• Title/Summary/Keyword: HPLC analysis methods

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Method validation for quantitative analyzing aflatoxin productivity in Aspergillus sp. isolated from soybean paste

  • SeongEui Yoo;WooSoo Jeong;Soo-Hwan Yeo;So-Young Kim
    • Food Science and Preservation
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    • v.30 no.1
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    • pp.28-41
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    • 2023
  • Non-aflatoxigenic Aspergillus oryzae and aflatoxigenic A. flavus cannot be clearly identified by partial sequencing of the internal transcribed spacer (ITS) and 18S ribosomal ribonucleic acid (18S rRNA) regions. This study aimed to compare the accuracy among three aflatoxin detection methods using ultra-performance liquid chromatography (UPLC), high-performance liquid chromatography (HPLC), and an enzyme-linked immunosorbent assay (ELISA) kit and to select the non-aflatoxigenic Aspergillus sp. isolated from soybean paste. All analytical methods were suitable according to the international standards of Codex Alimentarius FAO-WHO (CODEX) or the Ministry of Food and Drug Safety (MFDS). UPLC exhibited the best of limit of detection (LOD) and limit of quantification (LOQ). Based on UPLC, HPLC, and the ELISA kit assay, the P5 and P7 strains isolated from soybean paste had 1,663.49, 1,468.12, and >20 ㎍/kg and 1,470.08, 1,056.73, and >20 ㎍/kg, respectively, detected and re-identified as A. flavus. In contrast, the P3 and P4 strains (A. oryzae), which were detected below the MFDS standards in all assays, were confirmed as non-aflatoxigenic fungi. Among the methods evaluated for quantitative analysis of aflatoxin, UPLC and HPLC are superior in terms of accuracy, and the ELISA kit rapidly detects low concentrations of aflatoxin. Furthermore, this study demonstrates that any Aspergillus sp. isolated for use as a fermentation starter should be analyzed for potential aflatoxin production using UPLC and HPLC for accurate quantitative analysis or ELISA for the rapid detection of low-level concentrations of aflatoxin.

Component Analysis of Bee Venom from lune to September (월별 봉독의 함량 비교 분석)

  • Chu, Ching-Seng;Lee, Jang-Ho;Kim, Min-Ki;Cha, Bae-Chun;Kwon, Ki-Rok
    • Journal of Pharmacopuncture
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    • v.10 no.2 s.23
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    • pp.93-98
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    • 2007
  • Objectives : The aim of this study was to observe variation of Bee Venom content from the collection period. Methods : Content analysis of Bee Venom was rendered using HPLC method by standard melittin Results : Analyzing melittin content using HPLC, 478.97mg/g at june , 493.89mg/g at july, 468.18mg/g at August and 482.15mg/g was containing in Bee Venom at september. So the change of melittin contents was no significance from June to September. Conclusion : Above these results, we concluded carefully that collecting time was not important factor for the quality control of Bee Venom, restricted the period from June to September.

Comparative Analysis of Metabolites in Roots of Panax ginseng Obtained from Different Sowing Methods (파종 방법에 따른 고려인삼의 대사체 비교)

  • Yang, Seung Ok;Lee, Sung Woo;Kim, Young Ock;Lee, Sang Won;Kim, Na Hyun;Choi, Hyung Kyoon;Jung, Joo Yeoun;Lee, Dong Ho;Shin, Yu Su
    • Korean Journal of Medicinal Crop Science
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    • v.22 no.1
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    • pp.17-22
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    • 2014
  • Ginsenosides of roots in Panax ginseng were analyzed by metabolic-targeting HPLC using the partial least squares discriminant analysis (PLS-DA) and compared depending on sowing methods between direct seeding and transplanting method. Score plots derived from PLS-DA could identify the sowing method between the direct seeding and transplanting method in P. ginseng roots. The ginsenoside compounds were assigned as Rg1, Re, Rf, Rg2, Rb1, Rc, Rb2, Rb3, and Rd. Contents of Re, Rf, Rg2, Rb1, Rc, Rb3, and Rd of main roots produced from the transplanting method were relatively higher than those of samples produced from direct seeding method. Also, contents of Rg1, Re, Rf, Rg2, Rb1, Rc, Rb2, Rb3, and Rd of lateral roots from the transplanted samples were relatively higher than those of samples produced from direct seeding method. Therefore, HPLC with PLS-DA analysis can be a straightforward tool for identification of ginsenosides in main or lateral roots of P. ginseng obtained from two different seeding methods between direct and transplanting methods.

Analysis of Imidazoline Type Cationic Surfactants (Imidazoline계 양이온 계면활성제 분석)

  • Bak, Hong-Soon;Choi, Kyu-Yeol;Lee, Jae-Duk;Kim, Yeo-Kyung;Ahn, Ho-Jeong
    • Applied Chemistry for Engineering
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    • v.9 no.3
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    • pp.404-406
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    • 1998
  • Analysis for the imidazoline type cationic surfactants was performed by the gas chromatography(GC) and high performance liquid chromatography(HPLC). The composition of the alkyl chain distribution was investigated by GC after base/Acid hydrolysis of the imidazoline ring. The distribution of total alkyl chain was separated clearly by a Bondclone C18/NOVA-Pak C18 HPLC column using 50% acetonitrile in methanol containing 0.1M sodium perchlorate as a mobile phase. Alkyl chain distribution and average molecular weight of imidazoline type cationic surfactants were obtained based on these analytical methods. The agreement of results from GC and HPLC was good. The detection limit of imidazoline by HPLC method was 10ng without pretreatment.

