• Title/Summary/Keyword: HPLC pattern analysis

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Discrimination of Lonicera japonica and Lonicera confusa using chemical analysis and genetic marker

  • Ryuk, Jin Ah;Lee, Hye Won;Ko, Byoung Seob
    • The Korea Journal of Herbology
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    • v.27 no.6
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    • pp.15-21
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    • 2012
  • Objective : Lonicera japonica THUNB. a traditional herbal medicine, has been commonly used anti-inflammatory disease. It has been very complicated with respect to its sources on the market. The significant selection of medicine depends on its origin. However, it is difficult to discrimination criteria for confirming L. japonica authenticity using the senses. This study was performed to determine the discriminant analysis of L. japonica and L. confusa. Methods : The identification of L. japonica and L. confusa were performed by the classification and identification committee of the national center for standardization of herbal medicines. And we examined its differences using HPLC and genetic marker analysis. Results : The analytical pattern of High Performance Liquid Chromatography was determined from the corresponding peak curves ((E)-aldosecologanin, chlorogenic acid, luteolin 7-O-glucoside, sweroside). For L. japonica, additional unknown peaks were detected at 13.8 min, 20.6 min, and 36.9 min. And, we developed genetic marker using the the tRNA-Leu gene, trnL-trnF intergenic spacer and tRNA-Phe region of chloroplast DNA. By the method, 164 bp PCR product amplified from L. confusa was distinguished into L. japonica and L. confusa efficiently. Conclusion : Base on these results, two techniques provide effective approaches to distinguish L. japonica from L. confusa.

Analysis of the Content of Paeoniflorin in Peony Roots Cultivated on Kyeongbuk Area (경북지역(慶北地域)에서 재배중(栽培中)인 작약(芍藥)의 Paeoniflorin 함량분석(含量分析))

  • Kim, Tae Kang;Joo, Gil Jae;Chung, Jae Dong;Rhee, In Koo
    • Current Research on Agriculture and Life Sciences
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    • v.14
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    • pp.15-28
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    • 1996
  • The effective components of the peony cultivar, Euseongjagyag and Youngcheonjagyag, which were the major cultivars of peony in Kyeongbuk area, were determined with HPLC and TLC. The paeoniflorin content in the root of Euseongjagyag were more than that of Youngcheonjagyag. The root of Euseongjagyag contained much albiflorin and Youngcheonjagyag contained much oxypaeoniflorin in comparision with albiflorin and oxypaeoniflorin contents in both cultivars of peony. Paeoniflorin contents in accordance with peony prodution regions were ranged from 2.15% to 4.08%, and paeoniflorin content of local cultivar of Euseong and that of Geochang were approximatly the same but that of Youngcheon was the lowest. Paeoniflorin content in the 18 accessions of peony cultivar which were collected from Kyeongbuk area and harvested on November 1993, were ranged from 1.41% to 5.30%. The 18 accessions of peony were classified with the HPLC chromatogram pattern of peony root extract into the three groups which composed with Euseong peony group(9 accessions), Youngcheon peony group (4 accessions) and Punggi standard peony group(5 accessions). High content of paeoniflorin WaS contained in peony root harvested in May and November. but low content of paeoniflorin was contained in peony root harvested in March and September.

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The improvement of ginsenoside accumulation in Panax ginseng as a result of γ-irradiation

