• Title/Summary/Keyword: TLC analysis

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Bioethanol Production using a Yeast Pichia stipitis from the Hydrolysate of Ulva pertusa Kjellman (효모 Pichia stipitis를 이용한 구멍갈파래 가수분해 추출물로 부터 바이오 에탄올 생산)

  • Lee, Ji-Eun;Lee, Sang-Eun;Choi, Woon-Yong;Kang, Do-Hyung;Lee, Hyeon-Yong;Jung, Kyung-Hwan
    • The Korean Journal of Mycology
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    • v.39 no.3
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    • pp.243-248
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    • 2011
  • We studied the repeated-batch process for the bioethanol production from the hydrolysate of Ulva pertusa Kjellman using yeast Pichia stipitis, which is able to assimilate C6- and C5-monosaccharides. During 180-hour operations, the repeated-batch process was carried out stably, and the average bioethanol concentration reached 11.9 g/L from about 30 g/L of reducing sugar in the hydrolysate. Meanwhile, the bioethanol yields, based on the reducing sugar and the quantitative TLC analysis, were 0.40 and 0.37, respectively, which corresponded to 78.4% and 72.5% of theoretical value, respectively. Throughout the quantitative process analysis, it was also demonstrated that 39.67 g-bioethanol could be produced from 1 kg of dried Ulva pertusa Kjellman. In this study, we verified that the bioethanol production from the hydrolysate of Ulva pertusa Kjellman was feasible using a yeast Pichia stipitis, particularly during the repeated-batch operation.

A Study on Standardization of Shinbaro Pharmacopuncture Using Herbal Medicines Identification Test and HPLC-DAD (신바로 약침의 한약재 확인시험 및 HPLC-DAD를 통한 표준화 연구)

  • Lee, Jin Ho;Kim, Min Jeong;Lee, Jae Woong;Kim, Me Riong;Lee, In Hee;Kim, Eun Jee
    • Journal of Acupuncture Research
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    • v.32 no.2
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    • pp.1-9
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    • 2015
  • Objectives : The present study was an evaluation and standardization of herbal components in order to establish the efficacy and safety of Shinbaro pharmacopuncture. Methods : Among the raw materials of Shinbaro pharmacopuncture, the components Cibotii Rhizoma, Eucommiae Cortex, and Ledebouriellae Radix were assessed through ingredient verification experiments using thin-layer chromatography(TLC) and ultraviolet rays(UV) lamps. In addition, we standardized Acanthopanacis Cortex and Achyranthis Radix through validation using high performance liquid chromatograph-diode array detector(HPLC-DAD). Results : As result appeared a blue-white fluorescence under ultraviolet rays; changed to dark green after adding 1 % ferric chloride solution(due to Cibotii Rhizoma), and presented a yellow-green fluorescence when mixed with an ethyl ether under UV lamps by way of the ethyl ether layer, confirming Eucommiae Cortex. Ledebouriellae Radix was confirmed as dark brown spots at Rf values of 0.56 and 0.71 using TLC. Additionally, Acanthopanacis Cortex and Achyranthis Radix HPLC test results showed that linearity was $R^2{\geq}0.99$, and detection limit and quantitation limit were 0.23 to $1.29{\mu}g/mL$, and 0.71 to $3.90{\mu}g/mL$, respectively. Furthermore, precision and accuracy were confirmed to have relative standard deviation(RSD) values of 0.10 to 1.89 % and 96.19 to 103.72 %, respectively. Shinbaro pharmacopuncture did not have any overlapping or interference from other peaks in detection under the abovementioned analysis conditions. Conclusions : In conclusion, we confirmed that maintenance of Shinbaro pharmacopuncture validity was possible by means of quality control of Cibotii Rhizoma, Eucommiae Cortex, and Ledebouriellae Radix through ingredient identification and Acanthopanacis Cortex and Achyranthis Radix through high performance liquid chromatograph(HPLC) analysis. Further, we hope to contribute to the development strategy of herbal industry acupuncture.

Sugars in Korean and Japanese Pumpkin (한국산 호박 및 일본산 호박의 당 성분에 관한 연구)

  • 안용근
    • The Korean Journal of Food And Nutrition
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    • v.10 no.4
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    • pp.453-457
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    • 1997
  • Sugars in Korean and Japanese pumpkin were studied. The sugars in pumpkin were crushed and extracted by boiling for 30 min. Korean pumpkin was found to contain 0.41% of sucrose, 0.54% of fructose, 0.61% of glucose and 0.68% of starch. Japanese pumpkin was found to contain 2.60% of sucrose, 2.76% of fructose, 1.91% of glucose and 1.22% of starch. No other mono- and oligosaccharides were detected in the test of TLC and HPLC. Starch in Japanese pumpkin showed only signal of $\alpha$-1,4-glucosidic linkage by proton NMR analysis, and showed 86% of absorbance by iodine reaction compared with amylose(DP 117). These results indicated that starch in Japanese pumpkin is composed by only amylose. Pectin contents of Korean and Japanese pumpkin sowed 6.29% and 2.67%, respectively, as galacturonic acid by carbazole analysis.

