• 한국식품과학회지
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• 제31권1호
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• pp.30-35
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• 1999

기존의 PC나 TLC에 의한 타르색소의 정성은 많은 시간이 소요되고 회수율이 낮아 혼합색소의 경우 주 색소만이 검출되는 단점이 있고, 많은 표준용액이 동시에 전개되어질 수 없다는 단점이 있어 분 실험에서는 Polyamide와 Photodiode array 검출기가 장착된 HPLC를 이용하여 타르색소의 추출, 정제 및 정량을 여러 각도로 검토하여 정량분석의 최적 조건을 확립하였다. 검출 파장은 각 타르색소마다 고유의 최대흡수부가 있으나 동시분석을 위해서는 254 nm의 파장에서 가장 광범위한 타르색소의 검출이 가능하였으며 적색 계통은 $520{\sim}540\;nm$, 청색이나 녹색계통은 620 nm에서 최소검출한계가 낮음을 알 수 있었다. 따라서 타르색소의 분석에는 photodiode array detector가 장착된 HPLC를 이용함으로써 220 nm에서 800 nm까지의 넓은 영역의 파장에 걸쳐 일시에 분석하므로서 시간 절약과 함께 타르색소 피크마다 3차원 스펙트럼을 제공함으로서 실험의 정확도를 높일 수 있었다. 한편 Polyamide column을 사용하였을 때의 회수율을 측정한 결과 청색 1호가 가장 낮은 94.4%를 보였으며, 전체적으로 100% 부근의 아주 높은 만족할만한 회수율을 얻었다. 타르색소를 사용한 국내산 빙과류, 캔디류, 청량음료류를 시료로 하여 타르색소의 정량을 시도한 결과 허용의 타르색소는 검출되지 않았으며, 주로 적색 계통의 적색 2호와 40호가, 황색계통의 황색 4호, 5호가 많이 사용되고 있음을 알 수 있었고 타르색소의 사용량도 200 ppm을 초과하는 것은 발견되지 않았다. 한편 녹색은 황색 4호에 청색 1호를, 보라색은 적색과 청색을 혼합한 혼합색소를 사용되어지고 있는 것을 알 수 있었다.

• 한국버섯학회지
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• 제17권1호
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• pp.12-18
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• 2019

충청남도 부여군 석성면 양송이 재배 농가에서 양송이 수확 후 배지로부터 토양을 채취하여 auxin(IAA) 생성능이 뛰어난 세균 Yangsong-1 균주를 분리하였다. TLC 및 HPLC 분석을 통해 분리균이 생성한 IAA 농도를 확인한 결과, 0.2% L-tryptophan를 함유한 pH 7.0의 R2A broth 배지에 $35^{\circ}C$, 48시간 배양 시 최대 생성농도는 $126.33mg\;L^{-1}$이었다. 생리적 특성 및 계통학적특성 분석을 통해 분리균은 Gram 음성 간균인 Arthrobacter enclensis Yangsong-1로 동정되었다. 배양조건별 IAA의 생산능 비교 시, IAA 농도의 증가가 배양액의 pH 산성화에 기인함으로서 IAA 생성량과 pH 변화에는 부의 상관성이 있는 것으로 관측되었다. IAA 생성을 위한 전구물질로 알려진 L-tryptophan의 첨가효과는 0.2% 첨가 시 균 생육 및 IAA 생성량이 최대이었으며, 0.4% 이상 고농도 첨가 배지에서는 오히려 IAA의 생성이 저해되었다. 또한 분리균에 의한 식물생육촉진효과를 조사하기 위하여 수경재배 및 pot 재배를 통한 녹두발근 생검법과 상추발근 생검법을 수행한 결과, A. enclensis Yangsong-1의 배양액 접종 시 녹두발근 생검법에서는 대조구에 비해 발근수와 뿌리길이에서 약 1.5배의 뿌리신장효과를 보였고, 상추발근 생검법에서는 대조구에 비해 뿌리길이와 무게에서 약 1.9배의 뿌리신장효과를 보였다.

