• 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.

• 분석과학
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• 제7권2호
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• pp.245-251
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• 1994

혼합물 중에서 미량 존재하는 황산콘드로이틴나트륨을 정량하기 위하여, Chondroitinase ABC를 사용하여 효소반응을 시킨 후 얻어지는 ${\Delta}Di-6S$(2-acetamido-2-deoxy-3-0-(${\beta}$-D-gluluco-4-enepyranosyluronic acid)-6-0-sulf-D-galactose)를 지표성분을 설정하여 고성능 액체크로마토그래피(HPLC)를 이용하는 방법을 확립하였다. 230nm에서 흡광도를 측정하였고, 검출한계는 $1{\mu}g/ml$였다. 본 분석법을 재제에 적용하였을 때 캅셀제는 $100.01{\pm}1.58%$, 점안제는 $99.89{\pm}1.80%$로 재현성이 있는 결과를 나타내었다.

• 대한화학회지
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• 제35권6호
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• pp.612-616
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• 1991

Sodium dodecyl sulfate(SDS), chondroitin sulfate 그리고 L-${\alpha}$-lecithin vesicle의 각각의 수용액에서 methylene blue (MB)의 metachromatic 성질을 $18^{\circ}$C에서 $52^{\circ}$C까지 흡수분광법으로 측정했다. MB는 지질이 고농도일 때 vesicle 매트릭스에 뭉치를 형성했다. MB의 metachromasy는 vesicle의 상전이 온도에 무관하게 나타났으며, 이것은 색소가 vesicle의 친수성 표면에서 회합함을 의미한다. Vesicle에서 MB의 metachromasy 현상은 hexadecyltrimethylammonium bromide(CTAB)를 첨가할 때보다 SDS를 첨가할 때 매우 약하게 관측되었다. 이것은 vesicle 표면에서 계면활성제들의 끼어드는 위치가 다른 것으로 보이며, CTAB가 SDS보다 색소뭉치 사이에 더욱 잘 끼어드는 것으로 사료된다.

• KSBB Journal
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• 제18권3호
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• pp.180-185
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• 2003

GAGs의 추출 방법 중 효소의 사용으로 추출 효율의 상승과 원료수급의 문제점을 해결할 수 있었다. 그러나 효소 사용에 따른 생산단가의 상승과 단백질 함량의 증가에 따른 화장품 규격을 만족시킬 수 없음으로, 1/60 M sodium phosphate buffer로 105$^{\circ}C$로 열수 추출하는 것이 화장품 규격을 만족시키는 방법으로 나타났다. 이렇게 추출한 GAGs의 SO$_4$ 함량은 31.2%이고 회분의 함량이 22.2%로 GAGs 추출 향상을 위해 사용되어지는 sodium phosphate의 사용으로 회분 함량이 높아졌음을 알 수 있고 무기질 분석에서 Na의 함량이 3.0 mg%로 총 무기질의 47.6%를 차지하고 있었다. 시료를 HPLC로 분석하였을 때 chondroitin sulfate A, C와 동일한 분석시간을 나타내어 chondroitin sulfate의 함유 가능성을 나타내고, glucosamine과 galactose가 78.0% 존재하는 것을 당조성 분석을 통해 확인하였다. 일반 성분, HPLC분석, 당 조성분석과 아미노산 분석결과로 glucosamine과 galacturonic acid가 주가 되어 threonine으로 연결된 구조를 하는 GAGs라는 것을 알 수 있다. 화장품 원료 기준에 맞추기 위해서는 sulfate 함량이 35.0~45.0%, 단백질 함량은 14.0~22.0%로 유지해야하므로 제단백결과 5.0%, 10.0%, 20.0% TCA (w/v) 처리, 10.0%, 20.0%, 40.0% HCI (v/v) 처리, UF(ultra filtration)를 포함한 10.0% TCA (w/v), 20.0% SSA (w/v), 25.0% HCI (v/v) 처리 결과 5.0% TCA (w/v) 및 10.0% HCI (v/v)이 화장품 기준에 적합하며 경제적이며 효율적이라는 것을 알 수 있었다.

