1、 KSKSKSKSKSKSKSK KSKSKS KSKSK KSKS KSK KS KS H ISO 12228 KS H ISO 12228:2008 2008 8 6 http:/www.kats.go.krKS H ISO 12228:2008 : ( ) ( ) () () ( ) : (http:/www.standard.go.kr) ： ： 2003 8 30 ： 2008 8 6 2008-0410 ： ： ( 02-509-7266) (http:/www.kats.go.kr). 10 5 , . KS H ISO 12228:2008 i .1 1 1 2 1 3 .1
2、4 2 5 2 5.1 2 5.2 .2 5.3 .2 5.4 2 5.5 2 5.6 (TLC) .2 5.7 2 5.8 2 5.9 3 5.10 (silylating) 3 6 .3 6.1 .3 6.2 3 6.3 3 6.4 3 6.5 .3 6.6 3 6.7 .3 6.8 3 6.9 (reaction vials) .3 6.10 .3 6.11 3 6.12 .4 6.13 4 7 .4 8 4 9 4 9.1 .4 9.2 .4 KS H ISO 12228:2008 ii 10 .6 10.1 6 10.2 6 10.3 7 11 8 11.1 .8 11.2 r .8
3、 11.3 R8 12 .8 A( ) .11 B( ) 16 KS H ISO 12228:2008 Animal and vegetable fats and oils Determination of individual and total sterols contents Gas chromatographic method 1999 1 ISO 12228, Animal and vegetable fats and oils Determination of individual and total sterols contents Gas chromatographic met
4、hod , . ISO 12228 ISO/TC 34 (SC 11) . A B . 1 . 2 . . ( ) . KS H ISO 661, KS M ISO 3696, 3 . 3.1 , 7- (7-avenasterol)( 1 ) (%) 100 % . 3.2 KS H ISO 12228:2008 2 7- (7-avenasterol)( 1 ) , 100 g mg( mg/100 g) . 4 . . . . . . 5 , KS M ISO 3696 3 . 5.1 , c(KOH) 0.5 mol/L 5 mL 3 g (5.3) 100 mL . (straw-c
5、oloured) . 5.2 1.0 mg/mL (5.10 ) 5- -3- (5-cholestan-3-ol)( 1 2) . 5.3 95 %(V/V) 5.4 , 0.0630.200 mm, I . 5.5 . . 1.748 %(V/V) . . 5.6 (TLC) 20 cm20 cm, 0.25 mm . 5.7 / 1/1(V/V) 5.8 KS H ISO 12228:2008 3 1.0 mg/mL 5.0 mg/mL 5.9 5.10 (silylating) N- -N-( )- - N-methyl-N-(trimethylsilyl)-hepta-fluorob
6、utyramide(MSHFBA) 1 mL 1- (1-methyl imidazole) 50 L . . GLC 2 . 6 . 6.1 25 mL 50 mL . 6.2 (6.1) . 6.3 PTFE , (sintered glass frit) 100 mL (reservoir) . 25 cm, 1.5 cm 6.4 40 . 6.5 20 cm20 cm . 6.6 100 L . 6.7 (1053) . 6.8 TLC 6.9 (reaction vials) 0.3 mL PTFE , . 6.10 , . 6.11 KS H ISO 12228:2008 4 25
7、60 m, 0.20.25 mm, SE-54( 280300 ). 0.1 m 6.12 1 L 6.13 0.001 g , 0.000 1 g . 7 . KS H ISO 5555 1 . 8 KS H ISO 661 . 9 9.1 25 mL (6.1) 250 mg 1 mg . 100 g 100 mg 3 . . 9.2 9.2.1 (5.3) 20 mL (5.4) 10 g (6.3) . . 9.2.2 (9.1) (5.2) 1.00 mL . (5.1) 5 mL (anti-bumping granules) . (6.2) 15 . . (5.3) 5 mL .