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Simultaneous Analysis of Six Compounds in Yijin-tang by HPLC-PDA (HPLC-PDA을 이용한 이진탕 중 6종 성분의 동시분석)

  • Kim, Seong-Sil;Kim, Jung-Hoon;Shin, Hyeun-Kyoo;Seo, Chang-Seob
    • Herbal Formula Science
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    • v.21 no.1
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    • pp.71-79
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    • 2013
  • Objectives : Yijin-tang has been used in the treatment of gastrointestinal diseases such as irritable bowel syndrome, gastritis, and gastric ulcer. In this study, a high-performance liquid chromatography (HPLC) method was established for simultaneous analysis of six compounds, liquiritin, glycyrrhizin, hesperidin, 6-gingerol, homogentisic acid, and 3,4-dihydroxybenzaldehyde in Yijin-tang, a traditional Korean herbal medicinal preparation. Methods : A Gemini C18 column was used for the separation of six constituents at $40^{\circ}C$. The mobile phase using gradient elution consist of two solvent systems, 1.0% acetic acid in water (A) and 1.0% acetic acid in acetonitrile (B). The flow-rate was 1.0 mL/min and injection volume was $10{\mu}g$. The detector was a photodiode array (PDA) detector set at 254 nm and 280 nm. Results : The calibration curves of six compounds showed good linearity in various concentration ranges ($R^2{\geq}0.9997$). The limits of detection (LOD) and limits of quantification (LOQ) were 0.028-$0.192{\mu}g/mL$ and 0.093-$0.540{\mu}g/mL$, respectively. The RSD (%) of the intra and inter day validations were 0.03-0.84% and 0.05 -1.00%, respectively. Recovery was 96.14-01.90% and RSD (%) was less than 1.5%. Conclusions : The established simultaneous analysis methods will help management to improve the quality of Yijin-tang.

Identification of Anthocyanin from The Extract of Soybean Seedcoat

  • Park, Sun-Mi;Kim, Ji-Na;Dung, Tran-Huu;Do, Le Thanh;Thu, Do Thi Anh;Sung, Mi-Kyung;Kim, Jong-Sang;Yoo, Hoon
    • International Journal of Oral Biology
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    • v.36 no.2
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    • pp.59-64
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    • 2011
  • Anthocyanins are naturally occuring phytochemicals and the main components of the coloring of plants, flowers and fruits. They are known to elicit antioxidative, anti-inflammatory and cancer preventive activity. In this study, we investigated anthocyanins in black / yellow soybean seedcoats using different methods of detection - thin layer chromatography (TLC), capillary zone electrophoresis (CZE) and HPLC analysis. The anthocyanins in soybean seedcoats were extracted by five independent methods of extraction and the aglycons (anthocyanidins) of the corresponding anthocyanins were prepared by acid mediated hydrolysis. The anthocyanin / anthocyanidin in black soybean seedcoat showed characteristic TLC mobility, CZE electrophoretic retention and HPLC migration time while little of anthocyanins were detected from yellow soybean seedcoat. The extracted anthocyanins showed pH dependent retention time in CZE and spectral change in UV-Vis spectrum. HPLC analysis of the hydrolyzed extract of black soybean seedcoat identified the presence of four anthocyanidins. The major anthocyanin in black soybean seedcoat was cyanin (cyanidin-3-O-glucoside), with the relative order of anthocyanidin in cyanidin > delphinidin > petunidin > pelargonidin.

Development of Alternative Testing Methods without Hazardous Reagents used in Korean Pharmaceutical Codex (고시의약품 시험에 사용되는 유해시약 대체 시험법 개발)

  • Kim, Hee-Yun;Kang, Hyun-Kyung;Choi, Seon-Hee;Bang, Su-Jin;Han, Kyung-Jin;Choi, Sung-Hee;Kim, Jin-Hee;Lee, Hwa-Jung;Kang, Chan-Soon
    • YAKHAK HOEJI
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    • v.54 no.2
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    • pp.142-149
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    • 2010
  • Development of alternative testing methods for the replacement of hazardous reagents with less hazardous ones is strongly enforced because exposure of human and environment to hazardous reagents are restricted and hazardous reagents are gradually prohibited from using in various testing methods. Thus, in this study, we developed 8 monographs from the Korean Pharmaceutical Codex by substituting the use of the hazardous reagents including ICH class 1 such as benzene, chloroform and dioxane to the use of less toxic ones like ICH class 2 or 3 reagents. We also improved their qualification and quantification performance. Among 8 monographs, the 6 newly developed TLC methods for the identification of nifedipine, oxolamine citrate, ketoprofen lysinate, chlorquinaldol, retinol acetate, and riboflavin showed a clear spot of corresponding material without any interference in spite of the replacement with ICH class 2 or 3 reagents. For the quantification of domperidone and trimebutine, HPLC methods were developed for the substitution of UV/VIS spectrometry and titrimetry, respectively. These HPLC methods were validated for the linearity, recovery, reproducibility, and inter-laboratory variations. In conclusion, the newly developed methods could be expected to become valuable tools for revising the Korean Pharmaceutical Codex.