  • Kim, Dong Sub;Song, Mira;Kim, Sun-Hee;Jang, Duk-Soo;Kim, Jin-Baek;Ha, Bo-Keun;Kim, Sang Hoon;Lee, Kyung Jun;Kang, Si-Yong;Jeong, Il Yun
    • Journal of Ginseng Research
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    • v.37 no.3
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    • pp.332-340
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    • 2013
  • In this study, gamma rays were used to irradiate embryogenic calli induced from cotyledon explants of Panax ginseng Meyer. After the embryogenic calli were irradiated, they were transferred to adventitious roots using an induction medium; next, mutated adventitious root (MAR) lines with a high frequency of adventitious root formations were selected. Two MAR lines (MAR 5-2 and MAR 5-9) from the calli treated with 50 Gy of gamma rays were cultured on an $NH_4NO_3$-free Murashige and Skoog medium with indole-3-butyric acid 3 mg/L. The expression of genes related to ginsenoside biosynthesis was analyzed using reverse transcription polymerase chain reaction with RNA prepared from native ginseng (NG), non-irradiated adventitious root (NAR) and 2 MAR lines. The expression of the squalene epoxidase and dammarenediol synthase genes was increased in the MAR 5-2 line, whereas the phytosterol synthase was increased in the MAR 5-9 line. The content and pattern of major ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1) were analyzed in the NG, NAR, and 2 MAR lines (MAR 5-2 and MAR 5-9) using TLC and HPLC. In the TLC analysis, the ginsenoside patterns in the NG, NAR, and 2 MAR lines were similar; in contrast, the MAR 5-9 line showed strong bands of primary ginsenosides. In the HPLC analysis, compared with the NG, one new type of ginsenoside was observed in the NAR and 2 MAR lines, and another new type of ginsenoside was observed in the 2 MAR lines irradiated with gamma rays. The ginsenoside content of the MAR 5-9 line was significantly greater in comparison to the NG.

Safety Evaluation of Filamentous Fungi Isolated from Industrial Doenjang Koji

  • Lee, Jin Hee;Jo, Eun Hye;Hong, Eun Jin;Kim, Kyung Min;Lee, Inhyung
    • Journal of Microbiology and Biotechnology
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    • v.24 no.10
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    • pp.1397-1404
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    • 2014
  • A few starters have been developed and used for doenjang fermentation but often without safety evaluation. Filamentous fungi were isolated from industrial doenjang koji, and their potential for mycotoxin production was evaluated. Two fungi were isolated; one was more dominantly present (90%). Both greenish (SNU-G) and whitish (SNU-W) fungi showed 97% and 95% internal transcribed spacer sequence identities to Aspergillus oryzae/flavus, respectively. However, the SmaI digestion pattern of their genomic DNA suggested that both belong to A. oryzae. Moreover, both fungi had morphological characteristics similar to that of A. oryzae. SNU-G and SNU-W did not form sclerotia, which is a typical characteristic of A. oryzae. Therefore, both fungi were identified to be A. oryzae. In aflatoxin gene cluster analysis, both fungi had norB-cypA genes similar to that of A. oryzae. Consistent with this, aflatoxins were not detected in SNU-G and SNU-W using ammonia vapor, TLC, and HPLC analyses. Both fungi seemed to have a whole cyclopiazonic acid (CPA) gene cluster based on PCR of the maoA, dmaT, and pks-nrps genes, which are key genes for CPA biosynthesis. However, CPA was not detected in TLC and HPLC analyses. Therefore, both fungi seem to be safe to use as doenjang koji starters and may be suitable fungal candidates for further development of starters for traditional doenjang fermentation.

Plankton Community Composition Related to Marine Environmental Factors in Haengam Bay (행암만 해양환경요인과 플랑크톤 군집구조의 변동)

  • Kim, Jeong Bae;Hong, Sokjin;Lee, Won-Chan;Kim, Hyung Chul;Lee, Yong-Woo;Youn, Seok-Hyun;Cho, Yoonsik
    • Journal of Environmental Science International
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    • v.23 no.12
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    • pp.2015-2028
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    • 2014
  • We analyzed with HPLC (High Performance Liquid Chromatography) analysis photosynthetic pigments and environmental factors, microscopic observations of the phytoplankton and zooplankton in the seawater every month from February 2009 to November 2010 in Haengam Bay. The level of dissolved inorganic nutrients was the highest between July and September, when freshwater influx was at its peak, whereas chlorophyll a levels were the highest in April and August. Also, phytoplankton pigment concentration increased when dissolved inorganic nutrients are carried into nearshore waters by rainfall runoff. Based on identification of phytoplankton and photosynthetic pigments results, diatoms were mainly dominant while dinoflagellate populations increased at July and August 2009, May 2010. The zooplankton communities are dominated in terms of Noctiluca scintillans. The contribution of Noctiluca scintillans in 2010 accounts for approximately 77.3% of the total zooplankton. Distribution patterns over time of zooplankton in the seasonal distribution of phytoplankton showed a different pattern.

