• Title/Summary/Keyword: Palmitoleic acid

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Direct Analysis in Real Time Mass Spectrometry (DART-MS) Analysis of Skin Metabolome Changes in the Ultraviolet B-Induced Mice

  • Park, Hye Min;Kim, Hye Jin;Jang, Young Pyo;Kim, Sun Yeou
    • Biomolecules & Therapeutics
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    • v.21 no.6
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    • pp.470-475
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    • 2013
  • Ultraviolet (UV) radiation is a major environmental factor that leads to acute and chronic reactions in the human skin. UV exposure induces wrinkle formation, DNA damage, and generation of reactive oxygen species (ROS). Most mechanistic studies of skin physiology and pharmacology related with UV-irradiated skin have focused on proteins and their related gene expression or single-targeted small molecules. The present study identified and analyzed the alteration of skin metabolites following UVB irradiation and topical retinyl palmitate (RP, 5%) treatment in hairless mice using direct analysis in real time (DART) time-of-flight mass spectrometry (TOF-MS) with multivariate analysis. Under the negative ion mode, the DART ion source successfully ionized various fatty acids including palmitoleic and linolenic acid. From DART-TOF-MS fingerprints measured in positive mode, the prominent dehydrated ion peak (m/z: 369, M+H-$H_2O$) of cholesterol was characterized in all three groups. In positive mode, the discrimination among three groups was much clearer than that in negative mode by using multivariate analysis of orthogonal partial-least squares-discriminant analysis (OPLS-DA). DART-TOF-MS can ionize various small organic molecules in living tissues and is an efficient alternative analytical tool for acquiring full chemical fingerprints from living tissues without requiring sample preparation. DART-MS measurement of skin tissue with multivariate analysis proved to be a powerful method to discriminate between experimental groups and to find biomarkers for various experiment models in skin dermatological research.

Identification of mountain-cultivated ginseng and cultivated ginseng using UPLC/oa-TOF MSE with a multivariate statistical sample-profiling strategy

  • Xu, Xin-fang;Cheng, Xian-long;Lin, Qing-hua;Li, Sha-sha;Jia, Zhe;Han, Ting;Lin, Rui-chao;Wang, Dan;Wei, Feng;Li, Xiang-ri
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.344-350
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    • 2016
  • Background: Mountain-cultivated ginseng (MCG) and cultivated ginseng (CG) both belong to Panax ginseng and have similar ingredients. However, their pharmacological activities are different due to their significantly different growth environments. Methods: An ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS)-based approach was developed to distinguish MCG and CG. Multivariate statistical methods, such as principal component analysis and supervised orthogonal partial-least-squares discrimination analysis were used to select the influential components. Results: Under optimized UPLC-QTOF-MS/MS conditions, 40 ginsenosides in both MCG and CG were unambiguously identified and tentatively assigned. The results showed that the characteristic components of CG and MCG included ginsenoside Ra3/isomer, gypenoside XVII, quinquenoside R1, ginsenoside Ra7, notoginsenoside Fe, ginsenoside Ra2, ginsenoside Rs6/Rs7, malonyl ginsenoside Rc, malonyl ginsenoside Rb1, malonyl ginsenoside Rb2, palmitoleic acid, and ethyl linoleate. The malony ginsenosides are abundant in CG, but higher levels of the minor ginsenosides were detected in MCG. Conclusion: This is the first time that the differences between CG and MCG have been observed systematically at the chemical level. Our results suggested that using the identified characteristic components as chemical markers to identify different ginseng products is effective and viable.

Development of animal protein(feed for fry) utilizing the rumen ciliates (제1위 섬모충(rumen ciliates)을 이용한 동물성 단백질(치어용 사료) 개발)

