• Asian-Australasian Journal of Animal Sciences
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• 제19권4호
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• pp.601-604
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• 2006

The objective of this study was to investigate technical methods for extraction of mucopolysachharide-protein containing chondroitin sulfate from keel cartilage of chickens. The chemical composition of chicken keel cartilage was determined. For the preparation of mucopolysaccharide-protein from lyophilized chicken keel cartilage, hot water extraction and alcalase hydrolysis methods were examined. Results showed that the optimum condition of hot water extraction was incubation for 120 min with a yield of 40.09% and chondroitin sulfate content of 28.46%. For alcalase hydrolysis, the most effective condition was 2% alcalase in 10 volumes of distilled water for 120 min. The yield of hydrolysate was 75.87%, and chondroitin sulfate content was 26.61%. For further separation of chondroitin sulfate from the alcalase hydrolysate, which has a higher yield than that of hot water, 60% ethanol precipitation was performed. The yield of the ethanol precipitate was 21.41% and its chondroitin sulfate content was 46.31%. The hot water extract, alcalase hydrolysate and ethanol precipitate showed similar electrophoretic migration with standard chondroitin sulfate (chondroitin sulfate A), using cellulose acetate membrane electrophoresis. These results indicated that a significant amount of mucopolysaccharide-protein containing chondroitin sulfate could be acquired form chicken keel cartilage. Therefore, keel cartilage in chicken may provide an inexpensive source of chondroitin sulfate for commercial purposes.

• 한국식품영양과학회지
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• 제24권3호
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• pp.446-453
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• 1995

The improtant components of extracellular matrix(ECM) are collagen and chondroitin sulfate. The hydrophobic surface of collagen is one of the determining factors of diameter of collagen fiber and also is closely related to the aging phenomena. The controlling mechanism of the diameter of collagen fiber influenced by the interaction with chondroitin sulfate was evaluated using bis-ANS as a hydrophobic probe. Hydrophobic surface area of collagen molecule shielded by chondroitin sulfate was evaluated. Relative fluorescence intensity of collagen in thepresence of chondroitin sulfate was measured using bis-ANS as a hydrophobic probe. The fluorescence intensity decreased with the increase in chondroitin sulfate up to 3.8 chondroitin sulfate/collagen(mole/mole). Further increase in the ratio of chondroitin sulfate to collagen did not change the fluorescence intensity. Similar changes in the relative fluorescence intensity were observed for both rat tail and lathyrific rat skin collagen. The fluorescence intensity indicated by the binding between bis-ANS and hydrophobic sites of collagen was pH dependent, and the shielding effect of collagen-chondroitin sulfate interaction could not be detected at pH above 6.0. This is probably due to the charge repulsions caused by negative charged collagen molecules at higher pH.

• Bulletin of the Korean Chemical Society
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• 제5권6호
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• pp.249-253
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• 1984

Mutual diffusion coefficients of choline chloride were determined by using the diaphragm cell method in aqueous solutions of chondroitin sulfate A at $25^{\circ}C$. The diffusion coefficients of choline chloride in 0.1g/100ml, 0.5g/100ml and 1g/100ml respectively of chondroitin sulfate solutions were compared with those of binary systems of water-choline chloride. At low concentrations, the diffusion coefficients of the choline chloride in the presence of chondroitin sulfate were significantly smaller than the values obtained in the absence of chondroitin sulfate, indicating a strong interaction between these solutes. The effect of this interaction on the diffusion of choline ion is largest at higher chondroitin sulfate concentrations and at lower choline chloride concentrations. The influence of chondroitin sulfate is overcome at higher choline chloride concentrations. Self-diffusion coefficients of choline ion in the presence of chondroitin sulfate are also obtained. Excellent agreements were obtained between the experimental data and the calculated values obtained by using the Manning's equations. These observations suggest that the interaction between choline chloride and chondroitin sulfate involves primarily a long range electrostatic effect and there is no appreciable "condensation" or binding of choline ion to the chondroitin sulfate.

