• Title/Summary/Keyword: HPLC assay

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Anti-oxidative and Anti-inflammatory Activities of Fermented Turmeric (Curcuma longa L.) by Rhizopus oryzae (Rhizopus oryzae으로 발효한 울금의 항산화 및 항염효과)

  • Kim, Eun-Ju;Song, Bit-Na;Jeong, Da-Som;Kim, So-Young;Cho, Yong-Sik;Park, Shin-Young
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1315-1323
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    • 2017
  • Turmeric is a rhizomatous herbaceous perennial plant (Curcuma longa (CL)) of the ginger family, Zingiberaceae. A yellow-pigmented fraction isolated from the rhizomes of CL contains curcuminoids belonging to the dicinnamoyl methane group. Curcumin is an important active ingredient responsible for the biological activity of CL. However, CL is not usually used as a food source due to its bitter taste. The present study was designed to determine the effect of the CL fermented by Rhizopus oryzae (FCL) on pro-inflammatory factors such as nuclear factor ${\kappa}B$ ($NF-{\kappa}B$), tumor necrosis factor alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-induced RAW 264.7 cell line. The cell viability was determined by MTT assay. To evaluate the anti-inflammatory effect of FCL 80% EtOH extracts, IL-6 and $TNF-{\alpha}$ were measured by ELISA kit. Also, the amount of $NO/PGE_2/NF-{\kappa}B$ was measured using the $NO/PGE_2/NF-{\kappa}B$ detection kit and the iNOS/COX-2 expression was measured by Western blotting. The results showed that the FCL reduced NO, $PGE_2$, iNOS, COX-2, $NF-{\kappa}B$, IL-6 and $TNF-{\alpha}$ production without cytotoxicity. These results suggest that FCL extracts may be a developed the functional food related to anti-inflammation due to the significant effects on inflammatory factors.

Antioxidants in Serum and Induced Sputum of COPD Patients (만성폐쇄성폐질환 환자의 혈청 및 유도객담에서의 항산화제)

  • Park, Hyeon-Kwan;Yu, Young-Kwon;Kim, Kyu-Sik;Lim, Sung-Chul;Kim, Young-Chul;Park, Kyung-Ok
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.2
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    • pp.158-170
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    • 2001
  • Background : Although an oxidants and antioxidants imbalance has been considered in the pathogenesis of chronic obstructive pulmonary disease (COPD), there is a paucity of reports focussing on the smoking-induced changes of oxidants and antioxidants in COPD. Method : The concentration of antioxidants (ascorbic acid, uric acid, retinol, and $\alpha$- & $\gamma$-tocopherol) was measured in the serum and induced sputum of 30 healthy controls and 34 stable COPD patients using high performance liquid chromatography (HPLC). The inhibition of lipid peroxidation as an index of antioxidant capacity was measured in the serum by a TBA assay. Results : The serum concentration of ascorbic acid, $\alpha$-tocopherol, and retinol were significantly lower in the patients with COPD than in healthy controls ($484.8{\pm}473.3$ vs $1497.8{\pm}819.2\;pmol/L$, $48.38{\pm}17.34$ vs $73.96{\pm}26.29\;pmol/L$, p<0.001, and $9.51{\pm}8.33$ vs $15.01{\pm}5.88\;pmol/L$, p<0.05, respectively, mean$\pm$SD). However, there were little differences in the ascorbic acid and uric acid concentrations in the induced sputum between the COPD patients and the controls. The induced sputum to serum ratio of ascorbic acid was significantly higher in COPD patients compared with healthy control (0.375 vs 0.085, p<0.05). In the normal controls, the serum ascorbic acid concentration was lower in smokers than in nonsmokers ($1073{\pm}536$ vs $1757{\pm}845\;pmol/L$, p<0.05), but the level was still higher than that of the COPD patients (p<0.05). The serum retinol levels were correlated with $FEV_1$ in COPD patients (r=0.58, p<0.05). The products of lipid peroxidation were increased in normal smokers and COPD compared with norma1 nonsmokers ($115.56{\pm}19.93$ vs $120.02{\pm}24.56$ vs $91.87{\pm}20.71\;{\mu}mol/{\mu}mol$ Pi of liposome, p<0.05). Conclusion : Cigarette smoking may induce the dep1etion of serum antioxidants and this depletion of antioxidants is suggested to play a role in the pathogenesis of COPD.