Rapid Determination of Ascorbic Acid in Red Pepper Leaves by Near-Infrared Reflectance Spectroscopic Analysis (근적외 분광분석법에 의한 고춧잎의 Ascorbic Acid 함량 측정)

    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.3
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    • pp.393-398
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    • 1998
  • The loss of ascorbic acid in dried red pepper leaves prepare with different drying methods of air-, oven-, microwave oven-, and vacuum drying with blanching or without was determined by a HPLC method. Vacuum drying showed the least loss of ascorbic acid than the other drying methods. Additionally, the feasibility of near infrared reflectance spectroscopy(NIRS) to determine the contents of ascorbic acid in the red pepper leaves was studied. NIRS was found to be an efficient way of determining ascorbic acid contents in red pepper leaves, requiring only 30 seconds of an analytical time.

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The Difference of Ginsenoside Compositions According to the Conditions of Extraction and Fractionation of Crude Ginseng Saponins (추출 및 분획조건에 따른 인삼 조사포닌 중 ginsenoside 조성 차이)

  • Shin, Ji-Young;Choi, Eon-Ho;Wee, Jae-Joon
    • Korean Journal of Food Science and Technology
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    • v.33 no.3
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    • pp.282-287
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    • 2001
  • This study was carried out to investigate the difference of ginsenoside compositions in crude ginseng saponins prepared by five different methods including three new methods. Two known methods are hot methanol(MeOH) extraction/n-butanol(n-BuOH) fractionation and hot MeOH extraction/Diaion HP-20 adsorption/MeOH elution. Three new methods are hot MeOH extraction/cation AG 50W $absorption/H_2O$ elution/n-BuOH extraction, cool MeOH extraction/Diaion HP-20 adsorption/MeOH elution and direct extraction with ethyl acetate(EtOAc)/n-BuOH. Analysis of ginsenoside composition in the crude saponins by conventional HPLC/RI(Refractive Index) did not show great difference between methods except EtOAc/n-BuOH method. However, HPLC/ELSD (evaporative light scattering detector) employing gradient mobile phase afforded fine resolution of ginsenoside Rf, $Rg_1$ and $Rh_1$, and great difference of ginsenoside compositions between methods. LC/MS revealed that large amount of prosapogenins were produced during the pass through the cation exchange (AG 50W) column being strongly acidic. Six major ginsenosides such as $Rb_1,w;Rb_2,$ Rc, Rd, Re and $Rg_1$, 5 prosapogenins and one chikusetsusaponin were identified by LC/MS. A newly established HPLC method employing ODS column and gradient mobile phase of $KH_2PO_4/CH_3CN$ revealed that malonyl ginsenosides were detected only in the crude saponin obtained from cool MeOH extraction.

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HPLC Fractionation of Antioxidant Substances of E. hyemale Extract and Analysis of Indicator Components Using LC-MS (속새(Equisetum hyemale) 추출물의 항산화 물질의 HPLC 분획과 LC-MS를 이용한 지표성분 분석)

  • Song, Jin Hwa;Lee, Geo Lyong
    • Journal of Naturopathy
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    • v.10 no.2
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    • pp.108-113
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    • 2021
  • Background: Results on the action of antioxidants in extracts of Equisetum hyemale stems and roots have already been reported, but the antioxidant properties have not been analyzed. Purpose: This study was to determine the molecular structure of antioxidants in substances extracted from E. hyemale stems and roots. Methods: Component analysis was analyzed by HPLC and LC-MS after extraction with hot water and ethyl alcohol. Results: The HPLC chromatogram of stem and root extracts showed four significant peaks at a wavelength of 205 nm. Peak 1 at 280 nm is a typical simple phenolic type, and both peaks 2-4 near 280 nm and 370 nm are typical flavonoid glycosides. As for the antioxidant level of the extract by HPLC analysis, the sum of the peaks at 740 nm was the highest at 3,669 mAU in the 100% ethanol extract, 3,096 mAU in the 70% ethanol extract, and 2,868 mAU in the hot water extract. As a result of LC-MS analysis of the antioxidant extract, kaempferol-3-sophoroside-7-glucoside with a molecular weight of 772 da at peak 3, and kaempferol-3-sophoroside-8-glucoside with a molecular weight of 788 and 772 at peak 4 was identified. Conclusions: The above results show that two types of antioxidants were identified in the antioxidant extract of E. hyemale exrtracts. Therefore, the potential as a raw material for functional cosmetics has increased.