• Title/Summary/Keyword: HPLC analysis of flavonoid

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Analysis and Quantitative Distribution of Glycosided Flavonoids in Citruses and Korean Chung-pi (감귤류와 한국산 청피에 함유된 Glycosided Flavonoids의 분석과 정량적 분포)

  • Baik, S.O.;Bock, J.Y.;Chun, H.J.;Jeong, S.I.;Baek, S.H.;Oh, H.B.;Kim, I.K.
    • Analytical Science and Technology
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    • v.14 no.4
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    • pp.340-348
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    • 2001
  • Glycosided flabonoids (naringenin, naringin, narirutin, hesperidin, and hesperetin) in Citrus and Korea Chung-pi were isolated and analyzed with HPLC, GC-mass, UV and high resolution NMR. Contents of glycosided flavonoids were compared according to kinds of Citrus and fruit ripening periods. Major compound of Korean Chung-pi was hesperidin and minors were narirutin and hesperetin. Major compounds of Gisil were naringin, narirutin, naringenin and were narirutin, hesperidin in Gigak. Major compounds of milgam and orange were narirutin, hesperidin, and the contents of glycoside flavonoids decreased according to the age of maturity.

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HPLC Analysis of Some Flavonoids in Citrus Fruits (조생온주 밀감의 품종에 따른 Flavonoids 함량 분석에 관한 연구)

  • 이창환;강영주
    • Food Science and Preservation
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    • v.4 no.2
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    • pp.181-187
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    • 1997
  • New HPLC method was developed for determination of some flavonoids such as naringin, hesperidin, neohesperidin, rutin, quercitrin, naringenin, hesperetin and apigenin and their contents in citrus juice and citrus peel from citrus varieties Brown in Cheju. Detection was at 280nm and reverse phase ${\mu}$-Bondapak C-18 column was used. Water/methanol/acetic acid as the mobile phase was better than water/acetonitrile/acetic acid. Flavonoids were more stable in 20% n,n-dimethylformamide in methanol(20% DMF) than methanol and pH 12 adjusted by 1N-sodium hydroxide solution. Standard flavonoid solutions were injected three times consecutively and the reproduciability was 0.236 to 3.550%, Correlation coefficient of the calibration curve was 0.9946 to 0.9999. The exiraction efficiency of hesperidin from citrus peel was evaluated with different extraction method such as reflux, ultra-sonicating method, using three solvents (aqueous solutions with pH12 adjusted by 1N-sodium hydroxide, methanol and 20% DMF), respectively. The reflux for 4 hour in 20% DMF was the most efficient of the tested methods and solvents, and recovery percentage were 78.0∼130.0%. Flavonoids were determined in citrus juice. Naringin was 68.2mg/100$m\ell$ in Natsudaidai, Hesperidin were 85.6mg/100$m\ell$ in Sankyool and Neohesperidin was 25.3mg/100$m\ell$ in Dangyooja. Flavonoids were determined in citrus peel. Naringin was 110mg/g in Dangyooja, Hesperidin was 242mg/g in Hungjin and Neohesperidin was 87.9mg/g in Dangyooja.

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Quantitative Analysis of Flavonoids in the Unripe and Ripe Fruits and the Leaves of Four Korean Rubus species (한국산 4종 Rubus속 숙과, 미숙과 및 잎의 플라보노이드 함량 분석)

  • Kim, Min-Young;Choi, Moo-Young;Nam, Jung-Hwan;Park, Hee-Juhn
    • Korean Journal of Pharmacognosy
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    • v.39 no.2
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    • pp.123-126
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    • 2008
  • The five flavonoids, isoqueritrin, juglanin, astragalin and 2-O-trans-p-coumaroyl astragalin were quantitatively analyzed in the leaves, unripe- and ripe fruits of four Korean Rubus species including Rubus crataegifolius, R. pungens var. oldhami, R. parvifolius, and R. coreanus belonging to the Rosaceae family. The HPLC analysis demonstrated that flavonoids are more abundant in the leaves than in the fruits. In the four Rubus species, ripe fruits generally contained similar or more amount of flavonoids than in unripe fruits. In particular, the quantity levels of total flavonoid (49.81${\pm}$0.50 mg/g) and isoquercitrin (35.08${\pm}$0.37 mg/g) were highest in the leaf of R. crataegifolius. In addition, the content of the flavonoids were quite low in the unripe fruits of R. coreanus that is most widely used as Rubi Fructus.