  • Jee, Cha-ho;Hyun, Gong-yool
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.327-336
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    • 1995
  • This study was carried out to develop the animal protein(feed for fry) that was isolated, purified and lyophilized the rumen ciliates from the healthy rumen contents which have $10^5-10^6/g$ ciliates and were discarded in abattoirs. The rumen ciliates are non-pathogenic, anaerobic and the weight of this protozoa is 2% of rumen content. The rumen protozoan and bacterial proteins both have a biological value for rats of 80-81, which is higher than the 72 of brewer's yeasts. Furthermore, the true digestibility and net protein utility of the protozoan protein are 91 and 73, much higher than those of bacterial(74 and 60) or yeast(84 and 60) proteins. The amino acids of rumen protozoa is nutritionally superior than the others. The size of rumen ciliates is $30-200{\times}20-110{\mu}m$ and so we had isolated and purified the rumen ciliates from the rumen contents by the physical methods. The purified rumen protozoa was lyophilized with freezing dryer. The results of this experiment were as follows : 1. Population dynamics of protozoan ciliates in slaughtered rumens; % of samples which small ciliates were predominated was 82.5%(52/63) and that of large ciliates was 17.5%(11/63). 1) predominant species of small ciliates were Entodinium ovinum and E nanellum. 2) predominant species of large ciliates were Epidinium ecaudatum and Diploplastron affine. 2. The lyophilized rumen ciliates which were isolated and purified from 1 kg of rumen content at the pH 6.2-6.8 was about 7.0 gram. 3. The nutrient analysis of lyophilized rqmen ciliates(LRC) was as follows: 1) Proximate analysis of the LRC and the composition of fry feed; moisture 8.05%(below 10.0), protein 35.37%(45), fat 5.39%(4.5), fiber 1.23%(below 2.5), ash 2.25%(below 15.0), Ca 0.26%(below 2.0), P 0.14%(below 1.1), energy 4,608.11(fish meal 5000 cal/g) 2) Amino acids (% in crude protein) of the LRC and the rotifer(Brachionus plicatilis); Arg 5.19%(4.50), His 2.50%(1.55), Ile 5.29%(3.45), Leu 8.11%(5.85), Lys 10.34%(6.15), Met 2.25% (0.85), Phe 5.66%(3.80), Thr 5.14% (3.45), Val 4.18%(3.90), Ala 4.13%(3.35), Asp 13.26%(8.25), Glu 16.62%(9.20), Gly 4.23%(3.10), Pro 3.25%(5.05), Ser 4.85%(3.85), Tyr 5.04%(3.05) 3) Fatty acids(% in fat) of the LRC and the rotifer(biological feed ; Brachionus plicatilis); myristic acid(C14:0) 3.27%(0.3), myristoleic acid(C14:1) 0.83%(-), palmitic acid(C16:0) 39.11% (23.5), palmitoleic acid(C16:1) 2.81%(2.0), stearic acid(C18:0) 9.36%(5.6), oleic acid(C18:1) 25.54%(3.5), linoleic acid(C18:2) 15.05%(32.9), linolenic acid(C18:3) 1.74%(9.8). Judging from the above investigated results, the analytical data of proximate analysis, amino acids, fatty acids of the purified and lyophilized rumen protozoa are reasonable for the feed of freshwater fishes(fry and fingerling). But it was disappointed of our expectation that the crude protein of lyophilized rumen ciliates contains low percentage, it was thought that because of the small ciliates(starch digester) in beef cattle rumens which were administered the concentrated feed, is much difficult to isolate and purify than the large ciliates(fiber digester).

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Purification and Characterization of the Red Carotenoprotein from the Muscle of Blue Mussel, Mytilus edulis (진주담치 근육중의 적색 Carotenoprotein의 정제 및 특성)

  • YANG Huyn-Pil;LEE An-Jong;KIM Yong-Tae;KIM Se-Kwon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.5
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    • pp.482-494
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    • 1994
  • Most of carotenoprotein complexes have been extracted by using buffered solutions. However, in this study carotenoprotein from the muscle of Blue mussel(Mytilus edulis) was extracted by a detergent such as Triton X-100. It was purified and characterized by $20\%$ (w/v) $(NH_4)_2SO_4$, DEAE-cellulose ion exchange and Sephacryl S-300 gel filtration. The carotenoprotein(${\lambda}_{max}=462nm$) had an approximate M. W. of 372KDa(gel filtration). SDS-PAGE analysis of the carotenoprotein indicated the presence of four polypeptides of 60KDa($23.70\%$), 46.9KDa($9.14\%$), 26KDa($49.14\%$) and 13KDa($18.02\%$). Carotenoprotein denaturated by treatment with SDS to a final concentration of $0.2\%$ (w/v) caused a hypsochromic shift of ${\lambda}_{max}$ from 462nm to 456nm. The carotenoprotein contained lipids as structure units. The amino acid composition of the carotenoprotein contained large essential amino acid amounts of $62.8\%$, and the content of threonine($35.9\%$) was higher than other amino acids, but histidine, methionine and proline were not present. In the carotenoprotein, the major fatty acids were $C_{16:4},\;C_{16:0},\;C_{20:5}\;and\;C_{22:6}$. The percentages of polyunsaturated fatty acids($62.4\%$) were higher compared to other fatty acids(saturated fatty acids $19.6\%$, monounsaturated fatty acids $18.0\%$). Carotenoid was extracted from the carotenoprotein by acetone and it was separated into five different components by preparative TLC(benzene:petroleum ether:acetone=69:17:14). The major components of carotenoid were mytiloxanthin($74.79\%$) and 3,4,3'- trihydroxy-7',8'-didehydro-${\beta}$-carotene($18.26\%$), and they were at least presented as prosthetic groups of carotenoprotein.

