• Title/Summary/Keyword: UPLC

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How to Prepare Rehmanniae Radix Preparata Described in the 『Treasured Mirror of Eastern Medicine』 (『東醫寶鑑』의 熟地黃 제조방법)

  • Roh, Jong Seong;Yoon, Michung;Shin, Soon Shik
    • Herbal Formula Science
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    • v.24 no.1
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    • pp.17-30
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    • 2016
  • Objectives Rehmanniae Radix Preparata was prepared in the traditional Rehmanniae Radix Preparata preparation method set forth in the 『Treasured Mirror of Eastern Medicine(Donguibogam)』 with a view to measuring the contents of 5-Hydroxymethyl-2-furaldehyde(5-HMF) at individual stages of steaming and sundrying and identifying new chemical components.Methods Based on the traditional Rehmanniae Radix Preparata preparation method set forth in the 『Treasured Mirror of Eastern Medicine』, Rehmanniae Radix Preparata steamed and sundried once through nine times was prepared. Thereafter, 5-HMF contents were analyzed and new chemical components were identified in the Rehmanniae Radix Preparata using Waters HPLC e2695, 2640 detectors, a Waters Acquity UPLC system, and a Micromass Q-TOF Premier mass spectrometer.Results The Rehmanniae Radix Preparata preparation method set forth in the 『Treasured Mirror of Eastern Medicine』 is a unique preparation method in Republic of Korea different from that in China. In the first stage of the method, fresh Rehmanniae Radix Crudus was divided into high quality, medium quality, and low quality ones named Rehmanniae Radix Crudus (Caelum)(天黃), Rehmanniae Radix Crudus (Homo)(人黃), and Rehmanniae Radix Crudus (Terra)(地黃) respectively to use Rehmanniae Radix Crudus (Caelum) and Rehmanniae Radix Crudus (Homo) for preparation of juice while using Rehmanniae Radix Crudus (Terra) to make Rehmanniae Radix Preparata. In the second stage, Rehmanniae Radix Crudus (Caelum) and Rehmanniae Radix Crudus (Terra) were made into juice and Rehmanniae Radix Crudus (Terra) was soaked in the juice. In the third stage, among auxiliary materials, rice wine named Purum Vinum Oryzae(淸酒) brewed from sticky rice was sprinkled on Rehmanniae Radix Crudus (Terra) to the extent that Rehmanniae Radix Crudus (Terra) became wet. In the fourth stage, Rehmanniae Radix Preparata steamed in earthenware steamer was dried under natural sunlight. The contents of 5-HMF in Rehmanniae Radix Preparata steamed and sundried once through nine times were shown to be below 0.1% in all cases. Pomolic acid was identified as a new chemical component.Conclusions In conclusion, the Rehmanniae Radix Preparata preparation method set forth in the 『Treasured Mirror of Eastern Medicine』 is thought to be a unique preparation method in South Korea in which Rehmanniae Radix Preparata is completed through the first stage in which fresh Rehmanniae Radix Crudus collected from fields is divided into high, medium, and low quality ones and fresh Rehmanniae Radix Crudus juice is made, the second stage in which the high quality fresh Rehmanniae Radix Crudus is soaked in the fresh Rehmanniae Radix Crudus juice, the third stage in which the fresh Rehmanniae Radix Crudus is steamed, and the fourth stage in which the steamed Rehmanniae Radix Crudus is dried.

Content and Distribution of Flavanols, Flavonols and Flavanones on The Common Vegetables in Korea (국내산 채소류에 함유된 플라바놀, 플라보놀 및 플라바논에 대한 함량 및 분포 조사)

  • Shin, Jae-Hyeong;Kim, Heon-Woong;Lee, Min-Ki;Lee, Sung-Hyen;Lee, Young-Min;Jang, Hwan-Hee;Hwang, Kyung-A;Cho, Young-Sook;Kim, Jung-Bong
    • Korean Journal of Environmental Agriculture
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    • v.33 no.3
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    • pp.205-212
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    • 2014
  • BACKGROUND: This study focused on the contents of flavonoid compounds in vegetables. Generally vegetables have contributed to a healthy diet, arisen from contains a large amount of fiber and functional ingredients. And flavonoid compounds are one of major functional components in the vegetables. currently research of flavonoid contents does not enough, specially in the part of homegrown vegetable. METHODS AND RESULTS: Vegetable samples were purchased in domestic market. Sample extraction by methanol, distilled water, and formic acid based solvent. Also same solvent used for mobile phase in UPLC. Eleven types of flavonoid compounds were analyzed with same kind of external standard and one kind of internal standard (galangin) for quantification. Standard calibration curve presented linearity with the correlation coefficient $R^2$ > 0.98, analysed from 1 to 50 ppm concentration. The quantitative value and multivariate analysis results were derived from the Excel and SIMCA-P11. Overall, onion has largest amount(916.5 mg/100 g) of flavonoid and also other vegetables have has significant amount[Mugwort: 138.8, Galic stem:123.6 mg/100 g etc.] of flavonoid compounds. Edible portion of vegetables per share for simulating by SIMCA-P11, root vegetables has had difference with other vegetables according to distributions and amounts of flavonoid compounds. CONCLUSION: Optionally, the results from this experiment can use to select the material for flavonoid researches. And based on these results, if this experiment will be continuously complemented, and performed, could used in various fields.

