• Title/Summary/Keyword: HPLC quantification

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New Method for Simultaneous Quantification of 12 Ginsenosides in Red Ginseng Powder and Extract: In-house Method Validation

  • In, Gyo;Ahn, Nam-Geun;Bae, Bong-Seok;Han, Sung-Tai;Noh, Kil-Bong;Kim, Cheon-Suk
    • Journal of Ginseng Research
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    • v.36 no.2
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    • pp.205-210
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    • 2012
  • For quality control of components in Korean red ginseng powder and extract, a new method for simultaneous quantification of 12 ginsenosides ($Rg_1$, Re, Rf, $Rh_1$, $Rg_2$[S], $Rg_2$[R], $Rb_1$, Rc, $Rb_2$, Rd, $Rg_3$[S], and $Rg_3$[R]) was studied. Compared to the official method for quantification of marker substances (ginsenosides $Rg_1$ and $Rb_1$), the proposed methods were guaranteed by in-house method validation. Several criteria such as linearity, specificity, precision and accuracy were evaluated. For red ginseng powder, recovery (averaging 95% to 105%) was calculated, and analysis of variance was carried out to estimate the relative standard deviation (0.20% to 2.12%). For red ginseng extract, the average recovery rate was 90% to 99% and the relative standard deviation was 0.39% to 2.40%. These results indicate that the proposed method could be used in the laboratory for determination of 12 ginsenosides in red ginseng powder and extract. In addition, this method was found to be suitable for quality control of ginseng products and potentially offer time and cost benefits.

Simultaneous Analysis of Bangpungtongseong-san and Its Antioxidant Effect (방풍통성산의 동시분석 및 항산화 효능 연구)

  • Seo, Chang-Seob;Kim, Ohn Soon;Shin, Hyeun-Kyoo
    • Herbal Formula Science
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    • v.21 no.2
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    • pp.133-143
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    • 2013
  • Objectives : We carry out the simultaneous quantification for quality control of four components in Bangpungtongseong-san (BPTSS) sample. In addition, we assessed the antioxidant effects of BPTSS sample. Methods : The used column for separation and analysis of four compounds was Luna C18 column and column oven temperature was maintained at $40^{\circ}C$. The mobile phase for simultaneous determination consisted of two solvent systems, 1.0% acetic acid in water and 1.0% acetic acid in acetonitrile. High performance liquid chromatography-photodiode array (HPLC-PDA) method for analysis was performed at a flow rate of 1.0 mL/min with PDA detection at 254 and 280 nm. The injection volume was 10 ${\mu}L$. The antioxidant activities of BPTSS were evaluated by measuring free radical scavenging activities on 2,2'-Azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The inhibitory effects on low-density lipoprotein (LDL) oxidation were evaluated by the formation of thiobarbituric acid relative substances (TBARS) and relative electrophoretic mobility (REM). Results : Calibration curves were acquired with $r^2{\geq}0.9999$. The values of limit of detection (LOD) and quantification (LOQ) were 0.06-0.29 ${\mu}g/mL$ and 0.20-0.98 ${\mu}g/mL$, respectively. The amounts of geniposide, liquiritin, baicalin, and glycyrrhizin in BPTSS were 5.06, 7.33, 27.56, and 7.81 mg/g, respectively. The BPTSS showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction (RC50) against ABTS and DPPH radicals were 72.51 ${\mu}g/mL$ and 128.49 ${\mu}g/mL$. Furthermore, GMGHT reduced the oxidation properties of LDL induced by CuSO4. Conclusions : The established HPLC-PDA method will be helpful to improve quality control of BPTSS. In addition, BPTSS has potentials as therapeutic agent on anti-atherosclerosis.

Identification and quantification of anthocyanin pigments in colored rice

  • Kim, Min-Kyoung;Kim, Han-Ah;Koh, Kwang-Oh;Kim, Hee-Seon;Lee, Young-Sang;Kim, Yong-Ho
    • Nutrition Research and Practice
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    • v.2 no.1
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    • pp.46-49
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    • 2008
  • Anthocyanin pigments from varieties of black, red and wild rice were identified and quantified to evaluate their potential as nutritional function, natural colorants or functional food ingredients. Anthocyanin extraction was conducted with acidified methanol with 0.1M HCl (85:15, v/v) and identification of anthocyanin, aglycone and sugar moieties was conducted by comparison with purified standards by HPLC, Ultraviolet-Visible absorption spectrophotometer and paper chromatography. Black and wild rice showed three different types of pigments by HPLC whereas red rice variety did not show any anthocyanins. Out of three pigments detected, one (peak 2) was characterized as cyanidin-3-glucoside (C3G) by comparison of spectroscopic and chromatographic properties with an authentic standard, and another (peak 3) was tentatively identified as cyanidin-fructoside on the basis of spectroscopic properties with ${\lambda}_{max}$ of aglycone in 1% HCl methanol at 537 nm, electrospray ionization mass spectra with major ions at 449 and 287 m/z and chromatographic properties. But another pigment (peak 1) has not been characterized. The most abundant anthocyanin in black and wild rice was C3G.