• 셀메드
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• 제11권1호
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• pp.4.1-4.8
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• 2021

Background: The Unani system of medicine has been practised since centuries for the treatment of a range of diseases. In spite of their efficacy they have been widely criticised due to the lack of standardization and poor quality control. Standardization of Unani medicine is a valuable issue at the present because they are very prone to contamination, deterioration, adulteration and variation in composition due to biodiversity as well as careless collection. Objective: To Standardize and Development of HPTLC Fingerprinting of a polyherbal Unani formulation Qurs-e-Safa. Materials and methods: The conventional and modern analytical techniques were used to standardise Qurs-e-Safa. The study was carried into three different batches of Qurs-e-Safa prepared with its ingredients. The parameters studied are organoleptic, microscopic, physicochemical parameters, phytochemical screening, TLC, HPTLC profile, aflatoxin, microbial load and heavy metal analysis. Results and conclusion: Qurṣ-e-Sa'fa is dark yellow in colour and aromatic smell. Uniformity of diameter and weight variation were found to be 13 ± 0, and 524.7 ± 1.72 mg. friability, hardness and disintegration time of all 3 batches were found to be (0.0615 ± 0.004, 0.0885 ± 0.0047 and 0.0725 ± 0.0058), (3.5 ± 0.2886, 3.67 ± 0.1674 and 3.67 ± 0.1674) and (16 to 17 minutes). Extractive value were found to be maximum in distilled water (38.488 ± 0.20, 37.3824 ± 0.38 and 39.8177 ± 0.13) followed by alcohol (27.5406 ± 0.54, 27.5656 ± 0.32 and 26.9229 ± 0.25). Loss of weight on drying, pH, total ash, acid insoluble ash, qualitative test was set in. Phytochemical screening revealed the presence of Carbohydrates, Phenols, Resins, Proteins, Steroids, fixed oil and Flavonoids. The microbial load was found absent and heavy metals were within permissible limits. The data evolved from the study may serve as a reference to validate and also help in the quality control of other finished products in future research.

• Asian Pacific Journal of Cancer Prevention
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• 제16권16호
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• pp.7179-7188
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• 2015

Cancer is a major health obstacle around the world, with hepatocellular carcinoma (HCC) and colorectal cancer (CRC) as major causes of morbidity and mortality. Nowadays, there isgrowing interest in the therapeutic use of natural products for HCC and CRC, owing to the anticancer activity of their bioactive constituents. Boswellia serrata oleo gum resin has long been used in Ayurvedic and traditional Chinese medicine to alleviate a variety of health problems such as inflammatory and arthritic diseases. The current study aimed to identify and explore the in vitro anticancer effect of B. Serrata bioactive constituents on HepG2 and HCT 116 cell lines. Phytochemical analysis of volatile oils of B. Serrata oleo gum resin was carried out using gas chromatography-mass spectrometry (GC/MS). Oleo-gum-resin of B. Serrata was then successively extracted with petroleum ether (extract 1) and methanol (extract 2). Gas-liquid chromatography (GLC) analysis of the lipoidal matter was also performed. In addition, a methanol extract of B. Serrata oleo gum resin was phytochemically studied using column chromatography (CC) and thin layer chromatography (TLC) to obtain four fractions (I, II, III and IV). Sephadex columns were used to isolate ${\beta}$-boswellic acid and identification of the pure compound was done using UV, mass spectra, $^1H$ NMR and $^{13}C$ NMR analysis. Total extracts, fractions and volatile oils of B. Serrata oleo-gum resin were subsequently applied to HCC cells (HepG2 cell line) and CRC cells (HCT 116 cell line) to assess their cytotoxic effects. GLC analysis of the lipoidal matter resulted in identification of tricosane (75.32%) as a major compound with the presence of cholesterol, stigmasterol and ${\beta}$-sitosterol. Twenty two fatty acids were identified of which saturated fatty acids represented 25.6% and unsaturated fatty acids 74.4% of the total saponifiable fraction. GC/MS analysis of three chromatographic fractions (I,II and III) of B. Serrata oleo gum resin revealed the presence of pent-2-ene-1,4-dione, 2-methyl- levulinic acid methyl ester, 3,5- dimethyl- 1-hexane, methyl-1-methylpentadecanoate, 1,1- dimethoxy cyclohexane, 1-methoxy-4-(1-propenyl)benzene and 17a-hydroxy-17a-cyano, preg-4-en-3-one. GC/MS analysis of volatile oils of B. Serrata oleo gum resin revealed the presence of sabinene (19.11%), terpinen-4-ol (14.64%) and terpinyl acetate (13.01%) as major constituents. The anti-cancer effect of two extracts (1 and 2) and four fractions (I, II, III and IV) as well as volatile oils of B. Serrata oleo gum resin on HepG2 and HCT 116 cell lines was investigated using SRB assay. Regarding HepG2 cell line, extracts 1 and 2 elicited the most pronounced cytotoxic activity with $IC_{50}$ values equal 1.58 and $5.82{\mu}g/mL$ at 48 h, respectively which were comparable to doxorubicin with an $IC_{50}$ equal $4.68{\mu}g/mL$ at 48 h. With respect to HCT 116 cells, extracts 1 and 2 exhibited the most obvious cytotoxic effect; with $IC_{50}$ values equal 0.12 and $6.59{\mu}g/mL$ at 48 h, respectively which were comparable to 5-fluorouracil with an $IC_{50}$ equal $3.43{\mu}g/mL$ at 48 h. In conclusion, total extracts, fractions and volatile oils of B. Serrata oleo gum resin proved their usefulness as cytotoxic mediators against HepG2 and HCT 116 cell lines with different potentiality (extracts > fractions > volatile oil). In the two studied cell lines the cytotoxic acivity of each of extract 1 and 2 was comparable to doxorubicin and 5-fluorouracil, respectively. Extensive in vivo research is warranted to explore the precise molecular mechanisms of these bioactive natural products in cytotoxicity against HCC and CRC cells.

