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임선영한국해양대학교해양환경생명과학부 1. (Allium cepa L.),,,, [1], [2-4], [5], [6], [7,8], [9], [10], [11].,,,, [12]. (polyphenol), C, E, - quercetin, quercitrin, rutin. diphenlypropane(c 6 -C 3 -C 6 ),,,, [13], Miean Mohamed[14] 62. hydroxylation, glycosylation,,,, 5,000 [15]..,,,,,. ally propyl disulfide dially disulfide quercetin, isorhamnetin, kaempferol, rutin [5,16]. 377

[17].,,, [18,19].. Bakhsh [20], Sklan [21] Dawa [22], Kendler[18]. Makheja [23] Srivastava[24] thromboxane. Hughes [25], Dorsch [26]. Jurdi-Haldeman [27].,. Woo [28],, Kim Lee[3],, LDL- HDL-,. 60g [29]. 50g., [30~32].,,, [33].,,,.... 24. Kim [34] 378

, 13.29% 12.99% 10.78%. C, > >. [34,35].. [36,37]. 10% 10%. quercetin quercetin 0.01%, 6.5% [12]. 0.4~0.5%, 25~32% 50 [45]... 2. 2.1 재료및추출. (STVD-50, Sanya, Korea) 15~25, 15~40 mmhg 24.. -75 379

acetone methylene chloride 1 : 1 24. 2 40 rotary vacuum evaporator(n-1000, EYELA, Japan) aceton/methylene chloride (A+M). A+M methanol 2 methanol (MeOH). hexane, 85% aq. MeOH, butanol(buoh), water. dimethyl sulfoxide (DMSO). DMSO 0.05%. 2.2 일반성분및비타민 C 측정,,, AOAC [38]., (105 ), Kjeldahl, Soxhlet, 550. C HPLC(P580, Dionex, USA) [39]. 1g, 10% HPO 3 9ml 3,000rpm 15 20L HPLC C. column (u-bondapak C 18 ), 0.05M KH 2 PO 4 / acetonitrile(60 : 40) 1ml/min UV/VIS detector 254nm. L-ascorbic acid(sigma-adrich, USA). 2.3 총플라보노이드측정 1ml 10ml diethyleneglycol 1N NaOH 1ml 37 1 UV-visible spectrophotometer(helios Beta, Thermo Electron Corporation, USA) 420nm. rutin [38]. 2.4 총페놀측정 Folin-Denies [40]. 380

1mg 1ml 10 2ml 2 Folin 2ml 3 10% Na 2 CO 3 2ml 1. UV-visible spectrophotometer(helios Beta, Thermo Electron Corporation, USA) 750nm. tannic acid(sigma-aldrich, USA). 2.5 유리아미노산비교분석 10g 75% 100ml 80, 2 40 0.2M sodium citrate buffer(ph 2.2)(Merck, Germany) 5ml Sep-Pak C 18 0.45M membrane filter [41]. 2.6 색도측정 (CR-10, Konica Minolta Sensing, Inc., Japan), 3 total color difference(e) [42]. 2.7 암세포및암세포배양 () (HT-1080), (AGS) (HT-29). 100units/ml penicillin-streptomycin 10% FBS DMEM RPMI 1640 37, 5% CO 2 incubator(mco-15ac, Sanyo Electric Biomedical Co., Japan). 2. phosphate buffered saline(pbs) 0.05% trypsin-0.02% EDTA(Gibco Co., USA) 75mm 3 cell culture flask 10ml 6~7. 2.8 MTT assay 96 well cell culture plate 510 4 cells/ml 100L 37, 5% 381

CO 2 incubator 24 well 100L, PBS 100L. plate 37, 5% CO 2 incubator 24. MTT assay 3-(4,5-dimethylthiazol)-2,5- diphenyltetrazolium bromide(mtt) 5mg 1ml PBS, 10% FBS 9ml 100L 3~4 MTT. formazan well MTT. well formazan DMSO 100L 5~10 microplate reader(victor3, Perkin Elmer, USA) 540nm. MTT, well [43]. 2.9 세포내활성산소종 (reactive oxygen species) 측정 HT-1080 100units/ml penicillin-streptomycin 10% FBS DMEM cell culture dish 37, 5% CO 2 incubator. DCFH-DA(dichlorodihydrofluorescin diacetate) assay [44]. DCFH-DA(Sigma- Aldrich, USA) (dichlorofluorescein, DCF). 96 well cell culture plate 24, PBS 20M DCFH-DA well 37, 5% CO 2 incubator 20 pre-incubation. well 37, 5% CO 2 incubator 1, DCFH-DA PBS 500M H 2 O 2 DCF fluorescence excitation 488nm, emission 530nm microplate reader(victor3, Perkin Elmer, USA). (blank control) PBS, control 500M H 2 O 2, blank 500M H 2 O 2 PBS. 382

