creatinine) in children's urine were 25.5 for MEP, 130.3 for MnBP, 56.8 for MiBP, 19.5 for MEHP, 85.6 for 5-OH-MEHP and 83.1 for 5-oxo-MEHP, respectively. Levels of estimated daily intake of parent phthalate compounds (${\mu}g$/kg bw/day) were 0.8 for DEP, 5.0 for DnBP, 1.9 for DiBP and $8.9{\sim}14.2$ for DEHP, respectively. Estimated daily intake for DEP and DiBP were lower than those of other studies but the value for DEHP was higher than that of other study.

MTHFR, As3MT and GSTO1 Polymorphisms Influencing Arsenic Metabolism in Residents Near Abandoned Metal Mines in South Korea (폐금속광산 지역 주민의 비소 대사에 영향을 미치는 MTHFR, As3MT, GSTO1 유전자 다형성)

  • Surenbaatar, Ulziikhishig;Kim, Byoung-Gwon;Son, Hyun-Jin;Cho, Seong-Sik;Kim, Gwon-Min;Lim, Hyoun-Ju;Kwon, Jung-Yeon;Kim, Ki-Hwan;Hong, Young-Seoub
    • Journal of Environmental Health Sciences
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    • v.47 no.6
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    • pp.530-539
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    • 2021
  • Background: In South Korea, areas around abandoned metal mines are designated as regions with high arsenic (As) contamination. However, studies assessing urinary As exposure, As metabolism, and relevant genetic polymorphisms in residents of these metal mine areas are lacking. Objectives: To identify factors associated with As exposure and evaluate the effects of MTHFR, As3MT, and GSTO1 genetic polymorphisms on As metabolism in residents of abandoned metal mine areas by measuring urinary As species. Methods: Urinary As species (arsenite [As3+], arsenate [As5+], monomethyl arsonic acid, and dimethylarsinic acid) were isolated using high-performance liquid chromatography in combination with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). Four genetic polymorphisms (MTHFR A222V, MTHFR E429A, GSTO1 A140D, As3MT M287T) were analyzed in 144 residents of four areas around abandoned metal mines. Results: The study sample was comprised of 34.7% men and 65.3% women, with a mean age of 70.7±10.9 years. The urinary inorganic As concentration was higher among those consuming more than half locally produced rice (0.31 ㎍/L) than those consuming less than half such rice (0.18 ㎍/L). The urinary dimethylarsinic acid concentration was higher in the group that had consumed seafood in the past day (31.68 ㎍/L) than in those who had not (22.37 ㎍/L). Furthermore, individuals heterozygous in the MTHFR A222V and GSTO1 A140D polymorphism had higher urinary arsenic species concentrations than did individuals with a wild type or homozygous for the variant allele. Conclusions: Consumption of locally produced rice was associated with inorganic As exposure, whereas seafood consumption was associated with organic As exposure among residents of abandoned metal mine areas. There was no clear association between MTHFR A222V and GSTO1 A140D polymorphisms and As metabolism.

Monitoring of Benzoic Acid, Sorbic Acid, and Propionic Acid in Spices (향신료에서 유래되는 안식향산, 소브산, 프로피온산의 함유량 조사)

  • Yun, Sang Soon;Lee, Sang Jin;Lim, Do Yeon;Lim, Ho Soo;Lee, Gunyoung;Kim, MeeKyung
    • Journal of Food Hygiene and Safety
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    • v.32 no.5
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    • pp.381-388
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    • 2017
  • In this study, we investigated the levels of natural preservatives of benzoic acid, sorbic acid, and propionic acid in spices. The quantitative analysis was performed using high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) for benzoic acid and sorbic acid and gas chromatography-mass spectrometry (GC-MS) for propionic acid. The sample was extracted with ethanol using sonication, then centrifuged and evaporated to dryness and redissolved to 1 mL with ethanol to use for the instrumental analysis. The analytical method was validated based on linearity, recovery, limit of detection (LOD), and limit of quantification (LOQ). This method was suitable to determine low amounts of naturally occurring preservatives (benzoic acid, sorbic acid, and propionic acid) in various spices. Benzoic acid, sorbic acid, and propionic acid were found in 165 samples, 88 samples, and 398 samples, respectively from the total of 493 samples. The concentration of benzoic acid, sorbic acid, and propionic acid were ranged at ND-391.99 mg/L, ND-57.70 mg/L, and ND-188.21 mg/L in spices, respectively. The highest mean levels of benzoic acid, sorbic acid, and propionic acid were found in cinnamon (167.15 mg/L), basil leaves (22.79 mg/L), and white pepper (51.48 mg/L), respectively. The results in this study provide ranges of concentration regarding naturally occurring benzoic acid, sorbic acid, and propionic acid in spices. Moreover, the results may use to the case of consumer complaint or trade friction due to the inspection services of standard criteria for the preservatives of spices.

