• 제목/요약/키워드: DPC

검색결과 183건 처리시간 0.022초

옻나무 유래 Flavonoid처리가 흰쥐 Leydig 세포의 체외배양에서 Testosterone분비에 미치는 영향

  • 성환후;최선호;민관식;장유민;나천수;정일정
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 춘계학술발표대회
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    • pp.80-80
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    • 2001
  • 옻나무 유래 Flavonoid(F)는 항암 효과, 숙취해소 등의 효능이 있으며, 신장의 기능을 촉진하고 생식에도 영향을 준다고 알려져 있다. 그 주성분으로는 fustin, fisetin, sulfuretin, butein으로 추정되고 있다. 본 연구팀의 예비실험에서 성숙 흰쥐에게 F를 30일간 5mg씩 구강투여 하여 정소 중량 및 혈액 내 Testosterone 농도를 분석한 결과, 정소 중량이 증가되고 혈중 Testosterone의 농도도 대조구에 비해 유의적으로 높은 결과를 얻었다. 이에 본 연구의 목적은 옻나무 유래 F가 성숙 수컷 횐쥐의 생식 기능에 직접 미치는 영향을 알아보고자, 흰쥐 Leydig 세포를 분리, 회수하여 체외배양에서 Testosterone농도를 조사하였다. SD계 흰쥐(약 12주령, 체중 250g 전후)의 정소를 적출 한 후, Leydig 세포를 회수하기 위해 0.25% collagenase용액에 넣어 34$^{\circ}C$에서 15분간 진탕 수조에서 배양하였다. 배양된 정소조직은 D-MEM 배양액(10% FCS와 antibiotics 첨가)으로 2~3회 세척한 후 세포 수를 1$\times$$10^{6}$ cells/$m\ell$/well로 분주하여, F(20, 40, 80, 160ng), IGF-I(50, 100ng) 와 LH(10, 100ng) 용량별 각각 첨가하여 배양하였다. 각 처리 후, 배양시간(3, 6, 12, 24, 36시간)에 따라 배양액을 회수하여 COAT-A-COUNT(DPC, USA) kit를 이용하여 RIA방법에 의해 Testosterone을 분석한 결과는 다음과 같다. 대조구 Leydig 세포를 36시간까지 체외 배양하여 Testosterone의 농도를 조사한 결과, 24시간 배양구가 가장 높은 농도를 나타내었다. F를 단독 첨가하여 12시간 배양한 실험에서 F 80ng 첨가구에서 유의적으로 높은 농도를 보였다. LH 10ng 및 100ng 첨가구의 시간별 변화로는, LH 10ng 첨가구에서는 6~12시간 이후, LH 100ng 첨가구에서는 3~6시간 사이에서 유의적인 증가를 보였다. 또한, LH 10ng 첨가 실험에서는 LH 10ng+F 40ng에서 12시간 배양 시, 유의적으로 높은 Testosterone 분비를 확인하였으며, LH 100ng을 첨가하여 3시간 배양 시는 각 처리구 마다 높은 유의적 차이를 보였다. 한편, GH에 관여하는 IGF-I 첨가효과를 비교 검토한 결과, LH 100ng+IGF-I 100ng, 6시간 배양구에서 유의적으로 높게 나타내었다. 이상의 결과로, F 단독 처리 시의 적정량은 약 80ng이며, 흰쥐 정소 Leydig 세포의 Testosterone분비를 촉진하는 작용을 하는 것으로 사료되며, 특히 LH+F구에서 Testosterone분비를 더욱 향상시킴으로써 정소 Leydig 세포의 Androgen 생성을 촉진시키는 역할이 있는 것으로 보여진다 따라서 옻나무 유래 F는 포유동물의 생식기능에 중요하게 작용하는 것으로 사료된다.

