• Title/Summary/Keyword: LC-MS-MS

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Simultaneous Characterization of Sofalcone and Its Metabolite in Human Plasma by Liquid Chromatography -Tandem Mass Spectrometry

  • Han, Sang-Beom;Jang, Moon-Sun;Lee, Hee-Joo;Lee, Ye-Rie;Yu, Chong-Woo;Lee, Kyung-Ryul;Kim, Ho-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.26 no.5
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    • pp.729-734
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    • 2005
  • A sensitive and selective method for quantitation of sofalcone and its active metabolite in human plasma has been established using liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS). Plasma samples were transferred into 96-well plate using an automated sample handling system and spiked with 10 $\mu$L of 2 $\mu$g/mL $d_3$-sofalcone and $d_3$-sofalcone metabolite solutions (internal standard), respectively. After adding 0.5 mL of acetonitrile to the 96-well plate, the plasma samples were then vortexed for 30 sec. After centrifugation, the supernatant was transferred into another 96-well plate and completely evaporated at 40 ${^{\circ}C}$ under a stream of nitrogen. Dry residues were reconstituted with mobile phase and were injected into a $C_{18}$ reversed-phase column. The limit of quantitation of sofalcone and its metabolite was 2 ng/mL, using a sample volume of 0.2 mL for analysis. The reproducibility of the method was evaluated by analyzing 10 replicates over the concentration range of 2 ng/mL to 1000 ng/mL. The validation experiments of the method have shown that the assay has good precision and accuracy. Sofalcone and its metabolite produced a protonated precursor ion ([M+H]$^+$) of m/z 451 and 453, and a corresponding product ion of m/z 315 and 317, respectively. Internal standard ($d_3$-sofalcone and $d_3$-sofalcone metabolite) produced a protonated precursor ion ([M+H]$^+$) of m/z 454 and 456 and a corresponding product ion of m/z 315 and 317, respectively. The method has been successfully applied to a pharmacokinetic study of sofalcone and its active metabolite in human plasma.

Environmental Exposure to Tobacco-specific Nitrosamines in an Area Near a Fertilizer Plant (비료제조공장 인근 지역의 담배특이니트로사민 환경 노출)

  • Ha, Jae-Na;Yoon, Mi-Ra;Chang, Jun Young;Koh, Dohyun;Shin, Ho-Sang;Kim, Suhyang;Lee, Chul-Woo;Lee, Bo-Eun;Kim, Jeong-Soo
    • Journal of Environmental Health Sciences
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    • v.46 no.4
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    • pp.398-409
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    • 2020
  • Objectives: This study aimed to evaluate environmental exposure to tobacco-specific nitrosamines (TSNAs) by conducting an analysis of the concentration of TSNAs in deposited dust collected from a fertilizer plant and the surrounding village, a simulation of high-temperature drying of tobacco waste, and CALPUFF modeling. Methods: The raw materials of the products, deposited dust (inside and outside the plant and residential area), soil, and wastewater were sampled and the TSNA concentrations were analyzed by LC-MS/MS. As the plant was closed down before the investigation, simulation tests were conducted to confirm the substances discharged during high-temperature (300℃) drying of tobacco waste. CALPUFF modeling was performed to identify the area of influence due to exposure to TSNAs. Results: TSNAs were detected in organic fertilizers estimated to contain tobacco waste, deposited dust, and soil collected from inside and outside the plant. N'-nitrosonornicotine (NNN), 4-(methylnitrosoamino)-1-(3-pyridyl)-1-butanone (NNK), and N'-nitrosoanatabine (NAT) components were detected in five of 15 deposited dust samples collected from the residential area around the plant, while TSNAs were not detected in the five sampling points in the control area. Also, the simulation test for the high temperature drying of tobacco waste found emissions of TSNAs. The CALPUFF modeling results showed that the survey area was likely to be included in the area of influence of TSNA emissions from the plant. Conclusions: It is estimated that harmful tobacco ingredients such as TSNAs were dispersed in nearby areas due to the illegal use of tobacco waste as a raw material to produce organic fertilizers at the plant. These findings assume that the residents have been exposed to TSNAs and suggest that the need for the establishment of measures to manage environmental health.