Analysis of Active Components in Cirsium japonicum var. ussuriense Extracts and Their Effect on TGF-beta Induced Hepatic Stellate Cells Activation (엉겅퀴 추출물의 기능 성분 분석 및 TGF-beta에 의한 간 성상 세포 활성 억제 효과)

  • Kim, Seon-Young;Kim, Sang-Jun;Choi, Young Ji;Yu, Kang-Yeol;Chung, Chang-Ho;Shim, Jae-Suk;Jang, Seon-Il;Yu, Dong-Hyun;Jeong, Seung-Il
    • Korean Journal of Pharmacognosy
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    • v.44 no.2
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    • pp.110-117
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    • 2013
  • Cirsium japonicum (CJ) leaf (L) alcoholic extracts were investigated for analysis their active components (flavonoids and flavanolignans; silymarins) and inhibitory effect on transforming growth factor (TGF)-${\beta}$ induced hepatic stellate cells (HSCs, LX-2 cells) activation. The CJ root (R) extracts were also analyzed and compared with leaf extracts. Total flavonoid and polyphenol contents of the leaf extracts showed higher than those of the root extracts. The content of each flavonoid compound, which was analyzed by HPLC, in CJ-L extracts was also higher than in CJ-R extracts. The results of flavanolignans content in CJ-L and CJ-R extracts were consistent in flavonoid and polyphenol. We studied inhibitory effect of two extracts against TGF-${\beta}1$ induced HSCs activation. The CJ-L extracts significantly suppressed overexpression of profibrogenic factor, ${\alpha}$-smooth muscle actin and collagen-${\alpha}1$(I). The CJ-R extract also showed inhibitory effect on TGF-${\beta}1$ induced HSCs activation, but the efficacy was lower than in CJ-L extract. These results suggest that CJ-L may contribute to the fibrotic liver treatment.

Rapid separation of Capsicum annuum L. leaf extract using automated HPLC/SPE/HPLC coupling system (Sepbox system) and identification of α-glucosidase inhibitory active substances (자동화 HPLC/SPE/HPLC 시스템(Sepbox system)을 활용한 고추 잎 (leaf of Capsicum annuum L.) 추출물 분리 및 α-glucosidase 억제 활성 물질 탐색)

  • Kim, Min-Seon;Jin, Jong Beom;Lee, Jung Hwan;An, Hye Suck;Pan, Cheol-Ho;Park, Jin-Soo
    • Journal of Applied Biological Chemistry
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    • v.64 no.1
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    • pp.25-32
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    • 2021
  • Phytochemicals include plant-derived natural products that promote and improve the human metabolism and physiological activity, and there is a lot of research to find the value of the molecules is in progress. Likewise, we obtained 288 fractions of Capsicum annuum L. extract in less than 20 h using HPLC/SPE/HPLC coupling experiment through Sepbox system, an effective separation system to search for active substances in natural resources and ensure efficacy and reliability. Therefore, this experiment allowed rapid identification of biologically active molecules from the extract compared to traditional separation processes. Of the above fractions, eight fractions showed the α-glucosidase inhibitory (AGI) activity and subsequent LC-MS analysis revealed one of the active molecules as luteolin 7-O-glucoside. In addition, we proved the increase in AGI activity according to deglycosylation of flavonoid glycoside. Therefore, this study suggests that the Sepbox system can quickly separate and identify active components from plant extract, and is an effective technique for finding new active substances.