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Effects of α-, γ-, and δ-tocopherol on the oxidative stability of horse fat (마유(Horse Fat)의 산화안정성에 대한 α-, γ-, δ-토코페롤의 첨가 효과)

  • Park, Youn Hyung;Cho, Man Jae;Kim, Hyun Jung
    • Korean Journal of Food Science and Technology
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    • v.50 no.3
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    • pp.267-273
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    • 2018
  • Horse fat was extracted from fatty horse meat at $70^{\circ}C$ under vacuum conditions. The oxidative stability of horse fat was investigated by the addition of 0, 30, 60, and 150 mg/kg of ${\alpha}$-, ${\gamma}$-, or ${\delta}$-tocopherol during storage of 14 days at $65^{\circ}C$ in the dark. Changes of tocopherol concentration and fatty acid composition, peroxide value, and 2-thiobarbituric acid (TBA) value were analyzed during storage. The levels of the added tocopherols were found to decrease during storage. Unsaturated fatty acids contents of horse fat without tocopherol decreased from 60.87% to 57.22% after 14 days. The peroxide value and TBA value increased as storage time increased. The peroxide values of horse fat after addition of 0, 30, 60, and 150 mg/kg of ${\gamma}$-tocopherol were 43.75, 25.17, 20.87, and 15.41 meq/kg, respectively, and the TBA values were 7.87, 5.64, 4.43, and 4.23 mg malonaldehyde (MA)/kg, respectively, after 14 days. At the concentration of 150 mg/kg, both ${\gamma}$- and ${\delta}$-tocopherol impeded the oxidation of horse fat during storage.

Variation of Species and Nutrient Composition of Benthic Diatoms on Plate during Culture Periods I. Wando area (배양기간에 따른 파판 부착규조류 종조성과 영양성분 변화 I. 완도지역)