8、 (9.2.1) 5 mL . 50 mL (6.1) . 2 mL/min (5.3) 5 mL , (5.5) 30 mL KS H ISO 12228:2008 5 . (6.4) . . . 9.2.3 (5.5) 9.2.2 . (6.6) TLC (5.6) 2 cm . 3 cm . 1.5 cm 5 L TLC (5.8) . 100 mL (5.7) (6.5) . . . TLC (9.2.2) . . . 9.2.4 ( ) (5.9) . . ( 1 ) 2 mm 4 mm . . ( 4 mm 2 mm) . 3 (5- , 7- ). 0.5 mL . (5.5)
9、5 mL 3 . 1 mL (6.9) . . 9.2.5 (6.9) (5.10) 100 L . 105 15 . (6.10) . 9.2.6 2 . 2 . 1 . GC ： SE-54, 50 m, 0.25 mm, 0.10 m, H2, 36 cm/s, 1： 20, / 320 , 4 /min 240255 , 1 L. . 2 , , . , , , ( ) (blank run) . KS H ISO 12228:2008 6 10 10.1 (RT) / (RT) (RRT) . SE-54 (RRTC) (RRTB) RRT 1 . 1 RRT(9.2.6 1 ) (
10、 2 .) . RRT (GLC , ) 1 RRT . 10.2 i Ci . 100=AACii Ai ： i A ： ( 116) KS H ISO 12228:2008 7 1 RRT GLC (SE-54 ) RRTC RRTB1 (cholesterol) -5- -3- (cholest-5-en-3-ol) 1.00 0.44 2 (cholestanol) 5- -3- (5-cholestan-3-ol) 1.02 0.45 3 (brassicasterol) 24S-24- -5,22- -3- (24S-24-methyl cholesta-5,22-dien-3-o
11、l) 1.09 0.48 4 24- (24-methylene cholesterol) 24- -5,24- -3- (24-methylene cholesta-5,24 -dien-3-ol) 1.21 0.53 5 (campesterol) 24R-24- -5- -3- (24R-24-methyl cholest-5-en-3-ol) 1.23 0.54 6 (campestanol) 24R-24- -3- (24R-24-methyl cholestan-3-ol) 1.25 0.55 7 (stigmasterol) 24S-24- -5,22- -3- (24S-24
12、ethyl cholesta -5,22-dien-3-ol) 1.31 0.57 8 7- (7-campestrol) 24R-24- -7- -3- (24R-24 methyl cholest-7-en-3-ol) 1.38 0.59 9 5,23- (5,23-stigmastadienol) 24R,S-24- -5,23- -3- (24R,S-24-ethyl cholesta-5,23-dien-3-ol) 1.40 0.60 10 (clerosterol) 24S-24- -5,25- -3- (24S-24-ethyl cholesta-5,25-dien-3-ol)
13、1.42 0.62 11 (sitosterol) 24R-24- -5- -3- (24R-24-ethyl cholest-5-en-3-ol) 1.47 0.64 12 (sitostanol) 24R-24- -3- (24R-24-ethyl cholestan-3-ol) 1.50 0.65 13 5- (5-avenasterol) 24Z-24(28)- -5- -3- (24Z-24(28)-ethylidene cholest-5-en-3-ol) 1.52 0.66 14 5,24- (5,24-stigmastadienol) 24R,S-24- -5,24- -3-
14、(24R,S-24-ethyl cholesta-5,24-dien-3-ol) 1.59 0.69 15 7- (7-stigmastenol) 24R,S-24- -7- -3- (24R,S-24-ethyl cholest-7-en-3-ol) 1.65 0.72 16 7- (7-avenasterol) 24Z-24(28)- -7- -3- )(24Z-24(28)-ethylidene cholest-7-en-3-ol) 1.70 0.74 X (erythrodiol) 2.03 0.88 Y (uvaol) 2.17 0.95 17 (betulin) Lup-2029-
15、 -3,28- (lup-2029-ene-3,28-diol) 2.30 1.00 RRTC： = 1.00 RRTB： = 1.00 (sitosterol) - (-spinasterol) 7,22,25- (7,22,25-stigmastatrienol) . 24R-24- -7,25(27)- -3- (24R-24-ethyl cholesta-7,25(27)-dien-3-ol) 145,24- (5,24-stigmastadienol) . 10.3 (response factor) . FID . KS H ISO 12228:2008 8 S 100 g mg
16、. 100)(TBB=mAmAS mB ： (mg) )(A ： AB ： mT ： (g) (erythrodiol) (uvaol)( X Y) 7- ( 16) . 11 11.1 A . . 11.2 r (r) 95 % . . 11.3 R (R) 95 % . . 12 . ( ) KS H ISO 12228:2008 9 1 (triterpene) 2 (methyl sterols) 3 5- (5-sterols) 4 7- (7-sterols) 5 (betulin) 6 . ( 4 mm 2 mm). (band) Rf： 0.30, 7- 0.33, 5- 0.
