• Title/Summary/Keyword: HPLC Chromatography

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Purification and Characterization of Sulfated Polysaccharide Isolated from Hot Water Extract of Pachymeniopsis elliptica (Pachymeniopsis elliptica의 열수 추출물로부터 분리한 함황 다당류의 정제 및 특성)

  • Lee, Sun-Hee;Jun, Woo-Jin;Yu, Kwang-Won;Chun, Hyug;Shin, Dong-Hoon;Hong, Bum-Shik;Cho, Hong-Yon;Yang, Han-Chul
    • Korean Journal of Food Science and Technology
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    • v.32 no.5
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    • pp.1191-1197
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    • 2000
  • In the preliminary study, we investigated the anti-complementary activities of 62 extracts from Korean edible seaweeds. Of those, Pachymeniopsis elliptica showed the highest anti-complementary activity. Therefore, it was purified as follows; i) PE-1 by ethanol precipitation, ii) PE-1-C by ultrafiltration, iii) PE-1-CIV by DEAE-Toyopearl 650C, and iv) PE-1-CIV-ii by Sepharose CL-6B. The purified compound, PE-1-CIV-ii, was the complexed homogeneous polysaccharide (molecular mass: 780 kDa) with 82.9% of anti-complementary activity. Also, it contained a significant amount of sulfate group (30.5%), which indicated it as a sulfated algal polysaccharide. Its structural monosaccharides were galactose (44.3%), 3,6-anhydrogalactose (34.0%), glucose (8.2%), fucose (5.4%), xylose (5.2%) and rhamnose (2.9%). After the treatment of periodate on a sample, a significant decrease in anti-complementary activity was found, which was a characteristic of bioactive polysaccharides. And-tumor activity of PE-1-A, B and C was tested in the sarcoma-180 solid tumor model. The PE-1-C with the largest molecular mass (more than 300 kDa) showed 81% of inhibition on the solid tumors, suggesting that the anti-complementary activity was, at least in part, related to anti-tumor activity. Based upon these results, the purified polysacchardes could be an immunopotentiator in vivo.

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Protective effect of Gabjubaekmok (Diospyros kaki) extract against amyloid beta (Aβ)-induced cognitive impairment in a mouse model (아밀로이드 베타(amyloid beta)로 유도된 인지장애 마우스 모델에서 갑주백목(Diospyros kaki) 추출물의 인지기능 및 뇌 신경세포 보호 효과)

  • Yoo, Seul Ki;Kim, Jong Min;Park, Seon Kyeong;Kang, Jin Yong;Han, Hye Ju;Park, Hyo Won;Kim, Chul-Woo;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.51 no.4
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    • pp.379-392
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    • 2019
  • The current study investigated the effect of Gabjubaekmok (Diospyros kaki) ethanolic extract (GEE) on $H_2O_2$-induced human neuroblastoma MC-IXC cells and amyloid beta $(A{\beta})_{1-42}$-induced ICR (Institute of Cancer Research) mice. GEE showed significant antioxidant activity that was evaluated based on ABTS, DPPH scavenging activity, and inhibition of malondialdehyde (MDA) and acetylcholinesterase activity. Further, GEE inhibited ROS production and increased cell viability in $H_2O_2$-induced MC-IXC cells. Administration of GEE ameliorated the cognitive dysfunction on $A{\beta}$-induced ICR mice as evaluated using Y-maze, passive avoidance, and Morris water maze tests. Results of ex vivo test using brain tissues showed that, GEE protected the cholinergic system and mitochondrial functions by increasing the levels of antioxidants such as ROS, mitochondrial membrane potential (MMP), and adenosine triphosphate (ATP) against $A{\beta}$-induced cognitive dysfunction. Moreover, GEE decreasd the expression levels of apoptosis-related proteins such as $TNF-{\alpha}$, p-JNK, p-tau, BAX and caspase 3. While, expression levels of p-Akt and $p-GSK3{\beta}$ increased than $A{\beta}$ group. Finally, gallic acid was identified as the main compound of GEE using high performance liquid chromatography.

