• Animal cells and systems
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• 제7권1호
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• pp.75-79
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• 2003

Highly purified high performance thin layer chromatography (HPTLC) fractions containing a putative calcium influx factor (CIF) were prepared from the Jurkat cells and Xenopus oocytes in which $Ca^{2+}$ stores were depleted by thapsigargin treatment and from the yeast in which intracellular $Ca^{2+}$ stores were also depleted by genetic means. Microinjection of the fractions has been shown to elicit $Ca^{2+}$ dependent currents in Xenopus oocytes. The nature of the membrane currents evoked by the putative CIF appeared to be carried by chloride ions since the current was blocked by the selective chloride channel blocker 1 mM niflumic acid and its reversal potential was about -24 mV. Injection of the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N, N, N',N'-tetraacetic acid (BAPTA) eradicated the current activities, suggesting the current responses are entirely $Ca^2$-dependent. Moreover, the currents were sensitive to the removal of extracellular calcium, indicating the dependence on calcium entry through the plasma membrane calcium entry channels. CIF activities were insensitive to protease, heat, and acid treatments and to Dische-reaction whereas the activities were sensitive to nucleotide pyrophosphatase and hydrazynolysis. The fraction might have a sugar because it was sensitive to Molisch test and Seliwaniff's resorcinol reaction. From the above results, CIF as a small and stable molecule seems to have pyrimidine, pyrophosphate, and a sugar moiety.oiety.

• 원예과학기술지
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• 제33권3호
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• pp.403-408
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• 2015

색도는 고추의 품질을 결정하는 중요한 요인으로, 색도 측정을 위해 high-performance liquid chromatography(HPLC), thin layer chromatography(TLC), ASTA-20 방법 등이 활용되고 있다. 특히 ASTA-20 방법은 간단하고 정확하게 다수의 시료에 대한 색도 분석을 수행할 수 있다는 장점 덕분에 주로 사용된다. 하지만 전처리 과정에 시간이 많이 소요되고 아세톤과 같은 폐시약이 발생한다. 따라서 본 연구에서는 ASTA-20 방법을 대체하기 위하여 Vis/NIR 초분광 분석법을 활용한 빠른 색도 분석법을 개발하고자 하였다. ASTA-20 방법과 Vis/NIR 초분광 분석법의 상관관계를 분석하기 위하여 총 488점 고춧가루의 색도를 두 가지 방법으로 측정하였다. 이후 무작위로 선발한 366개의 시료를 이용하여 Vis/NIR 초분광 분석법으로 측정한 값으로부터 ASTA 값을 예측하는 부분최소자승법(PLS) 모델을 확립하였다. 모델 개발에 활용하지 않은 122개 시료의 ASTA 값을 PLS 모델을 이용하여 예측하고, ASTA-20 방법으로 측정한 값과 비교해본 결과, 매우 유의성 있는 상관관계($R^2=0.88$)를 나타내 Vis/NIR 초분광 분석법의 신뢰도를 확인할 수 있었다. 따라서 본 연구에서 개발한 간편하고 빠른 ASTA 값 측정 방법은 전처리 단계를 요구하지 않고, 30분 이내에 100개 시료에 대한 분석을 수행할 수 있어 다수의 고춧가루 색도를 빠르게 측정하는데 유용하게 사용될 것으로 기대한다.

• 한국식품영양학회지
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• 제27권3호
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• pp.532-537
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• 2014

