• Title/Summary/Keyword: HPTLC and HPLC

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The Application of High Performance Thin Layer Chromatography for Herbal Formula Standardization (한약처방의 표준화를 위한 HPTLC의 활용)

  • Choi, Min-Kyung;Kim, Hyeong-Geug;Wang, Jing-Hua;Son, Chang-Gue
    • The Journal of Korean Medicine
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    • v.32 no.4
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    • pp.68-74
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    • 2011
  • Objective: The purpose of this study is to expatiate on high performance thin layer chromatography (HPTLC) as a simple, easy and scientific method for evaluation and standardization of herbal formulae. Methods: Through retrieving HPTLC application for herbal formulae in the literatures, the current situation of HPTLC application, and potential as well as limitation of HPTLC as a standardization method for multi-herbal drug was studied. Results: HPTLC is a speedy, inexpensive and well-operable tool for possessing multi-capability on component identification, separation, quantification and purification compared to other methods, such as high performance liquid chromatography (HPLC), gas chromatography (GC). Conclusions: HPTLC is considered as an available and convenient method for quality control and standardization of multi-herbal drugs. Thereby, this method could be recommended to widely applicate in the traditional Korean medicine.

Isolation and Quantitative Analysis of Betulinic Acid and Alphitolic Acid from Zyziphi Fructus (대추로부터 베튜리닉 산과 알피톨릭 산의 분리 및 정량)

  • Bae, Gi-Hwan;Lee, Sang-Myeong;Lee, Eun-Sil;Lee, Jun-Seong;Gang, Jong-Seong
    • YAKHAK HOEJI
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    • v.40 no.5
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    • pp.558-562
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    • 1996
  • Betulinic acid and alphitolic acid, the triterpenoids of Zyziphi Fructus, were isolated with silica gel column chromatography and used as the standard substances for the analysis. The compounds were determined with HPLC and HPTLC. They were separated on reversed phase column (Nova-Pak C18) with 0.05M Na2HPO4-methanol (19:81) in HPLC and detected at 210nm. Separation on HPTLC precoated silica gel F254 plates was carried out with chloroform-methanol (6:1) and the separated compounds were reacted with p-anisaldehyde and detected at 540nm. The contents of betulinic acid and alphitolic acid in Zyziphi Fructus from four different regions in Korea were in the range of 2.9~3.8% and 3.2~3.9%, respectively.

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Quality Evaluation on Magnoliae Cortex (후박의 품질평가)

  • Bae, Ki-Hwan;Kim, Young-Ho;Won, Do-Hee;Lee, Jun-Sung;Kang, Jong-Seong
    • YAKHAK HOEJI
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    • v.41 no.4
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    • pp.407-413
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    • 1997
  • Magnolol and honokiol, the main components of Magnoliae Cortex, were isolated and used as the standard substances for the analysis. In order to determine the contents of magnolol and honokiol in Magnoliae Cortex originated from Korea, China and Japan, both HPLC and HPTLC methods are applied and compared with each other. The components were separated on C8 column with acetonitrile-water-acetic acid (50:50:1) in HPLC and detected at UV 294nm. The components separated on HPTLC precoated silica gel plate with chloroform-methanol (9:1) were detected directly on the plate at 254nm. The contents of magnolol and honokiol in Magnoliae Cortex were in the wide range of 0.01~2.8% and 0.005~0.8%, respectively, according to their purchase places. It is also applicable to the quality control of various preparation from Magnoliae Cortex.

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Antiproliferative effect of Schisandrae Fructus extract on PC-3 human prostate cancer cells (오미자(五味子) 추출물의 인간 전립선암 세포주 PC-3에 대한 성장 억제 효과)

  • Moon, Jung-Min;Seok, Ga-Hyeong;Cho, Su-In
    • The Korea Journal of Herbology
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    • v.27 no.4
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    • pp.17-23
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    • 2012
  • Objectives : Schisandrae Fructus (SF) has traditionally been used to balance level of body fluid and to strengthen kidney function. It has been reported that the SF extract has antioxidant, hepatoprotective, neuroprotective and anticancer effects. This study investigated an antiproliferative effect of SF extract on PC-3 human prostate cancer cells and analyzed active ingredients of SF extract qualitatively and quantitatively. Methods : We examined the antiproliferative effect of SF extract with MTT assay, DAPI staining and annexin-V/7-AAD double staining. The active ingredients of SF extract were identified by using HPTLC and HPLC/DAD system. Results : SF-chloroform fraction inhibited growth of PC-3 cells and changed the morphology of nucleus in a dose dependent manner. A dose-dependent apoptotic cell death was also measured by flow cytometry analysis. It was analyzed that SF-chloroform fraction contained more schizandrin than other fractions by using HPTLC and HPLC/DAD system. Conclusions : These results suggest that SF extract and schizandrin may be a potential chemotherapeutic agent for the control of PC-3 human prostate cancer cells.

