• Title/Summary/Keyword: Chromatographic methods

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Neuroprotective effects of antioxidant constituents isolated from Opuntia ficus-indica var. saboten Makino

  • K.J. Jung;Lee, E.H.;Kim, H.J.;Lee, J.Y.;Y.S. Song;Lee, Y.H.;J. Cho;Park, M.;Park, H.
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.63-63
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    • 2003
  • Opuntia ficus-indicavar. saboten Makino (Cactaceae) is a tropical or subtropical plant that has been widely used as folk medicine for the treatment of diabetes, asthma, burn, edema and gastritis. The purposes of the present study were to identify antioxidant constituents from fruits and stems of the plant cultivated in Cheju island, Korea, and examine their in vitro neuroprotective activities. Using a chromatographic fractionation method, ten chemical constituents were isolated from ethyl acetate extracts. By means of chemical and spectroscopic methods, those were identified as eight flavonoids such as kaempherol (a), quercetin (b), kaempferol 3-methyl ether (c), quercetin 3-methyl ether (d), narcissin (e), dihydrokaernpferol (f), dihydroquercetin (g) and erioclictyol (h), and two terpenoids such as 3-oxo-${\alpha}$-ionol-${\beta}$-d-glucopyranoside (i) and roseoside (j). Among the isolated compounds, comrounds c~e and h~j were those reported for the first titre from the plant. Compounds b, d and g showed DPPH free radical scavenging activities with IC$\sub$50/ values of 28, 19 and 31, ${\mu}$M respectively. Compounds d and g also inhibited iron-dependent lipid peroxidation with IC$\sub$50/ values of 2.4 and 3.5 ${\mu}$M. In a primary rat cortical neuronal cell culture system, compounds b, d and g inhibited xanthine/xanthine oxidase-induced (IC$\sub$50/ values of 18.2, 2.1 and 54.6 ${\mu}$M) and H$_2$O$_2$-induced (IC$\sub$50/ values of 13.6, 1.9 and 25.7 ${\mu}$M) cytotoxicities. In addition, compounds d and g inhibited NMDA-induced excitotoxicity by 21 and 33%, and only compound d inhibited growth factor withdrawal-induced apoptosis by 31% at a tested concentration of 3 ${\mu}$M. The results suggest that the antioxidant constituents with in vitroneuroprotective activities may serve as lead chemicals for the development of neuroprotective agent.

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The Development and Application of New Chromatographic Methods Using Smart Devices (스마트 기기를 활용한 새로운 크로마토그래피 분석법 개발 및 적용)

  • Jae Hwan Lee;Ye Geon Choi;Jae Jeong Ryoo
    • Journal of Science Education
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    • v.48 no.2
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    • pp.91-100
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    • 2024
  • The use of smart devices in science classes has brought about positive changes, such as increased student participation and more self-directed learning. Smart devices are increasingly being used in science classes, creating a need to develop lesson models that can stimulate students' interest and encourage active, self-directed learning in scientific inquiry and experimental activities. In smart education, smart devices and applications play a major role. However, in the "Mixture Separation" section of middle school science, chromatography focuses mainly on paper chromatography, which is not currently used in the field of actual research. This approach is not well-suited for students preparing for a new future society, and it is becoming obsolete due to curriculum revisions. Although chromatography can be used as an activity for career exploration, removing it is not convincing. The advantage of using thin-layer chromatography (TLC), which is employed in actual research, is that it is inexpensive and easy to use in classroom settings. In this study, we have developed a new, faster, and simpler analysis method for TLC that uses smart devices for both qualitative and quantitative analysis. We hope this method will enhance student engagement and facilitate small-scale learning by integrating smart devices into learning activities, making it a practical tool for actual school settings.