Estimation of the Daily Human Intake of Acrylamide (AA) Based on Urinary N-acetyl-S-(2-carbamoylethyl)-cysteine (AAMA) and the Contribution of Dietary Habits in South Korean Adults (요중 AAMA에 의한 한국 성인 아크릴아마이드(AA)의 하루섭취량 추정 및 기여 식습관에 대한 분석)

  • LEE, Jin-Heon;LEE, Kee-Jae;KANG, Hee-Sook
    • Journal of Environmental Health Sciences
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    • v.42 no.4
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    • pp.235-245
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    • 2016
  • Objectives: This study estimated the adult Korean daily intake of acrylamide (AA) and investigated its relationship with demographic, lifestyle and dietary habits by using urinary concentrations of N-acetyl-S-(2-carbamoylethyl)-cysteine (AAMA). Methods: Human data (n=1870) was collected in a nationwide cross-sectional biomonitoring program representing the population (18-69 years) residing in South Korea. Urinary AAMA was analyzed with a LC-MS/MS system. Daily intakes of AA were estimated using mass daily AAMA, which was calculated through urinary AAMA concentration and daily creatinine excretion. Statistical analysis was performed with SAS procedures for calculating geometric means, confidence intervals and the exponentiated beta coefficient of multiple linear regressions. Results: Daily intake of AA was estimated at $0.475{\mu}g/kg$ body weight (BW) per day (95% confidence interval (CI): 0.447-0.503). In the case of current smokers, AA intake was $0.957{\mu}g/kg$ BW per day (95% CI: 0.847-1.067), which was significantly higher than that of former smokers and never smoked (p<0.0001). The strong affecting factors were age (95% CI: 0.68-1.14; p=0.0180), education level (95% CI: 1.05-1.42; p=0.0163), body mass index (BMI) (95% CI: 1.00-1.82; p<0.0001), and smoking status (95% CI: 0.97-3.05; p<0.0001). Korean dietary habits increasing AA intake were coffee (p=0.0005), cup noodles (p=0.0010) and canned foods (p=0.0005). Meanwhile, foods decreasing AA intake were fresh fruit (p=0.0076), cooked beef (p=0.0335) and cooked pork (p=0.0147). Conclusion: The Korean daily intake of AA in adults was estimated to be similar with those found in developed countries. The factors increasing daily AA intake were coffee, cup noodles and canned foods, and decreasing factors were fresh fruit, cooked beef and cooked pork.

A Diagnostic Algorithm after Newborn Screening for 21-hydroxylase Deficiency (선천성 부신 과형성증(21-hydroxylase 결핍)의 신생아 선별 검사 후 진단 알고리즘)

  • Cho, Sung Yoon;Ko, Jung Min;Lee, Kyung-A
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.16 no.2
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    • pp.70-78
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    • 2016
  • 21-hydroxylase deficiency (21-OHD), most common form of congenial adrenal hyperplasia, is categorized into classical forms, including the salt-wasting (SW) and the simple virilizing (SV) types, and nonclassical (NC) forms based on the severity of the disease. Newborn screening for 21-OHD has been performed in Korea since 2006. $17{\alpha}$-hydroxyprogesterone (17-OHP) is a marker for 21-OHD and is measured using a radioimmunoassay or a fluoroimmunoassay. Premature and low birth weight infants are likely to give false positive 17-OHP findings, therefore, cutoff values for these infants should be determined based on gestational weeks or birth weight. ACTH simulation test is helpful when the 17-OHP shows equivocal increase, and it is gold standard for diagnosis of NC type. Recently, liquid chromatography linked with tandem mass spectrometry was developed for rapid, highly specific, and sensitive analysis of multiple analytes. Molecular analysis of CYP21A2 is useful for confirming diagnosis of mild SV or NC type, predicting prognoses, and genetic counseling. In order to make newborn screening for 21-OHD more efficient, early detection of boy with SW type, early determination of girl with ambiguous genitalia, detection of NC type, and overcoming of false positive in premature and low birth weight infants should be considered. Above all, early treatment should be started when the patient is suspected as having 21- OHD clinically before confirming the diagnosis to prevent adrenal crisis. Here, author reviewed recent articles of guideline and proposed guideline for 21-OHD.

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Regulatory mechanism of Angelica Gigas extract powder on matrix metalloproteinases in vitro and in vivo model (참당귀 추출분말이 in vitro and in vivo model에서 MMPs 조절 기전)

  • Kwon, Jin-Hwan;Han, Min-Seok;Lee, Yong-Moon
    • Analytical Science and Technology
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    • v.28 no.6
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    • pp.361-369
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    • 2015
  • The precise mechanism underlying the therapeutic efficacy of an extraction powder of Angelica gigas (AGE) for the treatment of degenerative osteoarthritis was investigated in primary cultured rabbit chondrocytes and in a monosodium-iodoacetate (MIA)-induced osteoarthritis rat model. The treatment with AGE (50 μg/mL) effectively inhibited NF-B activation. The anti-inflammatory mechanism was clarified by gelatin zymography and western blotting measurements of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) activities. The AGE (50 μg/mL) treatment significantly reduced MMP-9 activity. The constituents of AGE— decursinol, decursin, and decursinol angelate—were determined by LC-MS/MS after a 24 hr treatment of rabbit chondrocytes. The contents of the major products, decursin and decursinol angelate, were 3.62±0.47 and 2.14 ±0.36 μg/mg protein, respectively in AGE-treated (50 μg/mL) rabbit chondrocytes. An in vivo animal study on rats fed a diet containing 25, 50, and 100 mg/kg AGE for 3 weeks revealed a significant inhibition of the MMPs in the MIA-induced rat articular cartilage. The genetic expression of arthritic factors in the articular cartilage was examined by RT-PCR of collagen Type I, collagen Type II, aggrecan, and MMP (MMP3, MMP-9, MMP13). Specifically, AGE up-regulated the expression of collagen Type I, collagen Type II, and aggrecan and inhibited MMP levels at all tested concentrations. Collectively, AGE showed a strong specific site of action on MMP regulation and protected against the degeneration of articular cartilage via cellular regulation of MMP expression both in vitro and in vivo.