Immunochemical Determination of Plant Constituents (1990년 제23회 학술발표회 논문초록)

  • 성철기
    • Journal of the Korean Society of Tobacco Science
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    • v.12 no.1
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    • pp.29-38
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    • 1990
  • Until relatively recently plant scientists have made little use of immunological techniques. Now, however, more and more researchers are discovering the powder of these techniques for the screening of immunomodulators and for the detection, quantitative determination and localization of compounds in plant materials. Especially, the recent developments in the fields of plant biotechnology and plant genetic engineering make it even more important for forkers in the plant sciences to become acquainted with the more sophisticated methods. The possible methods include onestep purification of antigens, visualization in situ by immunocytochemis try and on polyacrylam ids gel s by ni trocellulose Western blotting and quantification by various immunoassay. Among them, in this reviews, the quantitative determination methods are to be reviewed. There are several kinds of methods for the quantitative determination of plant constituents such as colorimetry, TLC, GLC, DCC, UV derivatization, densitometry and HPLC. When the complexity of plant constituents is considered. densitometry and HPLC have many advantages in sensitivity and separation ability. After a 11 some advarltages of two methods meritiorled above, all of these methods have many disadvantages and inconveniences. Previous purification for the application of all these methods make them less sensitive and more tedious. Immunoassay can solve these problems in part. But immunoassay also has some limitations. Specificity of immunoassay, contrary, can be considered to be disadvantages. Including this the advantages and disadvantages of immunoassay are to be discussed.

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Development of Analytical Method of Biotin in Complex Drugs and Dietary Supplements Using HPLC-UV

  • Huh, Yoon-Young;Kang, Yun-Pyo;Choi, Yong-Seok;Park, Jeong-Hill;Kwon, Sung-Won
    • Journal of Pharmaceutical Investigation
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    • v.41 no.1
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    • pp.25-30
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    • 2011
  • Recently, Korean Food and Drug Administration (KFDA) has focused on developing quality control guidelines for all commercial products in Korea to enforce regulations, improve the quality control, and protect consumers by developing prevalently used and efficient analytical tools to determine and quantify target compounds. Because the Korean Pharmacopeia (KP) presents microbiological assays for biotin, which is laborious and time-consuming, this study is focused on applying HPLC-UV to detect and quantify biotin in complex drugs and dietary supplements like multi-vitamin. Biotin in complex drugs was extracted from methanol and analyzed using mobile phase with 10 mM potassium phosphate (monobasic, pH=3.0) in distilled water and acetonitrile. Gradient condition was used to successfully detect and quantify biotin within 20 minutes. Validation result for linearity was significant that average $r^2$ was 0.999 (n=3) and its relative standard deviation (RSD) was 0.0578% which was less than 2%. Using this method, quantification of biotin in complex drugs was completed successfully and recovery tests were finished that recovery percentage greater than 95% with relative standard deviation less than 2%.

Quantitative Analysis of Paeoniflorin from Paeonia lactiflora Using $^1H-NMR$

  • Yoo, Jong-Su;Song, Myoung-Chong;Ahn, Eun-Mi;Lee, Youn-Hyung;Rho, Yeong-Deok;Baek, Nam-In
    • Natural Product Sciences
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    • v.12 no.4
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    • pp.237-240
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    • 2006
  • Paeoniflorin, the major component of the root of Paeonia lactiflora, was quantitatively analyzed using $^1H-NMR$ spectrometry. The quantity of paeoniflorin was calculated by the ratio of the intensity of the signals (H-9, H-10, H-2', 6') to the aldehyde peak of the known amount of internal standard, 2,4,6-trihydroxybenzaldehyde. These results were compared with the conventional HPLC method. The contents of paeoniflorin in P. lactiflora, which were respectively calculated by H-9, H-10, H-2', 6' in the $^1H-NMR$ spectra and HPLC, were determined $2.60{\pm}0.07,\;2.44{\pm}0.09,\;2.77{\pm}0.12\;and\;2.46{\pm}0.16%$. The advantages of quantitative $^1H-NMR$ analysis are that can be analyzed to identify and quantify, and no reference compounds required for calibration curves. Besides, it allows rapid and simple quantification for paeoniflorin with an analysis time for only 20 min without any preprocessing.

High-Performance Liquid Chromatographic Analysis of Chrysin Derivatives on A $Nova-Pak^{\circledR}C_{18}$ Column

  • Kim, Kyoung-Soon;Shin, Joon-Su;Park, You-Mie;Lee, Sanghyun;Kim, Yang-Bae;Kim, Bak-Kwang
    • Archives of Pharmacal Research
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    • v.25 no.5
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    • pp.613-616
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    • 2002
  • A high-performance liquid chromatographic method has been developed for the separation and quantification of chrysin and synthetic chrysin derivatives (12 chrysin alkyl and 7 chrysin acyl derivatives). The chromatography was performed using a $Nova-Pak^{\circledR}C_{18}$ column. A RP-HPLC was performed by using a binary mixture (MeOH-10 mM H$_3$PO$_4$) as a mobile phase, and the column temperature was maintained at room temperature. A flow rate was 1.0 ml/min, and the effluent was monitored at a wavelenth of 280 nm. The retention times for chrysin acyl and alkyl derivatives were within 10 minutes and 20 minutes, respectively. The absolute recovery of samples were all over 96%. The detection limits were 0.1~18 ng at S/N = 3 ratio.