황백(Cortex Phellodendri: CP)은 황벽나무(Phellodendron amurense)의 건조된 수피로부터 얻어진다. 이 수피는 한국의 전통 한약제로서 설사, 황달, 무릎과 발의 통증, 요도관 및 피부 감염증에 폭넓게 사용되어 왔다. 이들 기능성 성분의 분리 및 정제는 박층 크로마토크래피, 컬럼 액체 크로마토크래피 및 HPLC와 같은 여러 분석법들이 동양의 약초연구에 이용되어 왔다. 본 연구는 CP로부터 berberine을 분리하기 위해 향류분배 크로마토크래피법(CPC)으로 효과적으로 수행하였다. 두 용매의 CPC 최적조성은 n-butanol: acetic acid: water(4:1:5 v/v/v)이었다. 이동상의 유속은 1,000 rpm 회전력에서 상승법으로 분당 3 mL 속도로 전개시켰다. CPC에서 분리된 분획분은 prep-HPLC로 정제하였다. $^1H$-NMR 스펙트럼은 4.10과 4.20 ppm에서 $3H(-OCH_3)$, 6.10 ppm에서 2H의 ($-OCH_2O-$) proton signal의 공명이 관찰되었다. 2개의 방향족 proton은 이중결합 패턴을 보였다. H-11과 H-12 doublet은 각각 7.98과 8.11에서 나타났다. $^{13}C$-NMR 스펙트럼에서는 C2와 C3의 methylenedioxy group($-OCH_2O-$), C9과 C10에 methoxy group($-OCH_3$)이 4개의 치환된 형태로 보였다. 분리 정제된 berberine의 화학구조는 $^1H$, $^{13}C$-NMR, ESI-MS 데이터 분석으로 확인하였다.

• 셀메드
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• 제11권4호
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• pp.21.1-21.9
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• 2021

Background: Unani System of Medicine (USM) has its origin to Greece. To ensure and develop the quality, authenticity of Unani drugs, standardization on modern analytical parameter is essential requirement for drugs. Objectives: The aimed of the present study was to develop a standard profile of "Qurṣ-e-Mafasil" by systematic study through authenticated ingredients, pharmacognostic identification followed by physicochemical, TLC, HPTLC fingerprinting analysis as per standard protocol. Material and Methods: In this study three batches of "Qurṣ-e-Mafasil" QM were prepared by standard method as per UPI had been followed by organoleptic properties of formulation such as appearance, color, odor, taste. Powder Microscopy and physicochemical studies were carried out such as Uniformity of weight, Friability, Disintegration time, hardness, LOD, ash vales and extractive values in like aqueous, alcohol & hexane. Further qualitative tests such as Thin-Layer Chromatography (TLC), and High-Performance Thin Layer Chromatography (HPTLC) studies were also carried out to develop fingerprint pattern of the alcoholic solvent extract of QM. Phytochemical screening was carried out in different solvent extracts such as alcoholic, aqueous and chloroform extracts to detect the presence phytoconstituents in the formulation QM. Heavy metals, Microbial Load Contamination and pesticidal residues were also determined. Results: Qurṣ-e-Mafasil showed tablet-like appearance, light brown colour, mild pungent odour and acrid taste. Uniformity of weight (mg), friability (rpm), and hardness (kg/cm) and disintegration time was ranged between (500 to 503), (0.0340 to 0.038), (8.40 to 8.67) and (4-5 minutes) respectively for the three batches. Loss in weight on drying at 105℃ was ranged between (8.3425 to 8.7346). Extracted values were calculated in distilled water ranged between (30.9091 to 31.4358), hexane (1.1419 to 1.4281), and alcohol (3.3352 to 3.3962). The ash values recorded were ranged between (3.7336 to 3.8378), and acid insoluble ash (0.5859 to 0.6112).

• Food Science and Biotechnology
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• 제17권2호
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• pp.302-307
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• 2008

Novel amylase inhibitors were synthesized via transglycosylation by Thermotoga maritima glucosidase (TMG). TMG hydrolyzes acarbose, acarviosine-glucose, and maltooligosaccharide by releasing $^{14}C$-labeled glucose from the reducing end of each molecule. When TMG was incubated with acarviosine-glucose (the donor) and glucose (the acceptor), two major transfer products, compounds 1 and 2, were formed via transglycosylation. The structures of the transfer products were determined using thin-layer chromatography (TLC), high-performance ion chromatography (HPIC), and $^{13}C$ nuclear magnetic resonance (NMR) spectroscopy. The results indicate that acarviosine was transferred to glucose at either C-6, to give a $\alpha-(1{\rightarrow}6$) glycosidic linkage, or at C-3, to produce an $\alpha-(1{\rightarrow}3$) glycosidic linkage. The transfer products showed a mixed-type inhibition against porcine pancreatic $\alpha$-amylase; therefore, they may be useful not only as inhibitors but also as acarbose transition-state analogs to study the mechanism of amylase inhibition.