3. 3.1 저온진공건조된양파분말의화학성분및생리활성 C C 1... 11.53% 7.15%. C, 6.27mg/100g 17.34mg/100g 9.55mg/100g. 1 Chemical composition of onion powder as affected by drying methods Components Raw Vacuum drying Freeze drying Moisture (%) 93.07 0.11 16.03 0.50 18.81 0.20 Carbohydrate (%) 0.65 0.01 62.08 0.20 56.92 0.16 Crude protein (%) 3.65 0.05 7.15 0.08 11.53 0.11 Crude fat (%) 0.55 0.001 1.18 0.32 1.48 0.29 Crude ash (%) 1.26 0.01 4.16 0.03 4.10 0.08 Vitamin C (mg/100 g) 6.27 17.34 9.55 2 phosphoethanolamine, L-aspartic acid, L-glutamic acid, L-alanine, L-citrulline, -aminobutryric acid. phosphoserine, phosphoethanolamine, L-valine, L-isoleucine. 383

2 Free amino acid contents of onion powder by different drying methods (ppm in sample) Amino acids Raw Vacuum drying Freeze drying Phosphoserine 7.9 148.1 57.4 Taurine - - - Phosphoethanolamine 720.2 7,870.4 1,588.3 Urea - - - L-Aspartic acid 119.1 323.3 2,060.3 L-Threonine 43.8 184.3 982.2 L-Serine 14.1 145.1 122.3 Asparagine - - - L-Glutamic acid 575.6 705.9 9,578.5 L--Aminoadipic acid - - - L-Proline - - - L-Glycine 14.2 208.5 540.5 L-Alanine 190.9 1,141.8 4,003.8 L-Citrulline 190.4 1,126.7 4791.7 L--Aminobutric acid - - - L-Valine 50.8 2,891.0 814.1 L-Cystine - - - L-Methionine - - - L-Isoleucine 23.1 754.7 429.4 L-Leucine 62.3 245.7 946.0 L-Tyrosine 27.5 150.2 570.9 L-Phenylalanine 37.5 355.7 621.6 -Alanine - - - -Aminobutyric acid - - - -Aminobutyric acid 2,476.5 37,739.9 50,412.0 NH 3 - - - L-Orinithine - - - L-Lysine 37.0 99.0 759.7 1-Mothyl-L-Histidine - - - L-Histidine 32.0 50.9 646.2 3-Methyl-L-Histidine - - - L-Carnosine - - - L-Arginine - - - Total 4,622.9 54,151.3 78,924.8 384

3. Hunter-L > >. Hunter-a 10.47. Hunter-b > >. 3 Color of onion powder as affected by drying methods Color Raw Vacuum drying Freeze drying L-value 58.47 3.86 67.70 1.01 75.87 1.42 a-value 3.27 2.11 10.47 0.25 1.47 0.12 b-value 19.70 0.44 25.60 0.44 28.53 0.71, C 4,. 93.1% 14.3% 4.6 45.3%, 0.36 5.0%. C, 6.27mg/100g. 19.67 951.1g/ml. 4 Chemical composition of onion flesh and peel Components Onion flesh Onion peel Moisture (%) 93.07 0.11 14.32 0.01 Carbohydrate (%) 4.60 0.05 45.29 0.06 Crude protein (%) 1.03 0.02 1.41 0.07 Crude fat (%) 0.15 0.001 1.11 0.01 Crude ash (%) 0.36 0.01 5.00 0.03 Vitamin C (mg/100 g) 6.27 - Total flavonoids (g/ml) 19.67 0.48 951.1 34.0 A+M MeOH 0.05mg/ml HT-1080. 500M H 2 O 2 control 385