Component Analysis of Suaeda asparagoides Extracts (나문재 추출물의 성분 분석)

  • Yang, Hee-Jung;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.3
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    • pp.157-165
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    • 2008
  • In the previous study, the anti-oxidant activity of oxtract/fraction of Sueada aspparagoides(SA) and the stability test for the cream containing SA extract were investigated respectively[1,2]. In this study, the components of SA extract were analyzed by TLC, HPLC, and LC/ESI-MS/MS, $^1H$-NMR. TLC chromatogram of ethyl acetate fraction of SA extract revealed 5 bands $(SA1{\sim}SA5)$. HPLC chromatogram of aglycone fractions obtained from deglycoylation reaction of ethyl acetate fraction showed 2 bands (SAA 2 and SAA 1), which were identified as quercetin (composition ratio, 16.88%) and kaempferol (83.12%) in the order of elution time. Among 5 bands of TLC chromatogram, 4 bands $(SA2{\sim}SA5)$ also were Identified as kaempferol-3-O-glucoside (SA 2), quercetin-3-O-glucoside (SA3), kaempferol-3-O-rutinoside (SA 4), quercetin-3-O-rutinoside (SA 5) by LC/ESI-MS/MSMS/MS. respectively. The spectrum generated for SAA 1 by LC/ESI-MS/MS in the negative ion mode also gave the ion corresponding to the deprotonated aglycone $[M-H]^-$ (285m/z), the $^1H$-NMR spectrum contained signals [${\delta}$ 6.19 (1H, d, J=1.8Hz, H-6), ${\delta}$ 6.44 (1H, d, J=1.8Hz, H-8), ${\delta}$ 6.92 (2H, d, J=9.0Hz, H-3', 5'), ${\delta}$ 8.04 (2H, d, J=9.0Hz, H-2', 6', thus SAA 1 was identified as kaempferol. SAA 2 yielded the deprotonated agycone ion $[M-H]^-$ (301m/z), $^1H$-NMR spectrum showed signals [${\delta}$ 6.20 (1H, d, J=2.0Hz, H-6), ${\delta}$ 6.42 (1H, d, J=2.0Hz, H-8), ${\delta}$ 6.90 (1H, d, J=8.6Hz, H-5'), ${\delta}$ 7.55 (1H, dd, J=8.6, 2.2Hz, H-6'), ${\delta}$ 7.69 (1H, d, J=2.2Hz, H-2', thus SAA 2 was Identified as quercetin. In conclusion, with the anti-oxidant activity and the stability test reported previously, component analysis of SA extracts could be applicable to new cosmeceuticals.

Identification and Quantification of Glucosinolates in Rapeseed (Brassica napus L.) Sprouts Cultivated under Dark and Light Conditions

  • Lee, Min-Ki;Arasu, Mariadhas Valan;Chun, Jin-Hyuk;Seo, Jeong Min;Lee, Ki-Teak;Hong, Soon-Taek;Kim, In Ho;Lee, Yong-Hwa;Jang, Young-Seok;Kim, Sun-Ju
    • Korean Journal of Environmental Agriculture
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    • v.32 no.4
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    • pp.315-322
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    • 2013
  • BACKGROUND: This study was performed for the identification and quantification of glucosinolate (GSL) contents in seven varieties of rapeseed (Brassica napus L.) sprouts cultivated under dark and light conditions. METHODS AND RESULTS: Crude glucosinolates (GSLs) were desulfated by treating with aryl sulfatase and purified using diethylaminoethyl sepharose (DEAE) anion exchange column. Individual GSLs were quantified using high-performance liquid chromatography (HPLC) with electrospray ionization-tandem mass spectrometry (ESI-MS/MS). Eleven GSLs including six aliphatic (progoitrin, sinigrin, glucoalyssin, gluconapoleiferin, gluconapin, and glucobrassicanapin), four indolyl (4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, and neoglucobrassicin) and one aromatic (gluconasturtiin) were identified based on the fragmentation patterns of MS spectrum. Aliphatic GSLs were noted as the predominant group with average 85.2% of the total contents. The most abundant GSLs were progoitrin which was ranged at $8.14-118.68{\mu}mol/g$ dry weight (DW). The highest total GSL amounts were documented in 'Hanra' ($146.02{\mu}mol/g$ DW) under light condition and 'Mokpo No. 68' ($86.67{\mu}mol/g$ DW) in dark condition, whereas the lowest was in 'Tamra' (30.13 and $14.50{\mu}mol/g$ DW) in both conditions. The sum of aliphatic GSLs attributed > 80% in all varieties, except 'Tamra' (67.7% and 64.9% in dark and light conditions, respectively) in the total GSL accumulation. Indolyl GSLs were ranged $2.41-15.73{\mu}mol/g$ DW, accounted 2.78-33.6% of the total GSLs in rapeseed varieties. CONCLUSION(S): These results provide valuable information regarding potential beneficial GSL contents individually. This study attempts to contribute to knowledge of the nutritional properties of the different varieties of rapeseed plants. These results may be useful for the evaluation of dietary information.