Analysis of Amperometric Response to Cholesterol according to Enzyme-Immobilization Methods (효소고정화 방법에 따른 콜레스테롤 검출용 바이오센서의 전류 감응도 분석)

  • Ji, Jung-Youn;Kim, Mee-Ra
    • Journal of the East Asian Society of Dietary Life
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    • v.21 no.5
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    • pp.731-738
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    • 2011
  • Cholesterol is the precursor of various steroid hormones, bile acid, and vitamin D with functions related to regulation of membrane permeability and fluidity. However, the presence of excess blood cholesterol may lead to arteriosclerosis and hypertension. Moreover, dietary cholesterol may affect blood cholesterol levels. Generally, cholesterol determination is performed by spectrophotometric or chromatographic methods, but these methods are very time consuming and costly, and require complicated pretreatment. Thus, the development of a rapid and simple analysis method for measuring cholesterol concentration in food is needed. Multi-walled carbon nanotube (MWCNT) was functionalized to MWCNT-$NH_2$ via MWCNT-COOH to have high sensitivity to $H_2O_2$. The fabricated MWCNT-$NH_2$ was attached to a glassy carbon electrode (GCE), after which Prussian blue (PB) was coated onto MWCNT-$NH_2$/GCE. MWCNT-$NH_2$/PB/GCE was used as a working electrode. An Ag/AgCl electrode and Pt wire were used as a reference electrode and counter electrode, respectively. The sensitivity of the modified working electrode was determined based on the amount of current according to the concentration of $H_2O_2$. The response increased with an increase of $H_2O_2$ concentration in the range of 0.5~500 ${\mu}M$ ($r^2$=0.96) with a detection limit of 0.1 ${\mu}M$. Cholesterol oxidase was immobilized to aminopropyl glass beads, CNBr-activated sepharose, Na-alginate, and toyopearl beads. The immobilized enzyme reactors with aminopropyl glass beads and CNBr-activated sepharose showed linearity in the range of 1~100 ${\mu}M$ cholesterol. Na-alginate and toyopearl beads showed linearity in the range of 5~50 and 1~50 ${\mu}M$ cholesterol, respectively. The detection limit of all immobilized enzyme reactors was 1 ${\mu}M$. These enzyme reactors showed high sensitivity; especially, the enzyme reactors with CNBr-activated sepharose and Na-alginate indicated high coupling efficiency and sensitivity. Therefore, both of the enzyme reactors are more suitable for a cholesterol biosensor system.

Two new triterpenoid saponins derived from the leaves of Panax ginseng and their antiinflammatory activity

  • Li, Fu;Cao, Yufeng;Luo, Yanyan;Liu, Tingwu;Yan, Guilong;Chen, Liang;Ji, Lilian;Wang, Lun;Chen, Bin;Yaseen, Aftab;Khan, Ashfaq A.;Zhang, Guolin;Jiang, Yunyao;Liu, Jianxun;Wang, Gongcheng;Wang, Ming-Kui;Hu, Weicheng
    • Journal of Ginseng Research
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    • v.43 no.4
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    • pp.600-605
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    • 2019
  • Background: The leaves and roots of Panax ginseng are rich in ginsenosides. However, the chemical compositions of the leaves and roots of P. ginseng differ, resulting in different medicinal functions. In recent years, the aerial parts of members of the Panax genus have received great attention from natural product chemists as producers of bioactive ginsenosides. The aim of this study was the isolation and structural elucidation of novel, minor ginsenosides in the leaves of P. ginseng and evaluation of their antiinflammatory activity in vitro. Methods: Various chromatographic techniques were applied to obtain pure individual compounds, and their structures were determined by nuclear magnetic resonance and high-resolution mass spectrometry, as well as chemical methods. The antiinflammatory effect of the new compounds was evaluated on lipopolysaccharide-stimulated RAW 264.7 cells. Results and conclusions: Two novel, minor triterpenoid saponins, ginsenoside $LS_1$ (1) and 5,6-didehydroginsenoside $Rg_3$ (2), were isolated from the leaves of P. ginseng. The isolated compounds 1 and 2 were assayed for their inhibitory effect on nitric oxide production in LPS-stimulated RAW 264.7 cells, and Compound 2 showed a significant inhibitory effect with $IC_{50}$ of $37.38{\mu}M$ compared with that of NG-monomethyl-L-arginine ($IC_{50}=90.76{\mu}M$). Moreover, Compound 2 significantly decreased secretion of cytokines such as prostaglandin $E_2$ and tumor necrosis factor-${\alpha}$. In addition, Compound 2 significantly suppressed protein expression of inducible nitric oxide synthase and cyclooxygenase-2. These results suggested that Compound 2 could be used as a valuable candidate for medicinal use or functional food, and the mechanism is warranted for further exploration.