Comparison of Analytical Methods for Saikosaponins in Bupleurum falcatum L. (자호(紫胡) 사이코사포닌 정량분석방법(定量分析方法) 비교(比較))

  • Kim, Kwan-Su;Lee, Seoung-Tack;Seong, Nak-Sui;Lee, Jung-Il;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.3
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    • pp.226-232
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    • 1995
  • Extraction methods and instrumental analytical conditions were compared to establish a fast and appropriate analytical method to determine saikosaponins in Bupleurum falcatum. Using HPLC, analysis of diene-saikosaponin treated with 2% acid was faster than that of saikosaponin itself. Among various extraction methods, extraction by standing in methanol at room temperature showed highest efficieny, and extraction with boiling methanol was shorter in analytical time and showed good chromatogram. And we could analyze many samples faster using HPTLC but the analytical accuracy was low. In extraction and analysis of saikosaponins, extraction with boiling methanol and acidic treatment was fast and easy analytical method. And for selecting useful lines in component breeding, we think TLC method was better.

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Analysis of Nutritional Composition and Phenolic Compound in Propolis Collected from Falseacacia and Chestnut Tree in Korea (국내산 아까시나무와 밤나무 유래 propolis의 영양성분 및 페놀성 화합물 분석)

  • Song, Hyo-Nam;Gil, Bog-Im
    • Korean Journal of Food Science and Technology
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    • v.34 no.4
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    • pp.546-551
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    • 2002
  • Nutritional composition and phenolic compounds of raw propolis collected from falseacacia (Robinia pseudoacacia L.) and chestnut tree (Castanea crenata), and their 70% ethanol extracts of propolis (EEP) were analyzed. Propolis had high crude lipid content, but no significant differences in general compositions in terms of collection area and plant origins. Mineral contents varied greatly depending on the plant origins, with falseacacia propolis showing the highest mineral content. Sixteen amino acids were analyzed, among which aspartic acid content was the highest at $328.4{\sim}410.6\;mg%$ and methionine the lowest at $0{\sim}21.1\;mg%$. Extraction yield for EEP was relatively high at $64.2{\sim}81.9%$, and total polyphenol and flavonoid contents were $13.9{\sim}23.7$ and $8.6{\sim}10.8%$, respectively. HPTLC and HPLC analysis on the phenolic compounds revealed the overall chromatographic patterns were almost equal, showing similar polyphenol compositions between the propolis. About 16 peaks were identified by HPLC analysis, among which 6 peaks of p-hydroxy benzoic acid, caffeic acid, ferulic acid, benzoic acid, cinnamic acid, and chrysin were identified.

HPLC SEPARATION AND QUANTITATIVE DETERMINATION OF GINSENOSIDES FROM PANAX GINSENG, PANAX QUINQUEFOLIUM AND FROM GINSENG DRUG PREPARATIONS

  • Soldati F
    • Proceedings of the Ginseng society Conference
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    • 1980.09a
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    • pp.59-69
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    • 1980
  • A new HPLC-method for separation and quantitative determination of ginsenosides in Panax ginseng, Panax quinquefolium and in pharmaceutical drug preparations is elaborated. A reversed-phase-system with ${\mu}Bondapak\;C_{18}$ column (3.9 mm $I.D.{\times}30\;cm$) using acetonitrile-water (30:70) 2 ml/min and acetonitrile-water (18:82) 4 ml/min is suitable for the base-line separation of $Rb_1,\;Rb_2,\;Rc,\;Rd,\;Rf,\;Rg_2,\;respectively\;Re,\;Rg_1$ in 30 minutes. The ginsenosides are directly detected at 203 nm (without derivatization) with the LC-55 or LC-75 spectrophotometer (Perkin-Elmer) at $100\%$ transmission. Detection limit is 300 ng at a signal-to-noise ratio of 10:1. The ginsenosides-peak identification is carried out with HPTLC (high performance thin layer chromatography), with MIR-IR (multiple internal reflection-IR-spectros-copy) and with FD-MS (field desorption mass spectrometry). The calibration curve of each ginsenoside has a correlation coefficient very near to 1. Relative standard deviation for quantitative determinations depends upon the amount of ginsenosides and is approximately 1\%$ for ginsenoside contents of 1\%$. This method is adaptable for routine analysis in quality control laboratories.

