• Bulletin of the Korean Chemical Society
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• 제34권12호
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• pp.3817-3824
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• 2013

Various separation modes in HPLC, such as anion exchange (AE), reversed-phase (RP), and affinity (AF) chromatography were examined for the separation of selenium species in human blood serum and urine. While RP- and AE-HPLC were mainly used for the separation of small molecular selenium species, double column AF-HPLC achieved the separation of selenoproteins in blood serum efficiently. Further, the effluent of AF-HPLC was enzymatically hydrolyzed and then analyzed with RP HPLC for selenoamino acid study. The versatility of the hybrid technique makes the in-depth study of selenium species possible. For quantification, post column isotope dilution (ID) with $^{78}Se$ spike was performed. ORC ICP/MS (octapole reaction cell inductively coupled plasma/mass spectrometry) was used with 4 mL $min^{-1}$ Hydrogen as reaction gas. In urine sample, inorganic selenium and SeCys were identified. In blood serum, selenoproteins GPx, SelP and SeAlb were detected and quantified. The concentration for GPx, SelP and SeAlb was $22.8{\pm}3.4\;ng\;g^{-1}$, $45.2{\pm}1.7\;ng\;g^{-1}$, and $16.1{\pm}2.2\;ng\;g^{-1}$, respectively when $^{80}Se/^{78}Se$ was used. The sum of these selenoproteins ($84.1{\pm}4.4\;ng\;g^{-1}$) agrees well with the total selenium concentration measured with the ID method of $87.0{\pm}3.0\;ng\;g^{-1}$. Enzymatic hydrolysis of each selenium proteins revealed that SeCys is the major amino acid for all three proteins and SeMet is contained in SeAlb only.

• Korean Chemical Engineering Research
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• 제56권3호
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• pp.376-380
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• 2018

강력한 항산화물질인 astaxanthin의 함량이 다른 천연 공급원에 비해 높아 astaxanthin 생산균주로 주목받고 있는 Haematococcus pluvialis는 상당한 두께의 견고한 세포벽을 가지고 있어, 세포 파쇄를 위해 많은 에너지가 소모되고 비용이 비싼 방법들이 이용되고 있다. 이에 H. pluvialis로부터 막자와 막자사발을 이용하여 astaxanthin을 손쉽게 효율적으로 추출하는 방법을 제시하였다. 막자와 막자사발을 이용하여 분쇄한 후 추출용매로 acetonitrile, acetone, methanol, dichloromethane : methanol (1:3, v/v), ethylacetate : ethanol (1:1, v/v)로 사용하여 비교하였을 때, acetone을 이용하였을 때 astaxanthin을 1.13~1.29 배 더 높은 효율로 추출할 수 있었다. 또한 acetone으로 H. pluvialis로부터 추출할 경우, 1차 추출로 H. pluvialis에 축적되어 있는 전체 astaxanthin의 96.7%를 회수할 수 있을 정도로 acetone은 astaxanthin 추출효율이 높았다. H. pluvialis가 세포내에 축적하는 astaxanthin은 축적 특성상 ester-형태의 astaxanthin로 다량 축적하므로, 추출물 내의 다양한 형태의 astaxanthin을 분리하기 위하여 농도 구배 시스템을 적용한 HPLC 분석을 수행하였다. H. pluvialis에 축적되어 있는 전체 astaxanthin 중 free astaxanthin이 45.9%이고, 나머지 54.1%는 ester-형태의 astaxanthin이었다.

• 한국식품영양학회지
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• 제20권4호
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• pp.440-449
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• 2007

The purpose of this study was to examine the effects of taurine supplementation on serum lipidperoxide(TBARS), a risk factor for cardiovascular disease. The subjects were 22 healthy middle-aged women(33 to 54 years). Serum lipids, thiobarbituric acid reactive substances(TBARS), and plasma taurine levels were measured before and after supplying 3 g of taurine per day for 4 weeks. Plasma taurine was analyzed by Dabsyl-Cl(4-dimethylamino azobenzen-4-sulfonyl-chloride) derivatization and reversed-phase HPLC. Serum TBARS was measured by the Yagi method. Daily dietary taurine intake was calculated by food frequency questionnaire method. The weight and height means of the 22 subjects were $57.9{\pm}5.2$ kg and $159.2{\pm}5.2$ cm, respectively. Their percent body fat and waist/hip ratio(WHR) were 26.8% and 0.84, respectively, which were slightly higher than the average for middle-aged Korean women. Serum TC, TG and LDL-C levels tended to decrease after taurine supplementation, but HDL-C was not changed. A positive correlation between plasma taurine and HDL-C was shown after taurine supplementation. The serum TBARS concentration was significantly decreased from $5.05{\pm}0.84nmol/d{\ell}$ to $4.17{\pm}0.64nmol/d{\ell}$ after taking taurine(p<0.01), and the plasma taurine concentration was significantly increased from $63.7{\pm}14.2{\mu}mol/{\ell}$ to $73.8{\pm}16.6{\mu}mol/{\ell}$ after taurine supplementation(p<0.05). The average dietary intake of taurine was $178.5{\pm}50.4$ mg/day, which is similar to the average daily taurine intake of Korean women. In conclusion, taurine is an effective nutrient that antagonizes TBARS levels. Therefore, this study suggests that a sufficient taurine intake may be an effective way to prevent cardiovascular disease such as atherosclerosis.