Changes in the constituents and UV-photoprotective activity of Astragalus membranaceus caused by roasting (황기의 볶음 조건에 따른 성분 및 자외선 광보호 활성 변화)

  • Park, Jeong-Yong;Lee, Ji Yeon;Kim, Hyung Don;Jang, Gwi Yeong;Seo, Kyung Hye
    • Journal of Nutrition and Health
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    • v.52 no.5
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    • pp.413-421
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    • 2019
  • Purpose: Astragalus membranaceus (AM) is an important traditional medicinal herb. Pharmacological research has indicated that AM has various physiological activities such as antioxidant, anti-inflammatory, immunoregulatory, anticancer, hypolipidemic, antihyperglycemic, and hepatoprotective activities. The bioactive substances responsible for the physiological activities in AM, including many antioxidant substances, change during the roasting process. This study investigated and compared the changes in the antioxidant constituents of AM caused by roasting. Methods: DPPH (1,1-diphenyl-2-picryl hydrazyl) and $ABTS^+$ (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) radical scavenging activities and their total phenolic content (TPC) were measured. High-performance liquid chromatography (HPLC) analysis was performed to confirm any changes in the isoflavonoids of roasted AM (R-AM),. The cell viability of UVB-induced HDF (Human dermal fibroblast) cells treated with AM and R-AM extracts was investigated. The comet assay was used to examine the inhibitory effects of R-AM extracts on DNA damage caused by oxidative stress. Results: The DPPH and $ABTS^+$ radical scavenging activities were $564.6{\pm}20.9$ and $108.2{\pm}3.1$ ($IC_{50}$ value) respectively, from the 2R-AM. The total phenol content was $47.80{\pm}1.40mg$ GAE/g from the 1R-AM. The values of calycosin and formononetin, which are the known isoflavonoid constituents of AM, were $778.58{\pm}2.72$ and $726.80{\pm}3.45{\mu}g/g$ respectively, from the 2R-AM. Treatment of the HDF cells with R-AM ($50{\sim}200{\mu}g/mL$) did not affect the cell viability. Furthermore, the R-AM extracts effectively protected against UVB-induced DNA damage. Conclusion: The findings of this study indicate that R-AM increases its isoflavonoid constituents and protects against UVB-induced DNA damage in HDF cells.

Contents of vitamin B9 (folate) and B12 (cobalamins) in commonly consumed seafood menus in Korea (한국인 상용 수산물 식단의 비타민 B9과 B12 함량)

  • Park, Eun-Young;Jeong, Bomi;Chun, Jiyeon
    • Journal of Nutrition and Health
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    • v.54 no.2
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    • pp.211-223
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    • 2021
  • Purpose: A total of 39 seafood menus were prepared according to the Korean standard recipe, and analyzed for vitamin B9 (folate) and B12 (cobalamins) contents, using validated applied analytical methods. The menus included Guk/Tang/Jjigae (boiled or stewed dishes, n = 10), Bokkeum (stir-fried dishes, n = 10), Jjim/Jorim (braised or steamed dishes, n = 7), Gui (baked or grilled dishes, n = 7), Twigim (deep-fried dishes, n = 2) and Muchim (dried or blanched-seasoned dishes, n = 3). Methods: The contents of vitamin B9 and B12 in all food samples were determined by the trienzyme extraction-Lactobacillus casei and immunoaffinity-high-performance liquid chromatography/photodiode array detection methods. Analytical quality control was performed in order to assure reliability of the analysis. Results: Accuracy (97.4-100.6% recoveries) and precision (< 6% relative standard deviations for repeatability and reproducibility) of vitamin B9 and B12 analyses were determined to be excellent. The vitamin B9 and B12 contents of the 39 seafood menus evaluated, varied in the range of 1.83-523.08 ㎍/100 g and 0.11-38.30 ㎍/100 g, respectively, depending on the ingredients and cooking methods. The vitamin B9 content was highest in Jomi-gim (523.08 ㎍/100 g), followed by Geonsaeu-bokkeum (128.34 ㎍/100 g) and Janmyeolchi-bokkeum (121.53 ㎍/100 g). Vitamin B12 was detected in all seafood menus, with highest level obtained in Kkomack-jjim (41.58 ㎍/100 g). The seaweed dish was found to have high levels of both vitamin B9 and B12. All assays were performed under strict quality control. Conclusion: Guk and Tang menus, which contain a large amount of water, were relatively lower in the vitamin B9 and B12 contents than the other menus. Bokkeum menus containing various vegetables were high in the vitamin B9 content, but the vitamin B12 content was dependent on the type of seafood used in the menu.