Major Metabolites Involved in Skin Blackening of 'Niitaka' Pear Stored under Cold Temperature (신고 배 저온 저장 중 발생하는 과피 흑변에 관여하는 주요 대사체)

  • Lee, Eun Jin
    • Horticultural Science & Technology
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    • v.32 no.3
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    • pp.359-365
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    • 2014
  • Oriental pear (Pyrus communis L. cv 'Niitaka') was stored at $0^{\circ}C$ for 5 months and major metabolites involved in blackening of the peel were analyzed by untargeted GC-MS and targeted HPLC methods. In this study, peels of sound and skin-blackened pears were analyzed and compared. Skin-blackened fruit was clearly characterized by a distinctive pattern in changes which included a decrease of malic acid, succinic acid, and ascorbic acid, while an increase of fumaric acid, threonine, and gluconic acid, which indicated both reduced metabolic activity and anti-oxidative capacity of the cells. Chlorogenic acid was a major phenolic compound and the peel of sound fruit showed high levels of free phenolic compounds compared than the peel of skin-blackened fruit which are believed to be related to oxidation of phenolics in skin-blackened tissue. The changes or profiling of major metabolites by targeted or untargeted analysis method could become a useful tool for understanding physiology, disorder mechanism, and identifying metabolic networks connecting primary and secondary metabolism in postharvest research.

Analysis of Related Compounds from Commercial Atenolol Raw Materials and Preparations by High-Performance Liquid Chromatography (HPLC를 이용한 시판 아테놀롤 원료 및 제품 중 유연물질의 분석)

  • Dong, Nguyen Thanh;Kang, Ji-Youn;Jung, Young-Hee;Lim, Eun-Hee;Hwang, Gwi-Seo;Kang, Chan-Soon;Kim, Eun-Jung;Kang, Jong-Seong
    • Journal of Pharmaceutical Investigation
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    • v.34 no.6
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    • pp.453-457
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    • 2004
  • Atenolol and related compounds found in raw materials and commercial products were analyzed by reversed-phase high-performance liquid chromatography. A mixed solution of phosphate buffer (3.4 g/l, pH 3.0), tetrahydrofurane and methanol (800:20:180, v/v/v) including sodium octanesulfonate (1 g/l) and tetrabutylammonium-hydrogensulfate (0.4 g/l) was used as mobile phase at the flow rate of 0.25 ml/min. Detection was carried out at UV 226 nm. Atenolol related compounds, such as bis ether, tertiary amine and blocker acid were identified by comparing the retention time of the standard. The within-day and between-day precisions of the separated compounds were less than 1.2% and 3.4%, respectively. The contents of related compounds of the tested samples were under the limit prescribed in the European Pharmacopoeia. The pattern of the related compounds showed that atenolol raw materials and products could be classified in three different groups, indicating that the materials originated from different source or treated in different way.

Analysis of Scutellaria baicaleinsis Georgi (Scutellariae Radix) by LC-DAD and LC-ESI/MS

  • Yu, Youngbob;Choi, Pil-Son;Koo, Sungtae;Chang, Suhwan
    • Korean Journal of Plant Resources
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    • v.31 no.6
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    • pp.652-659
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    • 2018
  • In this study, baicalin, as a marker substance of Scutellariae Radix, was quantitatively analyzed by a high performance liquid chromatography-photodiode array detector (HPLC-DAD). We identified wogonoside, baicalein, and wogonin in the Scutellariae Radix by a high performance liquid chromatography-electrospray ionization-mass spectrometer (HPLC-ESI-MS). The baicalin was separated on a Xterra C18 column ($5{\mu}m$, $4.6{\times}250mm$) using mobile phase consisting of 38% acetonitrile in 0.68% phosphoric acid. The baicalin spectrum in the Scutellariae Radix extracts was coincided by comparing with UV-visible spectrum (200-550 nm) of baicalin standard in the library. The amount of baicalin in Scutellariae Radix was 10.46%, which is higher than KFDA's guideline. The marker substances of Scutellariae Radix showed a strong base peak $[M]^+$ in the positive detection mode following as; baicalin (m/z; $271[MH^+-sugar]^+$, $447[M+H]^+$), wogonoside (m/z; $285[MH^+-sugar]^+$, $461[M+H]^+$), baicalein (m/z; $271\;[M+H]^+$), wogonin (m/z; $285[M+H]^+$). These results are consistent with the fragment pattern and molecular weight of standard components from literature.