• 한국축산식품학회지
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• 제21권4호
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• pp.367-373
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• 2001

MFGM 당단백질의 하나인 PAS-7을 GPC 및 affinity chromatography에 의해 정제하여 2-AB로 형광 표식한 후, anion-exchange column 및 reversed-phase column을 이용해 5개의 성분을 분리하였다. 그 중 가장 상대량이 많은 성분 e에 대하여 RAAM2000을 이용한 당쇄구조 해석을 실시하여, 성분 e는 RAAM2000 GPC에 의하여 4개의 성분으로 분리되어 각각 calibration standard 12.10, 8.88, 5.84 및 4.86GU의 용출위치에 검출되었다. 이 용출위치와 당쇄구조는 livrary의 component-7457과 일치하며, 12.2GU의 크기로 분자량은 1788로 판단되며 library의 당쇄와 약 85%의 확률로 일치했다. 그 결과 성분 e의 당쇄구조는 환원말단에 $\alpha$1-6결합된 fucose를 1개 함유하며, core부분의 비환원말단에 N-acetyllactosamine branch를 2개 함유한 전형적인 biantennary 당쇄구조인 것으로 추축되어, 이전 HPLC, acetolysis, sequential exoglycosidase 소화, NMR분석에 의해 보고된 성분 7N1A의 구조와 일치함으로써, OGS RAAM2000을 이용한 PAS-7의 당쇄구조 해석의 유용성이 증명되었다.

• Bulletin of the Korean Chemical Society
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• 제35권7호
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• pp.1956-1964
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• 2014

To provide the scientific corroboration of the traditional uses of Akebia quinata (Thunb.) Decne., a detailed analytical examination of A. quinata stems was carried out using a reversed-phase high performance liquid chromatography (RP-HPLC) method coupled to photodiode array detector (PDA) for the simultaneous determination of four phenolic substances; cuneataside D (1), 2-(3,4-dihydroxyphenyl)ethyl-O-${\beta}$-D-glucopyranoside (2), 3-caffeoylquinic acid (3) and calceolarioside B (4). Particular attention was focused on the main compound, 3-caffeoylquinic acid (3), which has a range of biological functions. In addition, 2-(3,4-dihydroxyphenyl)ethyl-O-${\beta}$-D-glucopyranoside (2) was considered as a discernible marker of A. quinata from its easy confuse plants. The contents of compounds 2 and 3 ranged from 0.72 to 2.68 mg/g and from 1.66 to 5.64 mg/g, respectively. The validation data indicated that this HPLC/PDA assay was used successfully to quantify the four phenolic compounds in A. quinata from different locations using relatively simple conditions and procedures. The pattern-recognition analysis data from 53 samples classified them into two groups, allowing discrimination between A. quinata and comparable herbs. The results suggest that the established HPLC/PDA method is suitable for quantitation and pattern-recognition analyses for a quality evaluation of this medicinal herb.

• Archives of Pharmacal Research
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• 제29권12호
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• pp.1164-1170
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• 2006

A triptolide-lysozyme (TP-LZM) conjugate was synthesized to achieve renal specific delivery and to reduce the side effects of triptolide. Triptolide was coupled to lysozyme through succinic via an ester bond with an average coupling degree of 1 mol triptolide per 1 mol lysozyme. The lysozyme can specifically accumulate in the proximal tubular cells of the kidney, making it a potential carrier for targeting drugs to the kidney. The structure of triptolide succinate (TPS) was confirmed by IR, $^{1}H-NMR$, MS and UV. The concentrations of triptolide in various samples were determined by reversed-phase high-performance liquid chromatography (HPLC). In this study, the physicochemical and stability profiles of TP-LZM under various conditions were investgated the stability and releasing profiles of triptolide-lysozyme (TP-LZM) under various conditions. In vitro release trails showed triptolide-lysozyme was relatively stable in plasma (less than 30% of free triptolide released) and could release triptolide quickly in lysosome (more than 80% of free triptolide released) at $37^{\circ}C$ for 24 h. In addition, the biological activities of the conjugate on normal rat kidney proximal tubular cells (NRK52E) were also tested. The conjugate can effectively reduce NO production in the medium of NRK52E induced by lipopolysaccharide (LPS) but with much lower toxicity. These studies suggest the possibility to promote curative effect and reduce its extra-renal toxicity of triptolide by TP-LZM conjugate.