Simultaneous Determination of Fructose, Glucose, and Sucrose in Honey and Commercial Drinks by GC and GC/MS (GC 및 GC/MS를 이용한 벌꿀 및 시판 음료 중의 Fructose, Glucose 및 Sucrose의 동시분석)

  • Yun, Jeong-Sik;Jeon, Hyun-Suk;Kim, In-Suk;Lee, Hee-Jin;Lee, Hye-Jeong;Hyun, Jae-Yeoul;Kim, Jong-Bae
    • Korean Journal of Food Science and Technology
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    • v.45 no.5
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    • pp.537-544
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    • 2013
  • This study was performed to establish method of simultaneous determination of fructose, glucose, and sucrose in honey and commercial vitamin drinks by GC and GC/MS. Optimum chromatographic separation of trimethylsilyl-oxime (TMSO) derivatives by GC was achieved on a DB-5 column. Calibration curves for fructose, glucose and sucrose TMSO derivatives by GC were linear in the range of 50-5000 ${\mu}g/mL$, and their $r^2$ values were 0.9999, The limit of detection and limit of quantification of fructose, glucose, and sucrose were 0.68, 0.47, and 0.53 ${\mu}g/mL$, respectively, and 2.27, 1.58, and 1.77 ${\mu}g/mL$, respectively. Average recoveries of fructose, glucose, and sucrose were 100.5, 101.0, and 99.7%, respectively. When the method was applied to 12 honey samples, the average concentrations of fructose, glucose and sucrose were $42.58{\pm}1.97%$, $27.74{\pm}1.16%$, and $0.79{\pm}0.52%$, respectively. The F/G ratio was $1.53{\pm}0.07$. For fructose and glucose contents, results from the GC analysis were similar to those from the HPLC analysis, but the sucrose content was different for each analysis method. We suggest that the GC method is more suitable than other official analytical methods for simultaneous determination of fructose, glucose, and sucrose in honey.

Determination of methamphetamine, 4-hydroxymethamphetamine, amphetamine and 4-hydroxyamphetamine in urine using dilute-and-shoot liquid chromatography-tandem mass spectrometry (시료 희석 주입 LC-MS/MS를 이용한 소변 중 메스암페타민, 4-하이드록시메스암페타민, 암페타민 및 4-하이드록시암페타민 동시 분석)

  • Heo, Bo-Reum;Kwon, NamHee;Kim, Jin Young
    • Analytical Science and Technology
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    • v.31 no.4
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    • pp.161-170
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    • 2018
  • The epidemic of disorders associated with synthetic stimulants, such as methamphetamine (MA) and amphetamine (AP), is a health, social, legal, and financial problem. Owing to the high potential of their abuse and addiction, reliable analytical methods are required to detect and identify MA, AP, and their metabolites in biological samples. Thus, a dilute-and-shoot liquid chromatography-tandem mass spectrophotometry (LC-MS/MS) was developed for simultaneous determination of MA, 4-hydroxymethamphetamine (4HMA), AP, and 4-hydroxyamphetamine (4HA) in urine. Urine sample ($100{\mu}L$) was mixed with $50{\mu}L$ of mobile phase consisting of 0.4 % formic acid and methanol and $50{\mu}L$ of working internal-standard solution. Aliquots of $8{\mu}L$ diluted urine was injected into the LC-MS/MS system. For all analytes, chromatographic separation was performed using a C18 reversed-phase column with gradient elution and a total run time of 5 min. The identification and quantification were performed by multiple reaction monitoring (MRM). Linear least-squares regression was conducted to generate a calibration curve, with $1/x^2$ as the weighting factor. The linear ranges were 2.0-200, 1.0-800, and 10-2500 ng/mL for 4HA and 4HMA, AP, and MA, respectively. The inter- and intraday precisions were within 6.6 %, whereas the inter- and intraday accuracies ranged from -14.9 to 11.3 %. The low limits of quantification were 2.0 ng/mL (4HA and 4HMA), 1.0 ng/mL (AP), and 10 ng/mL (MA). The proposed method exhibited satisfactory selectivity, dilution integrity, matrix effect, and stability, which are required for validation. Moreover, the purification efficiency of high-speed centrifugation was clearly higher than 6-15 % for QC samples (n=5), which was higher than that of the membrane-filtration method. The applicability of the proposed method was tested by forensic analysis of urine samples from drug abusers.