• 한국발생생물학회지:발생과생식
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• 제17권4호
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• pp.421-426
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• 2013

In this study, oocyte steroidogenesis are investigated in relation to oocyte development in the starry flounder, Platichthys stellatus, a marine multiple spawner. Vitellogenic (0.52 and 0.55 mm oocyte diameter) and mature oocytes (0.63, 0.66 and 0.71 mm oocyte diameter) were incubated in vitro in the presence of $[^3H]17{\alpha}$-hydroxyprogesterone ($[^3H]17{\alpha}$-OHP) as a precursor. Steroid metabolites were extracted from the incubated media and oocytes, the extracts were separated and identified by thin-layer chromatography (TLC), high performance liquid chromatography (HPLC) and gas chromatographymass spectrometry (GC-MS). The major metabolites produced from $[^3H]17{\alpha}$-OHP were androgens [androstenedione ($A_4$) and testosterone (T)] and estrogens [$17{\beta}$-estradiol ($E_2$) and estrone ($E_1$)] and progestins [$17{\alpha},20{\alpha}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\alpha}P$) and $17{\alpha},20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$)] in vitellogenic and mature oocytes. The results from this study suggest the potential roles of $E_1$ in the oocytes with diameter 0.52-0.71 mm, $17{\alpha}20{\alpha}P$ and $17{\alpha}20{\beta}P$ at the oocytes of 0.63, 0.66 and 0.71 mm.

• 한국식품과학회지
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• 제36권4호
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• pp.558-562
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• 2004

TLC를 이용하여 간편하고 신속하게 정량적인 이소플라본의 분석을 수행할 수 있는 조건을 개발하였다. 전개 용매로는 chloroform : methanol : water : acetic acid를 각각 60 : 30 : 10 : 0.5의 비율로 하였을 때 최적의 분리를 보였으며, 정량적인 분석은 UV에서의 흡광도를 densitometer를 사용하여 spot의 intensity를 계산함으로 가능하였다. HPLC분석법과의 비교 결과 $0.15-1.80{\mu}g/{\mu}L$의 범위에서 이소플라본을 정량적으로 분석할 수가 있었다. 또한 TLC를 이용하여 일반 콩과 식물에 포함되어 있는 이소플라본의 함량 측정 결과 HPLC의 결과와 상당히 근소한 값으로 일치하여 TLC방법으로 간편하고 빠르게 정량적인 측정이 가능함을 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제26권10호
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• pp.1687-1695
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• 2016

Ochratoxin A (OTA), a mycotoxin, contaminates agricultural products and poses a serious threat to public health worldwide. Microbiological methods are known to be a promising approach for OTA biodegradation because physical and chemical methods have practical limitations. In the present study, a total of 130 fungal isolates obtained from 65 traditional Korean meju (a fermented starter for fermentation of soybeans) samples were examined for OTA-biodegradation activity using thin-layer chromatography. Two fungal isolates were selected for OTA-biodegradation activity and were identified as Aspergillus tubingensis M036 and M074 through sequence analysis of the beta-tubulin gene. After culturing both A. tubingensis isolates in Soytone-Czapek medium containing OTA (40 ng/ml), OTA-biodegradation activity was analyzed using high-performance liquid chromatography (HPLC). Both A. tubingensis strains degraded OTA by more than 95.0% after 14 days, and the HPLC analysis showed that the OTA biodegradation by the A. tubingensis strains led to the production of ochratoxin α, which is much less toxic than OTA. Moreover, crude enzymes from the cultures of A. tubingensis M036 and M074 led to OTA biodegradation of 97.5% and 91.3% at pH 5, and 80.3% and 75.3% at pH 7, respectively, in a buffer solution containing OTA (40 ng/ml) after 24 h. In addition, the OTA-biodegrading fungi did not exhibit OTA production activity. Our data suggest that A. tubingensis isolates and their enzymes have the potential for practical application to reduce levels of OTA in food and feed.