• Journal of Ginseng Research
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• 제42권1호
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• pp.21-26
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• 2018

Background: Furosine (${\varepsilon}$-N-2-furoylmethyl-L-lysine, FML) is an amino acid derivative, which is considered to be an important indicator of the extent of damage (deteriorating the quality of amino acid and proteins due to a blockage of lysine and a decrease in the digestibility of proteins) during the early stages of the Maillard reaction. In addition, FML has been proven to be harmful because it is closely related to a variety of diseases such as diabetes. The qualitative analysis of FML in fresh and processed ginsengs was confirmed using HPLC-MS. Methods: An ion-pair reversed-phase LC method was used for the quantitative analysis of FML in various ginseng samples. Results: The contents of FML in the ginseng samples were 3.35-42.28 g/kg protein. The lowest value was observed in the freshly collected ginseng samples, and the highest value was found in the black ginseng concentrate. Heat treatment and honey addition significantly increased the FML content from 3.35 g/kg protein to 42.28 g/kg protein. Conclusion: These results indicate that FML is a promising indicator to estimate the heat treatment degree and honey addition level during the manufacture of ginseng products. The FML content is also an important parameter to identity the quality of ginseng products. In addition, the generation and regulation of potentially harmful Maillard reaction products-FML in ginseng processing was also investigated, providing a solid theoretical foundation and valuable reference for safe ginseng processing.

• 대한유전성대사질환학회지
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• 제15권2호
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• pp.59-64
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• 2015

사람의 신체내에서 주로 존재하는 lactate는 L-lactate로서 몇몇 선천성대사이상과 관련하여 증가된다. 최근 2형 당뇨병(type 2 diabetes)과 만성적인 염증성 위장질환(inflammatory bowel disease)과 관련하여 증가되는 D-lactate를 L-lactate와 분리해야 하는 요구도가 높아졌다. 이에 액체크로마토그래프-탠덤질량 분석기를 사용하여 소변 내 D-, L-lactate를 분리하는 방법을 확립하였으며, 분석시간과 정밀성, 정확성, 특이성 등에서 신뢰성 있는 방법임을 확인하였다.

• 한국식품조리과학회지
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• 제11권3호
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• pp.303-308
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• 1995

참죽나무(Cedrela sinensis A.Juss) 잎을 재료로 하여, 김치, 나물, 부각 등의 조리하였을 때 이 식물의 flavonoid 화합물인 quercitrin의 함량 변화를 HPLC에 의한 용매계 THF-dioxane-MeOH-HOAc-5% H/Sub 3/PO/Sub 4/H/Sub 2/O(145:125:50:20:2:658)에서 분석하였다. 참죽김치를 5$^{\circ}C$에서 저장하였을 때의 quercitrin 함량은 1일째 13.06%, 6일째는 10.99%, 10일째는 11.49%, 20일째는 7.21%이며, 김치를 2$0^{\circ}C$에 저장하였을 시는 저장 1일째 15.33%, 6일째 10.83%, 10일째 9.85%, 20일째 5.78% 함유하고 있다. 나물의 경우 주성분인 quercitrin의 함량은 13.06%, 부각은 5.03%이었다. 따라서 김치의 경우 저장 일수가 길어짐에 따라 quercitrin함량은 감소하는 경향을 나타내며, 나물의 경우에는 quercetrin함량의 큰 변화를 볼 수 없고, 부각으로 조리한 경우 quercitrin의 함량은 급격히 감소하였다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권12호
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• pp.4937-4944
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• 2015

MicroRNAs (miRNAs) are emerging as critical regulators in carcinogenesis and tumor progression. Recently, miR-99a has been reported as a tumor suppressor gene in various human cancers, but its functions in the context of anaplastic thyroid cancer (ATC) remain unknown. In this study, we reported that miR-99a was commonly downregulated in ATC tissue specimens and cell lines with important functional consequences. Overexpression of miR-99a not only dramatically reduced ATC cell viability by inducing cell apoptosis and accumulation of cells at G1 phase, but also inhibited tumorigenicity in vivo. We then screened and identified a novel miR-99a target, mammalian target of rapamycin (mTOR), and it was further confirmed by luciferase assay. Up-regulation of miR-99a would markedly reduce the expression of mTOR and its downstream phosphorylated proteins (p-4E-BP1 and p-S6K1). Similar to restoring miR-99a expression, mTOR down-regulation suppressed cell viability and increased cell apoptosis, whereas restoration of mTOR expression significantly reversed the miR-99a antitumor activity and the inhibition of mTOR/p-4E-BP1/p-S6K1 signal pathway profile. In clinical specimens and cell lines, mTOR was commonly overexpressed and its protein levels were statistically inversely correlated with miR-99a expression. Taken together, our results demonstrated for the first time that miR-99a functions as a tumor suppressor and plays an important role in inhibiting the tumorigenesis through targeting the mTOR/p-4E-BP1/p-S6K1 pathway in ATC cells. Given these, miR-99a may serve as a novel prognostic/diagnostic and therapeutic target for treating ATC.