Varied Flavonoid Contents of Citri Unshii Pericarpium Extracted from Several Traditional Herbal Prescriptions (진피의 복합처방에 따른 유효성분 추출율의 변화)

  • Kim Jin-Ju;Cho Sung-One;Lee Ho Young;Ha Hye-kyung;Oh Ju-hee;Yang In-cheol;Choi Hwan-Soo;Lee Je-Hyun
    • The Journal of Korean Medicine
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    • v.26 no.2 s.62
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    • pp.195-200
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    • 2005
  • Objecitves: Peel of Citrus unshiu (Rutaceae) has been used in traditional herbal prescriptions. Hesperidin and naringin contents of Citri Unshii Pericarpium were determined and compared to herbal prescriptions. Then, the different extractions from prescriptions are explained with traditional herbal theories. Methods: The herb and herbal prescriptions were extracted in boiled water. A quantitative analysis was made of hesperidin and naringin using HPLC with-reversed-phase CIS column and a UV detector at 204nm. Elution was carried out at 1.0ml/min with $38\%$ methanol. Results and Conclusions: The extract of Citri Unshii Pericarpium for 180 min was the better method for extracting naringin and hesperidin. The flavonoid contents in remedies are different among several traditional herbal prescriptions. The reason for the difference can be explained with traditional drug interaction theories.

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Isolation of Flavonoid Glycosides with Cholinesterase Inhibition Activity and Quantification from Stachys japonica

  • Nugroho, Agung;Choi, Jae Sue;Seong, Su Hui;Song, Byong-Min;Park, Kyoung-Sik;Park, Hee-Juhn
    • Natural Product Sciences
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    • v.24 no.4
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    • pp.259-265
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    • 2018
  • The three flavone glycosides, 4'-O-methylisoscutellarein 7-O-(6'''-O-acetyl)-${\beta}$-D-allopyranosyl(1${\rightarrow}$2)-${\beta}$-D-glucopyranoside (1), isoscutellarein 7-O-(6'''-O-acetyl)-${\beta}$-D-allopyranosyl(1${\rightarrow}$2)-${\beta}$-D-glucopyranoside (3), and isoscutellarein 7-O-${\beta}$-D-allopyranosyl(1${\rightarrow}$2)-${\beta}$-D-glucopyranoside (4) in addition to a flavonol glycoside, kaempferol 3-O-${\beta}$-D-glucopyranoside (astragalin, 2), were isolated from Stachys japonica (Lamiaceae). In cholinesterase inhibition assay, compound 1 significantly inhibited aceylcholinesterase (AChE) and butyrylcholinesterase (BChE) activities ($IC_{50}s$, $39.94{\mu}g/ml$ for AChE and $86.98{\mu}g/ml$ for BChE). The content of isolated compounds were evaluated in this plant extract by HPLC analysis. Our experimental results suggest that the flavonoid glycosides of S. japonica could prevent the memory impairment of Alzheimer's disease.

Antioxidative Activity and Component Analysis of Psidium guajava Leaf Extracts (구아바 잎 추출물의 항산화 활성과 성분 분석)

  • Yang, Hee-Jung;Kim, Eun-Hee;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.3
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    • pp.233-244
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    • 2008
  • In this study, the antioxidative effects, inhibitory effects on elastase and tyrosinase, and component analysis of Psidium guajava leaf extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities $(FSC_{50})$ of extract/fractions of Psidium guajava leaf were in the order: 50% ethanol extract $(7.05{\mu}g/mL)$ < ethyl acetate fraction $(3.36{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(3.24{\mu}g/mL)$. Reactive oxygen species (ROS) scavenging activities $(OSC_{50})$ of some Psidium guajava leaf extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activities were 50% ethanol extract $(OSC_{50},\;2.17{\mu}g/mL)$ < ethyl acetate fraction $(0.64{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(3.39{\mu}g/mL)$. Aglycone fraction showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of Psidium guajava leaf on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Psidium guajava leaf extracts suppressed photohemolysis in a concentration dependent manner $(1{\sim}10{\mu}g/mL)$, particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect ${\tau}_{50}\;107.5min\;at\;1{\mu}g/mL)$. Aglycone fraction obtained from the deglycosylation reaction of ethyl acetate fraction among the Psidium guajava leaf extracts, showed 1 band in TLC and 1 peak in HPLC experiments (360 nm). One component was identified as quercetin. TLC chromatogram of ethyl acetate fraction of Psidium guajava leaf extract revealed 5 bands and HPLC chromatogram showed 5 peaks, which were identified as quercetin 3-O-gentobioside (10.32%) , quercetin 3-O-${\beta}$-D-glucoside (isoquercitin, 13.30%), quercetin 3-O-${\beta}$-D-galactoside (hyperin, 11.34%), quercetin 3-O-${\alpha}$-L-arabinoside (guajavarin, 19.70%), quercetin 3-O-${\beta}$-L-rhamnoside (quercitrin, 45.33%) in the order of elution time. The inhibitory effect of Psidium guajava leaf extracts on tyrosinase were investigated to assess their whitening efficacy. Finally, their anti-elastase activities were measured to predict the anti-wrinkle efficacy in the human skin. Inhibitory effects $(IC_{50})$ on tyrosinase of some Psidium guajava leaf extracts was 50% ethanol extract $(149.67{\mu}g/mL)$ < ethylacetate fraction $(30.67{\mu}g/mL)$ < deglycosylated aglycone fraction $(17.10{\mu}g/mL)$. Inhibitory effects $(IC_{50})$ on elastase of some Psidium guajava leaf extracts was 50% ethanol extract $(6.60{\mu}g/mL)$ < deglycosylated aglycone fraction $(5.66{\mu}g/mL)$ < ethylacetate fraction $(3.44{\mu}g/mL)$. These results indicate that extract/fractions of Psidium guajava leaf can function as antioxidants in bioloigcal systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. And component analysis of Psidium guajava leaf extract and inhibitory activity on elastase of the aglycone fraction could be applicable to new functional cosmetics for smoothing wrinkles.

