• Title/Summary/Keyword: marker substances

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HPLC Analysis of Marker Substances in Taeumin Gamijowi-tang (태음인(太陰人) 가미조위탕(加味調胃湯)의 HPLC 분석)

  • Hwang, Young-Myung;Yu, Young-Beob;Kim, Jong-Youl;Koh, Byung-Hee;Lee, Eui-Ju
    • Journal of Sasang Constitutional Medicine
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    • v.18 no.1
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    • pp.147-154
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    • 2006
  • 1. Objectives This study was aimed to evaluate marker substances in Gamijowi-tang by high performance liquid chromatography(HPLC) 2. Methods The seperation was performed an AquasilC18(4.6X250mm)column by gradient elution with 0.1N H3PO4 in H2O - acetonitrile(0min 100:10,20min60:40, 40min 80:20, 60min 100:0) as the mobile phase at a flow-rate of 1.0 ml/min with detection at 190-800nm 3. Results Ephedrine and schizandrin were established as marker substances in Gamijowi-tang and its qualitative analysis was carried out. Analyzed was similarity of marker substances between in standard article and in Gamijowi-tang. The result of the analysis was that ephedrine and schizadrin showed 0.9999 of similarity, which meant they were really ephedrine and schizadrin. In addition, ephedrin content was 12.16mg, pseudoephedrine conrenr was 5.09mg, and schizandrin content was 26.6mg in 76g of Gamijowi-tang. 4. Conclusions Qualitative and quantitative analyses of ephedrine and schizandrin in Gamijowi-tang were conducted, and it was identified thar they could be used as marker substances in standardizing quality of Gamijowi-rang.

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Component Analysis and Toxicity Study of Combined Cultivated Wild Ginseng Pharmacopuncture (산양산삼복합약침의 표준화 및 급성독성시험)

  • Baek, Sang-hyun;Lee, In-hee;Kim, Min-jeong;Kim, Eun-jee;Ha, In-hyuk;Lee, Jin-ho;Le, Jae-woong
    • The Journal of Internal Korean Medicine
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    • v.36 no.2
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    • pp.189-199
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    • 2015
  • Objectives: The marker substances of cultivated wild ginseng pharmacopuncture that may not be detected during the process of steaming remain controversial. We developed a combined cultivated wild ginseng pharmacopuncture that contains all the marker substances. The aim of this experiment was to investigate the marker substances and test the toxicity of the combined cultivated wild ginseng pharmacopuncture. Methods: The marker substances were detected using HPLC. Intravenous injection toxicity studies were conducted at Medvill, an authorized institution for non-clinical studies, under the regulations of Good Laboratory Practice. We observed survival rates, abnormal behaviors, weight changes, gross findings in autopsy, blood biochemical properties, and histological abnormalities of organs such as the liver and kidney. Results: HPLC data showed that ginsenosides Rg1, Rb1, and Rg3 were detected at concentrations of 19.29, 47.64, and 3.02 μ g /ml, respectively. Administration of combined cultivated wild ginseng pharmacopuncture resulted in no dead animals or significant toxicological changes. Conclusions: The combined cultivated wild ginseng pharmacopuncture contains all the marker substances and is a relatively safe treatment medium. Further studies should be conducted to confirm the present findings.

Fingerprint of Marker Substances in Gami-Honghwa-Tang(KH-19) by HPLC-DAD (High Performance Liquid Chromatography-Diode Array Detector(HPLC-DAD)에 의한 가미홍화탕 (KH-19)의 지문 분석)

