• Title/Summary/Keyword: Spectrophotometric assay

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Screening of HMG-CoA Reductase Inhibitory Activity of Ethanol and Methanol Extracts from Cereals and Regumes (곡류 및 두류 추출물로 부터 HMG-CoA reductase 저해활성 검색)

  • Ha, Tae-Youl;Cho, Il-Jin;Lee, Sang-Hyo
    • Korean Journal of Food Science and Technology
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    • v.30 no.1
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    • pp.224-229
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    • 1998
  • A study was conducted to screen the inhibitory activity of 3-hydroxy-3-methylglutaryl coenzyme A(HMG-CoA) reductase, which is known to be rate-limiting enzyme in cholesterol bosynthesis, from the extracts of 80% methanol and 70% ethanol of cereals and regumes. The strongest inhibitory activity was shown in the ethanol extract of sorghum among the ethanol extracts. The inhibitory activity of HMG-CoA reductase of prosomillilet methanol extract was 73%, and highest among the methanol extracts. The inhibitory activity of 44.7% was observed in sorghum methanol extract. The methanol extracts of prosomillet and sorghum were further fractionated with hexane, chloroform, ethylacetate, butanol and water. HMG-CoA reductase inhibitory activity was shown in all fractions of prosomillet and sorghum methanol extracts. Hexan fraction of both prosomillet and sorghum had the strongest inhibitory activity among five fractions, and the inhibitory activity was increased compared to each crude extracts.

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Conjugated linoleic acid producing potential of lactobacilli isolated from goat (AXB) rumen fluid samples

  • Tyagi, Amrish Kumar;Kumar, Sachin;Choudhury, Prasanta Kumar;Tyagi, Bhawna;Tyagi, Nitin
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.8
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    • pp.1233-1241
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    • 2020
  • Objective: The present investigation was aimed to explore the potential of lactobacilli for conjugated linoleic acid (CLA) production, isolated from rumen fluid samples of lactating goats. Methods: A total of 64 isolates of lactobacilli were obtained using deMan-Rogosa-Sharpe (MRS) agar from rumen fluid of goats and further subjected to morphological and biochemical characterizations. Isolates found as gram-positive, catalase negative rods were presumptively identified as Lactobacillus species and further confirmed by genus specific polymerase chain reaction (PCR). The phylogenetic tree was constructed from the nucleotide sequences using MEGA6. Results: Out of the 64 isolates, 23 isolates were observed positive for CLA production by linoleate isomerase gene-based amplification and quantitatively by UV-spectrophotometric assay for the conversion of linoleic acid to CLA as well as gas chromatography-based assay. In all Lactobacillus species cis9, trans11 isomer was observed as the most predominant CLA isomer. These positive isolates were identified by 16S rRNA gene-based PCR sequencing and identified to be different species of L. ingluviei (2), L.salivarius (2), L. curvatus (15), and L. sakei (4). Conclusion: The findings of the present study concluded that lactic acid bacteria isolated from ruminal fluid samples of goat have the potential to produce bioactive CLA and may be applied as a direct fed microbial to enhance the nutraceutical value of animal food products.

Anti-Glyaction and Radical Scavenging Activities of Ligustri Fructus by Extraction Method (추출 방법에 따른 여정실의 최종당화산물 생성 저해 및 라디칼 소거 활성)

  • Jeong, Yun Hee;Kim, Seo Yoon;Jeong, Gyeong Han;Kim, Tae Hoon
    • The Korea Journal of Herbology
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    • v.33 no.4
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    • pp.95-100
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    • 2018
  • Objectives : Ligustri Fructus has been used since ancient times as a medicinal usages in folk medicines against antitumor purpose. Many biological active constituents have been identified from this biomass such as several terpenoids and lignans. In current study, the properties of antioxidant and anti-diabetic complications using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)($ABTS^+$) radicals scavenging, and advanced glycation end products (AGEs) inhibition assays were evaluated by different extraction methods of Ligustri Fructus. Methods : In present continuing research for development of bioactive natural ingredients, antioxidant and AGEs formation inhibitory capacities of Ligustri Fructus extracts using different organic solvents were prepared and the biological potentials were investigated using in vitro bioassays. Antioxidant properties were evaluated employing radical scavenging assays using DPPH and $ABTS^+$ radicals. In addition, the anti-diabetic complications effects of Ligustri Fructus extracts were tested via AGEs formation inhibitory assay. The total phenolic contents were determined using a spectrophotometric method. Results : All the tested extracts exhibited dose-dependent radical scavenging and AGEs formation inhibitory activities. Among the tested samples, hot water extract of Ligustri Fructus was showed the most potent activity with $IC_{50}$ value of $494.8{\pm}6.7{\mu}g/m{\ell}$ against DPPH radical scavenging assay. Also, $ABTS^+$ radical scavenging activity of hot water extract was higher than those of other extracts. In addition, AGEs formation inhibitory effects of each extacts and total phenolic contents were evaluated. Conclusions : These results suggested that Ligustri Fructus can be considered as a new effective source of natural antioxidant and anti-diabetic complications resources.