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Skin Whitening Effect of Ethyl Acetate Fraction of Adenophora triphylla var. japonica Sprout (잔대(Adenophora triphylla var. japonica)순 아세트산에틸 분획물의 피부 미백 효과)

  • Yoo, Seul Ki;Park, Seon Kyeong;Kang, Jin Yong;Kim, Jong Min;Park, Sang Hyun;Kwon, Bong Seok;Lee, Chang Jun;Kang, Jeong Eun;Park, Su Bin;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Plant Resources
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    • v.30 no.4
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    • pp.352-363
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    • 2017
  • To investigate skin-whitening effect of Adenophora triphylla var. japonica sprout extract, antioxidant activity, inhibitory effect on tyrosinase and melanin synthesis in B16/F10 melanoma cell were examined. Total phenolic content (246.25 mg GAE/g) and total flavonoid content (303.94 mg RE/g) of ethyl acetate fraction from Adenophora triphylla sprout (EFAT) showed the highest contents than other fractions (n-hexane, chloroform and distilled water). Antioxidant activities of EFAT has been evaluated using ABTS, DPPH radical scavenging activities, FRAP and inhibitory effect of lipid peroxidation. EFAT showed excellent radical scavenging activity and inhibitory effect on MDA production. Inhibitory effect of tyrosinase as a major enzyme of melanin synthesis was also measured. In these results, EFAT showed higher inhibitory effect against L-DOPA (51.27%) than L-tyrosine. $IC_{50}$ value on ${\alpha}-glucosidase$ was $41.93{\mu}g/ml$. In B16/F10 melanoma cells, EFAT inhibited melanin synthesis at $200{\mu}g/ml$ concentration (about 42% decrease). Finally, main physiological compounds of EFAT were identified as a rutin and a chlorogenic acid using high performance liquid chromatography.

Neuroprotective Effects of Modified Yuldahanso-tang (MYH) in a Parkinson's Disease Mouse Model (MPTP로 유도된 Parkinson's disease 동물 모델에서 열다한소탕 가감방 (MYH)의 신경 세포 보호 효과)

  • Go, Ga-Yeon;Kim, Yoon-Ha;Ahn, Taek-Won
    • Journal of Sasang Constitutional Medicine
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    • v.27 no.2
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    • pp.270-287
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    • 2015
  • Objectives To evaluate the neuroprotective effects of modified Yuldahanso-tang (MYH) in a Parkinson's disease mouse model. Methods 1) Four groups (each of 8 rats per group) were used in this study. 2) The neuroprotective effect of MYH was examined in a Parkinson's disease mouse model. C57BL/6 mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 30 mg/kg/day), intraperitoneal (i.p.) for 5 days. 3) The brains of 2 mice per group were removed and frozen at $-20^{\circ}C$, and the striatum-substantia nigra part was seperated. The protein volume was measured by Bradford method following Bio-Rad protein analyzing kit. Using mouse/Rat Dopamine ELISA Assay Kit. 4) The brains of 2 mice per group were separated and removed. TH-immunohistochemical was examined in the MPTP-induced Parkinson's disease mice to evaluate the neuroprotective effects of MYH on ST and SNpc. 5) Two mice out of each group were anesthetized and skulls were opened from occipital to frontal direction to take out the brains. The brains added TTC solution for 20 minutes for staining. 6) The water tank used for morris water maze test was filled with $28^{\circ}C$ water, and a round platform of 10cm in diameter was installed for mice to step on. The study was carried out once a day within 30 seconds, keep exercising to step on the platform in the pool. 7) The brains of two mice out of each group were fixed in 10% formaldehyde solution and paraphillin substance was infiltrated. They were fragmented by microtome, and observed under an optical microscope after Hematoxylin & Eosin staining. 8) A round acrylic cylinder with its upper side open was filled with clean water and depressive mouse models were forced to swim for 15 minutes. After 24 hours the animals were put in the same equipment for 5 minutes and were forced to swim. 9) The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of Neurotransmitters in MPTP-MYH group. Results 1) MYH possess Dopamine cell protective effect on MPTP-induced injury in striatum and substantia nigra pars compacta. 2) MYH inhibits the loss of tyrosine hydroxylase-immunoreacitive (TH-IR) cells in the striatum and substantia nigra pars compacta on MPTP-induced injury in C57BL/6 mice. 3) MYH possesses improvement effect on MPTP-induced memory deterioration in C57BL/6 mice through the reduction of prolongated Sort of lost time by MPTP injection using the Morris water maze test. 4) MYH possesses hippocampal neuron protective effect on MPTP-induced injury in C57BL/6 mice. 5) MYH possesses improvement effect on MPTP-induced motor behaviour deficits and depression in C57BL/6 mice through the reduction of prolongated losing motion by MPTP injection using the Forced swimming test. 6) MYH increases serotonin product amount on MPTP-induced injury in C57BL/6 mice. Conclusions This experiment suggests that the neuroprotective effect of MYH is mediated by the increase in Dopamin, TH-ir cell, Hippocampus and Serotonin. Furthermore, MYH essential oil may serve as a potential preventive or therapeutic agent regarding Parkinson's disease.