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Biological Function of Single Chain Glycoprotein Hormone Mutants

  • Min, Kwan-Sik;Chang, Yoo-Min;Chang, Sun-Hwa;Lee, Hyen-Gi;Lee, Yun-Gun;Chang, Won-Kyong;Cheong, Il-Cheong
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 춘계학술발표대회
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    • pp.54-54
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    • 2001
  • Human chorionic gonadotropin (hCG) is a member of the glycoprotein hormone family which includes FSH, hCG, TSH. These hormone family is characterized by a heterodimeric structure composed a common $\alpha$-subunit noncovalently linked to a hormone specific $\beta$-subunit. The correct conformation of the heterodimer is also important for efficient secretion, hormone-specific post-translational modifications, receptor binding and signal transduciton. To determine $\alpha$ and $\beta$-subunits can be synthesized as a single polypeptide chain (tethered-hCG) and also display biological activity, the tethered-hCG molecule by fusing the carboxyl terminus of the hCG $\beta$-subunit to the amino terminus of the $\alpha$-subunit was constructed and transfected into chinese hamster ovary (CHO-K1) cells. We also constructed C-terminal deletion mutants (D9l, D89, D88, D87, D86, D84, D83) of single chain hCG to determine the biological function (secretion, LH-activity, receptor binding, cAMP production) of these mutants. Between six and eight stably transfected pools of cells expressing wild type and mutant hCGs were selected for neomycin resistant. The hCGs secreted by the stably transfected cells into serum-free media were collected and quantified by radioimmunoassay, as described in protocol (DPC(hCG IRMA). LH activity was in terms of testosterone production and aromatase activity in primary cultured rat Leydig cells. The tethered-wthCG was efficiently secreted and showed similar LH-like activity to the dimeric hCG. The D83hCG mutant was not detected in this assay. It is suggest that hCG C-terminal part is very important for hCG secretion. Now, we checking the LH-like activity of these mutant hCGs. These data indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion.

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사람 모유두세포에서 코르티코트로핀분비인자에 의한 모발성장관련사이토카인의 발현 조절 (Corticotropin-Releasing Factor Down-Regulates Hair Growth-Related Cytokines in Cultured Human Dermal Papilla Cells)

  • 이은영;전지혜;이민호;이승호;김영호;강상진
    • 대한화장품학회지
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    • 제40권4호
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    • pp.413-421
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    • 2014
  • 코르티코트로핀분비인자(Corticotropin-releasing factor)는 스트레스 반응에 관여하는 호르몬으로, 최근 스트레스가 탈모와 같은 피부질환에 영향을 미친다는 보고들이 많아지고 있다. 보고에 따르면, 사람 모낭 배양에서 코르티코트로핀분비인자는 길이생장을 억제하며, 모낭의 조기퇴행을 유도하고 모기질각질형성세포(hair matrix keratinocyte)의 세포사멸을 촉진시킨다. 본 연구에서는 코르티코트로핀분비인자가 모발성장과 모주기조절에 핵심적으로 역할하는 모유두세포에 미치는 영향에 대해 알아보고자 했다. 시상하부-뇌하수체-부신축의 주요 스트레스호르몬들인 코르티코트로핀분비인자, 부신피질자극호르몬, 그리고 코르티솔을 사람 모유두세포에 처리하였다. 흥미롭게도, 코르티코트로핀분비인자가 모발성장과 관련된 사이토카인(KGF, Wnt5a, $TGF{\beta}-2$, Nexin)의 발현을 변화시키는 것을 관찰하였으며, 세포 내 cAMP의 수준을 증가시켰고, 수용체의 발현을 억제시켰다. 이러한 변화는 수용체의 길항제인 antalarmin과 astressin2B, 또는 PKA 억제제의 전처리로 인해 막을 수 있었다. 코르티코트로핀분비인자는 cAMP/PKA경로를 통해 POMC의 발현을 유도하는데, 사람 모유두세포에서도 이 호르몬의 처리가 POMC mRNA의 발현을 증가시키는 것을 확인할 수 있었으나 부신피질자극호르몬의 변화는 western blot으로는 확인할 수 없었다. 이러한 결과들을 바탕으로, 코르티코트로핀분비인자가 그 수용체를 통해 사람 모유두세포 내 모발성장 관련 사이토카인의 발현을 조절함을 확인하였으며, 이는 코르티코트로핀분비인자의 수용체 길항제가 스트레스성 탈모환자를 위한 치료제 혹은 화장품 소재로써 활용될 수 있음을 보여준다.