120, A+M ( 1). 2 hexane, 85% aq. MeOH, butanol(buoh), water 0.05mg/ml control. ( 2A), 85% aq. MeOH BuOH hexane water control. ( 2B) 85% aq. MeOH hexane water contorl. A+M 85% aq MeOH BuOH. Ra [5] butanol> ethyl acetate> ethyl ether> water. Kim Kim[45], 100,, 1 Inhibitory effect of acetone/methylene chloride (A+M, 0.05mg/ml) and methanol (MeOH, 0.05mg/ml) extracts of onion flesh and peel on production of cellular reactive oxygen species in HT-1080 cells. Control was treated with 500M H 2 O 2 and phosphate buffered saline. Blank was treated with phosphate buffered saline without H 2 O 2. 386

2 Inhibitory effect of solvent fractions (0.05mg/ml, A; Onion peel: B; Onion flesh) of extracts from onion flesh and peel on production of cellular reactive oxygen species in HT-1080 cells. Control was treated with 500M H 2 O 2 and phosphate buffered saline. Blank was treated with phosphate buffered saline without H 2 O 2..,, 387

. LDL thiobarbituric acid reactive substances(tbars) LDL TBARS scavanger quercetin LDL. Park Kim[46] 5% TBARS quercitin. Shon [47] -carotene-linoleate system BHT L-ascorbic acid. MTT assay. 3 acetone/methylene chloride (A+M) methanol (MeOH) 0.1, 0.5 1mg/ml AGS. A+M 0.1mg/ml 3 Inhibitory effect of acetone/methylene chloride (A+M) and methanol (MeOH) extracts from onion flesh and peel on the growth of AGS human gastric adenocarcinoma cells. *p<0.05, significantly different from control. #p<0.05, significant effect between onion flesh and peel. 388

AGS 0.5mg/ml 95%, 1mg/ml 96% (p<0.05). A+M A+M 1mg/ml 13%. MeOH A+M 1mg/ml 23 35%. 4 hexane, 85% aq. MeOH, BuOH, water 4 Inhibitory effect of solvent fractions of extracts from onion flesh and peel on the growth of AGS human gastric adenocarcinoma cells. *p<0.05, significantly different from control. #p<0.05, significant effect between onion flesh and peel. 389

5 Inhibitory effect of acetone/methylene chloride (A+M) and methanol (MeOH) extracts from onion flesh and peel on the growth of HT-29 human colon cancer cells. * p<0.05, significantly different from control. # p<0.05, significant effect between onion flesh and peel. 85% aq. MeOH 0.5mg/ml 90% (p<0.05). HT-29 ( 5), AGS A+M 0.5 1mg/ml 90% MeOH AGS 1mg/ml 44%. A+M MeOH 1mg/ml 28 21%. 6 hexane, 85% aq. MeOH, BuOH, water HT-29 85% aq. MeOH (p<0.05). 85% aq. MeOH 0.5mg/ml 93% BuOH 46 42% (p<0.05). 5 IC 50 A+M IC 50 AGS HT-29 0.07 0.05mg/ml MeOH IC 50 0.75 3.31mg/ml AGS. 85% aq. MeOH AGS IC 50 0.04 0.03mg/ml 390

. HT-29 AGS. water. A+M MeOH. Lee [19] 9.2 6 Inhibitory effect of solvent fractions of extracts from onion flesh and peel on the growth of HT-29 human colon cancer cells. *p<0.05, significantly different from control. #p<0.05, significant effect between onion flesh and peel. 391

25 50mg 6.3 6.1 32 34%, quercitin, detoxification enzyme. Dorant [48] allicin, Sankaranrayanan [49] case-control. AGS. 85% aq. MeOH. 5 IC50 values of onion flesh and peel extracts on human cancer cell lines IC 50 Concentrations (mg/ml) AGS HT-29 A+M extract Flesh 2.54 6.63 Peel 0.07 0.05 MeOH extract Flesh 2.53 5.09 Peel 0.75 3.31 Hexane fraction Flesh 0.50 2.84 Peel 0.59 0.91 85% aq. MeOH fraction Flesh 0.04 0.23 Peel 0.03 0.04 BuOH fraction Flesh 0.28 0.57 Peel 0.31 0.78 Water fraction Flesh 1.85 4.76 Peel 1.16 3.65 392