  • 김해영;허성범
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.141-142
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    • 2003
  • 본 연구는 전복의 유생사육을 목적으로 먹이생물인 부착미세조류의 파판 배양시 배양기간에 따른 종조성과 영양성분의 변화를 파악하기 위하여 수행되었다. 실험은 완도지역에서 봄철인 2~4월에 8주 동안 실시되었다. 배양을 위한 기초 조사로 배양지역 연안해수의 수온, 염분, pH, 총질소, 총인, Chlorophyll-a를 측정 하였다. 실험에 사용된 파판은 전복 종묘생산에 이용되는 규조배양용 파판과 틀을 이용하여 2주마다 파판을 수거한 뒤 부착된 미세조류를 부드러운 솔로 분리하여 원심분리한 후 지방산과 아미노산을 측정하였다. 각각의 시료는 광학현미경(Nikon)으로 동정한 후 Sedwick-rafter counting chamber를 이용하여 종별로 계수하고 이를 단위면적당 개체수 (cells/$\textrm{cm}^2$)로 환산하여 정량분석을 하였다. 실험 결과 봄철 완도지역의 평균 수온과 염분은 각각 15.4$^{\circ}C$와 33.88$\textperthousand$이었고 pH는 8.21이었다. 총질소와 총인은 각각 0.078mg/$\ell$, 0.018mg/$\ell$였고 chlorophyll-$\alpha$는 1.47$\mu$g/$\ell$ 였다. 부착미세조류의 아미노산 분석 결과는 배양 2주째의 총아미노산 양이 5.63%로 가장 낮았고, 4주는 6.50%, 6주 7.82% 8주 7.87%로, 6주까지 증가하다가 정체하는 경향을 보였다. 필수아미노산의 경우 총아미노산의 경향과 비슷하였고 2, 4, 6, 8주째의 양은 각각 2.75%, 3.16%, 4.01%, 3.96%였다. 필수 아미노산과 비필수아미노산의 비는 2, 4, 6, 8주 각각 0.95, .095, 1.05, 1.01의 결과를 보였다. 지방산의 경우 palmitic acid( $C_{16:0}$)와 palmitoleic acid( $C_{16:1}$)가 43.10~67.66%의 함량을 보였으나 4주째에는 검출되지 않았으며 EPA는 6주에서 10.97%를 보였고 나머지 구간에서는 검출되지 않았다. 부착미세조류의 biomass는 기간별로 2.4$\times$$10^3$ cells/$\textrm{cm}^2$, 7.8$\times$$10^3$ cells/$\textrm{cm}^2$ 25.1$\times$$10^3$ cells/$\textrm{cm}^2$, 44.3$\times$$10^3$ cells/$\textrm{cm}^2$ 이었다. 출현량은 총 64종으로 대부분이 부착규조류였으며 2주, 4주, 6주, 8주 각각 43종, 25종, 19종, 24종의 부착 규조류가 출현하였다. 그리고 우점종은 2주째 Nitzschia petitiana, Fragilia psedonana, 4주 Navicula mollis, Flagilia cylindrus, 6주 Flagilia capucaina, Achnanthes haukiana, 8주 Flagilia islandica, Flagilia cylindrus였다.s였다.

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Variations of Fish Community and Fish Density on Artificial Reefs (인공어초어장의 어류 군집상과 어획량 변동)

  • LEE Jeong-Woo;KANG Young-Shil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.5
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    • pp.535-548
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    • 1994
  • Community structures and fish densit were investigated on three different types of artificial reefs, dice, turtle artificial reef and tubes, constructed in the Korean waters. Variations of fish fauna according to type of artificial reef and the proper artificial reef in each area for optimizing harvest were discussed. Fish were captured by trammel gill net during May, June, September and November, 1988 and both identified and counted. Fourty-five fish species were found in the artificial reefs. Of these, Sebastes spp., Hexagrammos otakii, Pleuronectidae, Navodon modestus and Stephanolepis cirrhifer showed high occurrence-frequency. The dominant species groups were coastal settlement, demersal or rock fishes such as Pleuronectidae, Rajiformes, Stephanolepis cirrhifer, Navodon modestus, Hexagrammos otakii and Sebastes spp. in all of the Artificial reefs except the oceanic area of southern waters. Scomber japonicus was predominant in the oceanic area of southern waters. Composition of demersal, rock and pelagic fishes were different depending on the types of artificial reef. Dice artificial reefs were occupied by rock fish, on the other hand turtle artificial reefs were dominated by dermersal fish. Fish density was high at the dice artificial reef in all survey areas except the middle area of Eastern waters, with high fish density evident in the Tube artificial reef. Fish community structures were remarkably different between Dice and Turtle artificial reefs. The Tube artificial reef showed intermediate characteristics between the above two types of artificial reefs. The coastal areas of Southern waters and the middle and southern areas of Western waters revealed similar fish fauna. Results from the oceanic areas of Southern waters were well associated with the middle and southern areas of Eastern waters.

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Characterization of a Korean Domestic Cyanobacterium Limnothrix sp. KNUA012 for Biofuel Feedstock (토착 남세균 림노트릭스 속 KNUA012 균주의 바이오연료 원료로서의 특성 연구)