Feasibility of Coculture Method for Production of Chimeric Mice Using J1 Embryonic Stem Cells

  • Shin Hye-Jun;Park Sung-Sik;Kim Sun-Uk;Cho Sang-Mi;Han Ying-Hao;Kim Hyun-Sun;Kim Sang-Geun;Lee Dong-Seok;Yu Dae-Yeul
    • 대한의생명과학회지
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    • 제10권4호
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    • pp.391-395
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    • 2004
  • The demand for the production of gene-defective mice from embryonic stem (ES) cells is increasing to clarify decisive gene function in vivo. Although blastocyst injection is widely used to generate ES cell-mediated knockout mice, coculture method has been alternatively used because of several advantages, such as low cost and simple procedure. Thus, this experiment was designed to demonstrate the feasibility of the coculture method using J1 ES cells, which are known to be efficient for blastocyst injection. Eight-cell embryos were harvested from 2.5 days post-coitum (dpc), denuded with acid tyrode's solution, and transferred onto trypsinized J1 ES cells. Aggregation was carried out following two typical methods, which are simple coculture method and aggregation in groove prepared by aggregation needle. Successfully aggregated-embryos were developed to blastocysts for 24 h and transferred into uterus of pseudo-pregnant foster mother. Chimeric offspring was judged by coat pigmentation. In this study, we could obtain chimeric mice from all the two aggregation methods, but the chimera production efficiencies in coculture using groove were three times higher at least than those in the other group. In conclusion, these observations suggest that coculture method should be available for production of knockout mice from J1 ES cells. Presently, the germ-line transmission rates of the chimeras produced from the two methods are under investigation.

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위암에서 여러 종양억제유전자 부위의 이형접합성 소실과 현미 부수체 불안정성 (Loss of Heterozygosity and Microsatellite Instability at Multiple Tumor Suppressor Genes in Gastric Carcinomas)

  • 조용구;김창재;박조현;김영실;김수영;남석우;이석형;유남진;이정용;박원상
    • Journal of Gastric Cancer
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    • 제3권4호
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    • pp.214-220
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    • 2003
  • Purpose: The aim of this study was to investigate the frequency of loss of heterozygosity and the microsatellite instability at multiple tumor suppressor gene loci in gastric adenocarcinomas. Materials and Methods: Loss of heterozygosity and the microsatellite instability at several tumor suppressor gene loci were analyzed in 29 primary gastric carcinomas by using microdissection and the polymerase chain reaction. Results: Twenty-three ($79\%$) of the 29 cases demonstrated loss of heterozygosity at one or more loci. The frequency of loss of heterozygosity at the p53 locus was the highest ($63\%$) and those at the VHL, APC, p16, Rb, MEN1, BRCA1, DPC4, 3p21, and 16p13 region were $41\%,\;36\%,\;19\%,\;29\%,\;33\%,\;26\%,\;21\%,\;32\%,\;and\;11\%$, respectively. Compared with histological type, loss of heterozygosity was more common in diffuse-type gastric cancer (P<0.01). Interestingly, 9 of 10 tumors with allelic deletion at the p53 locus showed loss of heterozygosity at other tumor suppressor gene loci. The microsatellite instability was also detected in 6 ($20\%$) of the 29 cases at one or more loci. Conclusion: These data suggest that frequent loss of heterozygosity and the microsatellite instability at multiple tumor suppressor genes might be required for the development and the progression of gastric carcinomas and that p53 allelic loss may be the most frequent event in the development of gastric carcinomas.