Antioxidant Activity of Different Parts of Lespedeza bicolor and Isolation of Antioxidant Compound (싸리나무(Lespedeza bicolor) 부위별 추출물의 항산화 활성 및 항산화물질 분리)

  • Lee, Jae-Hak;Jhoo, Jin-Woo
    • Korean Journal of Food Science and Technology
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    • v.44 no.6
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    • pp.763-771
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    • 2012
  • In this study, total antioxidant properties of extracts from different parts of Lespedeza bicolor were determined using techniques of measuring 1,1-diphenyl-2-picryl hydrazyl/2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)-radical scavenging activity and total phenolic contents. The total antioxidant activities of leaf, stem and root extracts from various solvents (water, 50, 70, 100% ethanol, and hot-water) indicated that 50 and 70% ethanol extracts have high radical scavenging activities and phenolic contents. A systematic approach was used to determine the total antioxidant activity of different solvent fractions of the Lespedeza bicolor extracts, partitioning with chloroform, ethyl acetate, n-butanol, and water, and the ethyl acetate fraction was found to have the strongest antioxidant activity. Antioxidant assay-guided isolation was carried out to isolate potential antioxidant compounds. The ethyl acetate fraction of the leaf extract was subjected to silica gel, LH-20 and RP-18 column chromatography successively, and afforded compound 1, which was identified as eriodictyol by NMR and MS analysis, after which its antioxidant activity was determined.

Effect of gomchwi (Ligularia fischeri) extract against high glucose- and H2O2-induced oxidative stress in PC12 cells (PC12 신경세포에서 고당 및 과산화수소로 유도된 산화적 스트레스에 대한 곰취 추출물의 효과)

  • Park, Sang Hyun;Park, Seon Kyeong;Ha, Jeong Su;Lee, Du Sang;Kang, Jin Yong;Kim, Jong Min;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.48 no.5
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    • pp.508-514
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    • 2016
  • Effects of the ethyl acetate fraction from gomchwi (Ligularia fischeri) extract against high $glucose/H_2O_2-induced$ oxidative stress and in vitro neurodegeneration were investigated to confirm the physiological property of the extract. The ethyl acetate fraction of gomchwi extract showed the highest total phenolic contents than the other solvent fractions. An anti-hyperglycemic activity of the ethyl acetate fraction was evaluated using the ${\alpha}-glucosidase$ inhibitory assay, and the half maximal inhibitory concentration ($IC_{50}$) value for ${\alpha}-glucosidase$ was found to be $727.64{\mu}g/mL$. In addition, the ethyl acetate fraction showed excellent 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt radical scavenging activity, and inhibition of malondialdehyde production. The ethyl acetate fraction also decreased intracellular reactive oxygen species, whereas neuronal cell viability against high glucose/$H_2O_2$-induced cytotoxicity was found to be increased. Finally, 3,5-dicaffeoylquinic acid as a main phenolic compound in the ethyl acetate fraction was analyzed by high-performance liquid chromatography. These results suggest that gomchwi might be a good natural source of functional materials to prevent diabetic neurodegeneration.

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.