Nutrikinetic study of fermented soybean paste (Cheonggukjang) isoflavones according to the Sasang typology

  • Kim, Min Jung;Lee, Da-Hye;Ahn, Jiyun;Jang, Young-Jin;Ha, Tae-Youl;Do, Eunju;Jung, Chang Hwa
    • Nutrition Research and Practice
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    • v.14 no.2
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    • pp.102-108
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    • 2020
  • BACKGROUND/OBJECTIVES: In Oriental medicine, certain foods may be beneficial or detrimental based on an individual's constitution; however, the scientific basis for this theory is insufficient. The purpose of this study was to investigate the effect of body constitution, based on the Sasang type of Korean traditional medical classification system, on the bioavailability of soy isoflavones of Cheonggukjang, a quick-fermented soybean paste. SUBJECTS/METHODS: A pilot study was conducted on 48 healthy Korean men to evaluate the bioavailability of isoflavone after ingestion of food based on constitution types classified by the Sasang typology. The participants were classified into the Taeeumin (TE; n = 15), Soyangin (SY; n = 15), and Soeumin (SE; n = 18) groups. Each participant ingested 50 g of Cheonggukjang per 60 kg body weight. Thereafter, blood was collected, and the soy isoflavone metabolites were analyzed by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry. Ntrikinetic analysis of individual isoflavone-derived metabolites was performed. RESULTS: Our nutrikinetic analysis identified 21 metabolites derived from isoflavones in the blood samples from 48 healthy Korean men (age range, 21-29 years). Significant differences were observed in the time to maximum concentration (Tmax) and elimination half-life (t1/2) for nine metabolites among the three groups. The Tmax and t1/2 of the nine metabolites were higher in the SE group than in the other groups. Moreover, the absorption rates, as determined by the area under the plasma-level curve (AUC) values of intact isoflavone, were 5.3 and 9.4 times higher in the TE group than in the SY and SE groups, respectively. Additionally, the highest AUC values for phase I and II metabolites were observed in the TE group. CONCLUSIONS: These findings indicate that isoflavone bioavailability, following Cheonggukjang insgestion, is high in individuals with the TE constitution, and relatively lower in those with the SE and SY constitutions.

Bioequivalence of PirosTM Tablets to ActosTM Tablets (Pioglitazone HCl 15 mg) (액토스TM정(염산 피오글리타존 15 mg)에 대한 피로스TM정의 생물학적 동등성)