Evaluation of Antioxidant and Anti-diabetic Effects of Sappan Lignum by Extraction Method (추출방법에 따른 소목 심재의 항산화 및 항당뇨 활성 평가)

  • Hong, Young Ju;Jeong, Gyeong Han;Jeong, Yun Hee;Kim, Tae Hoon
    • The Korea Journal of Herbology
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    • v.32 no.6
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    • pp.1-7
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    • 2017
  • Objectives : The heartwood of Sappan Lignum has been used since ancient times as an ingredient in folk medicines against anti-bacterial and anti-anemia purposes. Many bioactive constituents have been derived from this biomass such as chalcones and homoisoflavonoids. In the current investigation, the antioxidant and anti-diabetic properties using DPPH and $ABTS^+$ radicals scavenging, ${\alpha}-glucosidase$, and advanced glycation end products (AGEs) inhibition assays were evaluated by different extraction methods of Sappan Lignum. Methods : In our continuing investigation for bioactive natural ingredients, the antioxidant and ${\alpha}-glucosidase$ inhibitory properties of Sappan Lignum extracts were prepared from different extraction methods and the biological efficacies were investigated in vitro. The antioxidant properties were evaluated employing radical scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) ($ABTS^+$) radicals. In addition, the anti-diabetic effects of Sappan Lignum extracts were tested via ${\alpha}-glucosidase$ and AGEs formation inhibitory assay. The total phenolic contents were determined using a spectrophotometric method. Results : All the tested samples showed dose-dependent radical scavenging and ${\alpha}-glucosidase$ inhibitory activities. Among the tested extracts, the 80% methanolic extract of Sappan Lignum was showed the most potent activity with an $IC_{50}$ value of $82.3{\pm}1.7{\mu}g/m{\ell}$ against DPPH radical scavenging assay. While, $ABTS^+$ radical scavenging activity of 80% methanolic extract was higher than those of other extracts. Also, ${\alpha}-glucosidase$ inhibitory and AGEs formation effects of each extacts and total phenolic contents were evaluated. Conclusions : These results suggested that Sappan Lignum can be considered as a new effective source of natural antioxidant and anti-diabetic materials.

Antihypertensive Properties of Dried Radish Leaves Powder in Spontaneously Hypertensive Rats (자연발증적 고혈압 흰쥐에서 무청 건조분말의 항고혈압 효과)

  • Kim, Bo-Ram;Park, Ji-Ho;Kim, Sun-Hee;Cho, Kang-Jin;Chang, Moon-Jeong
    • Journal of Nutrition and Health
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    • v.43 no.6
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    • pp.561-569
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    • 2010
  • The study aim was to investigate the antihypertensive effect after oral supplementation of dried radish leaves powder (DRLP). Angiotensin-converting enzyme (ACE) activity was measured by spectrophotometric assay. The systolic blood pressure (SBP) was measured in spontaneously hypertensive (SHR) and normotensive rats (Wistar) by the tail cuff method after a 4-week diet with DRLP at the level of 2.5% or 5%. The supplementation of DRLP decreased SBP of SHR although the 5% supplementation level did not show any more pronounced effect than the 2.5% level did. The decrease in the SBP observed for both 2.5% and 5% DRLP was accompanied by significant increases of the urinary Na and K excretion. The DRLP supplementation showed a potent ACE-inhibitory activity in pulmonary tissue from both hypertensive and normotensive rats. However, the DRLP supplementation did not affect the SBP in normotensive rats. These results indicated that DRLP exerted an antihypertensive effect in SHR due to the decreased ACE activity and increased urinary Na excretion.

Efficient Transformation of Trifolium repens L. Using Acetosyringone (Acetosyringone을 이용한 효율적인 White Clover의 형질전환)

  • TaeHoKwon
    • Korean Journal of Plant Resources
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    • v.10 no.2
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    • pp.107-113
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    • 1997
  • Transformants of White Clover(Trifolium repens L.) were efficiently produced from immature seed derived callus cocultivated with Agrobacterium twnefaciens LBA4404 harboring plant binary vector. pBI121, using acetosyringone. The mean frequencies of transformants on the two kanamycin-containing media were 16 to 19% when the immature seed-derived calli were infected with bacteria cultured in the presence of 100$\mu$M acetosyringone compared with 7% in media without acetosyringone. Transgenic white clover was subject to molecular analysis for integration into plant nuclear genome and expression of $\beta$-glucuronidase(GUS) gene. PCR and Northern blot analyses demonstrated that GUS gene was integrated into white clover nuclear genome and expressed into its mRNA. The expression of GUS gene into its protein was confirmed by spectrophotometric assay of GUS activity.