• 한국식품영양과학회지
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• 제23권6호
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• pp.945-949
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• 1994

Chondroitin sulfate (Chs) contents in edible snail , Achatina fulica Bowdich , andits processed meat extracts were determined by high-performance liquid chormatogrpahy(HPLC) and spectrophotometric method. Spectrophotometric method was based on the precipitation of acriflavine by ChS, and HPLC method was based on the detection of two unsaturated disaccharides, 2-acetamido-2-deoxy-3-O-($\beta$ -D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose($\Delta$Di-4S) and 2-acetamido-2deoxy-3-O-($\beta$-D-gluco-4-eepyranosyluronic acid)-6-O-sulfo-D-galactose ($\Delta$야-6S) librated from ChS byenzymeatic digestion with chondroitinase ABc. the ratio of 125$\mu$mol of sodium hydroxide to mg of ChS and 8$0^{\circ}C$ of reaction temperature were proper for alkaline hydrolysis to remove protein residue form ChS. In assay preparation for HPLC ethod, the iptimum concentration of the enzyme chondroitinase ABc was 0.15 unit per 50 $\mu\textrm{g}$ of ChS at a fixed reaction time (30 min) and pH 8.0 using Tris buffer. ChS content in edible snail was 177.6mg% by spectrophotometric method and 153.5mg% by HPLC method and those in the processed meat extract was 71.3mg% by spectrometric method ad 62.8mg% by HPLC method, respectively.

• Journal of Pharmaceutical Investigation
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• 제30권3호
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• pp.159-166
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• 2000

Kunitz-type soybean trypsin inhibitor (SBTI) and chondroitin sulfate (A, and C type) were conjugated using sodium periodate method. And the physicochemical, pharmacokinetic properties and immunogenecity of the conjugates (Chon-A-SBTI or Chon-C-SBTI) were characterized. We expected the conjugation using chondroitin sulfate to reduce the immunogenecity and to improve the pharmacological effect. As the results, the mean molecular weight of the conjugate highly increased. After I.V. injection of the radiolabeled conjugates or native SBTI into mice, it was found that native SBTI showed rapid elimination from plasma, whereas Chon-A-SBTI and Chon-C-SBTI were slowly eliminated. Organ distribution of the two agents at 30 min after I.V. injection was different : Chon-A-SBTI or Chon-C-SBTI accumulated to a large extent in the liver (13% in Chon-A-SBTI and 16% in Chon-C-SBTI), whereas native SBTI was taken up more rapidly by the kidney (107% dose/g of tissue) and excreated into the urine (26%). In addition we evaluated the therapeutic value of the conjugates by using the sublethal septic shock model caused by pseudomonal elastase and tested the immunogenecity by passive cutaneous anaphylaxis shock (PCA). The conjugates were more effective than native SBTI against pseudomonal elastase induced septic shock in guinea pig. In case of the conjugates, the pharmacological and therapeutic effect lasted over 3 hours long. In immunogenecity test, both of the conjugates showed the reduction of their immunogenecity, especially Chon-A-SBTI looked most effective.

• 한국가축번식학회지
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• 제16권1호
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• pp.1-6
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• 1992

These experiments were undertaken as a basic study to understand the role of cumulus cell on in vitro maturation and fertilization process with identifying the cumulus cell-secreted proteins. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and fast protein liquid chromatography(FPLC), the proteins of cumulus cells were identified. The results obtained in these experiments were summarized as follows ; 1. When the proteins secreted from cultured cell for 30 hours were separated by SDS-PAGE, there were a major band (>94,000) and other minor 2 bands with molecular weight ranging 30,000∼67,000 dalton. 2. In addition, the fractionations of these proteins by FPLC were idirectly shown that three bands were hyaluronic acid, chondroitin sulfate, and heparin.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제19권3호
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• pp.265-286
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• 1997