Quality Characteristics and Composition Profile of Traditional Doenjang and Manufactured Doenjang during Storage Time (저장기간에 따른 전통된장과 개량식된장의 이화학적 특성, 기능성분 및 대사체 분석)

  • Kang, Hee-Joo;Kim, Jin-Hee;Kim, Ri-Rang;Kim, Kang Sung;Hong, Sang-Phi;Kim, Min-Jung;Yang, Hye Jeong
    • The Korean Journal of Food And Nutrition
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    • v.29 no.5
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    • pp.785-794
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    • 2016
  • This research was conducted to evaluate quality changes in traditional Doenjang and manufactured Doenjang during a storage period of 8 weeks. Low-salt Doenjang and commercial Doenjang were purchased from different manufacturers and proximate analysis as well as changes in isoflavone, polyphenol, flavonoid contents of the samples were investigated using a mass spectrophotometer. The salinity of traditional Doenjang, low salt Doenjang, and commercial Doenjang were $13.2{\pm}1.15$, $7.17{\pm}2.74$, $10.67{\pm}0.35%$, respectively and the salt concentrations of the soybean pastes did not change during storage. After 8 weeks at $35^{\circ}C$, chromatic values of all the paste samples decreased somewhat, with traditional Doenjang exhibiting fewer changes as compared to manufactured Doenjang. Amino acid nitrogen, acidity, microbial population all tended to increase with time, although some samples showed fluctuations during the test period. Moreover, the total isoflavone contents of traditional Doenjang increased with storage time while that of manufactured Doenjang tended to decrease. The isoflavone aglycone was shown to be the highest in traditional Doenjang, while isoflavone glycoside was abundant in manufactured Doenjang. Total flavonoid contents showed similar trends regardless of samples; initial contents of total flavonoid was 0.6 mg/g which increased to more than twice to 1.4 mg/g at the end of storage period. Composition profile of Doenjang extracts was analyzed using UPLC-Q-ToF.

Identification and quantification of major malonyl ginsenosides isolated from Panax ginseng C.A. Meyer (인삼(Panax ginseng C.A. Meyer)로부터 Malonyl ginsenoside의 분리 및 정량분석)

  • Shin, Woo Cheol;Jung, Jiyun;Na, Hyeon Seon;Bo, Jeon Hwang;Kim, Hyoung-Geun;Yoon, Dahye;Choi, Bo-Ram;Lee, Young-Seob;Kim, Geum-Soog;Baek, Nam-In;Lee, Yi;Lee, Dae Young
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.375-384
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    • 2019
  • The root of Panax ginseng C.A. Meyer were extracted with 70% aqueous EtOH and the concentrates were partitioned into MeOH and H2O fractions using Diaion HP-20. The repeated SiO2 or octadecyl SiO2 column, and MPLC for the MeOH fraction led to isolation of four malonyl ginsenosides. The chemical structures of these compounds were determined as malonyl ginsenoside Rd (1) malonyl ginsenoside Rc (2) malonyl ginsenoside Rb2 (3) malonyl ginsenoside Rb1 (4) based on spectroscopic analyses including Nuclear magnetic resonance and HR-TOF/MS. The contents of malonyl ginsenoside Rb1 was highist as 5.44 mg/g of five years of ginseng. And malonyl ginsenoside Rd was lowest as 0.11 mg/g of six years of ginseng. Additionally, the malonyl ginsenoside Rd exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line.

Survey on Fusarium Mycotoxin Contamination in Oat, Sorghum, Adlay, and Proso Millet during the Harvest Season in Korea (귀리, 수수, 율무, 기장의 수확기에 발생하는 Fusarium 곰팡이독소 오염도 조사)