Pharmacokinetic Properties and Bioequivalence of Cefcapene Pivoxil Hydrochloride 75 mg in Korean Healthy Volunteers (건강한 한국인 피험자에서 Cefcapene Pivoxil Hydrochloride 75 mg 제제의 생물학적동등성시험)

  • Jeon, Ji-Young;Im, Yong-Jin;Hwang, Min-Ho;Kim, Yun-Jeong;Han, Su-Mi;Jo, Myeong-Jin;Kim, Hee-Sun;Kim, Sun-Young;Chae, Soo-Wan;Kim, Kang-Seog;Kim, Min-Gul
    • Korean Journal of Clinical Pharmacy
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    • v.22 no.1
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    • pp.9-12
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    • 2012
  • Background: Cefcapene pivoxil hydrochloride, is an ester-type oral cephem antibiotic. This study was performed to compare the pharmacokinetics and evaluate the bioequivalence of two cefcapene pivoxil hydrochloride 75 mg formulations. Method: In a randomized $2{\times}2$ crossover study, sixty healthy male volunteers were randomly assigned into two groups. After a single dose of 75 mg cefcapene pivoxil hydrochloride oral administration, blood samples were collected at specific time intervals from 0-12 hours. The plasma concentrations of cefcapene pivoxil hydrochloride were determined by LC-MS/MS. The pharmacokinetic parameters were determined from the plasma concentration-time profiles of both formulations. The pharmacokinetic parameters such as $AUC_{last}$, $AUC_{inf}$ and $C_{max}$, were calculated and the 90% confidence intervals for test/reference ratio for pharmacokinetic parameters were obtained by analysis of variance on logarithmically transformed data. Results: The mean value for $AUC_{last}$ in test and reference drug was $4053.1{\pm}876.5\;ng{\cdot}hr/mL$ and $3595.7{\pm}1029.1\;ng{\cdot}h/mL$, respectively. The mean value for $C_{max}$ in test and reference drug was $1324.9{\pm}321.4$ ng/mL and $1159.1{\pm}335.9$ ng/mL, respectively. The 90% confidence intervals of the $AUC_{last}$ and $C_{max}$ ratio for test drug and reference drug were log 1.09-log1.22 and log 1.09-log1.24, respectively. No adverse events were reported by subjects or found on analysis of vital signs or laboratory tests. Conclusion: This single dose study found that the test and reference products met the regulatory criteria for bioequivalence in these health volunteers. Both formulations were safe and well tolerated in 75 mg of cefcapene pivoxil hydrochloride.

Study on Monascus Strains and Characteristic for Manufacturing Red Yeast Rice with High Production of Monacolin K (Monacolin K 강화 홍국쌀 생산을 위한 균주 및 특성 연구)

  • Park, Ji-Young;Han, Sang-Ik;Seo, Woo Duck;Ra, Ji-Eun;Sim, Eun-Yeong;Nam, Min-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.2
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    • pp.167-173
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    • 2014
  • Red yeast rice (RYR) is the product of fermented yeast by Monascus strains on rice, and has recently become a popular dietary supplement as a traditional food in Asia. RYR contains monacolin K substances known to inhibit cholesterol synthesis as efficiently as statin drugs. To determine the optimal rice cultivar for manufacturing RYR, 7 rice cultivars (Goami, Goami2, Sangjuchalbyeo, Seolgaeng, Saegyejinmi, Yeonghojinmi and Chilbo) were fermented using two Monascus strains (M. ruber KCTC6122 and KCCM60141 of M. ruber) in this study. The monacolin K content of Sangjuchalbyeo were 47.24 ppm on KCTC6122 cultures and 117.03 ppm on KCCM60141 cultures, respectively. Other cultivars, especially Goami and Goami2, which had less content of monacolin K could not seem to ferment normally because those didn't show red color. These results imply that Sangjuchalbyeo can be optimal rice cultivar as a commercial RYR which is well fermented rice and has high content of monacolin K.