  • Yu Young-Beob;Yoon Yoo-Sik;Cho Gi-Ho
    • The Journal of Korean Medicine
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    • v.25 no.3
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    • pp.45-54
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    • 2004
  • Objectives : This study was aimed to evaluate marker substances in Gami-Honghwa-Tang (KH-19) by high performance liquid chromatography-photodiode array detector (HPLC-DAD). Gami-Honghwa-Tang is composed of nine crude herbs, Rehmanniae Radix Preparata, Angelicae Gigantis Radix, Cnidii Rhizoma, Paeoniae Radix, Corni Fructus, Moutan Cortex Radicis, Lycii Fructus, Carthami Flos and Glycyrrhizae Radix. Methods : The separation was performed on an Aquasil C18 (4.6×250mm) column by gradient elution with 0.05% TFA in H2O - 0.05% TFA in acetonitrile (0 min 100:0, 20 min 90:10, 40 min 70:30, 60 min 50:50, 80 min 0:100, 90 min 100:0) as the mobile phase at a flow-rate of 1.0 ml/min with detection at 190-800nm. Also we examined the contents for bacteria, pesticide residue and harmful heavy metals. Results : HPLC-DAD was employed to determine the quantities and the qualities of several marker substances such as 5­hydroxymethyl-2-furaldehyde (5-HMF), paeonol, loganin, paeoniflorin, glycyrrhizin, and decursin in the KH-19. There were no bacterial contents, pesticide residues, or harmful heavy metals. Conclusions : We suggest these results could be a useful evidence for quality control of KH-19. This method permits fingerprints of selected individual marker substances from herbal prescriptions without derivatization, multiple purification steps, or lengthy separation times.

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HPLC analysis of Gami-Samhwang-San and prediction of active compounds using QSAR (가미삼황산(加味三黃散) 분획물(SH-21-B)의 지표성분 정량과 구조활성상관(QSAR) 예측)

  • Yu, Young-Beob
    • Journal of Korean Traditional Oncology
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    • v.11 no.1
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    • pp.95-103
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    • 2006
  • Objective: Gami-Samhwang-San, a herbal prescription for obesity treatment, is composed of seven crude herbs such as Ephedrae Herba, Scutellariae Radix, Acori Gramineri Rhizoma, Polygalae Radix, Typhae Pollen, Armeniacae Semen, Nelumbo Folium. This study was aimed to evaluate marker substances in n-butanol fraction (SH-21-B) from Gami-Samhwang-San by high performance liquid chromatography (HPLC). And we predicted inhibition activity of major compounds of Gami-Samhwang-San using Quantitative Structure Activity Relationships (QSAR) Methods: The separation was performed on a YMC J,sphere-H80 CI8(250${\times}$4.6 mm I.D) column by gradient elution with $H_3PO_4$ buffers in acetonitrile as the moblie phase at a flow-rate of 1.0ml/min. Results: HPLC was employed to determine the quantities and the qualities of several marker substances such as ephedrine, pseudoephedirne, baicalin, ${\beta}-asarone$, tenuifoliside, naringenin, amygdalin and hyperoside in the SH-21-B. Conclusion: We suggest this results could be a useful evidence for quality control of SH-21-B.

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Characteristic Effects of Dangnyo-hwan for Diabetes Control Studied Using LC-MS/MS and ICP (LC-MS/MS 및 ICP를 이용한 당뇨환의 화학적 특성 분석)

  • In, Jeong-do;Im, Dai-sig;Moon, Seung-ho;Ki, Won-ill
    • The Journal of Internal Korean Medicine
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    • v.36 no.3
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    • pp.217-227
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    • 2015
  • Objectives: The primary aim of this study was to identify bioactive compounds in Dangnyo-hwan, a Korean herbal medicine, through instrumental analysis using LC-MS/MS and ICP, and investigate its potential use in diabetes treatment. Methods: The extract of Dangnyo-hwan has 12 medicinal herbs; these were compared with 18 marker substances selected from literature survey. Results: LC-MS/MS analysis could detect 9 of the 18 bioactive compounds: citruline, catalpol, berberine, ginsenoside Rb1, ginsenoside Rg1, oleanolic acid, β-sitosterol, mangiferin, and schizandrin. While harmful heavy metals such as As, Pb, Cd, Hg, Ni, and Cu were not present in high concentrations, Zn concentration was 4.2 mg in 100 g Dangnyo-hwan. Conclusions: Instrumental analysis such as LC-MS/MS and ICP was successfully used to identify bioactive compounds in Dangnyo-hwan. Detection of 9 bioactive substances and Zn from the herb medicine is a valuable finding, and suggests that Dangnyo-hwan is a candidate medicine for diabetes. Further investigations like in vitro assay, percent GPR 119 activity, and percent human DGAT-1 inhibition are underway.