  • Kang, Jong-Min;Lee, Myung-Jae;Seo, Ji-Hyung;Choi, Sang-Jun;Lee, Jin-Sung;Ryu, Ju-Hee;Tak, Sung-Kwon;Yim, Sung-Vin;Hong, Seung-Jae;Lee, Kyung-Tae
    • Journal of Pharmaceutical Investigation
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    • v.39 no.2
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    • pp.133-139
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    • 2009
  • The purpose of the present study was to evaluate the bioequivalence of two pioglitazone HCl tablets, $Actos^{TM}$, tablets (Lilly Korea. Ltd., Korea) as a reference drug and $Piros^{TM}$, tablets (Reyon Pharm. Co., Ltd., Korea) as test drug, according to the guideline of Korea Food and Drug Administration (KFDA). Twenty-four healthy male Korean volunteers received one tablet containing pioglitazone HCl 15 mg in a $2{\times}2$ crossover study. There was a one-week washout period between the doses. Plasma concentrations of pioglitazone were monitored for over a period of 36 hr after administration by using a high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). The area under the plasma concentration-time curve from time zero to 36 hr ($AUC_{0-36hr}$), maximum plasma drug concentration ($C_{max}$) and time to reach $C_{max}$ ($T_{max}$) were complied from the plasma concentration-time data. Analysis of variance (ANOVA) test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_{0-36hr}$ and $C_{max}$. The 90% confidence intervals of the $AUC_{0-36hr}$ ratio and the $C_{max}$ ratio for $Piros^{TM}$/$Actos^{TM}$. were log 0.8753-log 1.1286 and log 0.8669-log 1.1734, respectively. These values were within the acceptable bioequivalence intervals of log 0.80-log 1.25, recommended by KFDA. In all of these results, we concluded that the $Piros^{TM}$. tablet was bioequivalent to the $Actos^{TM}$. tablet, based on the rate and extent of absorption.

Identification of a Bioactive Compound, Violacein, from Microbulbifer sp. Isolated from a Marine Sponge Hymeniacidon sinapium on the West Coast of Korea (한국 서해안에 서식하는 주황해변해면에서 분리된 해양세균 Microbulbifer sp.으로부터 생리활성물질 비올라세인의 규명)

  • Won, Nam-Il;Lee, Ga-Eun;Ko, Keebeom;Oh, Dong-Chan;Na, Yang Ho;Park, Jin-Sook
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.124-132
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    • 2017
  • Microbial secondary metabolites of marine organisms are regarded as major sources of structurally and biologically novel compounds with numerous potential uses. Sponge-microbe associations are among the most interesting sources for exploring bioactive compounds. In this study, the bacterial strain Microbulbifer sp. (127CP7-12) was isolated from the Asian marine sponge Hymeniacidon sinapium collected at an intertidal zone on the west coast of Korea. Cultured bacteria produced a violet pigment, and optimal culture conditions for violet pigment production were investigated. Maximum production of the violet pigment from the strain culture was observed under the conditions of $25^{\circ}C$, pH 6.0, and 3% NaCl. Acetone provided better extraction of the pigment from fermented broth compared with ethanol and methanol. The proposed structure of the major component in the extracted crude pigment was determined via high-performance liquid chromatography, nuclear magnetic resonance, mass spectrometry, and UV spectra analyses, which showed that the metabolite was the promising bioactive compound violacein. This study describes the examination of marine bioactive materials from microbe-engaged metabolites and the ecological implications of the sponge-microbe association in a changing ocean.

Metabolomes and transcriptomes revealed the saponin distribution in root tissues of Panax quinquefolius and Panax notoginseng

  • Wei, Guangfei;Yang, Feng;Wei, Fugang;Zhang, Lianjuan;Gao, Ying;Qian, Jun;Chen, Zhongjian;Jia, Zhengwei;Wang, Yong;Su, He;Dong, Linlin;Xu, Jiang;Chen, Shilin
    • Journal of Ginseng Research
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    • v.44 no.6
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    • pp.757-769
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    • 2020
  • Background: Panax quinquefolius and Panax notoginseng are widely used and well known for their pharmacological effects. As main pharmacological components, saponins have different distribution patterns in the root tissues of Panax plants. Methods: In this study, the representative ginsenosides were detected and quantified by desorption electrospray ionization mass spectrometry and high-performance liquid chromatography analysis to demonstrate saponin distribution in the root tissues of P. quinquefolius and P. notoginseng, and saponin metabolite profiles were analyzed by metabolomes to obtain the biomarkers of different root tissues. Finally, the transcriptome analysis was performed to demonstrate the molecular mechanisms of saponin distribution by gene profiles. Results: There was saponin distribution in the root tissues differed between P. quinquefolius and P. notoginseng. Eight-eight and 24 potential biomarkers were detected by metabolome analysis, and a total of 340 and 122 transcripts involved in saponin synthesis that were positively correlated with the saponin contents (R > 0.6, P < 0.05) in the root tissues of P. quinquefolius and P. notoginseng, respectively. Among them, GDPS1, CYP51, CYP64, and UGT11 were significantly correlated with the contents of Rg1, Re, Rc, Rb2, and Rd in P. quinquefolius. UGT255 was markedly related to the content of R1; CYP74, CYP89, CYP100, CYP103, CYP109, and UGT190 were markedly correlated with the Rd content in P. notoginseng.