• 제목/요약/키워드: HPLC analysis methods

검색결과 485건 처리시간 0.035초

유산균 발효에 의한 사물탕들부터 노다케네틴의 분리 및 함량분석 (Analysis of Nodakenetin from Samultangs Fermented by Lactose Bactera Strains)

  • 김동선;노주환;조장원;마진열
    • 대한본초학회지
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    • 제27권1호
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    • pp.35-39
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    • 2012
  • Objectives : The purpose of this study was to investigate the changes in the contents of constituents in Samultang and its fermentations with 10 species of lactic acid bacteria. Methods : Ten strains of lactic acid bacteria, Lactobacillus casei, L. acidophilus, L. casei, L. plantarum, L. amylophilus, L. curvatus, L. delbruekil subsp. lactis, L. casei, B. breve, and B. thermophilum, were used for the fermentation of Samultang. The increased and decreased constituents were identified using HPLC/DAD and various liquid chromatographic techniques, and the structure was elucidated using NMR and MS. These compounds were quantitatively analyzed using an HPLC/DAD system. Results : A remarkably increased component was identified to be nodakenetin and a decreased component was determined to be nodakenin. The fermentation of the ten lactic acid bacteria demonstrated that the decomposable rate of these two compounds in fermented Samultang were different. Samultang fermented by L. plantarum showed the most remarkable changes. Conclusion : Nodakenetin was identified as bioconversion component after fermentation and L. plantarum was discovered the best bacteria to increase the component.

오미자(五味子) 추출물의 인간 전립선암 세포주 PC-3에 대한 성장 억제 효과 (Antiproliferative effect of Schisandrae Fructus extract on PC-3 human prostate cancer cells)

  • 문정민;석가형;조수인
    • 대한본초학회지
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    • 제27권4호
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    • pp.17-23
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    • 2012
  • Objectives : Schisandrae Fructus (SF) has traditionally been used to balance level of body fluid and to strengthen kidney function. It has been reported that the SF extract has antioxidant, hepatoprotective, neuroprotective and anticancer effects. This study investigated an antiproliferative effect of SF extract on PC-3 human prostate cancer cells and analyzed active ingredients of SF extract qualitatively and quantitatively. Methods : We examined the antiproliferative effect of SF extract with MTT assay, DAPI staining and annexin-V/7-AAD double staining. The active ingredients of SF extract were identified by using HPTLC and HPLC/DAD system. Results : SF-chloroform fraction inhibited growth of PC-3 cells and changed the morphology of nucleus in a dose dependent manner. A dose-dependent apoptotic cell death was also measured by flow cytometry analysis. It was analyzed that SF-chloroform fraction contained more schizandrin than other fractions by using HPTLC and HPLC/DAD system. Conclusions : These results suggest that SF extract and schizandrin may be a potential chemotherapeutic agent for the control of PC-3 human prostate cancer cells.

Major Components of Clinically used Bee Venom Pharmacopuncture

  • Jo, Na Young;Roh, Jeong Du
    • Journal of Acupuncture Research
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    • 제34권1호
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    • pp.31-38
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    • 2017
  • Objectives : The purpose of this study was to analyze the components of the clinically used bee venom (BV) pharmacopuncture. Methods : Two kinds of bee venom pharmacopuncture (BV-I and II), three kinds of separate purification BV (SPBV) pharmacopuncture (SPBV-I, II, and III), and apitoxin were investigated in this study. We performed a component analysis of melittin, apamin, and phospholipase $A_2$ using high-performance liquid chromatography (HPLC). Results :1. BV-I contained approximately 40% more melittin than BV-II did. 2. In the three separate purification BV pharmacopuncture, SPBV-I, SPBV-II, and SPBV-III, phospholipase $A_2$ content decreased remarkably. 3. The melittin content in SPBV-I increased by 5% compared to that in BV-I. 4. The amount of melittin in apitoxin was similar to that in SPBV-I. Conclusion : The compositions of the BV pharmacopuncture and separate purification BV pharmacopuncture changed depending on the collection method and concentration. Therefore, it is necessary to choose the most suitable BV for each specific medical treatment target. Furthermore, research into the composition of BV may be needed for its safe and effective use.