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cAMP-Dependent Signalling is Involved in Adenosine-Stimulated $Cl^-$ Secretion in Rabbit Colon Mucosa

  • Oh, Sae-Ock;Kim, Eui-Yong;Jung, Jin-Sup;Woo, Jae-Suk;Kim, Yong-Keun;Lee, Sang-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • 제2권4호
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    • pp.521-527
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    • 1998
  • An important property of the intestine is the ability to secrete fluid. The intestinal secretion is regulated by a number of substances including vasoactive intestinal peptide (VIP), ATP and different inflammatory mediators. One of the most important secretagogues is adenosine during inflammation. However, the controversy concerning the underlying mechanism of adenosine-stimulated $Cl^-$ secretion in intestinal epithelial cells still continues. To investigate the effect of adenosine on $Cl^-$ secretion and its underlying mechanism in the rabbit colon mucosa, we measured short circuit current ($I_{SC}$) under automatic voltage clamp with DVC-1000 in a modified Ussing chamber. Adenosine, when added to the basolateral side of the muocsa, increased $I_{SC}$ in a dose-dependent manner. The adenosine-stimulated $I_{SC}$ response was abolished when $Cl^-$ in the bath solution was replaced completely with gluconate. In addition, the $I_{SC}$ response was inhibited by a basolateral Na-K-Cl cotransporter blocker, bumetanide, and by apical $Cl^-$ channel blockers, dephenylamine-2-carboxylate (DPC), 5-nitro-2-(3-phenyl-propylamino)-benzoate (NPPB), glibenclamide. Amiloride, an epithelial $Na^+$ channel blocker, and 4,4-diisothiocyanato-stilbene-2,2-disulphonate (DIDS), a $Ca^{2+}-activated$ $Cl^-$ channel blocker, had no effect. In the mucosa pre-stimulated with forskolin, adenosine did not show any additive effect, whereas carbachol resulted in a synergistic potentiation of the $I_{SC}$ response. The adenosine response was inhibited by 10 ${\mu}M$ H-89, an inhibitor of protein kinase A. These results suggest that the adenosine-stimulated $I_{SC}$ response is mediated by basolateral to apical $Cl^-$ secretion through a cAMP-dependent $Cl^-$ channel. The rank order of potencies of adenosine receptor agonists was $5'-(N-ethylcarboxamino)adenosine(NECA)>N^6-(R-phenylisopropyl)adenosine(R-$ PIA)>2-[p-(2-carbonylethyl)-phenyl-ethylamino]-5'-N-ethylcarboxaminoadenosine(CGS21680). From the above results, it can be concluded that adenosine interacts with the $A_{2b}$ adenosine receptor in the rabbit colon mucosa and a cAMP-dependent signalling mechanism underlies the stimulation of $Cl^-$ secretion.

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갈색거저리 유충 추출물의 항산화 활성 및 모발 성장 촉진 효과 (The Antioxidant Activities and Hair-growth Promotion Effects of Tenebrio molitor Larvae Extracts (TMEs))

  • 백민희;서민철;김미애;윤은영;황재삼
    • 생명과학회지
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    • 제27권11호
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    • pp.1269-1275
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    • 2017
  • 최근 들어 곤충을 식품 및 바이오 소재로 이용한 연구가 활발히 진행되고 있다. 그러나 곤충을 이용한 모발 성장 효과에 대한 연구는 아직 미흡한 실정이다. 따라서 본 연구에서는 탈모 예방 및 모발 성장 효과를 가진 새로운 천연물 소재 개발을 위해 갈색거저리 유충 추출물의 항산화 활성 및 모발 성장 촉진 효과를 연구하였다. 갈색거저리 유충 추출물의 항산화 활성 평가를 위해서 DPPH 라디칼 및 아질산염 소거능을 측정하였다. 모발 성장촉진 효과를 측정하기 위해서는 인간 모유두세포(human dermal papilla cell)와 섬유아세포(fibroblast, NIH3T3 cell)를 이용하였으며 MTS assay를 통해 세포생존율 및 세포증식률을 측정하였다. 모유두세포에서 dihydrotesteone (DHT)에 의한 세포사 억제 효과를 확인하였으며, 섬유아세포에서는 tolbutamide (TBM)의 potassium channel blocker 역할에 의한 세포사 억제 효과를 확인하였다. DPPH radical 및 아질산염 소거능 측정 결과 갈색거저리 유충 추출물은 항산화 역할이 뛰어난 것으로 보고된 블루베리와 유사하거나 높은 정도의 항산화능을 가지는 것으로 확인되었다. In vitro 상에서 갈색거저리 유충 추출물을 48시간 동안 처리한 경우, 모유두세포와 섬유아세포의 세포증식을 218% 및 116%까지 증가시켰다. 또한, 모유두세포에서 DHT 처리에 의한 세포사가 갈색거저리 유충 추출물에 의해 억제되는 것을 확인하였으며, 섬유아세포에서는 potassium channel blocker인 TBM에 의해 세포생존율이 감소하였으나 갈색거저리 유충 추출물 처리 시 세포생존율이 정상군과 비슷한 정도로 회복되는 것을 확인하였다. 이상의 결과로부터 갈색거저리 유충 추출물을 이용한 모발성장 및 탈모방지 기능성 소재 개발 가능성을 확인하였다.

