• Title/Summary/Keyword: high performance liquid chromatography

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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.

Analysis of Flumeqnine in Meats (식육 중 항생제 flumequine의 분석)

  • Kuk, Ju-Hee;Ko, Yong-Seok;Kim, Yong-Hoon;Yoon, Chang-Yong;Sun, Nam-Kyu;Kim, Eun-Jung;Seo, Jee-Woo;Park, Ji-Won;Kang, Kil-Jin
    • Korean Journal of Food Science and Technology
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    • v.41 no.1
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    • pp.7-10
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    • 2009
  • An analytical method for the determination of flumequine in meats was developed and validated using high-performance liquid chromatography with fluorescence detection. The samples were mixed with sodium sulfate and extracted with ethyl acetate. After clean-up, the residues were dissolved in mobile phase. The calibration curves showed high linearity ($r^2$=0.9979) within the concentration range of 0.1-1.0 mg/kg. The limit of detection and limit of quantification were validated at 0.005 and 0.017 mg/kg, respectively. The recoveries in fortified meats ranged from 90.8 to 101.1%. The method was then validated in correspondence with the CODEX guidelines for flumequine residue in meats. Herein we monitored 150 samples of meats that were purchased in Korea (Seoul, Busan, Daegu, Daejeon, and Gwangju). Among the tested samples, flumequine was detected in 1 of beef and 1 of pork at levels in the range of 0.048-0.080 mg/kg. Overall, the flumequine residues in the tested samples were within the Maximun residue limit.

Determination of Free 4-hydroxyproline with Dansylchloride by HPLC in Human Urine (소변 중 4-hydroxyproline 분석에 관한 연구)

  • Lee, Keou-Weon;Cho, Young-Bong;Lee, Kyung-Jong
    • Journal of Preventive Medicine and Public Health
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    • v.35 no.4
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    • pp.282-286
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    • 2002
  • Objectives : The level of 4-hydroxyproline (4-Hyp) in human urine was measured using high performance liquid chromatography (HPLC) with a fluorescence detector. This method is useful for medical examinations and investigating the radicals induced by physical, chemical, mental stresses. This method is superior to many published several methods in terms of its low cost and ability to analyze many samples. Methods : The urine from workers in a tire manufacturing company (22 male pre- and post-shift workers) and 18 office-workers as controls were analyzed. Data concerning age, the cumulative drinking amount and the cumulative smoking amount was collected with a questionnaire. The optimum applied amount of dansyl-Cl, the optimum reaction temperature and time, the recoveries and the optimum pH of the eluent and buffer were determined.4-Hyp from human urine was derivatized with dansyl-Cl (dimethylamino-naphthalene-1-sulfonyl chloride) after removing the a-amino acid by a treatment with phthalic dicarboxaldehyde (OPA) and cleaned with Bond Elut C18 column. The 4-Hyp derivatives were separated on a reversed phase column by gradient elution with a phosphate buffer (5 mmol, pH 8.0) and acetonitrile, and detected by fluorescence measurements at 340 nm (excitation) and 538 nm (emission). Results : The detection limit for the urinary free 4-Hyp was $0.364{\mu}mol/l$. The recovery rate of 4-Hyp was 99.7%, and the effective pH of the phosphate buffer and borate buffer were 3.0 and 8.0, respectively. From statistical analysis, age, drinking and smoking did not affect the urinary free 4-Hyp in both the controls and workers. The range of urinary 4-Hyp in the controls, pre-shift, and post-shift workers were 0.33-16.44, N.D-49.06, and $0.32-56.27{\mu}mol/l$. From the pared-sample t-test, the urinary 4-Hyp levels in post-shift workers ($11.82{\pm}6.73\;nmmol/mg\;Cre$) were 2-fold higher than in pre-shift workers ($5.36{\pm}5.53\;nmol;/mg\;Cre$) and controls ($4.91{\pm}4.89\;nmol;/mg\;Cre$). Conclusions : This method was developed with high sensitivity, accuracy, and precision. The present method was effectively applied to analyze the urinary free 4-Hyp in both controls and workers.

