• 한국발생생물학회지:발생과생식
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• 제25권2호
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• pp.75-82
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• 2021

We studied steroid metabolites produced from red-spotted grouper ovarian follicles during maturation. Oocytes with 350-500 ㎛ diameter were in vitro incubated in the presence of [³H] 17α-hydroxyprogesterone as a precursor. Steroid metabolites were extracted from incubated media and oocytes. The extracts were separated and identified using thin layer chromatography, high performance liquid chromatography and gas chromatography-mass spectrometry. The identified metabolites were androstenedione (A₄), testosterone (T) and estrone (E₁). The metabolites of A₄ was dominant in all size of oocytes and it was the highest in 480 ㎛ diameter oocytes. The metabolites of two progestins, 17α,20β-dihydroxy-4-pregnen-3-one and 17α,20α-dihydroxy-4-pregnen-3-one were detected in the oocytes less than 480 ㎛ diameter although they were not identified definitely. In the oocytes of 480 ㎛ diameter, metabolite of progestin was the highest, and germinal vesicle (GV) was still in the middle of cytoplasm. In the oocytes of 500 ㎛ diameter, GV was began to migrate and the major metabolites were A₄ and E₁. The metabolite of E₁ was detected in all size of oocytes and it was higher than that of E₂. These results suggest that oocytes of 480 ㎛ diameter are the transitional stage involving steroidogenic shift to final oocyte maturation and potential function of E₁ during maturation process.

• 대한화장품학회지
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• 제35권1호
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• pp.11-17
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• 2009

스테로이드 함유 표시가 없는 화장품에서 항염증 효과가 있는 글루코코티코스테로이드인 베타메타손프로피오네이트 성분이 검출되었다. 이 화장품은 외용스프레이 및 샴푸로서 주성분으로 아연피리치온을 함유하는 것으로 표기되어 있었다. 화장품에서 스테로이드 구조와 활성을 갖는 물질의 존재를 확인하기 위하여 실리카겔 박층판을 이용한 박층크로마토그래프를 사용하였으며 이 성분을 분리하기 위해 high-performance liquid chromatography (HPLC)를 이용하여 확인 및 정량을 수행하였다. 분취용 HPLC를 이용하여 스테로이드를 함유한 것으로 판단되는 분획을 모은 다음 nuclear magnetic resonance (NMR) 및 mass spectrometry (MS)를 이용하여 스테로이드 성분을 확인하였다. 스테로이드 표준물질로 베타메타손 17-프로피오네이트 및 베타메타손 21-프로피오네이트를 합성하여 사용하였고 이 표준물질과 HPLC 크로마토그램을 비교하여 스테로이드 성분의 함량을 분석하였다. 이 방법으로 아연피리치온 제제와 같은 일부 시판 화장품에서 스테로이드 성분을 확인하였고 reversed-phase high-performance liquid chromatography (RP HPLC) 상의 유지시간 비교를 통하여 스테로이드 성분을 정량한 결과 시험한 총 8종의 화장품 시료 중 2개 제품에서 0.005 ${\sim}$ 0.02%의 베타메타손프로피오네이트가 검출되었다.

• FOCUS: LIFE SCIENCE
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• 제1호
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• pp.3.1-3.7
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• 2024

The article discusses the critical role of chromatography in the analysis and purification of proteins in biopharmaceuticals, emphasizing the importance of comprehensive characterization for ensuring their safety and efficacy. It highlights the use of Avantor® ACE® HPLC columns for the separation and purification of proteins, focusing on the analysis of intact proteins using reversed-phase liquid chromatography (RPLC) with fully porous particles. This article also details the application of different mobile phase additives, such as TFA and formic acid, and emphasizes the advantages of using type B ultra-pure silica-based columns for efficiency and peak shape in biomolecule analysis. Additionally, it addresses the challenges of analyzing intact proteins due to slow molecular diffusion and introduces the concept of solid-core (or superficially porous) particles, emphasizing their benefits over traditional porous particles for the analysis of therapeutic proteins. Furthermore, it discusses the development of Avantor® ACE® UltraCore BIO columns, specifically designed for the high-efficiency separation of large biomolecules, such as proteins, and demonstrates their effectiveness in achieving high-resolution separations, even for higher molecular weight proteins like monoclonal antibodies (mAbs). In addition, it underscores the complexity of analyzing and characterizing intact protein biopharmaceuticals, requiring a range of analytical techniques and the use of wide-pore stationary phases, operated at elevated temperatures and with relatively shallow gradients. It highlights the comprehensive range of options offered by Avantor® ACE® wide pore columns, including both fully porous and solid-core particles, bonded with a variety of complementary stationary phase chemistries to optimize selectivity during method development. The use of ultrapure and highly inert base silica is emphasized for enabling the use of lower concentrations of mobile phase modifiers without compromising analyte peak shape, particularly beneficial for LC-MS applications. Then the article concludes by emphasizing the significance of reversed-phase liquid chromatography and its compatibility with mass spectrometry as a valuable tool for the separation and analysis of intact proteins and their closely related variants in biopharmaceuticals.

