KS H ISO 14501-2006 Milk and milk powder-Determination of aflatoxin M1 content- Clean-up by immunoaffinity chromatography and determination by high-performance liquid chromatograph.pdf

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1、 KSKSKSKS SKSKSKS KSKSKS SKSKS KSKS SKS KS 2006 11 20 http:/www.kats.go.krKS H ISO 14501 M1 ( ) KS H ISO 14501： 2006 (2011 ) H ISO 14501： 2006 : ( ) ( ) ( ) ( ) : (http:/www.standard.go.kr) ： ： 2001 12 31 ： 2006 11 20 ： 2011 12 29 2011-0652 ： ： ( 02-509-7273) (http:/www.kats.go.kr). 10 5 , . ICS 67.

2、100.10 KS M1 ( ) H ISO 14501： 2006(2011 ) Milk and milk powder Determination of aflatoxin M1content Clean-up by immunoaffinity chromatography and determination by high-performance liquid chromatography 1998 1 ISO 14501 Milk and milk powder Determination of aflatoxin M1content Clean-up by immunoaffin

3、ity chromatography and determination by high-performance liquid chromatography . ISO 14501 ISO/TC 34 (SC 5), (IDF) (AOAC International) , . A . 1. M1 . , (WHO) . 2. , . 3. , ( 2 mol/L) , (pH 68 .). 4. , , , (decontamination) . (WHO) . 1. M1 . 0.08 g/kg , 0.008 g/L . , , . 2. . 2.1 M1 . M1 g/L g/kg .

4、 3. M1 . M1 H ISO 14501： 2006 2 . M1 HPLC M1 . 4. , . 4.1 M1 . M1100 ng( 50 mL 2 g/L ) , 4 ng( 50 mL 80 ng/L ) M1 80 % . (batch) 1 (4.1.1 4.1.2 ). 4.1.1 (5.4) M1 (4.5.2) 1.0 mL 20 mL (5.9) . (4.3) , 10 % (4.2.2) 10 mL . 40 mL , . , HPLC . . 4.1 . 4.1.2 (5.4) 0.005 g/mL M1 (4.5.3) 0.8 mL 10 mL , . ,

5、HPLC . . 4.1 . 4.2 HPLC . 4.2.1 25 % (4.2) 250 mL 750 mL ( ). 4.2.2 10 % (4.2) 100 mL 900 mL ( ). 4.3 4.4 0.51.0 %( ) . 4.5 M1 4.5.1 10 g/mL (nominal) M1 , . (5.11) 340 370 nm . 360 nm max A , Ci(g/mL) . ci=100 MA A： max M： M1 (M = 328 g/mol) ： ( = 1 995 m2 /mol) 4.5.2 (4.5.1) , H ISO 14501： 2006 3

6、0.1 g/mL . . 5 . 2 , 2 . 4.5.3 M1 M1 . 0.005 g/mL . (5.4) (4.5.2) 1 mL 20 mL (5.9) (4.3) , 10 % (4.2.2) 20 mL . 30 . . M10.05 ng, 0.1 ng, 0.2 ng 0.4 ng . (4.2.2) . 5. . 5.1 10 mL 50 mL . 5.2 , Vac-Elute (1) (1) Vac Elute Whatman . , ISO . 5.3 4 000g . 5.4 1 mL, 2 mL 50 mL . 5.5 250 mL . 5.6 100 mL .

7、 5.7 (302) , (502) 3537 . 5.8 Whatman No. 4(1) . 5.9 5 mL, 10 mL 20 mL . 5.10 HPLC 5.10.1 (pulse-free pump) 1 mL/min . 5.10.2 , 50500 L . 5.10.3 HPLC 35 m (octadecyl silicagel) , . 5.10.4 0.02 ng M1 , M1 365 nm 435 nm . 5.10.5 . 5.11 200400 nm , 1 cm . 5.12 0.01 g . 6. KS H ISO 707( 1) . . 7. 7.1 .