Evaluation of Biogenic Amines in Korean Commercial Fermented Foods (국내 유통 발효식품 중 biogenic amines 함량 분석)

  • Han, Gyu-Hong;Bahn, Kyeong-Nyeo;Son, Yeong-Wook;Jang, Mi-Ran;Lee, Chang-Hee;Kim, So-Hee;Kim, Dae-Byoung;Kim, Seon-Bong;Cho, Tae-Yong
    • Korean Journal of Food Science and Technology
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    • v.38 no.6
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    • pp.730-737
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    • 2006
  • High performance liquid chromatographic methods for the simultaneous determination of 12 biogenic amines were developed and contents of biogenic amines in 23 varieties of Korean commercial fermented food were analyzed. Dansyl derivatives of biogenic amine were very stable and had good peak resolution. Except agmatine, the recovery of biogenic amines from soybean paste with extraction of 0.1 N HC1 added biogenic amines to soybean paste was greater 85%. The calibration curve showed good linearity over a concentration range up to 50 ${\mu}g/mL$. In the determination of biogenic amine level in Korean commercial fermented foods, doenjang and chungkukjang (fermented soybean pastes), soy sauce, anchovy and pacific sand lance liquid jeotkal (fermented fish sauce products), and cabbage kimchi had high level biogenic amines. Especially, traditional doenjang had a histamine (HIS) level of 952.0 mg/kg, a tyramine (TYR) level of 1,430.7 mg/kg. Most cheese had low level of biogenic amines, but one Gouda cheese had a tyramine (TYR) level of 97.5 mg/kg. A low level of biogenic amines was detected in wines, beer, yoghurt, and sausage. Putrescine (PUT), cadaverine (CAD), tryptamine (TRY), histamine (HIS), tyramine (TYR), and 2-phenylethylamine (PHE) were mainly formed in fermented foods by the action of microorganism, so their levels were high with a range. On the other hand, spermidine (SPD), spermine(SPM), serotonin (SER), noradrenaline (NOR), and dopamine (DOP) were formed originally via biosynthesis with consequent low level.

Radiopharmceutical Factors in the Prepartion of $^{99m}Tc-HMPAO$ Images of the Brain (뇌스캔용 $^{99m}Tc-HM-PAO$의 방사성 동위원소표지에 영향을 미치는 인자에 대한 연구)