Quantitative Analysis of Marker Substances of Paeonia lactiflora by Solid Fermentation (작약의 고체발효에 따른 지표성분의 함량분석)

  • Lee, Ji-Hye;Um, Young-Ran;Park, Hwa-Yong;Lee, Jae-Hoon;Ma, Jin-Yeul
    • Korean Journal of Oriental Medicine
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    • v.15 no.2
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    • pp.119-124
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    • 2009
  • The purpose of this study was investigation of quantitative analysis of marker substances in Paeonia lactiflora extracts by solid fermentation. High performance liquid chromatography (HPLC) for the determination of albiflorin and paeoniflorin in P. lactiflora extracts by solid fermentation, the separation method was performed on C18 column ($250\;mm\;{\times}\;4.6\;mm$, $5\;{\mu}m$, RS tech) using gradient solvent mixtures of water-acetonitrile with photodiode array detector (230nm). The flow rate was 1.0 ml/min. Retention time of albiflorin and paeoniflorin was about 28.88, 31.92 min and linearity of calibration was showed good result(r2 = 0.9998, 0.9996), respectively. Content of albiflorin was $0.090\;{\pm}\;0.03%$ in P. lactiflora extract(control), $0.102\;{\pm}\;0.00%$ in P. lactiflora extract fermented with Paecilomyces japonica, $0.056\;{\pm}\;0.01%$ in P. lactiflora extract fermented with Ganoderma lucidum, $0.093\;{\pm}\;0.00%$ in P. lactiflora extract fermented with honey and $0.046\;{\pm}\;0.00%$ in P. lactiflora extract fermented with Nuruk. Content of paeoniflorin was $4.506\;{\pm}\;0.13%$ in control, $2.599\;{\pm}\;0.04%$ in P. lactiflora extract fermented with Paecilomyces japonica, $1.222\;{\pm}\;0.03%$ in P. lactiflora extract fermented with Ganoderma lucidum, $2.750\;{\pm}\;0.05%$ in P. lactiflora extract fermented with honey and $0.847\;{\pm}\;0.00%$ in P. lactiflora extract fermented with Nuruk, respectively. Content of the marker substances did not increase in all fermentation experiment group.

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Liposome Immunoassay for Bioactive Substances (리포좀을 이용한 생리활성물질의 면역학적 분석법)

  • Kim, Chong-Kook;Park, Kyung-Mi
    • Journal of Pharmaceutical Investigation
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    • v.24 no.4
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    • pp.201-215
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    • 1994
  • Recent development in the immunochemical technique has resulted in a new ultrasensitive analytical method known as liposome immunoassay (LIA). Liposome is a key element in performing liposome immunoassays, specifically designed to participate in immune reactions. A variety of markers can be encapsulated in liposomes and used as quantitative indicators of reactions. Liposome immunoassay based on agglutination, complement-mediated Iysis, cytolysin-mediated Iysis, detergent-mediated Iysis or destabilization of the liposomal membrane have been reviewed. The quantity of markers released from liposomes should be proportional to the concentration of the analytes. Therefore, liposomal agglutination and Iysis which are essential to liposomal Iysis are critically reviewed to provide a better understanding of liposome immunoassay. Based on the literature review of recent advances in liposome immunoassay for bioactive substances, this assay method may provide a convenient, specific and highly sensitive method for detecting and measuring trace amount of clinically relevant substances in the future.

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Acute Oral Toxicity Study of Standardized Gami-Honghwa-Tang (KH-19) in Rats and Beagle Dogs (HPLC로 표준화한 가미홍화탕(KH-19)의 랫드와 Beagle견에 대한 단회경구투여독성시험)