Validation and Uncertainty Evaluation of an Optimized Analytical Method Using HPLC Applied to Canthaxanthin, a Food Colorant (식품 색소 Canthaxanthin의 HPLC 최적 분석법 확인 및 타당성과 측정불확도 평가)

  • Suh, Hee-Jae;Kim, Kyung-Su;Hong, Mi-Na;Lee, Chan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.3
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    • pp.342-351
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    • 2016
  • This study was carried out to develop an optimized analytical method using high-performance liquid chromatography (HPLC) applied to canthaxanthin, which is not yet designated as a food colorant in Korea, as well as to perform validation and uncertainty evaluation of this method. Official methods of AOAC, UK, and Japan with HPLC-UV detection were evaluated for the analysis of canthaxanthin by comparison of linearity, resolution, selectivity, limit of detection (LOD), limit of quantitation (LOQ), accuracy, precision, recovery, inter-laboratory tests, and uncertainty measurement. The calibration curves showed high linearity with an $R_2$ value of over 0.999 for canthaxanthin standard solutions in all three official methods. The official method of Japan exhibited the best results in terms of resolution and selectivity, including the lowest LOD and LOQ. The average coefficients of variation were calculated as less than five of three institutes with a precision value less than 1, accuracy near 100%, and recovery ratio between $100{\pm}10%$. The expanded uncertainty for canthaxanthin was estimated to be $39.5{\pm}5.29mg/kg$ (95% confidence level, k=2), and the uncertainty of measurement was 13.4%. In this study, official methods of canthaxanthin were compared and the validities verified. The results will be further applied to establish an authorized analytical method for canthaxanthin in Korea.

Antioxidative activity of roasted Pueraria lobata root extracts (로스팅 칡 추출물의 항산화 활성)

  • Choi, Goo-Hee;Kim, Hyun Jung;Park, In-Jae;Kim, Bong-Gyun;Kim, Hoi-Yeong;Jeong, Jae-Hyun;Cho, Ju-Hyun
    • Food Science and Preservation
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    • v.24 no.3
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    • pp.440-445
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    • 2017
  • We evaluated the antioxidative activity of extracts of P. lobata root depending on roasting conditions. P. lobata roots were roasted at three different temperature at $150^{\circ}C$, $200^{\circ}C$, and $250^{\circ}C$ and three different time at 10 min, 20 min, and 30 min respectively. Roasted P. lobata root was extracted using water at $85^{\circ}C$ for 6 h and filtered using filter paper, followed by then evaporated ($12{\pm}0.3^{\circ}Brix$) and freeze-dried. The concentration of maker compound puerarin was determined using a high performance liquid chromatography system. 2 phenolic compounds, flavonoid contents, and antioxidant activities of the extract powder were evaluated. Puerarin contents, Phenolic compounds, and flavonoid contents of roasted P. lobata root were higher than those of unroasted P. lobata root. The results of DPPH and ABTS showed that roasted P. lobata root possessed higher antioxidant activity than unroasted P. lobata root. This study suggested that roasting process could be applied to P. lobata root in order to achieve its high quality and functionality.

Development and Validation of an Analytical Method for Betanine and Isobetanine in Processed Food Products Labeled with Beet Red

  • Kang, Hyun-Hee;Yun, Choong-In;Lee, Gayeong;Shin, Jae-Wook;Kim, Young-Jun
    • Journal of Food Hygiene and Safety
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    • v.36 no.5
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    • pp.376-381
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    • 2021
  • Red beet (Beta vulgaris L.) is a root vegetable and a popular functional food ingredient of dark red-purple appearance due largely to betacyanins, principally betanine (75-95%) and its isomer, isobetanine (15-45%). This study developed an analytical method for beet red in terms of betanine and isobetanine in processed food products labeled with beet red as a food additive. High Performance Liquid Chromatography-Diode Array Detector (HPLC-DAD) was used with a C18 column. Linearity, limit of detection (LOD), limit of quantitation (LOQ), accuracy, precision and uncertainty in measurement were calculated for method validation. Matrix-matched calibration was applied to the candy, ice cream, and cocoa product, respectively, and R2 was ≥0.9998, showing a high level of linearity. The LOD and LOQ were 0.16 to 0.32 and 0.48 to 0.97 mg/L, respectively. As a result of repeated intra-day and interday experiments to validate the accuracy and precision of the analytical method, the recovery rates were 96.0-103.1% and 100.0-102.2%, respectively and the RSD% was 0.5-3.3% and 0.9-3.8%, respectively. Moreover, the measurement uncertainty was estimated to be 1.71-12.43% depending on the matrix and the measured concentration. In this study, betanine and isobetanine were quantified (8.4-3,823.4 mg/kg) by applying the developed analytical method to processed food products (n= 26; e.g., candy, ice cream, and other processed foods) labeled with beet red as a food additive.