  • Lee, Mi Jeong;Wee, Chi-Do;Ham, Hyenheui;Choi, Jung-Hye;Baek, Ji Sun;Lim, Soo Bin;Lee, Theresa;Kim, Jeom-Soon;Jang, Ja Yeong
    • Journal of Food Hygiene and Safety
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    • v.35 no.1
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    • pp.13-22
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    • 2020
  • A total of 244 cereal samples (oat, sorghum, adlay, and proso millet) were collected from fields to examine the contamination of Fusarium mycotoxins in cereals during harvest season in 2017 and 2018. The contamination levels of deoxynivalenol (DON), nivalenol (NIV), and zearalenone (ZEA) were analyzed individually by using the immunoaffinity column clean-up method with ultra performance liquid chromatography, and fumonisins (FUM) were analyzed by using the QuEChERS method with liquid chromatography-mass spectrometry. Highest level of NIV contamination (120.0-3277.0 mg/kg) was observed in oat samples among the analyzed cereals. In the adlay samples, DON contamination was the highest (maximum level 730.0 ㎍/kg). The proso millet samples had a high frequency of detection of NIV and ZEA (61.5% and 57.9%, respectively), but the levels were low (average detection level of NIV, 75.6 ㎍/kg, for ZEA, 21.5 ㎍/kg). Among the cereal samples, sorghum had the highest contamination frequency of DON, ZEA, and FUM, and the co-occurrence of Fusarium mycotoxin was 70.0%, which was higher than the average of 29.9%. In order to safely manage Fusarium mycotoxin levels in cereals, continuous research on the development of contamination prevention technologies together with monitoring of mycotoxin contamination is needed.

Purification and Characterization of Antibacterial Compound Produced by Bacillus subtilis MJP1 (Bacillus subtilis MJP1이 생산하는 항세균 물질의 분리.정제 및 특성규명)

  • Yim, Eun-Jung;Yang, Eun-Ju;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.38 no.1
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    • pp.84-92
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    • 2010
  • Antibacterial compound from Bacillus subtilis MJP1 was purified using C18 Sep-Pak cartridge, ion exchange chromatography, and gel filtration chromatography. The purified antibacterial compound showed antibacterial activity against Listeria monocytogenes, Bacillus subtilis, Staphylococcus aureus subsp. aureus, and Enterococcus faecalis. The purified antibacterial compound was found to be stable at $100^{\circ}C$ for 5 min and in the pH range of 3.0~9.0, but it was unstable at pH 10.0. It was inactivated by proteinase K and pronase E, and heat treatment at $121^{\circ}C$ for 15 min, but it was stable with lipase and $\alpha$-amylase treatment, which indicated its proteineous nature. Ultra performance liquid chromatography and electrospray ionization tandem mass spectrometry analysis were used to identify the purified antibacterial compound and confirmed the existence of two peptides (3356.54 Da, 3400.5244 Da).

Utilization of [6]-gingerol as an origin discriminant marker influencing melanin inhibitory activity relative to its content in Pinellia ternata (반하(Pinellia ternata)에서의 [6]-gingerol 함량과 멜라닌 저해 활성에 영향을 미치는 원산지 판별 마커로의 활용)

  • An, Ju Hyeon;Won, Hyo Jun;Seo, Soo-Kyung;Kim, Doo-Young;Ku, Chang-Sub;Oh, Sei-Ryang;Ryu, Hyung Won
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.323-330
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    • 2016
  • Pinellia ternata Breitenbach, the natural medicinal plant of the Araceae family, is a perennial plant originated from the East Asia, but also widely distributed in Europe and North America. Its tuber is used as traditional medicine for treatment of various diseases such as vomiting, inflammation, and traumatic injury. Pharmacological studies revealed that P. ternata possesses anticonvulsant, anti-tumor, insecticidal, and cytotoxic activities. Despite being well-known as the useful medicinal plant, there is no reliable, standardized method for origin discrimination. Ultra performance liquid chromatography-photodiode array detector and quadrupole time of flight-mass spectrometry based metabolite-profiling was applied to explore significant metabolite for origin discrimination between Korean and Chinese P. ternata. One compound was isolated from Korean P. ternata using repeated ODS column chromatography by bioactivity guided fractionation, and determined as [6]-gingerol according to the results of spectroscopic data including nuclear magnetic resonance and MS. This compound was selected as cosmeceutical biomarker by fingerprints, and it was associated to melanin inhibitory effect determining its origin authenticity. Furthermore, the calibration curve of biomarker was prepared using validated method for the comparison of content between Korean and Chinese P. ternata. This is the report to address the selection and successful validation of the discriminant metabolite for confirmation of Korean P. ternata.

Chemical and bioactive comparison of flowers of Panax ginseng Meyer, Panax quinquefolius L., and Panax notoginseng Burk.