• 한국산업보건학회지
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• 제10권1호
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• pp.1-17
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• 2000

Carcinogen-DNA adduct analysis has potential for biomonitoring the earliest effects of exposure to many chemical carcinogens. They are the covalent reaction products of electrophiles and nucleophilic sites on DNA and the initial damage to DNA induced by many carcinogens. So many researchers begin to use them as biomarker for monitoring the earliest exposure of carcinogens and develop the effective analytical techniques about them. Randerath, Gupta and coworkers(1981, 1982) has also developed a $^{32}P$-postlabelling method as one among them. A major project for biomonitoring workers with carcinogen-DNA adducts is to develop non-invasive samples instead of tissues of target organs such as baldder and lung. This study use the exfoliated urothelial cells in urine for examine benzidine-DNA adducts. The content of exfoliated urothelial cells is not enough to significantly measure DNA content with spectrophotometer, and require the another way. So firstly washing the collected cells with PBS and 70% ethanol and centrifuge them for removing the crystals in urine, which block the isolation of DNA adducts. And then, measure the total nucleotide after $^{32}P$-postlabelling for calculating RAL. $[{\gamma}-^{32}P]ATP$ using for $^{32}P$-postlabelling, can synthesize with $[^{32}P]H_3PO_4$, and reagent and enzyme mixture (RM, EM), which is very economic in case of requiring a lot of them. Chromatography was composed of two steps. First step was to separate adduct ones from unadducted nucleotide, and secondary step was separate each adduct, which were performed with 4 kinds of solvents and different directions on TLC. With this procedure, we measure the DNA adducts in exfoliated urothelial cells of workers who were employed in benzidine and benzidine-dye company. RAL of adducts were $89.0{\times}10^7$ and $57.0{\times}10^7$ in them. In conclusion, we can significantly measure the DNA adduct in exfoliated urothelial cells by using the above $^{32}P$-postlabelling procedures, and use them to be biomonitoring workers who exposed carcinogens.

• 한국축산식품학회지
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• 제37권5호
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• pp.735-742
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• 2017

This study was conducted to isolate and characterize Paenibacillus sp. MBT213 possessing ${\beta}$-glucosidase activity from raw milk, and examine the enzymatic capacity on the hydrolysis of a major ginsenoside ($Rb_1$). Strain MBT213 was found to have a high hydrolytic ability on ginsenoside $Rb_1$ by Esculin Iron Agar test. 16S rDNA analysis revealed that MBT213 was Paenibacillu sp. Crude enzyme of MBT213 strain exhibited high conversion capacity on ginsenoside $Rb_1$ into ginsenoside Rd proven by TLC and HPLC analyses. The API ZYM kit confirmed that Paenibacillu sp. MBT213 exerted higher ${\beta}$-glucosidase and ${\beta}$-galactosidase activity than other strains. Optimum pH and temperature for crude enzyme were found at 7.0 and $35^{\circ}C$ in hydrolysis of ginsenoside $Rb_1$. After 10 d of optimal reaction conditions for the crude enzyme, ginsenoside $Rb_1$ fully converted to ginsenoside Rd. Ginseng roots (20%) were fermented for 14 d, and analyzed by HPLC showed that amount of ginsenoside $Rb_1$ significantly decreased, while that of ginsenoside Rd was significantly increased. The study confirmed that the ${\beta}$-glucosidase produced by Paenibacillus sp. MBT213 can hydrolyze the major ginsenoside $Rb_1$ and convert to Rd during fermentation of the ginseng. The ${\beta}$-glucosidase activity of this novel Paenibacillus sp. MBT213 strain may be utilized in development of variety of health foods, dairy foods and pharmaceutical products.