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Streptomyces endus YP-1이 생산하는 항암활성 물질의 분리 및 정제

  • Choi, Sung-Won;Kim, Byoung-Chan;Choi, Sun-Jin;Kim, Dong-Seob;Yeo, Ick-Hyun;Moon, Soon-Ok;Oh, Doo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.25 no.5
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    • pp.483-489
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    • 1997
  • Sulforhodamine B (SRB) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, RNA dot blot and Northern hybridization analysis were performed to screen microorganisms for the production of anticancer agent. Among microorganisms tested, strain YP-1 was selected for its cytotoxicity and ability to reduce the level of c-myc RNA. Strain YP-1 was identified as Streptomyces endus. The anticancer material produced by Streptomyces endus YP-1 was sequentially purified by solvent extraction, silica gel column chromatograpby, preparative TLC and preparative HPLC. The cancer material identified as azalomycin B by the instrumental analyses such as $^{1}$H-NMR, $^{13}$C-NMR, Mass, IR and UV absorption. It was colorless amorphous powder and its molecular weight was 1025.278. Azalomycin B, produced by Streptomyces endus YP-1, showed anticancer activity against several human cancer cell lines and reduction of c-Myc protein level in Colo320 DM cells which was determined by Western blot analysis.

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Polysaccharide Characteristics from Hot Water Extract of Aloe saponaria Callus (Aloe saponaria 캘러스의 열수 추출물 유래 다당의 특성)

  • Baek, Jin-Hong;Kim, Myung-Uk;Kang, Tae-Su;Hur, Won;Lee, Shin-Young
    • KSBB Journal
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    • v.24 no.1
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    • pp.59-64
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    • 2009
  • The callus formation from inferior leaf of Aloe saponaria was induced in M & S medium supplemented with 10-30 ${\mu}M$ NAA (${\alpha}$-naphthalene acetic acid) and 3-7 ${\mu}M$ kinetin under incubation in the dark at $25^{\circ}C$ for 6 weeks. The hot water extract ($100^{\circ}C$, 24 hrs) from cultured callus was obtained and the components analysis for the extract were examined to determine the callus can synthesized the bioactive component such as Aloe polysaccharide. The freeze dried extract contained the sugar of 53.2%, protein of 7.3%, ash of 18.5% and water of 21% (w/w). Two fractions (Fr-I and Fr-II) were obtained by Sepharose CL-4B gel permeation chromatography and Fr-I, major fraction was further purified with dialysis. From sugar analysis by TLC and GC, the purified Fr-I fraction consisted of glucose (77.6%), galactose (17.7%), mannose (4.7%, w/w) and uronic acid (trace). The molecular weight of purified Fr-I fraction determined by GPC was about 110 kDa.

Quantitative Analysis of Kirenol in Siegesbeckia glabrescens and S. pubescens by HPLC-UV (HPLC-UV에 의한 진득찰과 털진득찰의 Kirenol 정량분석)

  • Nugroho, Agung;Lee, Kyung-Tae;Park, Hee-Juhn
    • Korean Journal of Pharmacognosy
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    • v.43 no.4
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    • pp.286-290
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    • 2012
  • Many diterpenoids from Siegesbeckia species (Compositae) and their anti-inflammatory actions have been examined. In this research, high-performance liquid chromatography-ultraviolet spectrophotometer (HPLC-UV) method was used to compare the quantitative level of kirenol (ent-pimarane-type diterpenoid) in the aerial parts of Korean S. glabrescens and S. pubescens and the Chinese Siegesbeckiae Herba. Fingerprints of the two HPLC chromatograms of Korean S. glabrescens and S. pubescens were similar, but considerably different from Chinese Siegesbeckiae Herba. The content of kirenol in S. pubescens ($16.51{\pm}0.10$ mg/ml dry weight as mean${\pm}$RSD) was higher than S. glabrescens ($13.48{\pm}0.12$ mg/g dry weight). These values were considerably higher than the Chinese Siegesbeckiae Herba ($1.55{\pm}0.74$ mg/g dry weight). Thin layer chromatography (TLC) analysis demonstrated the containing of kirenol in the three plant materials, but the presence of siegeskaurolic acid (entkaurane-type diterpenoid) only in the Chinese Siegesbeckiae Herba.

Studies on the Analysis of Constituents of Deer Horn(II) -Analysis of gangliosides and free amino acids- (산지별 녹용(鹿茸)류의 성분분석 연구(II) -Ganglioside 및 유리 아미노산의 분석-)

  • Hong, Nam-Doo;Won, Do-Hee;Kim, Nam-Jae;Chang, Seung-Youb;Youn, Whang-Geum;Kim, Hae-Soo
    • Korean Journal of Pharmacognosy
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    • v.24 no.1
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    • pp.38-46
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    • 1993
  • Two kinds of gangliosides contained in deer horns were determined by integrating the peaks of TLC densitometry. Japanese deer horn originated from China showed the highest gangliosides among tested samples and the upper parts in deer horns showed higher gangliosides than the lower parts. In the case of graded samples, the best grade A showed the highest content and the worst grade E did the lowest content. Sixteen kinds of free amino acids were analyzed by auto amino acid analyzer. The lower region the deer horn was, the more the total content of free amino acids was and several kinds of amino acids were contained quite regularly.