Validation of an HPLC/UV Analysis Method for Cirsimaritin in Cirsium japonicum var. maackii (엉겅퀴에 함유된 Cirsimaritin 분석을 위한 HPLC 분석법 밸리데이션)

  • Lee, Ju Sung;Paje, Leo Adrianne;Rodriguez, Joyce P.;Kang, Ki Sung;Hahm, Dae-Hyun;Shim, Jae Suk;Choi, Yu-Jin;Lee, Sanghyun
    • Korean Journal of Pharmacognosy
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    • v.51 no.3
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    • pp.217-221
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    • 2020
  • Cirsium japonicum var. maackii (CM) has been used to treat certain medical conditions such as hemorrhage, hepatitis, and hypertension. Cirsimaritin was previously found as the major flavonoid in CM and is said to contribute to its pharmacological effects. There are currently no reports detailing the qualitative and quantitative detection of phytochemical indicators in the aerial parts of CM. Therefore, we developed a method to rapidly identify and quantify cirsimaritin in CM using HPLC/UV, and we optimized and validated this analytical method. The results showed good linearity in the concentration range tested (0.25-0.015 mg/mL, r2 ≥ 0.9999), accuracy (93.9-111.3%), and precision (RSD ≤ 0.59%). The developed method can therefore be used for the rapid evaluation of cirsimaritin in CM.

Antioxidant Activity and Component Analysis of Populus Tomentiglandulosa Extract (현사시나무 추출물의 항산화활성과 성분분석)

  • Choi, Sun-Il;Hwang, Seok-Jun;Lee, Ok-Hwan;Kim, Jong Dai
    • Korean Journal of Food Science and Technology
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    • v.52 no.2
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    • pp.119-124
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    • 2020
  • Populus Tomentiglandulosa (PT) is known for pharmacological effects against ischemia-injury and immune activity. This study aimed to investigate the nutritional components, total phenol and flavonoid contents, antioxidant activities of PT extract. Among the mineral contents, the K content (907.5 mg/100 g) was the highest in the PT extract. Vitamins C (6.1 mg/100 g) and nicotinic acid (3.1 mg/100 g) were also found in high amounts. Fructose (2.2%) and glucose (1.6%) were found as free sugars in the PT extract. The total phenolic and flavonoid contents of PT extract were 115.4±0.85 mg GAE/g and 20.9±1.14 mg QE/g, respectively. Results of HPLC analysis of PT extract identified catechin (9.1±0.27 mg/g), caffeic acid (4.1±0.57 mg/g), p-coumaric acid (2.1±0.49 mg/g), chlorogenic acid (1.6±1.86 mg/g), and gallic acid (1.4±0.35 mg/g), respectively. These results suggest that the PT extract possesses high nutritional component and antioxidant properties, which can be used as functional bioresources.