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Isolation and Structural Identification of Antioxidant Substances from Ethyl Acetate Extract of Conyza canadensis (망초(Conyza canadensis) Ethyl Acetate 추출물의 항산화성 물질의 분리와 동정)

  • Hyun Sook Song
    • Journal of Naturopathy
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    • v.12 no.1
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    • pp.7-15
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    • 2023
  • Background: As a result of analyzing the components of wild Conyza canadensis, it contains physiologically active ingredients, so it is necessary to identify the compound. Purposes: It was to study the compound's molecular structure; a previous study showed that C. canadensis contains antioxidant substances. Methods: The ultrasonic pulverized lysate of C. canadensis stem and leaves was first extracted with 90% methanol and then five organic solvents. Next, the extracts was fractionated by HPLC, LC/MS chromatography, and NMR analyzers identified the molecular structure. Results: 100 g of dry C. canadensis was sonicated in 90% methanol and concentrated under reduced pressure to 11.96 g of a crude extract. Then, this crude was extracted with five types of solvents to obtain 123.8 mg of n-hexane, 448.2 mg of dichloromethane, 1047.7 mg of ethyl acetate (EA), 2563.8 mg of butanol, and 7.04 g of water. The EA extracts were fractionated by LC-MS and then re-fractionated to obtain F1 to F20. Next, the F15 was further fractionated to obtain nine fine fractions. Finally, the F17 fraction was re-fractionated to obtain ten fine fractions. As a result of LC-MS and NMR spectrometer analysis of the F15-7, the structure of this compound was confirmed as 3,5-dicaffeoylquinic acid. As a result of examining the structures of the F17-4 and F17-5 fractions, Quercetin-3-o-β-galactose was identified. In addition, the form of the F17-10 was confirmed to be 1,3,4-tri-caffeoylquinic acid. Conclusions: This study demonstrated that C. canadensis contained phenolic antioxidants, and its utilization may be expected.

Development and Validation of a Simultaneous Analytical Method for 5 Residual Pesticides in Agricultural Products using GC-MS/MS (GC-MS/MS를 이용한 농산물 중 잔류농약 5종 동시시험법 개발 및 검증)

  • Park, Eun-Ji;Kim, Nam Young;Shim, Jae-Han;Lee, Jung Mi;Jung, Yong Hyun;Oh, Jae-Ho
    • Journal of Food Hygiene and Safety
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    • v.36 no.3
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    • pp.228-238
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    • 2021
  • The aim of this research was to develop a rapid and easy multi-residue method for determining dimethipin, omethoate, dimethipin, chlorfenvinphos and azinphos-methyl in agricultural products (hulled rice, potato, soybean, mandarin and green pepper). Samples were prepared using QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) and analyzed using gas chromatography-tandem mass spectrometry (GC-MS/MS). Residual pesticides were extracted with 1% acetic acid in acetonitrile followed by addition of anhydrous magnesium sulfate (MgSO4) and anhydrous sodium acetate. The extracts were cleaned up using MgSO4, primary secondary amine (PSA) and octadecyl (C18). The linearity of the calibration curves, which waas excellent by matrix-matched standards, ranged from 0.005 mg/kg to 0.3 mg/kg and yielded the coefficients of determination (R2) ≥ 0.9934 for all analytes. Average recoveries spiked at three levels (0.01, 0.1, 0.5 mg/kg) and were in the range of 74.2-119.3%, while standard deviation values were less than 14.6%, which is below the Codex guideline (CODEX CAC/GL 40).

The Effect of Various Culture Media on Histological Anatomy and Saikosaponin Content in Hairy Root Culture of Buplurum falcatum (시호 모상근 배양에서 배지성분에 따른 사이코사포닌 함량과 조직학적 형태의 차이)

  • Ahn, Jun-Cheul;Jeong, Young-Jae;Lee, Kyu-Bae;Kim, Ock-Tae;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.4
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    • pp.251-256
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    • 1999
  • Growth rate and saikosaponin content of Buplurum falcatum hairy root on MS and RCM basal media were measured. And using hairy root cultured in those media, it was investigated whether some correlation between differences in growth rate and saikosaponin content and histiological difference was or not. Results obtained showed that among media tested, MS and 3RCM medium which showed the faster growth in fresh weight and dry weight, respectively. And saikosaponin content of hairy root cultured in 3RCM was $2.2{\sim}5.5$ times higher than that of 1 year cultivated roots, whereas that in MS was extremely low. On the other hand, histological differences of root tips and about 1 cm region from root tip cultured in both MS and 3RCM were not prominent, however, in mature region, a lot of cells only in 3RCM contained densely stained vacuoles or organelles which probably contain tannin, suggesting that development of those has a correlation with biosynthesis of secondary metabolites containing saikosaponin.