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Characterization of γ-Aminobutyric acid(GABA) produced by a lactic acid bacterium from button mushroom bed

  • Lee, Yun-Seok;Song, Tae-Young;Kong, Won-Sik;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.11 no.4
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    • pp.181-186
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    • 2013
  • ${\gamma}$-Aminobutyric acid(GABA) is a four carbon non-protein amino acid that has several well-known physiological functions, such as a postsynaptic inhibitory neurotransmitter in the brain and induction of hypotensive and tranquilizer effects. A lactic acid bacterium was isolated from button mushroom bed, which is showing high GABA productivity by TLC or HPLC analysis. The strain was identified as Lactobacillus hilgardii by analysis of 16S rDNA gene sequence. When the maximum production of GABA by L. hilgardii was investigated with various concentration of monosodium glutamate, the yield of GABA reached to be 53.65 mM at 1% mono sodium glutamate (MSG) in flask cultivation. A Glutamate decarboxylase (GAD) enzyme, which was known to convert MSG to GABA, was purified from a cell-free extract of L. hilgardii and the molecular weights of purified GAD was estimated to 60,000 by SDS-PAGE. The optimum pH and temperature of GAD were at pH4.6 and at $37^{\circ}C$, respectively. The GAD activity was increased by the addition of sulfate ions such as ammonium sulfate, sodium sulfate and magnesium sulfate, indicating that the increase of hydrophobic interaction causes the increase of GAD activity.

Development of Alternative Testing Methods without Hazardous Reagents used in Korean Pharmaceutical Codex (고시의약품 시험에 사용되는 유해시약 대체 시험법 개발)

  • Kim, Hee-Yun;Kang, Hyun-Kyung;Choi, Seon-Hee;Bang, Su-Jin;Han, Kyung-Jin;Choi, Sung-Hee;Kim, Jin-Hee;Lee, Hwa-Jung;Kang, Chan-Soon
    • YAKHAK HOEJI
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    • v.54 no.2
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    • pp.142-149
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    • 2010
  • Development of alternative testing methods for the replacement of hazardous reagents with less hazardous ones is strongly enforced because exposure of human and environment to hazardous reagents are restricted and hazardous reagents are gradually prohibited from using in various testing methods. Thus, in this study, we developed 8 monographs from the Korean Pharmaceutical Codex by substituting the use of the hazardous reagents including ICH class 1 such as benzene, chloroform and dioxane to the use of less toxic ones like ICH class 2 or 3 reagents. We also improved their qualification and quantification performance. Among 8 monographs, the 6 newly developed TLC methods for the identification of nifedipine, oxolamine citrate, ketoprofen lysinate, chlorquinaldol, retinol acetate, and riboflavin showed a clear spot of corresponding material without any interference in spite of the replacement with ICH class 2 or 3 reagents. For the quantification of domperidone and trimebutine, HPLC methods were developed for the substitution of UV/VIS spectrometry and titrimetry, respectively. These HPLC methods were validated for the linearity, recovery, reproducibility, and inter-laboratory variations. In conclusion, the newly developed methods could be expected to become valuable tools for revising the Korean Pharmaceutical Codex.

Monitoring of Aflatoxin $B_1$ in Livestock Feeds Using ELISA and HPLC

  • Han Eun-Mee;Park Hee-Ra;Hu Soo-Jung;Kwon Ki-Sung;Lee Hyo-Min;Ha Mi-Sun;Kim Kyung-Mi;Ko Eun-Jung;Ha Sang-Do;Chun Hyang-Sook;Chung Duck-Hwa;Bae Dong-Ho
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.643-646
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    • 2006
  • Because of potential health hazards of aflatoxins for humans, the present study was conducted to monitor aflatoxin $B_1\;(AFB_1)$ in livestock feeds. A total of 249 samples of feeds collected in Korea were analyzed by DC-ELISA for qualitative analysis of $AFB_1$. Then, 27 samples that were verified to contain $AFB_1$ by DC-ELISA were quantitated by HPLC/FLD. HPLC/FLD analysis revealed that only one sample collected from a farm contained 11 ppb of $AFB_1$, whereas the other samples collected from feed companies did not contain $AFB_1$. The presence of $AFB_1$ was further confirmed by LC/MS analysis. TLC analysis indicated that the result of the DC-ELISA was most likely due to possible contamination of other mycotoxins rather than $AFB_1$. In conclusion, HPLC/FLD analysis following DC-ELISA is necessary for rapid and accurate detection of $AFB_1$.