  • Yeom, Mi-Kyoung;Kim, Sang-Eun;Lee, Dong-Soo;Chung, June-Key;Lee, Myung-Chul;Koh, Chang-Soon
    • The Korean Journal of Nuclear Medicine
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    • v.25 no.1
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    • pp.117-121
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    • 1991
  • Technetium-99m-hexamethylpropyleneamine oxime $(^{99m}Tc-HM-PAO)$ is a neutral-lipophilic chelate which is used for scanning cerebral blood flow. The labeling efficiencies of $^{99m}Tc-HM-PAO$ is known to be sensitive to the amount of pertechnetate added and the quality of the pertechnetate. Because of these factors, the manufacture recommends that HM-PAO kits be reconstituted with a maximum of 30 mCi pertechnetate which was eluted <4 hr earlier from a generator which had been eluted < 24 hr previously. So we measured the labelling efficiencies and the decomposition rate constant according to the amount of pertechnetate added, the volume of pertechnette added, and generator in-growth time. We used the 3-system chromatographic methods (paper & ITLC-SG chromatography) which analyzed the labelling efficiencies of the $^{99m}Tc-HM-PAO$. There was no significant difference in labelling efficiencies between variable pertechnetate acitvities added. ($39.9{\pm}4.9\;mCi:\;87.8{\pm}5.1\;(%)$, $60.8{\pm}5.0\;mCi:\;90.7{\pm}2.2\;(%)$, $79.0{\pm}6.0\;mCi:\;86.8{\pm}3.9\;(%)$, $106.6{\pm}11.6\;mCi:\;87.7{\pm}1.2\;(%)$, p>0.05) No significant difference in labelling efficiencies were found between pertechnetate of 4ml and 5ml. (4ml : $89.1{\pm}3.2(%)$, 5ml: $87.3{\pm}4.0(%)$, p>0.05). There was no difference between 1-6 and 10-48 hr of generator in-growth time. (1-6 hr: $87.8{\pm}4.0(%)$, 10-48 hr: $89.6{\pm}1.6(%)$, p>0.05) The mean value of decomposition rate constant was $0.196{\pm}0.097\;(hr^{-1})$, and there were no difference according to the amount of pertecnetate added and the volume of pertecnetate added, ($39.9{\pm}4.9\;mCi:\;0.208{\pm}0.059\;(hr^{-1})$, $60.8{\pm}5.0\;mCi:\;0.191{\pm}0.100\;(hr^{-1})$ $79.0{\pm}6.0\;mCi:\;0.192{\pm}0.118\;(hr^{-1})$, $106.6{\pm}11.6\;mCi:\;0.212{\pm}0.030\;(hr^{-1})$, p>0.05, 4 ml: $0.200{\pm}0.074\;(hr^{-1})$, Sml: $0.193{\pm}0.115\;(hr^{-1})$, p>0.05). In the case of using the first eluate, the labelling efficiency of $^{99m}Tc-HM-PAO$ W3S 82.1%. These data suggest that there were no significant alteration in labelling efficiency of $^{99m}Tc-HM-PAO$ according to the considerable range of pertechnetate activities and volume added, and generator in-growth time. Also, it was shown that one vial of HM-PAO kit supplied the $^{99m}Tc-HM-PAO$ which was used for 3-4 patients.

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Analytical Method Development for Determination of Silymarin by LC-MS/MS for Related Health Functional Foods (LC-MS/MS를 이용한 건강기능식품 중 실리마린 분석법 연구)

  • Oh, Mihyune;Lee, Jin Hee;Kim, Sang-A;Kim, Meehye
    • Journal of Food Hygiene and Safety
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    • v.33 no.2
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    • pp.124-130
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    • 2018
  • The Ministry of Food and Drug Safety (MFDS) is amending its test methods for the use of health functional foods (dietary food supplement), in order to establish regulatory standards and specifications in Korea. In this regard, we continue to pursue and perform our research on the analytical method development for the items being researched and reviewed. In this study, we have developed a sensitive and selective test method that could simultaneously separate and determinate six major bioactive flavonolignans in silymarin, which are based on the use of a liquid chromatographic-tandem mass spectrometry (LC-MS/MS). The standard calibration curves presented a linearity effect with the correlation coefficient ($r^2$) > 0.999. The limits of detection (LODs) and limits of quantitation (LOQs) were in the range of $0.3{\sim}9.0{\mu}g/L$ and $0.8{\sim}27.3{\mu}g/L$, respectively. The recovery results ranged between 96.2~98.6% at 3 different concentration levels, and its relative standard deviations (RSDs) were less than 5% as noted in this study. The proposed analytical method was characterized with a noted high resolution of the individual silymarin constituents, and the assay was fully validated as well. Our research can provide a significant scientific evidence that can be useful to amend the silymarin test method for the Health Functional Food Code.