Comparison of Chemical Constituents in Mung bean (Vigna radiata L.) Flour between Cultivation Regions and Seeding Dates

  • An, Yeon Ju;Kim, Mi Jung;Han, Sang Ik;Chi, Hee-Youn;Kwon, Chang;Kim, So Yeon;Yang, Yu Jin;Kim, Yun Ju;Moon, Hee Sung;Kim, Seung-Hyun;Chung, Ill-Min
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.4
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    • pp.457-467
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    • 2020
  • Legumes are one of the largest families of crop plants and are widely consumed and produced for their nutritional and commercial benefits. Mung bean (Vigna radiata L.) is a legume crop that contains various functional compounds ; moreover, it has strong antioxidant properties and is becoming an increasingly important food crop. However, most previous studies on mung beans have focused on their primary metabolites. In this study, we investigated the composition and contents of phenolic compounds, fatty acids, soyasapogenol and tocopherol in mung beans cultivated in different regions and cultivated at different seeding dates. Material analysis was conducted using the following methods: LC-MS/MS, GC-FID and HPLC-ELSD. In total, 57 different samples were analyzed. Thirteen phenolic compounds were detected in mung beans. Of these, vitexin and isovitexin were the most abundant compounds, accounting for approximately 99% of phenolic compounds. The difference in phenol compounds according to the seeding dates of mung bean was not statistically significant. The total fatty acid content in beans was the highest in Pyeongchang. Significant differences in total fatty acid content were found according to the cultivation regions. Crops grown in Sohyeon and Dahyeon showed the highest soyasapogenol B content in the Suwon region, and these were the lowest in Jeonju. The total tocopherol content of beans cultivated in Dahyeon and Sohyeon was the lowest and highest in Pyeongchang. Soyasapogenol B and total tocopherol content were not significantly different according to seeding dates. This study was conducted to obtain basic data for the cultivation of mung beans with a high content of various functional materials in terms of regional specialization and optimal seeding time.

Functional characterization of domestic and foreign green tea cultivars at different harvest periods (채취시기가 다른 국내외 녹차잎의 기능성분 함량, 뇌세포 생존 및 대사 효소 활성 조절 효과 조사)

  • Lee, Bang-Hee;Jeon, Sae Hyun;Jeong, Hana;Choi, Jung;Kim, Young-Min;Yang, Kwang-Yeol;Nam, Seung-Hee
    • Korean Journal of Food Science and Technology
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    • v.52 no.5
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    • pp.427-434
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    • 2020
  • This study was performed to compare nutritional compounds and physiological functions of five domestic and imported green tea cultivars at three time points. The five cultivars were compared for theanine, γ-aminobutyric acid, and catechin content by LC-MS/MS and HPLC. Furthermore, the five tea cultivars were functionally characterized with respect to antioxidant activity, brain cell protective effect, and inhibitions of α-glucosidase and HMG-CoA reductase activities. Among green tea cultivars, Chamnok had the highest content of catechins (198 mg/g DW), theanine (11.89 mg/g DW), and tannin (23.6 mg/g DW). Considering functional properties, Chamnok treatment resulted in the maximum viability of brain cells and reduced the cortisol content of SH-SY5Y cells. The inhibition of α-glucosidase and HMG-CoA reductase was the strongest following Chamnok treatment (72.9% and 69.8%, respectively). These results indicate that Chamnok could be optimal for consumption or favorable processing owing to its high nutritional compounds, such as theanine and catechin, and remarkable brain cell protective effects.

A study of the metabolites for 7-keto-DHEA-acetate in human urine (II) (뇨시료에 함유된 7-keto-DHEA-acetate의 대사체에 관한 연구 (II))

  • Kim, Yunje;Lee, Jinhee
    • Analytical Science and Technology
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    • v.17 no.5
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    • pp.401-409
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    • 2004
  • The 7-keto-DHEA-acetate is converted to 7-keto-DHEA, a metabolite of DHEA, and similar to its metabolism. We studied the metabolite M3, M4, and M5 of 7-keto-DHEA-acetate. The estimated molecular weight of M3 and M4 was 304 which were supposed to have more 3 hydroxyl and/or ketone groups. We could know that M3 is the 7-OH-DHEA which has the hydroxyl groups on 3 and 7-carbon and a ketone group on 17-carbon. In case of M4, it is the 7-oxo-diol metabolite which has the hydroxyl groups on 3 and 17-carbon and a ketone group on 7-carbon. The M5 was supposed that the molecular weight is 320 and has the three hydroxyl groups on 3, 6, and 16 carbon and the ketone group on 17-carbon. After dosing, 7-OH-DHEA showed the maximum urine flow in human urine after 5 hr and decreased rapidly. But we could find it until 58 hr why is a higher remaining substance.