A Study on the Influence of Digital Experience and Purchase in the 4th Industrial Revolution : Focusing on Differences between Satisfied, Neutral, and Dissatisfied Groups

  • Jung, Sang Hee;Lee, Sang-Jik
    • Journal of Information Technology Applications and Management
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    • 제26권4호
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    • pp.51-69
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    • 2019
  • One of the most considerate phenomena of the era of the Fourth Industrial Revolution is the use of digital devices. Digitalization is rapidly advancing through all areas of industry and life. Customer journey with digitalization is looking totally different from previous customer journey. The research targets were users of fashion, automobiles, cosmetics and online shopping malls. We analyzed 300 people for each valid questionnaire. The results of the study are as follows. First, it has been proven that digital experience affects positive (+) impact on purchasing intention and positive (+) impact on recommending intention and negative impact (-) on switching intent and subsequently affects positive impact (+) to purchase and incase of switching intent, negative impact (-) to purchase. Unlike traditional methods such as SPC(Service Profit Chain), the Digital experience to Purchase process Chain (DPC) has been identified to be suitable in the digital age. Second, the digital satisfied group (5 score-very satisfaction) has shown same result as above. However the digital neutral group (even though 4 score- satisfaction in five-point scale), specially in a highly competitive industry, has different from the satisfied group and 3 score-normal is same as dissatisfied group. It means that this group is that If there is a high level of attractiveness of substitute goods, there is a high possibility of switching them. It has supported Jones and Sasser [1995] that there have been two types of loyalty of true long-term loyalty and what we call false loyalty in the highly competitive industry zone which is commoditization or low differentiation, many substitutes, low cost of switching. Identifying true loyalty and false loyalty is crucial to establishing a customer experience strategy. it is necessary to actively utilize long-term digital experiences strategy to increase the total satisfaction of digital experience through all of customer purchasing journey in order to enhance the digital customer experience. It is difficult to see the effect as a one-time event. It should be scaled over the entire customer purchase process over a long period of time, which can positively affect purchase intention, recommendation intention, and conversion intention. This is also why it is difficult for second-runners to overtake first-runners in a short period.

보리와 옥수수 잎의 색소, 단백질 함량 및 관계II 활성에 미치는 ${SO}^2$의 영향 (Effects of Sulfur Dioxide on Pigments, Frotein Content and Photosystem II Activity of Barley and Corn Leaves)