  • Hong, Ji Won;Jo, Seung-Woo;Kim, Oh Hong;Jeong, Mi Rang;Kim, Hyeon;Park, Kyung Mok;Lee, Kyoung In;Yoon, Ho-Sung
    • Journal of Life Science
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    • v.26 no.4
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    • pp.460-467
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    • 2016
  • A filamentous cyanobacterium, Limnothrix sp. KNUA012, was axenically isolated from a freshwater bloom sample in Lake Hapcheon, Hapcheon-gun, Gyeongsangnam-do, Korea. Its morphological and molecular characteristics led to identification of the isolate as a member of the genus Limnothrix. Maximal growth was attained when the culture was incubated at 25℃. Analysis of its lipid composition revealed that strain KNUA012 could autotrophically synthesize alkanes, such as pentadecane (C15H32) and heptadecane (C17H36), which can be directly used as fuel without requiring a transesterification step. Two genes involved in alkane biosynthesis-an acyl-acyl carrier protein reductase and an aldehyde decarbonylase-were present in this cyanobacterium. Some common algal biodiesel constituents-myristoleic acid (C14:1), palmitic acid (C16:0), and palmitoleic acid (C16:1)-were produced by strain KNUA012 as its major fatty acids. A proximate analysis showed that the volatile matter content was 86.0% and an ultimate analysis indicated that the higher heating value was 19.8 MJ kg−1. The isolate also autotrophically produced 21.4 mg g−1 phycocyanin-a high-value antioxidant compound. Therefore, Limnothrix sp. KNUA012 appears to show promise for application in cost-effective production of microalga-based biofuels and biomass feedstock over crop plants.

Comparison of the Chemical Compositions of Korean and Chinese Safflower Flower(Carthamus tinctorius L.) (한국산과 중국산 홍화꽃의 화학적 성분조성비교)

  • 박금순;박어진
    • Korean journal of food and cookery science
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    • v.19 no.5
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    • pp.603-608
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    • 2003
  • In approximate composition, crude protein, lipid, ash, crude fiber, and N-free extract constituted 14.70%, 3.10%, 6.90%, 18.20%, and 57.10%, respectively, in Korean safflower flowers, compared to 12.60%, 2.70%, 5.80%, 16.40% and 62.50%, respectively, in Chinese safflower flowers. This indicated that Korean safflower flowers surpassed their Chinese counterparts except in terms of N-free extract. Free sugars such as fructose, glucose, and sucrose were proven to be dominant in both domestic and Chinese safflower flowers, while little xylose was contained. For content of polyphenolic compound, Korean safflower flowers contained 13.85% water soluble extract and 9.70% MeOH extract, compared to 9.39% and 7.04%, respectively, for the Chinese variety, confirming the higher levels in the Korean variety. For fatty acids, (Ed- the following results are not presented in ratio form) saturated fatty acids and unsaturated fatty acids comprised 6.80% and 93.20% in Korean safflower flowers and 16.0% and 84.0% in Chinese safflower flowers, respectively. Linoleic, oleic, and palmitoleic acids comprised 75.30%, 11.60%, and 3.40% in Korean safflower flowers, and 66.70%, 11.20%, and 6.10% in the Chinese variety, respectively. Of amino acids, essential amino acids comprised 46.67% in Korean safflower flowers and 36.79% in the Chinese variety. Moreover, total essential amino acids in Korean safflower flowers were higher than those of their counterparts. Non-essential amino acid comprised 65.17% in the Korean variety and 54.49% in the Chinese. In terms of mineral content, Korean safflower flowers contained more Ca, Cu, Fe and Mn than those of China, while Chinese safflower flowers contained more A1, Ba, Mg, K, Na, Zn, Sr and P.

Nutrient Composition of Benthic Diatom Cultured on Plate at Four Areas in Spring (봄철 해역별 파판배양 부착규조류의 영양성분)