The Biochemical Studies on Stored Soy-sauce (저장(貯藏)간장의 생화학적(生化學的) 연구(硏究))

  • Chang, Chi-Hyun
    • Applied Biological Chemistry
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    • v.9
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    • pp.9-27
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    • 1968
  • Studies were carried out in order to elucidate chemical components and microflora in three types of soy-sauce, 12-year aged soy-sauce prepared by improved method. 7-year aged and 20-year aged soy-sauce prepared by ordinary method. They results are summarized as follows: 1. The followings are found to be the important factors affecting the quality of soy sauce. a. Organic acids, reducing sugars and free amino acids were increased in the course of storage. b. In the aged soy-sauces under study non-volatile organic acid increased while volatile organic acid decreased and the total acidity was dependent only upon tie latter. c. It was found that suit concentration decreased during the storage. 2. The results of investigation of microflora in the stored soy-sauce are shown as follows. Soy-sauce Improved Ordinary Microbe 12-Y. 20-Y. 7-Y. Aerobic bacteria colony/1ml. 6 123 2 Halophilic lactic acid bacteria colony/1ml. 4 6 10 Osmophilic yeast colony/1ml. $828{\times}10^4$ 248 - b. In the stored soy-sauces, aerobic bacteria are incapable of growing due to drop in pH value and the influence of salt concentration. c. Halophilic lactic acid bacteria are incapable of growing due to drop in pH value, even the salt concentrations decreased during the storage. d. Osmophilic yeast are still growing in low pH value and in the decreasing salt concentration during the strage. 3. The results of amino acid analysis by paper partition chromatographic and calorimetric methods are shown as follows. a. Fourteen kinds of amino acid and thirteen amino acids were detected in the soy-sauce of 12-year aged improved soy-sauce and 7-year aged and 20-year aged ordinary one, respectively. b. The contents of aspartic acid, glutamic acid, serine, valine, leucine, lysine, histidine and methionine increased in the 20-year aged ordinary soy-sauce compared to the 7-year aged one. On the other hand those of alanine, tyrosine, phenylalanine and cystine decreased. 4. The results of sugar analysis by paper chromatography are as follows. a. In the 12-year aged improved soy-sauce, galactose, glucose, arabinose, xylose, rhamnose, maltose and an unknown were detected, and their amounts were in the above order except maltose and an unknown. b. Both in the 7 and 12-year aged ordinary soy-sauces, galactose, arabinose, xylose, glucose and rhamnose were detected and the amounts of the sugars were in the above order. c. In the non-aged ordinary soy-sauce, glucose was not detected but detected from 7-year and 20-year aged ordinary soy-sauce. 5. The results of organic acid analysis by paper chromatography were as follows. a. As volatile acids, acetic, propionic and butyric acids were detected in the 7-year aged ordinary soy-sauce. On the other hand in both the 20-year aged ordinary soy-sauce and the 12-year aged improved ones, only acetic acid was abundant while propionic and butyric acids were round in trace. It was found that propionic and butyric acids, as the unpleasant flavor components, decreased during the storage. b. In the ordinary soy-sauce, citric acid were produced during the storage and lack, malic and tartaric acids increased in the course of aging while succinic, glycolic, fumaric and malonic acids were shown to decrease. Glutaric and oxalic acids disappeared. Citric acid was produced also in the improved soy-sauce, but lactic, tartaric, succinic, malic, and glycolic acids decreased, while both malonic and glutaric acids disappeared. From the above results the citric acid production was considered to be a favorable factor for the taste. c. In the aged soy-sauces, pyruvic, α-ketoglutaric and probably acetoacetic and oxaloacetic acids (both in trace) were present and their amounts were in the above order. All of the α-keto acid abruptly decreased during the storage.

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