  • 정화숙
    • Journal of Plant Biology
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    • 제25권3호
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    • pp.135-151
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    • 1982
  • This investigation was carried out to clarify the changes of pigments and soluble protein, and photosystem II activity in the leaves of barley (${SO}_2$-sensitive) and corn (${SO}_2$-resistant) seedlings induced by the ${SO}_2$ fumigation (10, 50ppm). The pH changes of the leaf extract, the content of sulfite and sulfate, the activities of catalase, peroxidase, and polyphenoloxidase were compared in the leaves of barley and corn seedlings induced by ${SO}_2$ fumigation. The results are summarized as follows: An appreciable effect of pH change of leaf extract by ${SO}_2$ fumigation was observed in barley leaves (pH 6.10 to 5.18), but only a small change occurred in corn leaves (pH 5.66 to 5.50). The same pattern of pH changes was recorded when the solution of 0.2N HCl was added to leaf extract, providing lower buffering capacity of the barley leaves than corn leaves. After 2 hours of exposure to 10 ppm ${SO}_2$, the contents of ${SO}^{2-}_3$ and ${SO}^{2-}_4$ were increased in barley leaves, while only ${SO}^{2-}_4$ increased in corn leaves. After fumigation with 10ppm ${SO}_2$ for 2 hours, barley leaves showed significant decreases in activities of catalase, to 17% peroxidase, to 58%, and polyphenoloxidase, to 88%. Corn leaves showed increases in activities of peroxidase, to 136%, and polyphenoloxidase, to 128%. Absorption spectra of pigments obtained from ${SO}_2$-fumigated leaves were gradually decreased with the fumigation time increases, but the decrease was more significant in barley leaves. Fumigation with 50ppm ${SO}_2$ for 2 hours induced the greatest decomposition in carotenoid, followed by chlorophyll a and then chlorophyll b in barley leaves. The ratio of chlorophyll a/b was decreased from 4.1 to 3.6 in barley leaves, but in corn leaves it was maintained almost a constant level(4.9-4.8). The rate of decomposition of chlorophyll and carotenoid in corn leaves was very slow than those in the barley leaves. Fumigation with 50 ppm ${SO}_2$ for 2 hous, decreased the protein content of barley leaves to 59%, and that of corn leaves to 89%, and the extent of decrease in protein content was greater than that of pigments in barley and corn leaves. The rate of DCIP9dichlorophenol indophenol) photoreduction in ${SO}_2$-fumigated leaves was decreased to 18 and 67% in barley and corn leaves, respectively. However, DCIP photoreduction was considerably recovered about 32 and 92% with the addition of DPC(diphenylcarbazide) as an exogenous electron donor in barley and corn leaves, respectively.

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전처리방법에 따른 불용성 6가 크롬 분석에 관한 연구 (Pretreatment Effect on the Analysis of Insoluble Hexavalent Chromium)

  • 이은정;노재훈;원종욱;전미령;조명화;김치년
    • 한국산업보건학회지
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    • 제6권2호
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    • pp.292-300
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    • 1996
  • Hexavalent chromium($Cr^{+6}$) compounds are considered to be particularly hazardous, primarily because of the associated risk of allergic reaction and cancer. The analytic method of hexavalent chromium such as the s-diphenylcarba-zide(DPC) method and all ether previously used methods are often made uncertain due to significant interferences from organic components. This report can provide a technique for the more rapid and simple determination of total hexavalent chromium. than other currently using methods. The s-diphenylcarbazide method proposed by the U.S. National Institute for Occupational Safety and Health has low recovery rate(15.67 - 48.20%) due to interference, iron chloride and nickel chloride. A microwave oven technique has high recovery rate(about 70%) of insoluble hexavalent chromium. For the difference of ionic charges of $Cr^{+3}$-ethylenediamine tetraacetic acid(EDTA) chelate and $CrO_4{^{-2}}$, we could detect them simultaneously by ion exchanged high performance liquid chromatography. The confirmation of $Cr^{+3}$ and $Cr^{+6}$ were checked by fraction collector and flameless atomic absorption spectrometer. We observed that the small amount of hexavalent chromium is converted to trivalent chromium due to enhancement of chromium reduction by $Fe^{+3}$ or $Ni^{+2}$. As a result of this study, on the analysis of insoluble hexavalent chromium with microwave oven was used for, it may be better and more precise analysis after pretreatment by 2% NaOH-3% $Na_2CO_3$ and then analysis UV-spectrophotometer. It should be done for various studies on insoluble hexavalent chromium on the basis work environmental monitoring so called welding, painting etc.

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