• 한국식품과학회지
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• 제42권4호
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• pp.377-382
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• 2010

ADC는 밀가루개량제로 사용기준이 설정되어 있는 식품첨가물이나 이의 사용량과 잔류량을 확인할 수 있는 시험분석법이 확립되어 있지 않은 상태임으로 사전 사후 안전관리를 위해서 ADC의 분해산물인 biurea를 밀가루 및 빵류에서 분석 가능한 방법을 확립하고자 하였다. 식품 중 ADC의 분해산물인 biurea 분석을 위하여 컬럼, 이동상 조건, 전처리 조건, 기기조건 등을 검토하여 LC/MS/MS를 이용한 분석법을 확립하였으며, 개발된 분석법의 회수율, 분석법의 유효성 검증 등을 검토하였다. 회수율은 94.3-112.5%로 양호하였으며, 검출한계(LOD)는 0.003 mg/L의 농도이었고, 정량한계(LOQ)는 0.01 mg/L의 농도이었다. 확립된 분석법에 의해 밀가루 등 51건, 빵 등의 가공식품 59건에 대하여 biurea 함량을 분석한 결과 밀가루 1건에서 2.76 mg/kg의 농도로 검출되었고(검출율: 2%), 빵 등 가공식품에서는 16건이 검출(검출율: 27%)되었으며 검출농도범위는 0.19-18.01 mg/kg(평균: 3.79 mg/kg)이었다. ADC의 사용기준인 밀가루 1 kg당 45mg에 비하여 매우 낮은 값을 나타내었다. 본 연구를 통하여 확립된 식품 중 biurea 분석법으로 밀가루, 빵류 등의 가공식품에 대한 ADC의 사후관리에 기여할 것으로 본다.

• 분석과학
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• 제30권1호
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• pp.10-19
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• 2017

Anabolic steroids have similar structures to testosterone, both of which promote the growth of muscle mass and increase strength. However, the side effects of anabolic steroid use may lead to heart attacks or strokes. Additionally, the excessive use of steroids inhibits the production of the sex hormones in the body via a negative feedback loop, which results in testicular atrophy in males and amenorrhea in females. Currently, the method of choice used to test for the presence of anabolic steroids is GC-MS. However, GC-MS methods require chemical derivatization of the steroid sample to ensure compatibility with the analytical method; therefore, analysis of many different samples is difficult and time consuming. Unlike GC-MS, the liquid chromatography-quadrupole-time of flight mass spectrometry (LC-Q-TOF-MS) method is suitable for many samples. Twenty-two different anabolic steroids were analyzed by LC-Q-TOF-MS with various collision energies (CE). Accurate mass spectral data were obtained using a Q-TOF-MS equipped with an electro-spray ionization source and operated in the positive MS/MS mode for several classes of steroids that are often the targets of testing. Based on the collected data, fragmentation pathways were carefully elucidated. The high selectivity and sensitivity of the LC-Q-TOF-MS instrument combined with these fragmentation pathways offers a new approach for the rapid and accurate screening of anabolic steroids. The obtained data from the 22 different anabolic steroids will be shared with the scientific community in order to establish a library to aid in the screening of illegal anabolic steroids.

• 생약학회지
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• 제38권2호통권149호
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• pp.123-127
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• 2007

Aristolochic acid (AA) included in the plants Aristolochiaceae have been well known to be nephrotoxic and carcinogenic inducer and to cause renal disease such as Chinese Herb Nephropathy (CHN). In this study, we used a high performance liquid chromatopaphy-mass spectrometry (HPLC-MS) under the positive ion detection mode for the quantitative change of aristolochic acid-I and-II (AA-I and AA-II) in Aristolochiaceae (Aristolochia contorta Bunge, Aristolochia debilis Sieb. et Zucc., Aristolochia fangchi Wu), some related plants (Cocculus trilobus De candolle, Inula helenium Linne, Saussurea lappa Clarke), and its prescriptions (防己茯笭湯, 定喘散) with or without processing. Here, the processing methods and prescriptions in oriental medicine were generally used to alleviate toxicity or alter property of herbal medicines. However, the concentrations of AA-I and AA-II were highly determined in processed material extracts rather than unprocessed those, not measured in some related plants. Also, the concentrations of AA-I and AA-II even at the prescriptions mixed the plants of Aristolochiaceae were detected to range from 0.73 to 2.53 ppm. Thus, the present results suggest that the content of AA-I and AA-II contained to plants of Aristolochiaceae was not reduced by the processing methods or prescriptions which can induce the physico-chemical change and pharmacological transformation in traditional herbal medicines.

