최은옥인하대학교식품영양학과 1. 1,200, %, 50%,, -linolenic acid. (linseed oil; 38%)..,, ( 1998),, (, 1998).,., -3/ -6,,,,. 433

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1 최은옥인하대학교식품영양학과 1. 1,200, %, 50%,, -linolenic acid. (linseed oil; 38%)..,, ( 1998),, (, 1998).,., -3/ -6,,,,. 433

2 ,,.,, -3/ -6,,,, 들깨의발아 cm (30) 12, 36, , , 36, 48 <50mm, 50 < 100mm, >100mm ( 1). 2.2 발아들깨의지질, 토코페롤, 인지질, 폴리페놀함량분석 (2:1, v/v) Folch(Folch J et al 1957) (, perilla seed lipids ) (TLC)., Pre-coated Kieselgel 60F254 TLC plate(merck Co., Darmstadt, Germany) (90 : 1 : 1, v/ v/v) 5% imaging densitometer(model GS-700 Biorad, Hercules, Calif., USA). 14% BF 3 - GC -3/ -6 (Kim and Choe 2008). Supelcowax 10 capillary column (30m0.53mM, 1.0M thick; Bellefonte, Pa, USA) flame ionization detector Younglin M600L gas chromatograph, oven, injector, and detector 230, 280, ml. (High perf ormance liquid 434

표 1 Sprouted perilla seeds 단계 Germination time (h) Length of bud (mm) 0 0 0 1 12 < 50 2 36 3

3 표 1 Sprouted perilla seeds 단계 Germination time (h) Length of bud (mm) < Picture < chromatography, HPLC)를 사용하여 구하였다(Lee and Choe 2003). 발아들깨 지질 0.1g을 n-헥산 1ml에 녹이고 hydrophobic 0.2μm membrane filter로 여과한 후, 20μl를 HPLC(Younglin 9100 HPLC System, Seoul)에 주입하였다. 컬럼은 u-porasil( mm, 10μm ID, Waters, Milford, MA, USA) 을 사용하였고, 이동상으로는 n-헥산과 이소프로판올의 혼합용액(99.8 : 0.2, v/v)을 사용하여 분 당 2.0ml의 속도로 용출시켰다. 이때 형광검출기(G1321A, Agilent 110 series, Germany)의 파장은 excitation 290nm, emission 330nm이었다. 발아들깨 지질의 토코페롤 함량은 표준 토코페롤 검량곡 선(그림 1)을 이용하여 구하였다. 발아들깨 지질에 함유된 인지질 함량은 Nzai와 Proctor(1998)의 방법을 이용하여 얇은막크로마 토그래피법(Thin layer chromatography)과 덴시토미터법(Densitometry)을 사용하여 구하였다. 발아 발아에 따른 들깨 지질과 유용성분의 변화 및 들기름의 산화안정성 435

4 1 Calibration curve for tocopherol determination. 436

5 (95 : 5, v/v) 0.1ml TLC, (75 : 25 : 3, v/v/v) TLC chamber. 2g 1 chamber 5, 5% (Model GS-700 Imaging densitometer, BIO-RAD, Hercules, CA, USA), PC PE ( 2). Lee (2007). 3g n- 10ml (60 : 40, v/v) 6ml ,000rpm, ml 5ml volumetric flask, 1ml 5ml volumetric flask. Folins 2 Calibration curve for phospholipid determination. 437

6 3 Calibration curve for polyphenol determination. Ciocaltau 0.3ml 3 Na 2 CO 3 0.5ml 5ml. 1 UV-Visible spectrophotometer 725nm. caffeic acid ( 3). 2.3 발아에의한들깨지질의산화평가 (2:1, v/v) Folch conjugated dienoic acid (AOCS Ti 1a-64) (AOCS Cd 8-53). 2.4 발아들깨유의헥산추출 120g n- 500ml mixer 40 2, Buchner funnel, 40 n- (, perilla seed 438

4 Preparation of samples. oil ). 2.5 발아들깨유의자동산화 7g (20ml). ( 4), 60 incubator 4,. 2. 2.6 발아들깨유의산화정도, 토코페롤, 인지질, 폴리페놀함량분석 conjugated dienoic acid (AOCS Ti 1a-64) (AOCS Cd 8-53).

7 4 Preparation of samples. oil ). 2.5 발아들깨유의자동산화 7g (20ml). ( 4), 60 incubator 4, 발아들깨유의산화정도, 토코페롤, 인지질, 폴리페놀함량분석 conjugated dienoic acid (AOCS Ti 1a-64) (AOCS Cd 8-53). 5% cupric acetate (ph 6.0~6.2) (H-500R, Kokusan ensinki CO., Ltd., Tokyo, Japan) 1,470g UV-Visible spectrophotometer(hp 8453, Hewlett Packard, Wilmington, DE, USA) 715nm (Lowry and Tinsley 1975) oleic acid ( 5, Absorbance=0.0039oleic acid (ppm) ; r 2 = ).,, (HPLC, TLC 439

8 5 Calibration curve for free fatty acid contents determination. with densitometry, spectrophotometry) 발아에따른들깨성분의변화 (2:1, v/v) Folch (dry basis) 35.31~37.59% ( 2). (>90%) (Neutral lipids, NL), ( 6, 3).. 440

2 Total lipid contents of perilla seeds affected by germination periods Germination period (h) Yield (%) 0 35.311.24 12 37.591.28 36 37.150.96 48 37.350.