Characterization of degradation products of the Balsalazide by Mass spectrometry: Optimization of stability-indicating HPLC method for separation and quantification of process related impurities of Balsalazide

  • Chilakabattina Naga Narasimha Babu;Ch. Srinivasa Reddy;Bhagya Kumar Tatavarti;M. Radha Madhavi;Venkateswara Rao Anna
    • Analytical Science and Technology
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    • v.37 no.1
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    • pp.25-38
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    • 2024
  • The study aimed to investigate a novel approach by utilizing liquid chromatography (LC) and liquid chromatography-mass spectrometry (LC-MS) to separate, identify and characterize very nominal quantities of degradation products (DPs) of balsalazide along with its process related impurities without isolation from their reaction mixtures. The impurities along with balsalazide were resolved on spherisorb ODS2 (250×4.6 mm, 5.0 ㎛) column at room temperature using 0.2 M sodium acetate solution at pH 4.5 and methanol in the ratio of 55:45 (v/v) as mobile phase pumped isocratically at 1.0 mL/min as mobile phase and UV detection at 255 nm. The method shows sensitive detection limit of 0.003 ㎍/mL, 0.015 ㎍/mL and 0.009 ㎍/mL respectively for impurity 1, 2 and 3 with calibration curve liner in the range of 50-300 ㎍/mL for balsalazide and 0.05-0.30 for its impurities. The balsalazide pure compound was subjected to stress studies and a total of four degradation products (DPs) were formed during the stress study and all the DPs were characterized with the help of their fragmentation pattern and the masses obtained upon LC-MS/MS. The DPs were identified as 3-({4-[(E)-(4-hydroxyphenyl) diazenyl]benzoyl}amino)propanoic acid (DP 1), 4-[(E)-(4-hydroxyphenyl)diazenyl] benzamide (DP 2), 5-[(E)-(4-carbamoylphenyl)diazenyl]-2-hydroxybenzoic acid (DP 3) and 3-({4-[(E)-phenyldiazenyl]benzoyl}amino)propanoic acid (DP 4). Based on findings, it was concluded that, the proposed method was successfully applicable for routine analysis of balsalazide and its process related impurities in pure drug and formulations and also applicable for identification of known and unknown impurities of balsalazide.

Quantitative Comparison of Ginsenosides and Nitrogen Compounds in Korean Ginsengs and Related Origin (고려인삼류와 유사생약중 ginsenoside 및 질소화합물의 함량 비교)

  • Shin, Jae-Young;Park, Hee-Juhn;Lim, Sang-Cheol;Jung, Won-Tae
    • Korean Journal of Pharmacognosy
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    • v.27 no.1
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    • pp.6-14
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    • 1996
  • To differentiate the quality of Korean ginseng from those of other habitats, the quantitative analysis of free amino acids(FAA) and total amino acids(TAA) in addition to ginsenoside Rb1 and Rg1 was carried out using amino acid analyzer and HPLC, respectively. FAA pattern in Korean ginseng was much different from that of Panax notoginseng. The difference in total content of FAA was also found that Korean ginseng contained 26.3-39.8mg/g while Panax notoginseng contained 6.5mg/g. This FAA content had a tendency to increase with the age of radix. The contents of FAA and TAA in Korean ginseng(6 years old) from Kumsan were shown to be the highest than other ginseng origins tested. The content in the 6 years Panax ginseng from China was about same with that of 4 years Korean ginseng of Kumsan. However, regarding to gisenoside Rbl and Rg1, which have been accepted as the characteristic components of Panax ginseng-Panax notoginseng showed considerably higher content than those of any other ginseng origin.

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