Selection of Functional Lentinula edodes Strains with High Ergothioneine Content using Mono-mono Hybridization (단핵균주간 교잡에 의한 에르고티오네인 함량이 높은 기능성 표고 균주 선발)

  • Kim, Min-Jun;Jeong, Yeun Sug;Jang, Yeongseon;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.49 no.4
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    • pp.507-514
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    • 2021
  • Lentinula edodes have been cultivated extensively not only in South Korea but also in other Asian countries. In terms of taste and nutrition, it is a valuable mushroom that is comparable to other mushrooms. L. edodes contains various physiologically active compounds that promote human health. One compound, ergothioneine, exerts a powerful antioxidant function that inhibits cellular senescence. As it is not biosynthesized by plants or animals, L. edodes is very important for ergothioneine supply. The L. edodes cultivars Bambithyang (NIFoS 3404) and Sansanhyang (NIFoS 3420) of the National Institute of Forest Science (NIFoS) had higher ergothioneine content than the other cultivars, and 11 strains were obtained using mono-mono hybridization between them. The strains were inoculated into sawdust media to obtain fruiting bodies, and the ergothioneine content of each hybrid strain was confirmed using high-performance liquid chromatography (HPLC) analysis. Among the 11 strains, NIFoS 5108 had a higher ergothioneine content than the parental strains, Bambithyang (NIFoS 3404) and Sansanhyang (NIFoS 3420). This study revealed that it is possible to develop cultivars with improved specific functions using mono-mono hybridization.

Oral Bioavailability for Sub-micron Particle Curcumin (TheracurminCR-033P) and Absorption Rate by Product Composition in Healthy Human Volunteers (서브마이크론 입자 형태 커큐민의 생체 흡수율 및 제품 조성에 따른 흡수율 동등성 연구)

  • Park, Hee Jung;Takahashi, Tsukasa;Ozawa, Hitomi
    • Food Engineering Progress
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    • v.21 no.3
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    • pp.273-279
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    • 2017
  • Curcumin have various health-beneficial properties in numerous studies. However, its bioavailability is low due to its limited intestinal uptake and rapid metabolism. This study aimed to evaluate the pharmacokinetics of newly developed sub-micron particle curcumin with increased water dispersibility (Theracurmin(R) CR-033P). Plasma curcumin levels were measured at 0, 1, 2, 4, 8 h after Theracurmin(R) CR-033P intake using high-performance liquid chromatography. For analyzing pharmacokinetics of Theracurmin(R) CR-033P, eighteen healthy subjects were recruited and received Theracurmin(R) CR-033P at a single oral dose containing curcumin 30 mg. $C_{max}$ was 28.14 ng/ml, and the area under the curve for 8 h was estimated to be 104.36 ng/ml. Based on the area under the plasma concentration (AUC), the bioavailability of sub-micron particle curcumin was higher 22-, 35-, 28-fold than native curcumin in men, women, and all subjects, respectively. For comparing by formulation, seven healthy subjects were recruited and received two type of treatment: (1) existing dosage form 300 mg (contained curcumin 30 mg) ${\times}$ 3 capsule, (2) high dosage form 300 mg (contained curcumin 90 mg) ${\times}$ 1 capsule + placebo 300 mg ${\times}$ 2 capsule. In the cross-over study, there was no significant differences in $C_{max}$ and AUC of plasma curcumin. In conclusion, submicron particle curcumin with increased water dispersibility significantly improved its oral bioavailability and women absorbed curcumin more effectively than men. Different formulation of Theracurmin(R) CR-033P has shown equivalent to the reference in terms of pharmacokinetics.