9 2 Total lipid contents of perilla seeds affected by germination periods Germination period (h) Yield (%) Thin layer chromatogram of perilla seed lipid. 3 Composition of perilla seeds lipids affected by germination periods Germination period (h) Neutral lipids Glycolipids Phospholipids (94.35) 1) 0.02 (0.02) 7.01 (5.64) (95.03) 1.52 (1.54) 3.37 (3.42) (94.01) 2.12 (2.12) 3.88 (3.87) (89.47) 3.02 (2.88) 8.03 (7.65) 1) density (relative %). Folch ( 7). 8., Folch (~460nm), 0, nm 441

7 Color of the lipids extracted from perilla seeds affected

seeds affected by germination periods ( 0 h, 12 h, 36 h, 48

10 7 Color of the lipids extracted from perilla seeds affected by germination periods (step 0; 0 h, step 1; 12 h, step 2; 36 h, step 3; 48 h). 8 Absorption spectrum of the lipids extracted from perilla seeds affected by germination periods ( 0 h, 12 h, 36 h, 48 h).. (Fakourelis et al 1987),. Folch (>60%), (12~14%), (6~7%) (1~2%). 4 0, 12, 36, 48,., -6/

11 4 Fatty acid composition (relative %) of total lipids of perilla seeds affected by germination periods Germination period (h) Palmitic acid Stearic acid Oleic acid Linoleic acid Linolenic acid Tocopherol contents of perilla seed lipids affected by germination periods Germination period (h) Tocopherol content (mg/kg lipid) total Polyphenol contents of perilla seed lipids affected by germination periods Germination period (h) Polyphenol content (mg/100g lipid) , Folch 732~874ppm, (>78%)., - ( 5). 0 12, 36, ppm 36.9, 42.4, 46.9ppm ( 6). 443

12 7 Phosphatidylcholine (PC) and phosphatidylethanolamine contents (mg/kg lipid) of perilla seed lipids affected by germination periods Germination period (h) Phosphatidylcholine Phosphatidylethanolamine Peroxide (POV) and conjugated dienoic acid (CDA) values of perilla seed lipids affected by germination periods Germination period (h) POV (meq/kg oil) CDA (%) phosphatidylcholine phosphatidylethanolamine 112, 105ppm ( 7). Folch 8, 발아들깨유의자동산화안정성 37~38% Folch POV CDA 9, 9. Folch POV CDA., Folch 444

13 9 Peroxide value (meq/kg) and conjugated dienoic acid contents (%) of the perilla seed oil affected by germination periods during oxidation in the dark at 60 Germination period (h) Oxidation time (d) Peroxide value Conjugated dienoic acid a1) ab c b c a b c b c a b c b c c d d d d a a a a a b a a a a a ab b b b b c b 1) Different letters mean significant differences among the values at = c b.. 60 POV CDA. POV., 12, , POV CDA. POV 445

14 9 Peroxide value (a) and conjugated dienoic acid value (b) of the perilla seed oil affected by germination periods during oxidation in the dark at 60 (germination period ; 0 h, ; 12 h, ; 36 h, ; 48 h). 10 Regression equations 1) for the peroxide value of the perilla seed oil and oxidation time at 60 in the dark Germination time (h) a b r ) Peroxide value (meq/kg) = aoxidation time(day) + b, r: correlation coefficient., 12(0.543meq/kg/day) 48(0.672meq/kg/day), (1.522meq/kg/day) ( 10) meq/kg/day., lipase. 12,, 48, 0, 36 ( 11), ( 10). 446

15 11 Free fatty acid contents of the hexane-extracted perilla oil affected by germination periods Germination period (h) Free fatty acid contents (%, oleic acid) Relation between free fatty acid contents of the perilla seed oil. 12 Folch 발아들깨유의자동산화중토코페롤, 인지질, 폴리페놀의함량변화 0, 12, 36, , 952, 1018, 1100ppm Folch ( 732, 831, 839, 874ppm).,

16 12 Fatty acid compositions of the perilla seed oil during oxidation at 60 under dark Germination period (h) Oxidation time (d) Relative content (%) C16 : 0 C18 : 0 C18 : 1 C18 : 2 C18 : ( 11) ,. 448