  • Yu Young-Beob;Sung Hyun Jea;Yoon Yoo-Sik
    • Toxicological Research
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    • v.21 no.1
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    • pp.63-70
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    • 2005
  • Gami-Honghwa-Tang (KH-19), a herbal prescription for reducing the side effect of radiotherapy, is composed of eight crude herbs such as Rehmanniae Radix Preparata, Angelicae Gigantis Radix, Cnidii Rhizoma, Paeoniae Radix, Corni Fructus, Moutan Cortex Radicis, Lycii Fructus, Carthami Flos, and Glycyrrhizae Radix. In this study, marker substances in KH-19 were analyzed by high performance liquid chromatography-diode array detector (HPLC-DAD) and safety evaluation of standardized KH-19 was evaluated by good laboratory practices (GLP) guideline of Korea Food and Drug Administration. HPLC-DAD was employed to determine the quantities and the qualities of several marker substances such as 5-hydroxymethyl-2-furaldehyde (5-HMF), paeonol, loganin, paeoniflorin, glycyrrhizin, and decursin in the KH-19. In acute oral toxicity study on rat, transient inhibition of body weight was shown, but change in general symptom was not detected. No dead animal was observed up to 5,000 mg/kg in both male and female animals. In acute oral toxicity study on Beagle dog, transient vomiting, diarrhea, anorexia, and body weight reduction were observed, However, no dead animal was observed up to 2,000 mg/kg in both male and female animals.

Analysis of Scutellaria baicaleinsis Georgi (Scutellariae Radix) by LC-DAD and LC-ESI/MS

  • Yu, Youngbob;Choi, Pil-Son;Koo, Sungtae;Chang, Suhwan
    • Korean Journal of Plant Resources
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    • v.31 no.6
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    • pp.652-659
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    • 2018
  • In this study, baicalin, as a marker substance of Scutellariae Radix, was quantitatively analyzed by a high performance liquid chromatography-photodiode array detector (HPLC-DAD). We identified wogonoside, baicalein, and wogonin in the Scutellariae Radix by a high performance liquid chromatography-electrospray ionization-mass spectrometer (HPLC-ESI-MS). The baicalin was separated on a Xterra C18 column ($5{\mu}m$, $4.6{\times}250mm$) using mobile phase consisting of 38% acetonitrile in 0.68% phosphoric acid. The baicalin spectrum in the Scutellariae Radix extracts was coincided by comparing with UV-visible spectrum (200-550 nm) of baicalin standard in the library. The amount of baicalin in Scutellariae Radix was 10.46%, which is higher than KFDA's guideline. The marker substances of Scutellariae Radix showed a strong base peak $[M]^+$ in the positive detection mode following as; baicalin (m/z; $271[MH^+-sugar]^+$, $447[M+H]^+$), wogonoside (m/z; $285[MH^+-sugar]^+$, $461[M+H]^+$), baicalein (m/z; $271\;[M+H]^+$), wogonin (m/z; $285[M+H]^+$). These results are consistent with the fragment pattern and molecular weight of standard components from literature.

Comparison of Serum Fucose Levels in Leukoplakia and Oral Cancer Patients

  • Rai, Narendra Prakash;Anekar, Jayaprasad;Shivaraja, Shankara YM;Divakar, Darshan Devang;Al Kheraif, Abdulaziz Abdullah;Ramakrishnaiah, Ravikumar;Sebastian, Roopa;Raj, AC;Al-Hazmi, Ali;Mustafa, habil Mohamed
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7497-7500
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    • 2015
  • Background: Tumor markers, designated as a broad group of substances produced by malignancies, could be in the form of biochemical substances, immunological substances, cell surface changes and genetic alterations. Cancer, a disorder of cellular behavior is characterized by alteration of serum glycoproteins. L-fucose, a hexose, which is the terminal sugar in most of the plasma glycoproteins, may be useful as a tumor marker for the detection, monitoring and prognostic assessment of malignancies. The aim of the study was to ascertain the role of serum fucose as a biomarker for early detection of oral cancer and to compare serum fucose levels in healthy controls, leukoplakia and oral cancer patients. Materials and Methods: The study included 60 (100.0%) subjects, who were grouped as 20 (33.3%) control subjects, 20 (33.3%) squamous cell carcinoma patients and 20 (33.3%) leukoplakia patients. Fucose estimation was done using UV-visible spectrophotometry based on the method as adopted by Winzler using cysteine reagent. The results were analyzed statistically using ANOVA with Bonferroni post hoc tests. Results: Results showed a high significance in serum fucose in oral squamous cell carcinoma (OSCC) and leukoplakia subjects compared to normal controls. There was a gradual increase in the values noted from control to leukoplakia and to squamous cell carcinoma. Conclusions: Estimation of serum fucose may be a reliable marker and can be used as an effective diagnostic biomarker in oral squamous cell carcinoma patients.