KS I ISO 20179-2006 Water quality－Determination of microcystins－Method using solid phase extraction(SPE) and high performance liquid chromatography(HPLC) with ultraviolet(UV) detec.pdf

1、 KSKSKSKS SKSKSKS KSKSKS SKSKS KSKS SKS KS 2006 10 31 http:/www.kats.go.krKS I ISO 20179 (SPE) (HPLC)/(UV) KS I ISO 20179： 2006 (2011 )I ISO 20179： 2006 : ( ) ( ) KIST ( ) : (http:/www.standard.go.kr) ： ：2006 10 31 ：2011 12 26 2011-0623 ： ： ( 02- 509-7270) (http:/www.kats.go.kr). 10 5 , . ICS 13.060

2、.50 KS I ISO (SPE) (HPLC)/(UV) 20179： 2006(2011 ) Water quality Determination of microcystins Method using solid phase extraction(SPE) and high performance liquid chromatography(HPLC) with ultraviolet(UV) detection 2005 1 ISO 20179 Water quality Determination of microcystins Method using solid phase

3、 extraction(SPE) and high performance liquid chromatography(HPLC) with ultraviolet(UV) detection . . . , . (NaClO) . . . . , . 1. ( ) . MCYST RR, MCYST YR MCYST LR . 1 , . MCYST LR 1 g/L (WHO) (SPE) . 2. . , . ( .) . KS M ISO 3696： 2005 ISO 5667 4 Water quality Sampling Part 4： Guidance on sampling

4、from lakes, natural and man-made I ISO 20179： 2006 2 ISO 5667 5 Water quality Sampling Part 5： Guidance on sampling of drinking water from treatment works and piped distribution systems 3. . APCI (atmospheric pressure chemical ionization) MCYST (microcystin) MCYST LR crocystin containing leucine(L)

5、and arginine(R) MCYST RR microcystin containing two arginine(R) units MCYST YR microcystin containing tyrosine(Y) and arginine(R) SIM (selected ion monitoring) SEC (size exclusion chromatography) 4. ( ) . ( / ) . , . ( ). . 0.1 % (TFA) / (90/10 ) SPE . 238 nm UV DAD (RP HPLC) . 5. , KS M ISO 3696： 2

6、005 3 . 5.1 CH3OH, HPLC 5.2 CH3CN, HPLC 5.3 TFA, CF3COOH 5.4 SPE , / 20/80 5.5 / 75/25 5.6 SPE 0.1 % TFA / 90/10 5.7 100 mL 1 g , Na2S2O3( 5H2O ) . = 10 g/L ( Na2SO3 63 mmol) 5.8 1 mol/L , NH4OH 5.9 100 g ( ) , 0.05 g MCYST LR 80 % MCYST RR MCYST YR 70 % . SPE / , . C 18 . . (16.9 %), (54 m), (% C 3

7、33 I ISO 20179： 2006 3 g/m2), (3 mL), (500 mg). , SPE . SPE 2 . 5.10 HPLC (A) 1 000 mL 800 mL (5.2) 500 L TFA(5.3) , 1 000 mL . HPLC . . 3 . 5.11 HPLC (B) 1 000 mL 800 mL 500 L TFA(5.3) , 1 000 mL . HPLC . . 2 . 5.12 HPLC () 1 HPLC min HPLC (A) 0.05 % TFA(5.10) % HPLC (B) 0.05 % TFA(5.11) % mL/min 0

8、 30 70 0.31.0 10 35 65 0.31.0 40 70 30 0.31.0 42 100 0 0.31.0 44 100 0.31.0 46 30 70 0.31.0 55 30 70 0.31.0 5.13 . 5.14 . 5.14 (stock solution) , (5.1) . (5.1) 1 cm 220250 nm . (1) , g /L i . dMA=iimaxi000 1 (1) Amax： Mi： (g/mol) i： (5.1) L/(mol cm) d： (cm) 1 000： (g /mL) Mi i 2 . I ISO 20179： 2006

9、4 2 MCYSTLR, YR RR (, 238 nm) Mig mol 1iL/mol 1 cm 1 LR 994 39 800 YR 1 044 39 800 RR 1 037 39 800 1 , . HPLC MCYST LR, MCYST YR MCYST RR / , 20/80( , 5.4 ) . 10 g/mL . 5.15 (5.4) 2.5 g/mL MCYST LR, YR RR . 16 . , . , . , . 5.16 3 (5.14) 1 mL . 1 000 L 3 (5.4) (vial) . 3 g/mL MCYST LR, YR, RR (5.14)