• 한국환경과학회지
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• 제25권5호
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• pp.645-654
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• 2016

For the purpose of reuse the wasted by-products from the skate process to the health functional food or medicinal material, chondroitin sulfate was extracted from the skate cartilage with the method of hydrolysis with protease enzyme, and the contents of chondroitin sulfate and hydrolyzed protein were measured qualitatively and quantitatively. The effects of chondroitin sulfate on body weight or liver weight changes, hepatotoxicity elimination and anti-inflammatory actions were measured from in vivo test with feed-treated mice. From the hydrolytic extraction of skate cartilage with the mixture of 1% alcalase and 1% protease for 4 hours, the extraction yield of chondroitin sulfate was about 32.55%. The content and molecular weight of chondroitin sulfate was 26.63% and $2.85{\times}10^5Da$., respectively and the content ratio of chondroitin sulfate to protein was measured to 1 to 2.76 with gel permeation chromatography. For the odor component, trimethylamine decreased about 30% but almost not ammonia from chondroitin sulfate with the treatment of activated carbon. From the feeding chondroitin sulfate to mice, the control effect of body and liver weights decrease was measured, anti-inflammatory action and hepatotoxicity elimination action were also measured. From these results, process operation conditions for manufacturing of chondroitin sulfate were suggested.

• 한국식품위생안전성학회지
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• 제14권1호
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• pp.45-54
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• 1999

The purpose of this study was to examine the increased bone loss caused by ovariectomy through monitoring the concentrations of the collagen and the pyridinoline crosslinks of collagen. The ovariectomized rats treated for 8 weeks, were divided at random into two or three groups of 10. Ovariectomies were carried out from the saline-treated group (Ovx), the estrogentreated group (Ovx+ES) and chondroitin sulfate-treated group (Ovx+CS). Sham operations were performed on the sham-operated group (Sham). Ovx+ES and Ovx+CS groups showed the remarkably increased collagen and pyridinoline amount in the bone and cartilage compared to Ovx group. And as the result of the measurement of SOD, Catalase and GPx which are antioxidant enzyme, SOD and Catalase activities in Ovx group were much higher than in Sham group. But they were significantly decreased in Ovx+CS group. Based on these results, it is supposed that estrogen and condroitin sulfate can enhance collagen synthesis and affect the pyridinoline formation in collagen fibrils through stimulating lysyl oxidase activity. And it is also thought that chondroitin sulfate can inhibit aging by reducing antioxidant enzyme.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.791-795
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• 2001

This study was conducted to develop a new biomaterial to be used for skin whitening. The melanin elimination effect of chondroitin sulfate and phelinus linteus mushroom in rabbit back skin were evaluated. Rabbit dorsum was exposed to chronic UV irradiation(320nm) once daily for 30 days after initial melanin injection (100mg/kg). And then, chondroitin sulfate and phelinus linteus mushroom at dose of 0.7g for 30days were applied on the zone. The dorsal skin was histologically examined. Furthermore, we investigated free-radical extinction effect, antioxidation and tyrosinase activity inhibition effects. The histological study indicated that chondroitin sulfate and phelinus linteus mushroom decreasd melanine pigment significantly. As a result, chondroitin sulfate and phelinus linteus mushroom have a remarkable effect on the skin whitening by melanin elimination.

• 한국산학기술학회논문지
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• 제10권4호
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• pp.865-871
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• 2009

콘드로이틴황산은 상어연골로부터 알카리방법과 효소적 방법에 의해 추출하였다. 추출에 있어서 알카리의 농도, 효소인 알카라제의 농도, 온도 등에 따른 최적 추출조건을 규명하였으며 콘드로이틴 황산용 UF 막으로 막분리하여 순수한 콘드로이틴황산을 얻었다. 얻어진 콘드로이틴 황산의 분자량을 GPC로 측정하였을 때 분자량이 27만 Da 이었다.