Development and Validation of Analytical Method and Antioxidant Effect for Berberine and Palmatine in P.amurense (황백의 지표성분 berberine과 palmatine의 분석법 개발과 검증 및 항산화 효능 평가)

  • Jang, Gill-Woong;Choi, Sun-Il;Han, Xionggao;Men, Xiao;Kwon, Hee-Yeon;Choi, Ye-Eun;Park, Byung-Woo;Kim, Jeong-Jin;Lee, Ok-Hwan
    • Journal of Food Hygiene and Safety
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    • v.35 no.6
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    • pp.544-551
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    • 2020
  • The aim of this study was to develop and validate a simultaneous analytical method for berberine and palmatine, which are representative substances of Phellodendron amurense, and to evaluate the antioxidant activity. We evaluated the specificity, linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) of analytical methods for berberine and palmatine using high-performance liquid chromatography. Our result showed that the correlation coefficients of the calibration curve for berberine and palmatine exhibited 0.9999. The LODs for berberine and palmatine were 0.32 to 0.35 µg/mL and the LOQs were 0.97 to 1.06 µg/mL, respectively. The inter-day and intra-day precision values for berberine and palmatine were from 0.12 to 1.93 and 0.19 to 2.89%, respectively. The inter-day and intra-day accuracies were 98.43-101.45% and 92.39-100.60%, respectively. In addition, the simultaneous analytical method was validated for the detection of berberine and palmatine. Moreover, we conducted FRAP and NaNO2 scavenging activity assays to measure the antioxidant activities of berberine and palmatine, and both showed antioxidant activity. These results suggest that P.amurense could be a potential natural resource for antioxidant activity and that the efficacy can be confirmed by investigating the content of the berberine and palmatine.

Analysis of Components in the Different Parts of Lythrum salicaria L. (털부처꽃의 부위별 성분 분석)

  • Kim, Hee-Young;Park, Yea-Jin;Lee, Ju-Yeon;Kim, Ki-young;Shin, Su;Choi, Min-Woo;Hong, Eun-Jin;Kim, Min-jeong;Yeo, Sujung;Park, In-hwa;Jerng, Ui Min;An, Hyo-Jin;Cha, Yun-Yeop
    • The Korea Journal of Herbology
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    • v.37 no.5
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    • pp.89-96
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    • 2022
  • Objectives : This research was performed to analyze the components in the different parts of Lythrum salicaria L. and to compare which parts of L. salicaria L. are appropriate for food development. Methods : L. salicaria L. was extracted in 20% EtOH at 100 ℃ for 4 hours. Cytotoxicity was investigated in 3T3-L1 cells after treatment of 10-500 ㎍/ml L. salicaria L. for 24 hours. Total polyphenol content (TPC) was estimated using 1 N Folin-ciocateu reagent. 2,2-Diphenyl-1-picryhydrazyl (DPPH) radical scavenging activity was estimated using DPPH reagent and gallic acid. The chemical composition was analyzed by high-performance liquid chromatography (HPLC). 1) Results : The half maximal inhibitory concentration (IC50) in the extracts of the whole plant, aerial parts, and root parts was 350 ㎍/ml, over 500 ㎍/ml, and 150 ㎍/ml, respectively. The TPC in the extracts of the whole plant, aerial parts, and root parts was 527.1 mg/g, 422.6 mg/g, and 781.1 mg/g, respectively. The averages of vitexin contents in the aerial parts, and root parts were 256.7 ± 154.9 ㎍/g and 266.1 ± 63.2 ㎍/g, respectively. The averages of TPC in the leaves, roots, flower stalks and stems were 224.0 ± 53.7 tannin acid (TA) mg/g, 221.8 ± 70.2 TA mg/g, 249.8 ± 34.4 TA mg/g, and 67.7±8.9 TA mg/g, respectively. The averages of DPPH radical scavenging activity in the leaves, roots, flower stalks, and stems were 282.01 ± 43.3 gallic acid equivalent (GAE) 𝜇mole/g, 260.16 ± 44.1 GAE 𝜇mole/g, 288.0 ± 9.3 GAE 𝜇mole/g, and 97.6 ± 10.7 GAE 𝜇mole/g, respectively. Conclusions : There were no significant differences in the content of components or antioxidant activity in the aerial parts compared to those in the whole plant of L. salicaria L. Furthermore, the root parts had low extract yield, cytotoxicity, and quality control problems, therefore our results suggest that the use of the aerial part of L. salicaria L. would be the most appropriate for food development.