  • 김해영;허성범
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.143-144
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    • 2003
  • 봄철 해역에 따른 부착미세조류의 영양성분 변화를 알아보기 위하여 본 실험을 수행하였다. 실험은 울진, 부안, 완도, 부산 지역에서 봄철인 2~4월 동안 실시되었으며 실험간의 오차를 줄이기 위해 실험 시작일의 차이는 최대 10일 이내로 하였다. 배양을 위한 실험해역의 기초 조사로 배양지역 연안해수의 수온, 염분, pH, 총질소, 총인, Chlorophyll-a의 측정을 하였고 각 지역의 부착미세조류를 수확하여 영양분석을 하였다. 실험에 사용된 파판은 전복 종묘생산에 이용되는 규조배양용 파판과 틀을 이용하였고 실험기간은 총 8주 동안 실시하였다. 2주마다 파판을 수거한 뒤 부착된 미세조류를 부드러운 솔로 분리하여 원심 분리한 후 지방산과 아미노산의 측정을 하였다. 또 각 시료의 일정량을 Sedwick-rafter counting chamber를 이용하여 검경ㆍ계수한 후 이를 단위면적당 개체수(cells/$\textrm{cm}^2$)로 환산하여 biomass를 구하였다. 봄철 지역간의 수온의 범위는 13.9~16.6$^{\circ}C$를 보였으며 울진이 가장 높고 부안지역이 가장 낮았다. 염분은 부안지역의 염분이 평균 2$\textperthousand$ 이상 낮았으며 그 외의 지역은 큰 차이가 없었다. pH는 7.77~8.21의 범위를 보이고 있었으며 질산염 농토는 부산이 0.265mg/$\ell$로 최대값을 보였고 울진이 0.032mg/$\ell$로 제일 낮았다. 인산염 농도는 완도지역이 0.018mg/$\ell$로 최대치를 보였고 부안은 검출되지 않았다. Chlorophyll-a의 농도는 울진지역이 0.66mg/$\ell$으로 제일 낮았으며 부산이 2.69mg/$\ell$, 부안지역이 2.27mg/$\ell$ 로 높은 값을 보였다. 봄철 지역에 따른 부착미세조류의 아미노산 분석결과는 울진이 실험지역 중 제일 높은 총 아미노산과 필수아미노산의 함량을 나타냈으며 각 지역별로 울진은 4주에 20.3%, 부안은 8주째 8.1%, 완도는 8주째 4.7%, 부산은 6주째 10.4%로 서로다른 시기에 높은 아미노산의 함량을 나타냈다. 실험지역 부착미세조류의 지방산 결과를 보면 전 실험기간동안 지방산 중 대부분은 $C_{18}$이하의 포화 또는 불포화 지방산이 주를 이루고 있었으며 palmitic acid( $C_{16:0}$)와 palmitoleic acid( $C_{16:0}$)의 양이 전체 지방산 함량중 37.47~75.94%로 대부분을 차지하고 있었다. 그리고 n-3HUFA(high unsaturated fat쇼 acids)인 EPA는 부안지역이 2주째 20.85%로 전체 지역중 함량이 제일 높았고, 울진은 실험기간중 6주가 14.76%로 제일 높은 함량을 보였다. 완도는 6주에만 10.97%의 함량을 보였으며 부산지역의 시료에서는 EPA의 검출이 없었다. 봄철 각지역에 설치된 파판에서의 부착미세조류 출현량을 보면 2주째의 부착미세조류량은 부안이 8.3$\times$$10^3$ cells/$\textrm{cm}^2$로 가장 높았고 부산이 0.6$\times$$10^3$ cells/$\textrm{cm}^2$로 가장 낮았다. 울진의 경우 0.7$\times$$10^3$ cells/$\textrm{cm}^2$로 부산과 비슷하게 낮은 출현량을 보였고 완도는 2.4$\times$$10^3$ cells/$\textrm{cm}^2$였다. 4주째의 출현량은 울진지역이 471.1$\times$$10^3$ cells/$\textrm{cm}^2$로 가장 높았으며 부안, 완도, 부산 순으로 출현량이 낮았고 특히 부산은 1.2$\times$$10^3$ cells/$\textrm{cm}^2$로 가장 낮았다. 6주째의 출현량은 울진 지역이 368.6$\times$$10^3$ cells/$\textrm{cm}^2$로 가장 높았으며 부안 89.8$\times$$10^3$ cells/$\textrm{cm}^2$, 완도 25.1$\times$$10^3$ cells/$\textrm{cm}^2$, 부산 5.9$\times$$10^3$ cells/$\textrm{cm}^2$ 순이었다. 8주째의 경우에도 울진, 부안, 완도, 부산이 각각 255.8$\times$$10^3$ cells/$\textrm{cm}^2$, 157.8$\times$$10^3$ cells/$\textrm{cm}^2$, 44.3$\times$$10^3$ cells/$\textrm{cm}^2$, 1.1$\times$$10^3$ cells/$\textrm{cm}^2$로 4주, 6주의 경우와 같은 경향을 보였다.보였다.

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