Bone morphogenetic proteins(BMPs) are a group of transforming growth factor beta(TGF-${\beta}$)-related factors and multifunctional proteins, especially the only known biologic factors capable of inducing endochondral bone formation at an extraskeletal site. This study was performed to investigate the effect of the partially purified porcine BMP(pBMP) at an ectopic site. PBMP was partially purified from porcine bone matrix and its activity was monitored by an in vivo bioassay. The purification method utilized extraction of the bone-inducing activity with 4M guanidine, followed by chromatography on heparin-Sepharose. Active fractions were assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. And the fractions were reconstituted with inactive insoluble collagenous bone matrix from rats, acid soluble type I collagen from rat tail and chondroitin-6-sulfate sodium salt and implanted into the pectroralis muscle pouches of Sprague-Dawley rats. And the carrier complex was implanted on the opposite side as control. The rats were sacrificed at the day of 1st, 3rd, 5th, 7th, 11th, 14th and 21st after implantation and examined histologically, radiologically and biochemically. And alkaline phosphatase activity and calcium content were used as indices of bone formation. The results were as follows ; 1. Active fractions were localized in a zone between 31 and 40 KDa on SDS-PAGE. 2. The implanted 3.0mg of the partially purified pBMP induced cartilage and bone in the muscle tissue of rats through an endochondral ossification process. 3. Inactive insoluble bone matrix, type I collagen and chondroitin-6-sulfate have functioned as carriers for pBMP, but revealed some foreign body reactions. 4. Soft X-ray didn't reveal significant change between the experimental and the control group. 5. The alkaline phosphatase activities in the experimental group of 5th, 7th, 11th, 14th and 21st were increased significantly compared with control (p<0.01) with the peak in the group of 11th day. 6. With time, the calcium content of the experimental group increased. And the calcium contents in the experimental group of 11th, 14th and 21st were increased significantly compared with control (p<0.01).

• 한국미생물·생명공학회지
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• 제21권4호
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• pp.381-391
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• 1993

As the first stage of development of an artificial skin, fibroblasts were cultured in the collagen matrices to make a living dermal equivalent. Mouse embryonic fibroblasts were incorporated into a collagen matrices on plastic dishes containing concentrated DMEM culture media supplemented with sodium bicarbonate, hepes, antibiotics and fetal bovine serum. As the growth stimulation components, glycosaminoglycans were added: hyaluronic acid, chondroitin sulfate, heparin, chitosan were incorporated into the media at a concentration of either 1% or 5% w/w/ to collagen in order to investigate the effect on development of dermal equivalent. After the few days of incubation, gel matrics were contracted and firm dermal equivalent were formed. And the keratinocytes were cultured on top of dermal equivalent and make a three dimensional artificial skin tissue.

• 동의생리병리학회지
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• 제21권5호
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• pp.1285-1290
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• 2007

The present study aims to identify and characterize genes that cause differen genes between non-responders and responders to electroacupuncture (EA) on mechanical allodynia following peripheral nerve injury. Under sodium pentobarbital anesthesia, animals were subjected to unilateral transection of the superior caudal trunk at the level between S1 and S2 spinal nerves. EA stimulation (2Hz, 0.3 ms, 0.2-0.3 mA) was delivered to Zusanli (ST36) for 30 min 2 weeks after the surgery. The degree of mechanical allodynia was assessed quantitatively by touching the tail with von Frey hair (2.0 g) at 10 min intervals. The rats, which showed an EA-induced decrease of response frequencies under 10 %, were classified as non-responders and those displaying an EA-induced decrease of response frequencies 20 % or more were classified as responders. Results from oligonucleotide microarray, to which cDNAs from the spinal dorsal horn (DH) were applied, showed that hemoglobin beta chain complex and chondroitin sulfate proteoglycan-5 decreased and limbic system-associated membrane protein increased in the non-responder group, whereas calcium-independent alpha-Iatrotoxin receptor homolog-3 increased in the responder group. These results suggest that The functional abnormality of molecules regulating cell adhesion, intracellular signal transduction and cell differentiation in the spinal DH may be involved in the anti-allodynic effect of EA.