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Development of an Analytical Method for the Determination of Dexamethasone in Bovine Milk Using Liquid Chromatography Coupled to Tandem Mass Spectrometry (LC-MS/MS를 이용한 우유 중 덱사메타손의 잔류 분석법 개발)

  • Cha, Chun-Nam;Park, Eun-Kee;Yoo, Chang-Yeul;Lee, Sung Joong;Son, Song-Ee;Kim, Suk;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.32 no.5
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    • pp.418-423
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    • 2017
  • An analytical method for the determination of dexamethasone (DM) in bovine milk samples was developed and validated using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Milk samples were extracted by the liquid-liquid extraction based on acetonitrile. The chromatographic separation was achieved on a reverse phase $C_{18}$ column with gradient elution using a mobile phase of 0.1% formic acid in 95% acetonitrile. The procedure was validated according to the Ministry of Food and Drug Safety guideline determining accuracy, precision, limit of detection (LOD), and limit of quantification (LOQ). Mean recoveries of DM from spiked milk samples (25, 125, and 1,250 ng/mL) were 98.9-109.6%, and the relative standard deviation was between 1.7 and 4.4%. Linearity in concentration range of 12.5-1,250 ng/mL was obtained with the correlation coefficient ($r^2$) of 0.9997. LOD and LOQ for the investigated DM were 0.15 and 0.5 ng/mL depending on milk samples, respectively. This method was reliable, sensitive, economical and suitable for routine monitoring of DM residues in bovine milk.

Quantitative Analysis and Qualification of Amitrole Using LC/ESI-MS (LC/ESI-MS를 이용한 아미트롤의 정성확인 및 정량분석)

  • Park, Chan-Koo;Eo, Soo-Mi;Kim, Min-Young;Sohn, Jong-Ryeul;Mo, Sae-Young
    • Analytical Science and Technology
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    • v.17 no.2
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    • pp.117-129
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    • 2004
  • Amitrole in environment, difficult to be analyzed by GC or GC/MS due to high polarity and low volatility, was analyzed by LC/ESI/MS in the study. Maximum peak intensity of amitrole in LC/MS/ESI mass spectrum is m/z 85 of protonated molecular ion $(M+H)^+$ with 30V of cone voltage at SIR mode. It was confirmed that ratios between main ion of amitrole, 85 of protonated molecular ion, and m/z 58 fragmented ion of amitrole, had increased corresponding with the increment of cone voltage from 20V to 70V. The isotope molecular weight of amitrole was $86([M+H])^+$ at LC/MS analysis and the mass spectrum ratio between 85 mass and 86 mass was not different by the change of concentration but similar to theoretical ratio(less than 10% standard deviation).The linearity of standard calibration curve under same condition with sample treatment method had $y=1.09354e^6X+26947.2$ and $r^2=0.99$. Recovery rates in water and soil samples were 77.64~83.44% and 71.11~79.44%, respectively. Reliability of the analysis was performed with 5 repeated measurements at each level of standard concentration and the result showed that relative standard deviation was less than 10%; therefore, the extraction and analysis method in the study suggested that it would be reliable to measure amitrole in water and soil media.

Prevalence of Metabolic Syndrome and Assessment of Food·Nutrient Intakes among Adult Visitors of a Public Health Center in Korea (일부 보건소 내원자의 대사증후군 발현과 식품 및 영양소 섭취 실태)