• 한국미생물·생명공학회지
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• 제44권2호
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• pp.156-162
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• 2016

본 연구에서는 신규 해양성 한천분해균을 분리하고, 이 균주가 생성하는 한천분해효소의 특성을 조사하였다. 경상남도 남해군 미조면 연안에서 채취한 해수를 Marine agar 2216 배지에 도말하여 한천분해세균을 선별하였다. 선택된 한천분해균주는 16S rDNA 염기서열분석을 통해 Maribacter 속 세균과 99% 유사하여 Maribacter sp. SH-1으로 명명하였다. 세포외로 분비되는 agarase는 Maribacter sp. SH-1 균주 배양액에서 획득하였으며, 이를 이용하여 특성을 조사하였다. Maribacter sp. SH-1 균주의 한천분해효소는 20, 30, 40, 50 및 60℃ 에서 각각 56, 62, 94, 100, 8%의 상대활성을 나타냈으며, pH 5, 6, 7 및 8에서 각각 15, 100, 60, 21%의 상대활성을 나타냈다. 세포외 agarase는 50℃ , pH 6인 20 mM Tris-HCl buffer를 사용하는 조건에서 최대활성(231 units/l)을 보였다. 한천이 sol 상태로 존재하는 50℃에서 최적활성을 보여 이 효소는 응용 가능성이 높다고 할 것이다. 효소 활성은 20, 30 및 40℃에서 30분 동안 열처리하였을 때 약 90% 이상의 상대활성을 보였다. TLC 분석 결과, Maribacter sp. SH-1 균주의 한천분해효소는 한천올리고당인 neoagarohexaose (34.8%), neoagarotetraose (52.2%) 및 neoagarobiose (13.0%)를 생성하는 것으로 보아 β-agarase로 확인되었다. 따라서 Maribacter sp. SH-1 균주와이 균주가 생산하는 β-agarase는 보습효과, 미백효과, 세균성장 억제 혹은 전분노화 방지 등의 기능을 가지는 한천올리고당의 생산에 유용할 것으로 기대된다.

• Journal of the Korean Wood Science and Technology
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• 제34권6호
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• pp.61-71
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• 2006

국내산 주요 참나무류인 신갈나무, 상수리나무, 떡갈나무, 졸참나무, 갈참나무 및 상수리나무 잎의 추출성분의 구조를 규명하고 수종 상호간의 성분의 특성 및 상호 연관성 등을 조사하였다. 신갈나무에서는 gallic acid, (+)-catechin, (-)-epicatechin, (+)-gallocatechin, kaempferol, a stragalin, astragalin-6"-O-gallate, isoquercirin, isoquercitrin-6"-O-gallate 및 myricetin 등 10종의 화합물이 단리되었고 상수리나무에서는 gallic acid, kaempferol과 quercetin이 떡갈나무에서는 gallic acid, (+)-catechin, (-)-epicatechin, (+)-galocatechin, (-)-epigallocatechin, kaempferol, quercetin, guajaverin 및 tamarixin 등 9종의 화합물이 단리되었으며 갈참나무에서는 gallic acid, caffeic acid, astragalin, quercetin과 isoquercitrin이 졸참나무에서는 gallic acid, (+)-catechin, (-)-epicatechin, kaempferol, quercitrin, isoquercitrin 및 myricetin 등 7종의 화합물이 단리되었고 굴참나무에서는 gallic acid, (+)-catechin, astragalin, astagalin-6"-O-gallate와 isoquercitrin 등 5종의 화합물이 단리되었다. Gallic aicd는 모든 수종에서 단리되어 참나무류 잎 추출성분의 지표 화합물로 이용될 수 있다.