  • Li, Fang;Lv, Chongning;Li, Qiao;Wang, Jing;Song, Dan;Liu, Pengpeng;Zhang, Dandan;Lu, Jincai
    • Journal of Ginseng Research
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    • v.41 no.4
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    • pp.487-495
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    • 2017
  • Background: Although flowers of Panax ginseng Meyer (FPG), Panax quinquefolius L. (FPQ), and Panax notoginseng Burk. (FPN) have been historically used as both medicine and food, each is used differently in practice. Methods: To investigate the connection between components and enhancing immunity activity of FPG, FPQ, and FPN, a method based on a rapid LC coupled with quadrupole time-of-flight MS and immunomodulatory activity study evaluated by a carbon clearance test were combined. Results: According to quantitative results, the ratio of the total content of protopanaxatiol-type ginsenosides to protopanaxadiol-type ginsenosides in FPN was 0, but ranged from 1.10 to 1.32 and from 0.23 to 0.35 in FPG and FPQ, respectively. The ratio of the total content of neutral ginsenosides to the corresponding malonyl-ginsenosides in FPN ($5.52{\pm}1.33%$) was higher than FPG ($3.2{\pm}0.64%$) and FPQ ($2.39{\pm}0.57%$). The colorimetric analysis showed the content of total ginsenosides in FPQ, FPG, and FPN to be $13.75{\pm}0.60%$, $17.45{\pm}0.42%$, and $12.45{\pm}1.77%$, respectively. The carbon clearance assay indicated that the phagocytic activity of FPG and FPQ was higher than that of FPN. A clear discrimination among FPG, FPQ, and FPN was observed in the principal component analysis score plots. Seven compounds were confirmed to contribute strongly by loading plots, which may be the cause of differences in efficacy. Conclusion: This study provides basic information about the chemical and bioactive comparison of FPG, FPQ, and FPN, indicating that protopanaxtriol-type ginsenosides and malonyl-ginsenosides may play a key role in their enhancing immunity properties.

Chemical transformation and target preparation of saponins in stems and leaves of Panax notoginseng

  • Wang, Ru-Feng;Li, Juan;Hu, Hai-Jun;Li, Jia;Yang, Ying-Bo;Yang, Li;Wang, Zheng-Tao
    • Journal of Ginseng Research
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    • v.42 no.3
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    • pp.270-276
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    • 2018
  • Background: Notoginsenoside Ft1 is a promising potential candidate for cardiovascular and cancer disease therapy owing to its positive pharmacological activities. However, the yield of Ft1 is ultralow utilizing reported methods. Herein, an acid hydrolyzing strategy was implemented in the acquirement of rare notoginsenoside Ft1. Methods: Chemical profiles were identified by ultraperformance liquid chromatography coupled with quadruple-time-of-flight and electrospray ionization mass spectrometry (UPLC-Q/TOF-ESI-MS). The acid hydrolyzing dynamic changes of chemical compositions and the possible transformation pathways of saponins were monitored by ultrahigh-performance LC coupled with tandem MS (UHPLC-MS/ MS). Results and conclusion: Notoginsenoside Ft1 was epimerized from notoginsenoside ST4, which was generated through cleaving the carbohydrate side chains at C-20 of notoginsenosides Fa and Fc, and vinaginsenoside R7, and further converted to other compounds via hydroxylation at C-25 or hydrolysis of the carbohydrate side chains at C-3 under the acid conditions. High temperature contributed to the hydroxylation reaction at C-25 and 25% acetic acid concentration was conducive to the preparation of notoginsenoside Ft1. C-20 epimers of notoginsenoside Ft1 and ST4 were successfully separated utilizing solvent method of acetic acid solution. The theoretical preparation yield rate of notoginsenoside Ft1 was about 1.8%, which would be beneficial to further study on its bioactivities and clinical application.

Antioxidant activities of ethanol extracts from barley sprouts (새싹보리 에탄올 농도별 추출물의 산화방지 활성)

  • Chae, Kyu Seo;Ryu, Eun Hye;Kim, Ki Deok;Kim, Yong-Suk;Kwon, Ji Wung
    • Korean Journal of Food Science and Technology
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    • v.51 no.5
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    • pp.486-491
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    • 2019
  • Phenolic compounds and antioxidant activities of ethanol extracts from barley sprouts were evaluated in this study. Barley sprouts were extracted using water and ethanol in various concentration (25, 50, and 75%) using reflux extraction methods. Ultra performance liquid chromatography (UPLC) analysis showed that barley sprouts are mainly composed of rutin, gallic acid, ferulic acid, and ${\rho}$-coumaric acid. The 75% ethanol extracts had higher total polyphenol contents ($44.01{\pm}1.32mg/g$) and total flavonoid contents ($102.96{\pm}2.49mg/g$). 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity ($EC_{50}$ value: $1.65{\pm}0.02mg/mL$) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical scavenging activity ($EC_{50}$ value: $1.67{\pm}0.02mg/mL$) of the 75% ethanol extracts of barley sprouts were found to be the most effective. The 75% ethanol extracts of barley sprouts exhibited a strong reducing activity and ferric reducing antioxidant activity. As a result, the 75% ethanol extracts of barley sprouts showed stronger antioxidant activity than other extracts.