  • Jeong, Won-Hoon;Jin, Bok-Hee;Hwang, Eun-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.2
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    • pp.205-212
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    • 2012
  • This study was performed to investigate the prevalence of metabolic syndrome (MS) and assess nutrient intake levels for the purpose of improving MS risk factors. The participants in this study were 512 adults consisting of 271 men and 241 women aged 30 and over, who visited a public health center for a medical check up. The diagnosis of MS subjects was adapted from the NCEP-ATPIII guidelines and the WHO Asia-Pacific Area criteria for obesity. The MS group was defined as subjects displaying three or more risk factors, and the non MS group was defined as those displaying two or less risk factors. A dietary survey was conducted using the 24-hour recall method. The number of subjects displaying MS syndrome factors was 158 (30.9%), broken down into, 89 men and 69 women. Regarding risk factors in the MS group, the prevalence of waist circumference was 40.5%, hypertension 34.2%, hyperglycemia 31.0%, low HDL-cholesterol 24.7%, and hypertriglycemia 19.6%. BMI, sistolic blood pressure, blood glocose, blood triglyceride, and blood HCL-cholesterol of the MS group were significantly higher compared to the non MS group. Male subjects in the MS group reported high intakes of cereals, sugar, fruits, meat and poultry, oil and fats, and beverages and total food intake was significantly higher compared to the non MS group. Women in the MS group reported high intakes of meat and poultry, milk and dairy products, beverages, and seasonings, and total food intake was higher compared to the non MS group. Dietary diversity score (DDS) was 3.82~4.04, which was not significant among the groups. In men, dietary variety score (DVS) was 16.3 in the MS group and 19.4 in the non MS group, whereas in women, the DVS was 15.2 in the non MS group and 17.0 in the MS group. In GMVDF pattern, 11111 pattern was 30.7%, followed by 01111 for men and 11101 for women. Calorie, fat, and cholesterol intakes in men as well as, calorie, fat, and folate intakes in women in the MS group were higher compared to the non MS group. Intakes of protein, P, Fe, Na, vitamin $B_1$, vitamin $B_2$, niacin, vitamin E, and Zn were higher than the KDRIs. On the other hand, intakes of Ca, K, fiber, vitamin $B_2$, and vitamin C were below the KDRIs. Intakes of lipids, animal food, Na, and cholesterol in the MS group were higher compared to the non MS group, whereas intake of dietary fiber was lower. Our results indicate that continuous, systematic nutritional education program must implemented to reduce the risk factors associated with MS.

Quantitative Analysis of the Fifteen Constituents in Hyangso-San by LC-MS/MS (LC-MS/MS를 이용한 향소산 중 15종 성분의 정량분석)

  • Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
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    • v.47 no.4
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    • pp.381-388
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    • 2016
  • Hyangso-san is a traditional herbal medicine that consists of the seven herbal medicines, Cyperi Rhizoma, Perillae Folium, Atractylodis Rhizoma, Citri Unshius Pericarpium, Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma Crudus, and Allii Fistulosi Bulbus. Hyangso-san has long been clinically used to treat the influenza, including headache, ferver, chills, and pantalgia. In this study, we were performed the simultaneous analysis of the 15 marker compounds (liquiritin apioside, liquiritin, ferulic acid, naringin, hesperidin, rosmarinic acid, liquiritigenin, kaempferol, glycyrrhizin, nobiletin, 6-gingerol, elemicin, atractylenolide III, nootkatone, and atractylenolide I) in Hyangso-san using liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS/MS). Column for the separation of the 15 ingredients was used a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) at $45^{\circ}C$ by using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient condition. Identifications of all analytes were performed using a Waters ACQUITY TQD LC-MS/MS system. The flow rate and injection volume were 0.3 mL/min and $2.0{\mu}L$, respectively. Correlation coefficient of the calibration curve was ${\geq}0.9958$. The values of limits of detection and quantification of the 15 components were 0.002-4.29 and 0.01-12.88 ng/mL, respectively. The result of an analysis using the established LC-MS/MS method, kaempferol and atractylenolide I were not detected, while other 13 compounds were 0.08-56.87 mg/g in lyophilized Hyangso-san sample.

Validation of LC-MS/MS Method for Analysis of Paralytic Shellfish Toxins in Shellfish and Tunicates (LC-MS/MS를 이용한 패류 및 피낭류 중 마비성 패류독소 분석법의 유효성 검증)

  • Cho, Sung Rae;Kim, Dong Wook;Yu, Hean Jae;Cho, Seong Hae;Ryu, Ara;Lee, Ka Jeong;Mok, Jong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.2
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    • pp.174-180
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    • 2020
  • The mouse bioassay has been used widely for the monitoring of paralytic shellfish toxins (PSTs) in many countries. However, this method shows low sensitivity and high limit of detection (LOD), as well as it cannot confirm toxic profiles. Recently, LC-MS/MS method was studied for the quantitative of PSTs, however, the method has any problems with unstable retention times by ionization suppression caused by high salt concentration in shellfish extracts. To establish an alternative method for PSTs analysis, we tried to original LC-MS/MS methods adding desalting operation using amorphous graphitized polymer carbon solid-phase extraction cartridges. The method validation was conducted to determine linearity, limit of detection, limit of quantification (LOQ), accuracy, and precision in quantifying PSTs. The correlation coefficients for all tested PSTs maintained over 0.999. The LODs and LOQs for all PSTs were about 0.19-1.05 ㎍/kg and 0.58-3.18 ㎍/kg, respectively. The accuracies for PSTs were 95.4-107.7% for saxitoxin group, 97.1-100.9% for gonyautoxin group, 99.0-100.8% for N-sulfocarbamoyl toxin group, and 96.8-104.6% for decarbamoyl toxin group. These results indicate that the modified LC-MS/MS method was appropriate for analyzing the PSTs in shellfish and tunicates.