• Food Science and Biotechnology
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• 제14권5호
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• pp.645-650
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• 2005

To reduce ammonia-like odor in chondroitin sulfate, longnose skate (Rasa rhina) cartilage was processed by washing, autoclaving, and alkali pretreatments. Content of total volatile basic nitrogen (TVB-N), index of ammonia-like odor, of raw skate cartilage without pretreatment was 254 mg/100 g, whereas those of skate cartilage pretreated with washing and autoclaving increased to 630 and 636 mg/100 g, respectively. TVB-N of skate cartilage pretreated with sodium hydroxide sharply decreased to 15 mg/l00 g at optimal condition of 0.12 M and 3.6 volume of NaOH, as determined by surface response methodology of central composite design for optimization. Alkali pretreatment resulted in 97.6% deodorizing. Washing and autoclaving pretreatments had almost no effect on the yield of chondroitin sulfate (approximately 30%), whereas decreased to 16.0% after alkali pretreatment, showing chondroitin sulfate of skate cartilage as chondroitin sulfate C.

• 한국산학기술학회논문지
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• 제17권6호
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• pp.9-17
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• 2016

Alcalase와 protamex의 두 가지 단백질 분해효소를 사용하여 홍어연골로부터 콘드로이틴 황산을 분해추출하고, 몇가지 물리화학적 공정을 적용하여 고순도의 콘드로이틴 황산을 얻기 위한 정제방법을 검토하였다. 2% alcalase로 홍어연골을 가수분해하고 $40^{\circ}Brix$로 농축한 추출물의 수율은 23.3%이었으나, 에탄올로 1회 및 2회 추가 정제한 경우 각각 8.47%, 3.37%로 얻어졌다. 1% alcalase와 1% protamex로 혼합 사용하여 추출분해하고, $40^{\circ}Brix$로 농축한 다음 1회 에탄올 정제한 경우는 추출수율이 16.62%로 측정되어 단백질 분해효소를 혼합한 경우가 더 높은 수율을 보였다. 추출된 콘드로이틴 황산의 함량은 에탄올 용매비에 따라 39.88~45.08%의 범위로 측정되었으며, alcalase만 사용시에는 용매비 1:1에서 42.92%로 가장 높게 측정되었고, alcalase와 protamex를 혼합 사용한 경우에는 용매비 1:2에서 45.08%로 가장 높았으며, 에탄올에 의한 농축물의 정제시 교반 시간은 2시간이 효과적이었다. 콘드로이틴 황산의 GPC (Gel Permeation Chromatography) 측정 결과 alcalase와 protamex를 혼합 사용한 경우 에탄올 정제 횟수에 따라 콘드로이틴 황산의 분자량과 순도는 각각 11만~31만 Da.과 24.87%~49.92%의 범위로 측정되었으며, 한외여과를 통하여 분자량 약 11만 Da., 최고 순도 53.93%의 콘드로이틴 황산을 얻을 수 있었다. 콘드로이틴 황산의 냄새 강도는 에탄올 정제만으로는 33%, 활성탄 처리와 에탄올 정제를 병행한 경우 38%의 감소효과가 얻어졌으며, 활성탄 처리와 2회의 에탄올 정제시 암모니아는 52.1%, TMA (trimethylamine)는 37.89%의 탈취효과가 있었으나, 냄새성분의 충분한 제거를 위해서는 추가적인 물리화학적 처리가 요구되었다.

• 생약학회지
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• 제40권2호
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• pp.109-117
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• 2009

We examined the purity of six heparin samples by using heparinase, chondroitinase, $^{1}H-NMR$, and polyacrylamide gel electrophoresis. To obtain high molecular weight contaminants from heparin samples, heparinase I - digested samples were subjected to the exhaustive microcon filtration. The filtration process removed heparin-derived di- and oligosaccharides effectively. By combining chondroitinase ABC treatment and strong anion exchange - high performance liquid chromatography, the result showed all six samples contained chondroitin sulfate as a contaminant ranging from 1.3 to 14.9%. Among them, sample S3 showed the highest content of 14.9%, which was further analyzed by chondroitinase AC treatment to confirm chondroitin sulfate B (dermatan sulfate). $^{1}H-NMR$ chemical shifts of N-acetyl groups clearly suggested the existence of chondroitin sulfate B (sample S3) and oversulfated chondroitin sulfate (samples S2 and S4) as contaminants. In addition, polyacrylamide gel electrophoresis was useful for qualitative detection on the sample's purity. These results suggest that the tools of heparinase I and chondroitinase ABC in combination with NMR spectroscopy would give very useful information for investigation of heparin contaminants such as oversulfated chondroitin sulfate and dermatan sulfate in heparin samples.