17、35, 0.45, 0.53 1 TLC (9.2.3 9.2.4) KS H ISO 12228:2008 10 1 . 9.2.6 1 . 2 (A), (B) (C) GLC KS H ISO 12228:2008 11 A ( ) , 1996/97 (Deutsches Institution Normung： DIN) KS A ISO 5725 1 KS A ISO 5725 2 . A.1 mg/kg mg/kg 2 000 14 13 2 1 887 4 000 14 14 2 3 785 8 000 14 13 2 7 585 (sr) (r) % 66.75 186.89
18、 3.54 165.62 463.73 4.37 174.64 489.00 2.30 (sR) (R) % 212.20 594.17 11.25 542.38 1 518.66 14.33 762.67 2 135.48 10.05 A.2 ： % % 1.0 14 13 2 0.8 0.5 14 12 2 0.4 0.5 14 14 2 0.4 (sr) (r) % 0.15 0.41 19.54 0.05 0.13 11.53 0.08 0.22 18.33 (sR) (R) % 0.35 0.97 45.89 0.21 0.58 52.43 0.15 0.41 34.11 KS H
19、ISO 12228:2008 12 A.3 ：(brassicasterol) % % 0.1 14 13 2 0.1 0.1 14 13 2 0.09 10 14 13 2 10.8 (sr) (r) % 0.17 0.48 135.59 0.068 0.190 80.289 0.14 0.38 1.26 (sR) (R) % 0.19 0.52 147.53 0.089 0.249 104.940 0.26 0.72 2.39 A.4 ： 24- % % 5 14 12 2 3.5 10 14 11 2 9.0 40 14 13 2 37.5 (sr) (r) % 0.50 1.40 14
20、.15 0.21 0.58 2.31 0.19 0.54 0.51 (sR) (R) % 0.52 1.45 14.67 0.26 0.72 2.88 0.35 0.97 0.93 A.5 ： % % 0.1 14 9 2 0.03 0.1 14 10 2 0.1 0.10.5 14 12 2 0.2 (sr) (r) % 0.024 0.066 84.853 0.03 0.09 35.13 0.06 0.16 32.78 (sR) (R) % 0.059 0.165 212.132 0.13 0.37 147.45 0.18 0.50 106.00 KS H ISO 12228:2008 1
21、3 A.6 ： % % 1 14 13 2 1.4 10 14 13 2 8.1 0.5 14 13 2 0.3 (sr) (r) % 0.20 0.55 13.81 0.11 0.31 1.37 0.05 0.15 15.51 (sR) (R) % 0.22 0.61 15.22 0.25 0.69 3.03 0.10 0.29 30.87 A.7 ： 7- % % 0.1 14 13 2 0.1 1 14 14 2 1.6 0.5 14 14 2 0.3 (sr) (r) % 0.11 0.31 81.11 0.15 0.43 9.38 0.05 0.15 17.40 (sR) (R) % 0.21 0.59 156.63 0.91 2.56 56.21 0.15 0.43 50.10 A.8 ： 5,23- % % 0.5 14 14 2 0.7 1 14 14 2 1.2 0.5 14 14 2 0.5 (sr) (r) % 0.07 0.20 9.63 0.14 0.39 11.64 0.06 0.16 11.94 (sR) (R) % 0.57 1.59 77.37 0.94 2.62 78.57 0.15 0.42 31.69 KS H ISO 12228:2