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Pharmacognostical Evaluation of Andrographis stenophylla C. B. Clarke

  • Vamsadhara, C.;Bharathi, R. Vijaya
    • Natural Product Sciences
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    • v.13 no.3
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    • pp.241-246
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    • 2007
  • Andrographis stenophylla C. B. Clarke., (Acanthaceae) is an erect glabrous undershrub with very narrow leaves and stems from a stout rootstock, the corolla pale with dark red stripes. The plant grows in hills of about 1200 meters height in South India. No scientific work reports are available with regard to this plant. The present study, thus deals with the detailed pharmacognostical evaluation of the plant A. stenophylla using light and confocal microscopy, WHO recommended physico-chemical determinations and authentic phytochemical procedures. The physico-chemical, morphological and histological parameters presented in this paper may be proposed as parameters to establish the authenticity of A. stenophylla and can possibly help to differentiate the drug from its other species.

Characterization of Phospholipid and Fatty Acid Composition in the Amp 1-4 Mutant Compared to Wild-Type Arabidopsis thaliana

  • Nam, Im-Sook;Hong, Yong-Geun;Hwang, In-Hwan;Cho, Moo-Je;Pak, Yun-Bae
    • BMB Reports
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    • v.32 no.1
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    • pp.6-11
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    • 1999
  • To understand the function of phospholipids and their fatty acid composition on the morphological changes in the amp 1-4 mutant of Arabidopsis, the mutant was compared to the wild-type Arabidopsis by TLC, HPTLC, phosphorous assay, HPLC, and GC. In the mutant, phosphatidylethanolamine (PE) was increased 5-fold and phosphatidylglycerol (PG) was decreased 1.2-fold (nmol phosphorous/g tissue). Inositol phospholipids showed a generally increased trend ranging from 1.4-to 3.0-fold (nmol inositol/g tissue). When fatty acid composition of the mutant was compared to the wild-type, linoleic (18:2) and linolenic (18:3) acids of phosphatidylcholine (PC) and PG were decreased but palmitoleic acid (16:1) and oleic acid (18:1) of PC was increased 2.5- and 2.1-fold (mol%), respectively. In galactolipids, myristic acid (14:0) of monogalactosyl-diacylglycerol (MGDG) were increased 5.8-fold (mol%). Among the inositol phospholipids, lysophosphatidylinositol (L-PI) and phosphatidylinositol 4,5-bisphosphate ($PIP_2$) showed 4-and 1.9-fold (mol%) increase of 16:1, respectively. These results suggest that the increase of PE, the decrease of PG, the increase of inositol phospholipids, and the altered fatty acid composition are related to the phenotypic changes affecting the morphological features, and might cause different physiological changes in the amp 1-4 mutant compared to wild-type Arabidopsis.

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Royal jelly enhances migration of human dermal fibroblasts and alters the levels of cholesterol and sphinganine in an in vitro wound healing model

  • Kim, Ju-Young;Kim, Young-Ae;Yun, Hye-Jeong;Park, Hye-Min;Kim, Sun-Yeou;Lee, Kwang-Gill;Han, Sang-Mi;Cho, Yun-Hi
    • Nutrition Research and Practice
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    • v.4 no.5
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    • pp.362-368
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    • 2010
  • Oral administration of royal jelly (RJ) promotes wound healing in diabetic mice. Concerns have arisen regarding the efficacy of RJ on the wound healing process of normal skin cells. In this study, a wound was created by scratching normal human dermal fibroblasts, one of the major cells involved in the wound healing process. The area was promptly treated with RJ at varying concentrations of 0.1, 1.0, or 5 mg/ml for up to 48 hrs and migration was analyzed by evaluating closure of the wound margins. Furthermore, altered levels of lipids, which were recently reported to participate in the wound healing process, were analyzed by HPTLC and HPLC. Migration of fibroblasts peaked at 24 hrs after wounding. RJ treatment significantly accelerated the migration of fibroblasts in a dose-dependent manner at 8 hrs. Although RJ also accelerated the migration of fibroblasts at both 20 hrs and 24 hrs after wounding, the efficacy was less potent than at 8 hrs. Among various lipid classes within fibroblasts, the level of cholesterol was significantly decreased at 8 hrs following administration of both 0.1 ug/ml and 5 mg/ml RJ. Despite a dose-dependent increase in sphinganines, the levels of sphingosines, ceramides, and glucosylceramides were not altered with any concentration of RJ. We demonstrated that RJ enhances the migration of fibroblasts and alters the levels of various lipids involved in the wound healing process.