물리적 가공법을 이용한 강황으로부터 Curcuminoid의 고체상추출 (Solid-Phase Extraction of Curcuminoid from Turmeric Using Physical Process Method)

  • 이광진;양혜진;정상원;마진열
    • 생약학회지
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    • 제43권3호
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    • pp.250-256
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    • 2012
  • In order to extract the curcuminoid such as curcumin, demethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC) in turmeric (Curcuma longa), solvent extraction methods (dipping and ultrasonic extraction method) and solid-phase extraction (SPE) were used. RP-HPLC (reverse-phase high-performance liquid chromatography) and TLC (thin-layer chromatography) were used for identification and analysis the three curcuminoid. From the experimental results, it is evident that the percentage of curcuminoid extracted from turmeric by ultrasonic extraction method was higher than dipping method. The percentage of curcumin extracted from turmeric by pure methanol was higher than any aqueous methanolic composition. Moreover, the total peak area of three curcuminoid was above 92% in RP-HPLC using solid-phase extraction. These results will form a database for investigating the constituents of natural products and the resources of pharmaceutical, nutrition, and cosmetic products.

Clq-coated ELISA법을 이용한 정맥용 면역글로불린제제의 항보체성 측정 (Clq-Coated Microtitre Enzyme-linked Immunosorbent Assay for Measuring the Anticomplementary Activity of Intravenous Immunoglobulin Preparations)

  • 강혜나;김순남;신광훈;허숙진
    • 약학회지
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    • 제45권6호
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    • pp.656-663
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    • 2001
  • The quality of an intravenous immunoglobulin preparation (IVIG) is reflected by the degree of nonspecific activation of complements, the so-called anticomplementary activity (ACA). ACA of aggregates in IVIG was investigated using method by the European Pharmacopoeia and Clq-coated microtiter enzyme-linked immunosorbent assay (ELISA). Both the EP method and the ELISA method showed a dose response curve with the amount of complements bound increasing with the percentage content of aggregates in immunoglobulin standard. The correlation between the two tests was good (r=0.96, r=0.99). However, the correlation was not found when the ACA (EP method) of IVIG product was compared with its aggregate percentage. These results emphasize that the method of aggregate formation affects ACA and that estimation of the percentage distribution of aggregates by HPLC may not reflect ACA. In analysing WIG product for Clq binding activity test with the ELISA, the result by using Protein A-HRP correlated with aggregate percentage (r=0.84). But the correlation decreased (r=0.48) when the result used Protein A-AP(having poorer sensitivity than HRP) was compared with aggregate percentage. As a result, some variation between the two methods, due to differences in assay principles, is to be expected. However, ELISA technique has the advantage in that it is easier to perform, more precise and less subject to reagent variability, and is the more suitable screening method than HPLC analysis.

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Alpha-Tocopherol Contents of Peanut Seeds Depend on Varieties and Processing Methods

  • Doo, Hong-Soo
    • 한국자원식물학회지
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    • 제20권6호
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    • pp.553-557
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    • 2007
  • The purpose of this study is to establish an extraction and analysis method for $\alpha$-tocopherol, and to then distinguish among varieties. The $\alpha$-tocopherol contents of 22 varieties of peanut seeds were analyzed by HPLC. Peanut seeds of cv. Palkwang were processed in two ways, by boiling and roasting. The $\alpha$-tocopherol contents of the two types of peanuts were compared with fresh seeds without any processing. $\alpha$-Tocopherol was detected at a retention time of 2.95 minutes. Five standards of $\alpha$-tocopherol covering a range of $20{\sim}100{\mu}g/ml$ were made up in 2% isopropyl alcohol/n-hexane and analyzed in duplicate. The $\alpha$-tocopherol contents differed according to extraction temperature. The contents were $85{\mu}g/g$ or less at $10^{\circ}C\;and\;20^{\circ}C$ and $94{\mu}g/g$ at $30^{\circ}C$, but they were decreased at $40^{\circ}C$ or higher. The $\alpha$-tocopherol contents in 22 peanut varieties were $61.36{\sim}96.80{\mu}g/g$ according to variety. Fresh peanuts contained $106.7{\mu}g/g$ of $\alpha$-tocopherol, while boiled peanuts contained $108.8{\mu}g/g$ of $\alpha$-tocopherol, and roasted peanuts contained $109.2{\mu}g/g$ of $\alpha$-tocopherol.

Liquid Chromatographic Determination of Etofenprox Residues in Foods with Mass-Spectrometric Confirmation