Comparative of the Nutritional Status and Correlation of Cardiovascular Disease in Type Ⅱ Diabetes Mellitus Patients with Metabolic Syndromes (제2형 당뇨병환자에서 대사증후군 동반에 따른 영양소 섭취상태 비교 및 심혈관질환과의 관련성)

  • Lim, Hee-Sook;Kim, Soon-Kyung
    • Journal of Nutrition and Health
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    • v.41 no.4
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    • pp.327-340
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    • 2008
  • This study was designed to investigate the correlation between nutrition status and cardiovascular disease in type 2 diabetes patients with metabolic syndrome. The subjects were 66 patients and divided into Non MS (a group without metabolic syndrome, n = 37) and MS (a group with metabolic syndrome, n = 29). The percentage of patients accompanying metabolic syndrome was 43.9% and family history such as DM, skipping meal and eating speed were higher in MS (P < 0.05) The average values of BMI, body fat (%), waist circumference were significantly higher in patients of MS than that of Non MS. For hamatological values, MS showed higher FRS, HOMA-IR, LDL-Cholesterol, CRP. Percentage of FRS was 21.63% in MS that is relatively higher in comparison with 16.81% in Non MS. Moreover, the incidence of cardiovascular disease appeared 13.8% in MS that is higher than 2.7% in Non MS. The intake of sodium and vitamin E were higher, but the intake of fat, vitamin A and zinc were lower in MS than in Non MS. Close correlations were elucidated among FRS, occurrence of cardiovascular disease, weight, waist circumference, Total-Cholesterol, LDLCholesterol, sodium in both groups. In conclusion, cardiovascular disease risk factors would be higher in type 2 diabetes patients with metabolic syndrome and there were distinctive patterns that were associated with hamatological values, nutrition intake risk factors. This result should be considered when designing nutrition study and intervetion programs.

Histological Validation of Cardiovascular Magnetic Resonance T1 Mapping for Assessing the Evolution of Myocardial Injury in Myocardial Infarction: An Experimental Study

  • Lu Zhang;Zhi-gang Yang;Huayan Xu;Meng-xi Yang;Rong Xu;Lin Chen;Ran Sun;Tianyu Miao;Jichun Zhao;Xiaoyue Zhou;Chuan Fu;Yingkun Guo
    • Korean Journal of Radiology
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    • v.21 no.12
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    • pp.1294-1304
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    • 2020
  • Objective: To determine whether T1 mapping could monitor the dynamic changes of injury in myocardial infarction (MI) and be histologically validated. Materials and Methods: In 22 pigs, MI was induced by ligating the left anterior descending artery and they underwent serial cardiovascular magnetic resonance examinations with modified Look-Locker inversion T1 mapping and extracellular volume (ECV) computation in acute (within 24 hours, n = 22), subacute (7 days, n = 13), and chronic (3 months, n = 7) phases of MI. Masson's trichrome staining was performed for histological ECV calculation. Myocardial native T1 and ECV were obtained by region of interest measurement in infarcted, peri-infarct, and remote myocardium. Results: Native T1 and ECV in peri-infarct myocardium differed from remote myocardium in acute (1181 ± 62 ms vs. 1113 ± 64 ms, p = 0.002; 24 ± 4% vs. 19 ± 4%, p = 0.031) and subacute phases (1264 ± 41 ms vs. 1171 ± 56 ms, p < 0.001; 27 ± 4% vs. 22 ± 2%, p = 0.009) but not in chronic phase (1157 ± 57 ms vs. 1120 ± 54 ms, p = 0.934; 23 ± 2% vs. 20 ± 1%, p = 0.109). From acute to chronic MI, infarcted native T1 peaked in subacute phase (1275 ± 63 ms vs. 1637 ± 123 ms vs. 1471 ± 98 ms, p < 0.001), while ECV progressively increased with time (35 ± 7% vs. 46 ± 6% vs. 52 ± 4%, p < 0.001). Native T1 correlated well with histological findings (R2 = 0.65 to 0.89, all p < 0.001) so did ECV (R2 = 0.73 to 0.94, all p < 0.001). Conclusion: T1 mapping allows the quantitative assessment of injury in MI and the noninvasive monitoring of tissue injury evolution, which correlates well with histological findings.