10、 L 1 000 L (5.4) L MCYST LR MCYST YR MCYST RR 1 20 940 0.2 0.2 0.2 2 40 880 0.4 0.4 0.4 3 100 700 1.0 1.0 1.0 4 200 400 2.0 2.0 2.0 5 300 100 3.0 3.0 3.0 5.17 200 L (5.15 ) 500 mL . ( ) . MCYST LR, YR MCYST RR 1 g/L . 6. 6.1 . . (： , I ISO 20179： 2006 5 ) . 6.2 . 6.3 12 m . 47 mm . . 6.4 SPE 500 mL

11、6.5 SPE 6.6 4 000 , (RCF) 10 000 g . . 6.7 (probe) 60 W, 20 kHz . 6.8 6.9 . ： 3050 , ： 20 ： 99.996 % 6.10 1 0.45 m . ( ). . . 6.11 6.12 HPLC . 6.12.1 HPLC 0.31.0 mL/min . 6.12.2 HPLC (35 ) . 6.12.3 520 L . 6.12.4 HPLC (：C18) MCYST LR, YR RR 35 m 24.6 mm, 250 mm . . 70 000200 000 hPa (1 0152 900 psi)

12、 . 6.12.5 (UV)/(PDA : Phto Diode Array) = 238 nm . PDA 200300 nm . (LOD) 0.1 ng/L( = 3) ( ) (LOQ) 0.2 ng/L ( = 6) . 7. 7.1 ISO 5667 4 ISO 5667 5 , (4 8 ) 48 . SPE (5.9) . . , 4 mL (5.1) . 4 mL . 10 mL/min I ISO 20179： 2006 6 , . 7.2 7.2.1 / (7.4) . 7.2.2 ( ： 50100 mL) (6.3) . , 50 mL 1 . . (7.4 ) .

13、(7.3 ) , HPLC (7.6 ) (7.5 ) . , , g/g . 7.3 (1 , .) 3 mL (5.5) 3 . (probe)(6.7) (6.8) 2 . , 10 4 000 . , 1 mL (40 ). 500 L (5.4) , 5 . 7.4 (SPE) , pH 5.08.0 . pH , (5.3) (5.8) . 500 L (5.7) 500 mL . , 5 . 5 mL (5.1) , 10 mL/min ( ) (7.1 ) . 4 mL (5.4) . 4 mL (vial) 2.0 mL SPE (5.6)( .) . 40 . 500 L

14、(5.4) , 5 . HPLC(7.6) . 7.5 (SPE) (7.3) (7.1) , 500 L (5.4) . , 4 mL (5.4) . ： 4 mL SPE (5.6) 2.0 mL . 40 . 500 L (5.4) I ISO 20179： 2006 7 5 . 900 L (5.4) 100 L (7.3) . , (SEC)3, 4 . 57 . 7.6 (HPLC) , . 35 HPLC (5.12) (6.12.4) . ( , ) . MCYST RR, MCYST YR MCYST LR , . 200300 nm . PDA B . 7.7 (5.16)

15、 238 nm( ) . (520 L) (5.16) 3 (MCYSTRR, MCYST YR MCYST LR) . 0.23.0 g/mL . ( 2) (y x ) ( ) . bmxy += (2) b： y ( ) m： (mL/g) x： x (g/mL) y： y ( ) . , . (5.4) HPLC . 7 . 7.8 , (5.17) . ( ). 7.9 (2) , (3) (g/L) . sampddissMCYST,)(VmVby= (3) MCYST, diss： (g/L) Vd： SPE (mL) Vsamp： SPE (L) y, b, m： (2) I

16、ISO 20179： 2006 8 7.10 (2) (4) (g/mL ) . samexpartMCYST,)(VmVfby= (4) MCYST,part： (g/ mL) Vex： (mL) Vsam： (mL) f： cell y, b, m： (2) . 7.11 (3) (4) . . , 20 % . MCYST RR, MCYST YR MCYST LR UV ( B.2 ) . MCYST LR . MCYSTLR . 2 g/L . . ( / ), . , . , 2 . 95 % . , 2 (CVR)( C.1 ) . U . . , . 8. C . C.1 .

17、9. . . a) b) c) 7.11 d) ( ) I ISO 20179： 2006 9 e) f) g) h) , I ISO 20179： 2006 10 A() A.1 PDA MCYST RR, YR LR . , (ESI APCI) (MS) . , UV/ PDA UV/PDA . A.2 MS MCYST RR(m/z = 519.8, m = , z = ) MCYST YR (m/z = 1 045.5) MCST LR(m/z = 995.6) (SIM) m/z = 500 600( MCYSTs) 9001 200( MCYSTs) . ( ) . MCYSTs A.1 . MCYST LR . A.1 ( M r) No. Mr 1 909 MCYST-LA 2 923 MCYST-Laba 2a 9