• Nutrition Research and Practice
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• 제8권5호
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• pp.509-515
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• 2014

BACKGROUND/OBJECTIVES: The root of Vitis amurensis Ruprecht, a sort of wild-growing grape, has been used in oriental medicine for treatment of skin ailments; however, its dermatological activity is not sufficiently understood. The aim of this study was to investigate tyrosinase inhibitory and anti-melanogenic activities of V. amurensis Ruprecht root methanol extract (VARM) in B16F10 mouse melanoma cells and to attempt to isolate and identify the active compound issued from VARM. MATERIALS/METHODS: Anti-melanogenic activity of VARM was analyzed in ${\alpha}$-melanocyte stimulating hormone (MSH)-stimulated B16F10 cells through evaluation of antioxidative activity as well as inhibited tyrosinase activity and melanin contents compared with those of kojic acid and arbutin. After anti-melanogenic analysis of VARM, serial fractionation, nuclear magnetic resonance (NMR), and thin layer chromatorgraphy (TLC) were applied for identification of active compounds contained in VARM. RESULTS: VARM significantly inhibited oxidative stress and tyrosinase activity and attenuated ${\alpha}$-MSH-induced melanin production in B16F10 cells. For isolation of active compounds, VARM was fractionated using a series of organic solvents, including dichloromethane ($CH_2Cl_2$), ethyl acetate (EtOAc), and n-butanol (n-BuOH). Among fractions showing anti-melanogenic activity, the CH2Cl2 fraction induced the most potent attenuation of melanogenesis without cytotoxicity and the major compound in the $CH_2Cl_2$ fraction was identified as betulinic acid. Betulinic acid isolated from the $CH_2Cl_2$ fraction of VARM significantly attenuated ${\alpha}$-MSH-induced melanogenesis in a dose dependent manner, which was stronger than that of arbutin used as a positive control. CONCLUSIONS: These results indicate that VARM inhibits oxidative stress, tyrosinase activity, and ${\alpha}$-MSH-induced melanogenesis in B16F10 cells, due primarily to the active compound, betulinic acid, in the $CH_2Cl_2$ fraction.

• 한국산업보건학회지
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• 제6권1호
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• pp.28-37
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• 1996

Benzidine, an aromatic amine used primarily in the manufacture of azo dyes, is recognized as a urinary bladder carcinogen in humans. In rats, mice, and hamsters, chronic exposure to benzidine resulted in tumors of the liver. The present study was undertaken to suggest analyzing the metabolites of benzidine with the optimal condition, identify the metabolites of benzidine, and observe time variance of the metabolites in the isolated perfusated rat liver. N-acetylbenzidine was synthesized by acetylation of benzidine with acetic anhydride and separated by thin layer chromatography(TLC) and high performance liquid chromatography(HPLC). To analysis benzidine and the metabolites of benzidine, HPLC operating condition has been optimized by means of preliminary experiment. The mobile phase consisted of acetonitrile(37%) in phosphate buffer, flow rate maintained at 1.0 ml/min. Optimal detective conditions were electrochemicaldetector(ECD) at 0.75 V for benzidine and N-acetylbenzidine and ultravioletdetector(UVD) at 287 nm for N,N'-diacetylbenzidine. The separation system was composed of a guard column and a separation column(Polymer C18, $4.6{\times}250cm$) at a temparature of $40^{\circ}C$. The perfusion system was equilibrated for 30 minutes before addition of benzidine to the perfusate. Samples of the perfusate were collected at time intervals(0, 10, 20, 30, 60, 90, 120 min) during the 2 hour perfusion. Before analyzing samples by HPLC/ECD/UVD, samples had been treated with sep-pak. Samples of perfusate analyzed by HPLC/ECD/UVD and the metabolites of benzidine in the isolated perfused rat liver were N-acetylbenzidine and N,N'-diacetylbenzidine. Benzidine metabolized over 60% during the initial 30 minutes of perfusion, extensively by 1 hour, and was undetectable in the perfusate. N-acetylbenzidine increased by 30 minutes of perfusion, declined. N,N'-diacetylbenzidine increased the 0-90 minutes period, remained constant during the 90-120 minutes period.