Quantification of Quercetin in Different Parts of Onion and Its DPPH Radical Scavenging and Antibacterial Activity

  • Kim, Su-Jeong;Kim, Gun-Hee
    • Food Science and Biotechnology
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    • v.15 no.1
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    • pp.39-43
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    • 2006
  • Levels of quercetin in different parts of onion were investigated using high performance liquid chromatography (HPLC) and liquid chromatography mass spectrometry (LC/MS) suitable for use with functional food material. Two main peaks were observed on HPLC chromatograms from the extracts of the skin, and the outer, middle, and core parts of onion. Using LC/MS, peak 1 was tentatively identified as quercetin monoglucoside at m/z 466.4, and peak 2 as quercetin with [M]+ at m/z 303.3. The levels of quercetin in the skin, and the outer, middle and core parts of the plant were 16.83,2.67,0.95, and 0.35 mg/g, respectively. In the study of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, skin, the nonedible part, contained the highest amount of quercetin, compared to the other edible parts, and showed the highest DPPH radical scavenging activity. Levels of quercetin and DPPH radical scavenging activity increased from core to skin. All parts of onion exhibited the strongest antibacterial activity only against Staphylococcus aureus and Vibro parahaemolyticus. Antibacterial activities of onion exhibited that S. aureus was more sensitive than V. parahaemolyticus. Among the four onion extracts, the middle part showed the strongest inhibitory activity against S. aureus but all onion extracts showed similar antibacterial activities against V. parahaemolyticus.

Anthocyanin Analysis of Pressure-extracted Korean Blueberry Juice and in vitro Anti-inflammatory in RAW267.4 Cell line (국산 블루베리 착즙액의 안토시아닌 분석 및 RAW267.4 세포주에서의 항염효과)

  • Choi, Moon-Hee;Jeon, Young-Jin;Shin, Hyun-Jae
    • KSBB Journal
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    • v.30 no.4
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    • pp.191-196
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    • 2015
  • Blueberry juice possesses rich-procyanidins and - anthocyanidin, comprised a group of with numerous health benefits such as protection against coronary heart disease, detoxification, and obesity. Blueberry (Vaccinium virgatum) juice extracts were analyzed and separated by an HPLC method for the purpose of the separation and quantification in polyphenolic groups. In specific HPLC conditions, a binary mobile phase consisting of formic acid: water (10:90, v/v, solvent A) and formic acid: water: acetonitrile (10:60:30, v/v/v, solvent B) was utilized and it is detected at 546 nm wavelength. The phenolic contents of the extracts are determined using Folin-Ciocalteu phenol reagent. In order to test anti-inflammation activity assay, after producing nitric oxide (NO) in lipopolysaccharide activated RAW 264.7 cells, at concentration of $20-500{\mu}g/mL$ it reduced to NO production at a dose-dependent manner. Importantly, cytotoxicity assay with up to $500{\mu}g/mL$ of the extract from blueberry juice showed ~100% cell viability for RAW264.7 cell line. Therefore, Korean blueberry juice might have potential as anti-oxidant and antiinflammation agents.

Allelopathy and Quantification of Causative Allelochemicals in Sweet Potato

  • Chon, Sang-Uk
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.5
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    • pp.402-406
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    • 2003
  • Greenhouse and laboratory studies were conducted to determine the allelopathic potentials of extracts or residues from sweet potato (Ipomoea batatas L. (Lam). The extracts applied on filter paper in a Petri dish bioassay significantly inhibited root growth of alfalfa. Aqueous leachates at 40g dry tissue $\textrm{L}^{-1}$ (g $\textrm{L}^{-1}$) from leaves showed the highest inhibition against alfalfa, and followed by stems and roots. Alfalfa root growth was significantly inhibited by methanol extracts of the same plants as the concentration increased. The effect of residue incorporation into soil on seedling growth of com, soybean, barnyard grass and eclipta was examined in the greenhouse, and results showed that the leaf residues at 200g $\textrm{kg}^{-1}$ by plant parts inhibited shoot dry and root dry weights of test plants by 60-80%. By means of HPLC, causative allelopathic substances present in plant parts of sweet potato "Sinyulmi" were identified as coumarin, trans-cinnamic acid, o-coumaric acid, p-coumaric acid, and chlorogenic acid. Total content of these compounds for leaves extracts were detected as the greatest amount in EtOAc fraction, especially trans-cinnamic acid was the greatest component. These results suggest that sweet potato plants have herbicidal potentials, and that their activities exhibit differently depending on plant parts.ant parts.