8、H ISO 14501： 2006 4 , . , , , ( ) , . . 7.2 7.2.1 (5.7) 3537 . (5.8) ( .) 4 000g 15 . 50 mL 7.4 . 7.2.2 10 g 0.1 g 250 mL (5.5) , 50 50 mL . , 50 (5.7) 30 . 20 100 mL (5.6) . (5.8) 4 000g 15 . 50 mL , 7.4 . 7.3 50 mL (5.1) (4.1) , (5.2) . 7.4 (7.2.1 7.2.2) 50 mL 50 mL (5.1) , (5.2) 23 mL/min . 50 mL

9、 10 mL . 10 mL . 10 mL (4.2) 4 mL M1 . 60 . . (5.9) . (4.3) 30 Ve 50500 L . , . Ve 10 Vf (5005 000 L) . M1 10 % HPLC , 90 % . 7.5 7.5.1 (4.2.1) HPLC . M1 HPLC (4.2.1) . (4.2.1) (5.10.3) , ( 25 cm, 4.6 mm) 1 mL/min . 3 mm 0.5 mL/min . M1 M1 . H ISO 14501： 2006 5 7.5.2 . M1 . 5 % . M1 . M1 . 7.5.3 M1

10、0.05 ng, 0.1 ng, 0.2 ng 0.4 ng . M1 . 7.5.4 (7.4) (Vi) HPLC . M1 . . , 5 . M1 , (ng) . M1 , HPLC . 8. 8.1 8.1.1 M1 (wm) g/L , . wm = mAVVV 1if mA： M1 M1 (ng) Vi： (L) Vf： (L) V： (mL) 8.1.2 g/L , . 8.2 8.2.1 M1 (wp) g/kg , . wp = mAmVV 1if m： (7.4) 50 mL (g) mA： M1 M1 (ng) Vi： (L) Vf： (L) 8.2.2 g/kg ,

11、 . 9. A . . H ISO 14501： 2006 6 10. . H ISO 14501： 2006 7 A() 16 ( 1 %) ( 28 %) . , M1 . 0.080.6 g/kg , 860 ng/L . KS A ISO 5725 1 KS A ISO 5725 2 , A.1 . 7 8 . A.1 1 2 3 4 5 (1) 12 4 13 11 14 (ng/kg) 81 150 80 202 580 , r (ng/kg) 23 60.1 15 27 203 , R (ng/kg) 52 98 41 61 310 (%) 9.9 14.0 6.8 4.7 12

12、.5 (%) 23 22.7 18.3 10.8 19.1 (1) Cochran / Grubbs . H ISO 14501： 2006 8 1 KS H ISO 707： 2006 2 KS A ISO 5725 1： 2002 ( ) 1： 3 KS A ISO 5725 2： 2002 ( ) 2： 4 Laboratory decontamination and destruction of aflatoxins B1, B2, G1and G2in laboratory wastes. Castegnaro M., Hunt D.C., Sansone E.B., Schulle

13、r P.L., Siriwardana M.G., Telling G.M., Van Egmond H.P. and Walker E.A. IARC Scientific Publication No. 37, Int. Agency for Research on Cancer(WHO), Lyon(France), 1980, 59p 5 Laboratory decontamination and destruction of carcinogens in laboratory wastes： some mycotoxins. Castegnaro M., Barek J., Fre

14、my J.M., Lafontaine M., Miraglia M., Sansone E.B. and Telling G.M., IARC Scientific Publication No. 113, Int. Agency for Research on Cancer(WHO), Lyon(France), 1991, 63p 6 Commission of the European Community. Community Bureau of Reference, BCR. The certification of aflatoxin M1in three milk powder

15、samples, CRM No. 282, 284, 285. van Egmond H.P and Wagstaffe P.J., Report and Addendum report, EUR 10412, 1986 7 IDF collaborative study on determination of aflatoxin M1in milk powder, using immunoaffinity columns. L.G.M.T. Tuinstra, Roos A.H., and van Trijp J.M.P., RIKILT Report, 92, 1992, p.14 8 L

16、iquid chromatographic determination of aflatoxin M1in milk powder using immunoaffinity columns for cleanup： Interlaboratory study. Tuinstra L.G.M.T., Roos A.H. and van Trijp J.M.P., J. A.O.A.C int 1993, 76 (6), pp. 1248 1254 M1 ( ) 153787 1 92 3(13) (02)26240114 (02)2624 01489 http:/ KSKSKSSKSKS KSKS SKS KSKS SKSKS KSKSKSKorean Agency for Technology and Standards http:/www.kats.go.kr KS H ISO 14501： 2006 Milk and milk powderDetermination of aflatoxin M1contentClean up byimmunoaffinity chromatography anddetermination by high performanceliquid chromatographyICS 67.100.10