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Inhibitory effect of Capparis zeylanica Linn. on acetylcholinesterase activity and attenuation of scopolamine-induced amnesia

  • Chaudhary, Amrendra Kumar;Solanki, Ruchi;Singh, Vandana;Singh, Umesh Kumar
    • CELLMED
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    • v.2 no.2
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    • pp.19.1-19.6
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    • 2012
  • $Capparis$ $zeylanica$ Linn. a 'Rasayana' drug is used for its memory enhancing effects in the traditional Ayurvedic system of medicine. The aim of this study was to evaluate acetylcholinesterase (AChE) inhibitory and memory enhancing activities of $Capparis$ $zeylanica$ Linn. The$in-vitro$ and $ex-vivo$ models of AChE inhibitory activity were used along with Morris water maze test to study the effect on memory in rats. The anticholinesterase effect of methanolic and aqueous extracts of $Capparis$ $zeylanica$ was measured by spectrophotometric Ellman method at 0.1, 0.3, 1.0, 3.0, 10 and 30 mg/ml and brain monoamine oxidase (MAO-A and MAO-B) activity was assessed by Naoi's method. The results $in-vitro$ and $ex-vivo$ AChE assay revealed that methanolic and aqueous extracts of $Capparis$ $zeylanica$ inhibit AChE activity, whereas these extracts did not alter MAO activity at any concentration tested as compared to moclobemide and L-deprenyl. The results indicate that $Capparis$ $zeylanica$ improves scopolamine-induced memory deficits through inhibition of AChE activity, and not by direct MAO inhibition.

Determination of Buprenorphine in Raw Material and Pharmaceutical Products Using Ion-pair Formation

  • Amanlou, Massoud;Khosravian, Peghah;Souri, Effat;Dadrass, Orkideh Ghorban;Dinarvand, Rasoul;Alimorad, Mohammad Massoud;Akbari, Hamid
    • Bulletin of the Korean Chemical Society
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    • v.28 no.2
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    • pp.183-187
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    • 2007
  • A simple and sensitive extractive spectrophotometric method has been described for the determination of buprenorphine either in raw material or in pharmaceutical formulations. The developed method is based on the formation of a colored ion-pair complex (1 : 1 drug/dye) of buprenorphine and bromocresol green (BCG) in buffer pH 3 and extracting in chloroform. The extracted complex shows absorbance maxima at 415 nm. Beer's law is obeyed in the concentration range of 1.32-100.81 μ g mL-1. The proposed method has been applied successfully for the determination of drug in commercial sublingual tablets and injectable dosage form. No significant interference was observed from the excipients commonly used as pharmaceutical aids with the assay procedure.

Antioxidant and Antiproliferative Activities of Methanolic Extracts from Thirty Korean Medicinal Plants

  • Choi, Young-Min;Gu, Ja-Bi;Kim, Myung-Hee;Lee, Jun-Soo
    • Food Science and Biotechnology
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    • v.17 no.6
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    • pp.1235-1239
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    • 2008
  • To study the health promoting effects of medicinal plants, 30 medicinal plants commonly available in Korea have been evaluated for their antioxidant compounds and antioxidant and antiproliferative activities. Total polyphenolics and flavonoids in the methanolic extracts were measured by spectrophotometric methods and 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging activities and chelating effects have been determined for antioxidant activities. Moreover, the effects of medicinal plants on cell proliferation of intestinal (Caco-2) and pituitary (GH3) tumor cells were investigated using thiazolyl blue terazolium bromide (MIT) assay. The methanolic extracts of Pueraria thunbergiana and Artemisiae asiatria contained the highest total polyphenolic and flavonoid contents, respectively. P. thunbergiana exhibited the highest antioxidant activities. A. asiatria showed the strongest antiproliferative activity against Caco-2 and Ponciruc trifoliata Rafin and Lophathrum gracile Bronghiart exhibited the highest activities against GH3. Although there was positive correlation between ABTS radical scavenging activity and polyphenolic contents ($R^2=8189$), no relationship was found between antiproliferative and antioxidant activities.

High Glucose Potentiates the Alloxan-induced Cytotoxicity in Cultured Rat Insulinoma Cells (흰쥐 인슐린종세포에서 고농도 포도당의 Alloxan 독성 증강 효과)

  • 이병래;차종희;박재윤;고춘남;박평심
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.5
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    • pp.875-880
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    • 2000
  • Reactive oxygen species are produced under diabetic conditions and possibly cause various forms of tissue damage in patients with diabetes mellitus. The aim of this study was to examine the effects of high glucose on the alloxan-induced beta cell injury. The insulinoma (RINm5F) cells were clutured either with high glucose (22.2 mM) or normoglucose (5.6 mM) in RPMI 1460 media for 3 days. The SOD activities were determined by spectrophotometric assay and nitroblue tetrazolium (NBT) stain. The effects of high glucose on the cytotoxicity of alloxan were also investigated in RINm5F cells and the cells viability were determined by 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT) methods. Results showed that the CuZn-SOD activity was decreased but Mn-SOD activity was increased significantly in RINm5F cells cultured with high glucose (22.2 mM) media. The cytotoxicity of alloxan was increased by high glucose compared with normoglucose in RINm5F cells. Diethyl-dithiocarbarmate (DDC), as inhibitor of CuZn-SOC, also potentiate the alloxan-induced cytotoxocity in RINm5F cells. These results suggest that, in RINm5F cells, short term culture with high glucose media decreases Cu-Zn-SOD activity and the decreased activity of CuZn-SOD many one of the causative factors of beta-cell injury induced by high glucose.

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