• 한국환경과학회지
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• 제23권7호
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• pp.1253-1268
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• 2014

This study was investigated twenty two hazardous chemicals compounds for effluents of nine sewage treatment plants (STPs) and one waste water treatment plant (WWTP) in the Nakdong Ri-ver Basin. They are eleven phthalates(DMP, DEP, DIBP, DBP, BEEP, DNPP, DHP, DCP, DEHP, DNOP, Dinonyl phthalate, seven aliphatic hydrocarbons(n-Tridecane, n-Tetradecane, n-Pentadecan-e, n-Hexadecane, n-Heptadecane, n-Octadecane, n-Nonadecane, Isoquinoline, 2-Chloropyridine, 2-N-itrophenol, and Benzophenone. The twenty two compounds were analyzed by gas chromatograp-hy mass spectrometry (GC/MS) with liquid-liquid extraction (LLE). Twenteen of twenty two subs-tances were detected. They were DMP, DEP, DIBP, DBP, DEHP, n-Tetradecane, n-Pentadecane, n-Heptadecane, n-Octadecane, n-Nonadecane, Isoquinoline and Benzophenone. Among these, DEHP, DEP and Benzophenone were most frequently observed. They were obtained as $ND{\sim}36.881{\mu}g/L$, $ND{\sim}0.950{\mu}g/L$, $ND{\sim}2.019{\mu}g/L$, respectively. When the substances were calculated the average concentration at 10 points, the maximum average detection concentration was investigated at the Dalseocheon STP.

• 한국생물정보학회:학술대회논문집
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• 한국생물정보시스템생물학회 2005년도 BIOINFO 2005
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• pp.91-95
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• 2005

In the multidimensional protein identification technology of high-throughput proteomics, we use one-dimensional gel electrophoresis and after the separation by two-dimensional liquid chromatography, the sample is analyzed by tandem mass spectrometry. In this study, we have analyzed the Pseudomonas Putida KT2440 protein. From the protein identification, the protein database was combined with its reversed sequence database. From the peptide selection whose error rate is less than 1%, the SEQUEST database search for the tandem mass spectral data identified 2,045 proteins. For each protein, we compared the molecular weight calibrated from 1D-gel band position with the theoretical molecular weight computed from the amino acid sequence, by defining a variable MW$_{corr}$ Since the bacterial proteome is simpler than human proteome considering the complexity and modifications, the proteome analysis result for the Pseudomonas Putida KT2440 could suggest a guideline to build the protocol to analyze human proteome data.

• Journal of Microbiology and Biotechnology
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• 제26권10호
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• pp.1781-1789
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• 2016

Glargine insulin is a long-acting insulin analog that helps blood glucose maintenance in patients with diabetes. We constructed the pPT-GI vector to express prepeptide glargine insulin when transformed into Escherichia coli JM109. The transformed E. coli cells were cultured by fed-batch fermentation. The final dry cell mass was 18 g/l. The prepeptide glargine insulin was 38.52% of the total protein. It was expressed as an inclusion body and then refolded to recover the biological activity. To convert the prepeptide into glargine insulin, citraconylation and trypsin cleavage were performed. Using citraconylation, the yield of enzymatic conversion for glargine insulin increased by 3.2-fold compared with that without citraconylation. After the enzyme reaction, active glargine insulin was purified by two types of chromatography (ion-exchange chromatography and reverse-phase chromatography). We obtained recombinant human glargine insulin at 98.11% purity and verified that it is equal to the standard of human glargine insulin, based on High-performance liquid chromatography analysis and Matrix-assisted laser desorption/ionization Time-of-Flight Mass Spectrometry. We thus established a production process for high-purity recombinant human glargine insulin and a method to block Arg (B31)-insulin formation. This established process for recombinant human glargine insulin may be a model process for the production of other human insulin analogs.

• 한국잡초학회지
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• 제14권2호
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• pp.156-162
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• 1994

수수 줄기에 함유된 타감물질(他感物質)중 가장 강한 활성을 나타내는 물질을 rapid chromatography, flash flow column chromatography로 분리하고 thin layer chromatography와 HPLC로 정제하여 GC-MS로 분석하였다. Butanol, acetic acid 및 water의 용매 조합을 달리하여 타감물질을 분리한 결과 butanol (8) : actic acid (1) : water (1) 분획에서 억제효과가 가장 크게 나타났으며, flash flow column chronatography와 TLC로 분리 한 결과 가장 활성을 나타내는 물질은 Rf 0.71에서 나타났으며 HPLC로 순수분리한 결과 Rt 20.40min에서 elution되었다. 이 물질은 주황색을 띄며 methanol에 용해성이 있었다. 정제된 물질을 GC-MS로 분석한 결과 예상되는 물질은 1-methyl-1-(2-propinonyl)-hydrazine, 1-aziridineethanol, 5-chloro-2-pentanone, 2-(methylseleno)-ethanamine 중한 물질일 것으로 추측되었다.