Validation of an Analytical Method for Deacetylasperulosidic acid, Total Sugar and Monosaccharide Analysis in Fermented Morinda citrifolia Polysaccharide Powder (발효노니 다당체 분말의 deacetylasperulosidic acid, 총당 및 단당류 분석법 검증)

  • Kwon, Heeyeon;Choi, Jisoo;Kim, Soojin;Kim, Eunmin;Uhm, Jihyun;Kim, Bokyung;Lee, Jaeyeon;Kim, Yongdeok
    • Journal of Food Hygiene and Safety
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    • v.37 no.4
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    • pp.216-224
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    • 2022
  • This study was aimed at validating the analysis methods for deacetylasperulosidic acid (DAA), total sugar, galacturonic acid, glucose, and galactose, which are the indicator components of fermented Morinda citrifolia polysaccharide extract (Vitalbos). We modified the previously reported methods for validating the analytical methods. The specificity, linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) were measured using phenol-sulfuric acid method and high-performance liquid chromatography (HPLC). The retention time and spectrum of the standard solution of Vitalbos coincided, confirming the specificity. The calibration curve correlation coefficient (R2), of five indicator components, ranged from 0.9995-0.9998, indicating excellent linearity of 0.99 or more. The intra-day and inter-day precision range of the assay was 0.14-3.01%, indicating a precision of less than 5%. The recovery rate was in the range of 95.13-105.59%, presenting excellent accuracy. The LOD ranged from 0.39 to 0.84 ㎍/mL and the LOQ ranged from 1.18 to 2.55 ㎍/mL. Therefore, the analytical method was validated for DAA, total sugar, galacturonic acid, glucose, and galactose, in Vitalbos. The indicator component content in Vitalbos was determined using a validated method. The contents of DAA, total sugar, galacturonic acid, glucose, and galactose were 2.31±0.06, 475.92±5.95, 72.83±1.05, 71.63±2.44, and 67.30±2.31 mg/g of dry weight, respectively. These results suggest that the developed analytical method is efficient and could contribute to the quality control of Vitalbos, as a healthy functional food material.

Survey of Daily Caffeine Intakes from Children's Beverage Consumption and the Effectiveness of Nutrition Education (어린이들의 음료를 통한 카페인 섭취량 실태조사 및 영양교육에 따른 효과 평가)

  • Kim, Sung-Dan;Yun, Eun-Sun;Chang, Min-Su;Park, Young-Ae;Jung, Sun-Ok;Kim, Dong-Gyu;Kim, Youn-Cheon;Chae, Young-Zoo;Kim, Min-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.6
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    • pp.709-720
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    • 2009
  • This study was conducted to identify daily caffeine intakes in beverages for elementary school children and to evaluate its effectiveness after nutrition education. The caffeine contents of 140 commercial beverages were analysed by high performance liquid chromatography-ultraviolet detector (HPLC-UV) and information about their consumption were obtained by surveying 267 children. Researchers gave nutrition education to the children, who were 6 to 11 years old and attended 9 classes of 3 elementary schools, by lecture, Powerpoint file and moving picture. Their preference and intake amount on beverages were investigated by questionnaire before and after nutrition education. The order on caffeine contents was coffee ($33.8{\pm}2.4{\sim}49.1{\pm}5.6\;mg/100\;mL$)> coffee milk ($10.6{\pm}3.3\;mg/100\;mL$)> cola ($6.0{\pm}2.4\;mg/100\;mL$)> green black oolong tea drink ($6.0{\pm}2.4\;mg/100\;mL$)> chocolate milk and chocolate drink ($1.6{\pm}0.7{\sim}1.7\;mg/100\;mL$)> black ice tea mix ($1.3{\pm}1.7\;mg/100\;mL$). The order on children's preference was carbonated drink and fruit and vegetable drink (27%)> sports drink (26%)> processed cocoa mix (7%)> milk (6%)> vitamin & functional drink (3%)> green tea drink (2%)> black tea drink and coffee (1%). The average daily caffeine intakes except tea drink was $5.9{\pm}11.2$ mg/person/day ($0.17{\pm}0.32$ mg/kg bw/day), ranged from $0.0{\sim}80.5$ mg/person/day for children. The sources of caffeine were coffee 57% (3.4 mg/person/day), coffee milk 20% (1.2 mg/person/day), carbonated drink 15% (0.9 mg/person/day), chocolate milk and chocolate drink 6% (0.4 mg/person/day), and vitamin & functional drink 2% (0.1 mg/person/day). After nutrition education, the preference of carbonated drink, coffee, vitamin drinks & functional drink was decreased significantly (p<0.05, p<0.05, p<0.01) and the intakes of carbonated drink, chocolate milk & chocolate drink, and vitamin & functional drink were also decreased significantly (p<0.01, p<0.05, p<0.01). This study has shown that nutrition education influences the preference and the intake behavior of caffeinated beverages.