3.1 건조붉은양파의화학성분및생리활성, C 17.95% 80% ( 6).,, 62.96, 8.74, 0.70 4.04%. C, 15mg/100g 96mg/100g. (34.29mg/ml) 39.05mg/ml. 6 Chemical composition of red onion Flesh red onion Dried red onion Moisture (%) 90.96 0.08 1) 17.95 0.03 Carbohydrate (%) 7.46 0.01 62.96 0.03 Crude protein (%) 0.55 0.01 8.74 0.03 Crude fat (%) 0.06 0.001 0.70 0.01 Crude ash (%) 0.35 0.01 4.04 0.03 Vitamin C (mg/100 g) 15.03 96.00 Total phenol (mg/ml) 34.29 2.06 39.05 2.38 1) Values are Mean S.E. 7 L-aspartic acid, L-glutamic acid, L-alanine, L-leucine, -aminobutryric acid. phosphoserine, phosphoethanolamine, L-aspartic acid, L-alarine, L-citrulline, L-valine, L-leucine, L-phenylalanine, -aminobutryric acid, L-histidine. 393

7 Free amino acid contents of red onion (ppm in sample) Amino acids Raw Vacuum drying Phosphoserine 4.9 877.0 Taurine - - Phosphoethanolamine 30.1 2,343.2 Urea - - L-Aspartic acid 167.9 604.4 L-Threonine 33.3 129.5 L-Serine 25.8 150.6 Asparagine - - L-Glutamic acid 123.8 755.2 L--Aminoadipic acid - - L-Proline - - L-Glycine 4.4 260.1 L-Alanine 82.8 1,184.7 L-Citrulline - 760.7 L--Aminobutric acid - - L-Valine 44.4 794.8 L-Cystine - - L-Methionine - - L-Isoleucine 19.4 72.2 L-Leucine 118.9 705.7 L-Tyrosine 46.6 99.3 L-Phenylalanine 20.8 387.4 -Alanine - - -Aminobutyric acid - - -Aminobutyric acid 3,437.9 28,479.9 NH 3 - - L-Orinithine - - L-Lysine 32.9 57.2 1-Mothyl-L-Histidine - - L-Histidine - 66.2 3-Methyl-L-Histidine - - L-Carnosine - - L-Arginine - - Total 4,194.0 37,728.1 394

8. Hunter-L Hunter-b. Hunter-a 5.87. 8 Color of red onion powder Color Raw Vacuum drying L-value 50.37 1.58 77.87 0.86 a-value 16.43 1.86 5.87 0.65 b-value 1.50 0.46 4.60 0.53 acetone/methylene chloride (A+M) methanol (MeOH) 0.1 5mg/ml HT-1080 9. A+M 0.25mg/ml HT-1080 2.5mg/ml 96%, 5mg/ml 98% 9 Inhibitory effect of acetone/methylene chloride (A+M) and methanol (MeOH) extracts of red onion powder on the growth of HT-1080 human fibrosarcoma cells Treatments A+M extract MeOH extract *p<0.05, significantly different from control. Concentration Inhibition IC 50 (mg/ml) (%) (mg/ml) 0.1 6 0.25 24* 0.5 42* 2.5 96* 5 98* 0.1 2 0.25 5 0.5 4 2.5 21* 5 50* 10 89* 0.94 5.46 395

(p<0.05). MeOH, MeOH 5 10mg/ml 50 89% (p<0.05). A+M MeOH IC 50 0.94 5.46mg/ml A+M MeOH. hexane, 85% aq. MeOH, butanol(buoh), water HT-1080 ( 10), 5mg/ml water 86% 10 Inhibitory effect of solvent fractions of extract from red onion powder on the growth of HT-1080 human fibrosarcoma cells Treatment Hexane fraction 85% aq. MeOH fraction BuOH fraction Water fraction *p<0.05, significantly different from control. Concentration Inhibition IC 50 (mg/ml) (%) (mg/ml) 0.1 32* 0.25 67* 0.5 88* 2.5 92* 5 93* 10 98* 0.1 8 0.25 7 0.5 10 2.5 86* 5 86* 10 98* 0.1 3 0.25 5 0.5 5 2.5 67* 5 94* 10 97* 0.1 8 0.25 11* 0.5 15* 2.5 44* 5 43* 10 86* 0.19 1.51 2.39 5.58 396