17 13 Tocopherol contents of the perilla seed oil affected by germination periods during oxidation under dark at 60 Germination period (h) Oxidation time (d) Tocopherol (mg/kg oil) Total (100) 1) (100) (100) (100) (97.01) (94.40) (97.91) (94.64) (90.67) (91.83) (85.03) (91.66) (87.13) (85.96) (82.21) (86.00) (87.36) (83.67) (79.76) (83.90) (100) (100) (100) (100) (95.68) (94.47) (100.50) (94.79) (94.96) (95.45) (98.11) (95.38) (94.13) (96.02) (97.91) (95.64) (64.86) (94.05) (91.84) (87.81) (100) (100) (100) (100) (54.09) (95.42) (96.46) (85.83) (50.67) (90.38) (99.71) (81.26) (43.19) (82.62) (87.09) (73.51) (35.41) (79.79) (86.81) (69.56) (100) (100) (100) (100) (96.61) (93.89) (96.77) (94.55) (94.76) (92.92) (96.20) (93.38) (92.33) (93.86) (96.12) (93.53) (81.17) (82.77) (95.15) (82.54) 1) tocopherol content, ppm (retention % based on 0 oxidation time). 449

18 11 Relative retention (%) of (a) -, (b) -, (c) -, and (d) total tocopherols in the perilla seed oil affected by germination periods during oxidation in the dark at 60 (germination period ; 0 h, ; 12 h, ; 36 h, ; 48 h). 60 POV 15 (r>0.7). - POV. 450

19 14 Regression analysis for tocopherol retention (%) of the perilla seed oils affected by germination periods during oxidation under dark at 60 Germination period (h) Tocopherol Regression parameters 1) a b r Total Total Total Total ) Tocopherol retention (%) = a oxidation time + b, r: correlation coefficient. 15 Regression analysis between peroxide value (meq/kg) and tocopherol contents (ppm) in the perilla seed oil during oxidation under dark at 60 Tocopherol Germination period (h) Regression parameters 1) a b r

20 15 Tocopherol Germination period (h) Regression parameters 1) a b r Total ) Peroxide value (meq/kg) = a Total tocopherol content (ppm) + b, r: correlation coefficient. 16 Phosphatidylcholine (PC) and phosphatidylethanolamine (PE) contents of the perilla seed oil affected by germination periods during oxidation under dark at 60 Germination period (h) Oxidation time (d) PC PE (100) (100) (89.97) (95.05) (81.43) (92.72) (100) (100) (87.62) (91.65) (86.14) (88.21) (100) (100) 2 nd (0.00) (85.53) 4 nd (0.00) (89.35) (100) (100) (61.03) (91.83) (54.08) (82.95) 0, 12, 36, 48 Folch ( 16) PE PC 36 PC. 0, 12, 36, 48 (0.53~1.36mg/100g oil) Folch (3.09~4.69mg/100g oil). Folch - 452

21 17 Polyphenol (mg/100g oil) contents of the perilla seed oil during oxidation in the dark at 60 Germination period (h) Oxidation time (d) Polyphenol (100.00) (89.05) (84.39) (100.00) (63.72) (64.41) (100.00) (71.30)) (60.98) (100.00) (91.71) (92.84) ,, -3/ -6,,,,,,.,.,,,. linolenic acid oleic, stearic acid.,,. POV CDA

22 . 36.,,, AOCS Official methods and recommended practices of the American Oil Chemists Society. 4 th ed. AOAC Press, Champaign, IL., USA. Fakourelis N, Lee EC, Min DB Effects of chlorophyll and -carotene on the oxidation stability of olive oil. J. Food Sci. 52: Folch J, Lees M, Sloane-Stanley GH A simple method for the isolation and purification of total lipid from animal tissues. J Biol Chem 226: Kim H, Choe E Effects of egg yolk powder addition to the flour dough on the lipid oxidation development during frying. LWT-Food Sci. Technol. 41: Lee J, Choe E Lipid oxidation and tocopherol contents change in full-fat soy flour during storage. Food Sci Biotechnol. 12: Lee J, Kim M, Choe E Effects of carrot powder in dough on the lipid oxidation and caretene content of fried dough during storage in the dark. Journal of Food Science 69(5): C411-C414. Lee J, Lee Y, Choe E Temperature dependence of the autoxidation and antioxidants of soybean, sunflower, and olive oil. European Food Research and Technology A 226: Lowry RR, Tinsley IJ Rapid colorimetric determination of free fatty acids. J. Am. Oil Chem. Soc. 53: Nzai JM, Proctor A Phospholipids determination in vegetable oil by thin layer chromatography and imaging densitometry. Food Chem. 63: , : ,

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21(4)-02.fm J. Fd Hyg. Safety 21(4), 204 212 (2006) ŸŸ w» ƒ, ƒ, w ½ Á Á½ ¼Á½ Á Á½ Ášy Á» y Quantitative Analysis of Acid Value, Iodine Value and Fatty Acids Content in Sesame Oils by NIRS Jae-Kwan Kim, Myung-Jin Lee,