Secondary Metabolites and Morphological Diversity in the Leaves of Perilla Landrace from Korea

  • Assefa, Awraris Derbie;Sung, Jung-Sook;Jeong, Yi-Jin;Lee, Ho-Sun;Rhee, Ju-Hee;Hur, On-Sook;Noh, Jae-Jong;Ro, Na-Young;Hwang, Ae-Jin;Lee, Jae-Eun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.64-64
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    • 2019
  • Screening and identification of genetic resources based on their phytoconstituents and morphological characters potentially provide baseline data for researchers, breeders, and nutraceutical companies who wish to formulate a nutrient-dense diet and health beneficial supplement. Thus, we evaluated the amount of total phenolic content and major phenolic compounds; examined if phenolic compounds could be used as distinguishing factors for perilla genetic resources; and investigated the relation between some quantitative and qualitative morphological characters with the contents of phenolic compounds in 360 accessions obtained from National Agrobiodiversity Center gene bank, Jeonju, Korea. Total phenolic content (TPC) was estimated using Folin-Ciocalteu colorimetric assay. Individual phenolic compounds were determined using an Ultra-High Performance Liquid Chromatography system equipped with Photodiode Array detector. Considerable variations were observed in TPC (7.99 to 117.47 mg GAE/g DE), rosmarinic acid (RA) (ND to 19.19 mg/g DE), caffeic acid (CA) (ND to 0.72 mg/g DE), apigenin-7-O-diglucuronide (ADG) (ND to 1.24 mg luteolin equivalent (LUE)/g DE), scutellarein-7-O-glucuronide (SG) (ND to 4.32 mg LUE/g DE), and apigenin-7-O-glucuronide (AG) (ND to 1.60 mg LUE/g DE). RA was the most dominant phenolic compound in most accessions (95.3%) followed by SG. The adaxial leaf color was light green, green and dark green in 13.8%, 65.0%, and 21.1 % of the accessions, respectively. 78.8% of the accessions had light green color at the abaxial side with the remaining being described as green. Most of the accessions (96.9%) were cordate shape, the remaining being eclipse. Intensities of green pigment at abaxial and adaxial leaf surfaces were correlated with contents of individual phenolic compounds and TPC whereas leaf length and width had no correlation with TPC, CA and RA, and negatively correlated with ADG, AG, and SG. Leaf shape was not related with content of phenolic compounds, color of leaves, or the length or width of leaves. Accessions IT57426, IT157434, IT267710, and IT267712 which contained relatively high contents of TPC and major phenolic compounds (RA and SG) could be used for further research in breeding and bioassay test. Our study result showed the contents of total phenolics and individual phenolic compounds along with the morphological characters could be useful distinguishing factors for perilla genetic resources.

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Differential Expression of Isoflavone Biosynthetic Genes in Soybean During Germination (콩 발아기간 중 isoflavone 생합성 유전자 발현 변이)

  • Lim, Jin-Su;Kim, Seo-Young;Kim, Yong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.66 no.4
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    • pp.365-374
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    • 2021
  • Soybean isoflavones are essential secondary metabolites synthesized through the phenylpropanoid pathway, and they play vital roles in human health. Isoflavone content is a complex quantitative trait controlled by multiple genes, and the genetic mechanisms underlying isoflavone biosynthesis remain largely unknown. Therefore, the present study analyzed the content of isoflavone and expression of six key genes involved in its biosynthesis (i.e., CHS6, HID, IF7GT, IF7MaT, GmIMaT1, and GmIMaT3) during soybean seed germination. Isoflavone content was quantified using high-performance liquid chromatography, and isoflavone biosynthetic gene expression was analyzed using quantitative real-time PCR. Two cultivars, namely 'Daepung2ho' and 'Pungsannamulkong', which are high- and low-isoflavone cultivars, respectively, were used. Isoflavone accumulation gradually increased with the progression of the germination period. As such, malonyl glucosides accounted for over 80% of the total content, whereas acetyl glucosides were present at trace amounts. Transcriptional analysis of isoflavone biosynthetic genes demonstrated expression patterns parallel to isoflavone content; however, there was no clear correlation between isoflavone content and gene expression. Moreover, most isoflavone biosynthetic genes showed different expression patterns depending on the individual gene or genotypes. Among the tested genes, HID showed consistently higher expression, except at 3 days after germination, and its expression was upregulated in 'Daepung2ho' but downregulated in 'Pungsannamulkong'. In addition, all tested genes exhibited different expression patterns between cotyledons and hypocotyls and responded differently to the germination period. These findings suggest that the expression levels of isoflavone biosynthetic genes are not consistent with the germination period and appear to be genotype-dependent.