• 한국수산과학회지
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• 제48권4호
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• pp.483-488
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• 2015

We studied oocyte steroidogenesis in the blackfin flounder Glyptocephalus stelleri as a region-specific species, in the East Sea of Korea during the spawning season. Maturing oocytes (0.76, 0.82, 0.88, and 0.91 mm in oocyte diameter) were incubated in vitro in the presence of [$^3H$] $17{\alpha}$-hydroxyprogesterone ($[^3H]17{\alpha}$-OHP) as a precursor. Steroid metabolites were extracted from the incubated medium and oocytes, and the extracts were separated and identified by thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC) and gas chromatographymass spectrometry (GC/MS). The major metabolites produced from $[^3H]17{\alpha}$-OHP were androgens [androstenedione (A4) and testosterone (T)] and estrogens [$17{\beta}$-estradiol (E2) and estrone (E1)] and progestins [$17{\alpha},20{\alpha}$-dihydroxy-4-pregen-3-one ($17{\alpha}20{\alpha}P$) and $17{\alpha}20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$)] in maturing oocytes. The metabolic rate of $17{\alpha}20{\beta}$ was elevated (29.04%) in oocytes measuring 0.88 mm (nucleus migration stage following the induction of germinal vesicle breakdown), but was very low in oocytes measuring 0.76, 0.82, and 0.91 mm (0.42, 0.67, and 2.62%, respectively). From these results, we suggest that $17{\alpha}20{\beta}P$ acts as a maturation-inducing steroid in the blackfin flounder.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 국제심포지움
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• pp.20-45
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• 2002

In this study, we demonstrated the in vitro and in vivo formation of carcinogen-lipid adduct and its correlation with DNA or protein adducts. The lipids from serum or hepatocyte membranes of Spragu-Dawley rats. human serum, and standard major lipids were in vitro reacted with benzo[a]pyrene(BP) and BP metabolites. 7,8-Dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]-pyrene(BPDE-I), an ultimate carcinogenic form of BP, was covalently bound to triglyceride(TG). BPDE-I-TG adducts isolated by thin-layer chromatography (TLC) were further detected by high performance liquid chromatography(HPLC). TGs, including triolein, tripalmitin and tristearin, showed positive reactions with BPDE-I. However, cholesterol, phospholipids(Phosphatidylcholine, phosphatidyl-ethanolamine, phosphatidyl-inositol and sphingomyelin) and nonesterified fatty acids(palmitic acid, oleic acid, linoleic acid and stearic acid) did not react with BPDE-I. In addition, other BP metabolites (BP-phenols and -diols) did not react with TG, which TG appeared to be the most reactive lipid yet studied with respect to its ability to form an adduct with BPDE-I. There was a clear-cut dose-respect to its ability to form an adduct with BPDE-I-lipid adduct in vitro between TG and [1,3-3H]BPDE-I. In an animal study, BPDE-I-TG was also formed in the serum of rats orally treated with BP(25 mg/rat). Also, obvious correlations between [3H]BP related-biomolecule adducts (DNA, protein) or lipid damage and the BPDE-I-TG adduct were obtained in various tissues of mice i.p. treated with [3H]BP. These data suggest that TG can form an adduct with BPDE-I, as do other macromolecules (DNA, RNA, and protein). Therefore, a carcinogen-lipid adduct would be a useful biomarker for chemical carcinogenesis research and cancer risk assessment.