(p<0.05), hexane IC50 0.19mg/ml. 11 HT-29, HT-1080 A+M MeOH HT-29. 2.5 5mg/ml A+M 91 97% (p<0.05), IC 50 0.02mg/ml. MeOH 5 10mg/ml 58, 70% IC50 3.43mg/ ml HT-29 A+M (p<0.05). 12 hexane, 85% aq. MeOH, BuOH, water HT-1080 water 5mg/ml 88%. hexane 85% aq. MeOH IC 50 0.17 0.16mg/ml. HT-1080 HT-29 A+M MeOH. water HT-1080 HT-29. Rho Han[6] NCI-H522 NCI-H596 NCI-H522 48%, NCI-H596 25% NCI-H596, Lee 11 Inhibitory effect of acetone/methylene chloride (A+M) and methanol (MeOH) extracts of red onion powder on the growth of HT-29 human colon cancer cells Treatments A+M extract MeOH extract *p<0.05, significantly different from control. Concentration Inhibition IC 50 (mg/ml) (%) (mg/ml) 0.1 51* 0.25 59* 0.5 59* 2.5 91* 5 97* 0.1 35* 0.25 37* 0.5 37* 2.5 41* 5 58* 10 70* 0.02 3.43 397

[19] 20mg 55%, 25 50mg 32 34%. 12 Inhibitory effect of solvent fractions of extract from red onion powder on the growth of HT-29 human colon cancer cells Treatments Hexane fraction 85% aq. MeOH fraction BuOH fraction Water fraction *p<0.05, significantly different from control. Concentration Inhibition IC 50 (mg/ml) (%) (mg/ml) 0.1 37* 0.25 67* 0.5 80* 2.5 96* 5 98* 10 99* 0.1 37* 0.25 70* 0.5 88* 2.5 88* 5 88* 10 87* 0.1 16* 0.25 25* 0.5 36* 2.5 58* 5 95* 10 94* 0.1 29* 0.25 26* 0.5 28* 2.5 48* 5 52* 10 84* 0.17 0.16 2.15 4.41 398

A+M MeOH 0.01 0.05mg/ml HT-1080. 7 A+M MeOH 500M H 2 O 2 control 120. hexane, 85% aq. MeOH, BuOH, water 8 HT-1080 0.05mg/ ml 85% aq. MeOH. water. A+M, MeOH. Shon Park[50],. Shahidi Wanasundara[51] phenolic hydroxyl groups. Shon [47], 2. Jeong [52] C 19.20 28.34mg/100g C C. C. Kim [53] -carotene-linoleate system butylated hydroxy toluene(bht) L-ascorbic acid. 20~30%, FRAP(Ferric reducing ability of plasma), [54]. in vitro in vivo thiobarbituric acid reactive 399

7 Inhibitory effect of extracts of red onion powder on production of reactive oxygen species in HT-1080 cells. 1) Control was treated with 500M H 2 O 2 and phosphate buffered saline. 2) Blank was treated with phosphate buffered saline without H 2 O 2. 3) A+M 0.01, acetone with methylene chloride extract 0.01mg/ml; MeOH 0.01, methanol extract 0.01mg/ml (A). 4) A+M 0.05, acetone with methylene chloride extract 0.05mg/ml; MeOH 0.05, methanol extract 0.05mg/ml (B). 400

8 Inhibitory effect of solvent fractions of red onion powder on production of cellular reactive oxygen species in HT-1080 cells. 1) Control was treated with 500M H 2 O 2 and phosphate buffered saline. 2) Blank was treated with phosphate buffered saline without H 2 O 2. 3) Hexane 0.01, hexane fraction 0.01mg/ml; 85% aq. MeOH 0.01, 85% aqueous methanol fraction 0.01mg/ml; BuOH 0.01, butanol fraction 0.01mg/ml; water 0.01, water fraction 0.01mg/ml (A). 4) Hexane 0.05, hexane fraction 0.05mg/ml; 85% aq. MeOH 0.05, 85% aqueous methanol fraction 0.05mg/ml; BuOH 0.05, butanol fraction 0.05mg/ml; water 0.05, water fraction 0.05mg/ml (B). 401

substances(tbars) [55].. 4.... C. 48. A+M MeOH 0.05mg/ml (HT-1080). 85% aq. MeOH BuOH 85% aq. MeOH. A+M MeOH, (HT-29) (AGS). 85% aq. MeOH AGS IC 50 0.04 0.03mg/ml. 80% C. HT-1080 HT-29 A+M MeOH, hexane IC 50 0.19 mg/ml HT-1080. A+M, MeOH, hexane, 85% aq. MeOH, BuOH, water 500M H 2 O 2 control, 120. hexane 402

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