  • Lee, Young-Deuk;Kwon, Chan-Hyeok;Kwon, Ki-Sung
    • 한국환경농학회지
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    • 제30권4호
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    • pp.432-439
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    • 2011
  • BACKGROUND: An official analytical method was developed to determine etofenprox residues in agricultural commodities using high-performance liquid chromatography (HPLC). METHODS AND RESULTS: The etofenprox residue was extracted with acetone from representative samples of five raw products which comprised rice grain, apple, mandarin, cabbage, and soybean. The extract was then serially purified by liquid-liquid partition and Florisil column chromatography. For rice and soybean samples, acetonitrile/n-hexane partition was additionally coupled to remove nonpolar lipids. Reversed phase HPLC using an octadecylsilyl column was successfully applied to separate etofenprox from co-extractives. Intact etofenprox was sensitively detected by ultraviolet absorption at 225 nm. Recovery experiment at the quantitation limit validated that the proposed method could apparently determine the etofenprox residue at 0.02 mg/kg. Mean recoveries from five crop samples fortified at three levels in triplicate were in the range of 93.6~106.4%. Relative standard deviations of the analytical method were all less than 10%, irrespective of crop types. A selected-ion monitoring LC/mass spectrometry with positive atmospheric-pressure chemical ionization was also provided to confirm the suspected residue. CONCLUSION(s): The proposed method is simple, rapid and sensitive enough to be employed in routine inspection or monitoring of agricultural products for the etofenprox residue.

Analysis of Anti-adipogenic Constituents of Cordyceps militaris Using High Performance Liquid Chromatography-Diode Array Detection in Different Samples: Comparison with Anti-adipogenic Activity

  • Liu, Qing;Hong, In-Pyo;Han, Sang-Bae;Hwang, Bang-Yeon;Lee, Mi-Kyeong
    • Natural Product Sciences
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    • 제18권3호
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    • pp.171-176
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    • 2012
  • We previously isolated cordycepin, guanosine and tryptophan from Cordyceps militaris as antiadipogenic constituents. For the quality control of C. militaris for anti-adipogenic activity, simultaneous analytical method using high-performance liquid chromatography (HPLC)-diode array detection (DAD) was developed and validated. Quantitation of these compounds in various Cordyceps samples from different sources and various extraction methods were conducted using developed method. Our study shows that natural Cordyceps and host insect possess higher content than cultured ones and fruiting bodies, respectively. The content of cordycepin showed great difference in different C. militaris samples whereas trytophan content was similar in tested samples. Addition of water to extraction solvent greatly increased the yield of guanosine and tryptophan. High temperature and longer extraction time increased yield of guanosine, whereas the content of trytophan was decreased in high temperature during extraction with water. Extraction using ultrasonic apparatus slightly increased extraction efficiency. Cordycepin, however, has little variation in different extraction method tested. Strong anti-adipogenic activity was observed in the samples that contain all the three constituents. Taken together, quantitation of these compounds using developed analytical method might provide basic requirement for the anti-adipogenic activity of C. militaris.

후박의 품질평가 (Quality Evaluation on Magnoliae Cortex)

  • 배기환;김영호;원도희;이준성;강종성
    • 약학회지
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    • 제41권4호
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    • pp.407-413
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    • 1997
  • Magnolol and honokiol, the main components of Magnoliae Cortex, were isolated and used as the standard substances for the analysis. In order to determine the contents of magnolol and honokiol in Magnoliae Cortex originated from Korea, China and Japan, both HPLC and HPTLC methods are applied and compared with each other. The components were separated on C8 column with acetonitrile-water-acetic acid (50:50:1) in HPLC and detected at UV 294nm. The components separated on HPTLC precoated silica gel plate with chloroform-methanol (9:1) were detected directly on the plate at 254nm. The contents of magnolol and honokiol in Magnoliae Cortex were in the wide range of 0.01~2.8% and 0.005~0.8%, respectively, according to their purchase places. It is also applicable to the quality control of various preparation from Magnoliae Cortex.

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Determination of L-Carnitine in Infant Powdered Milk Samples after Derivatization

  • Park, Jung Min;Koh, Jong Ho;Kim, Jin Man
    • 한국축산식품학회지
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    • 제41권4호
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    • pp.731-738
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    • 2021
  • Herein, a novel analytical method using a high-performance liquid chromatography-fluorescence detector (HPLC/FLD) is developed for rapidly measuring an L-carnitine ester derivative in infant powdered milk. In this study, solid-phase extraction cartridges filled with derivatized methanol and distilled water were used to effectively separate L-carnitine. Protein precipitation pretreatment was carried out to remove the protein and recover the analyte extract with a high recovery (97.16%-106.56%), following which carnitine in the formula was derivatized to its ester form. Precolumn derivation with 1-aminoanthracene (1AA) was carried out in a phosphate buffer using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) as the catalyst. Method validation was performed following the AOAC guidelines. The calibration curves were linear in the L-carnitine concentration range of 0.1-2.5 mg/L. The lower limit of quantitation and limit of detection of L-carnitine were 0.076 and 0.024 mg/L, respectively. The intra- and interday precision and recovery results were within the allowable limits. The results showed that our method helped reduce the sample preparation time. It also afforded higher resolution and better reproducibility than those obtained by traditional methods. Our method is suitable for detecting the quantity of L-carnitine in infant powdered milk containing a large amount of protein or starch.