1、 KSKSKSKSKSKSKSK KSKSKS KSKSK KSKS KSK KS KS H ISO 20369 , KS H ISO 20369:2012 2012 9 19 http:/www.kats.go.krKS H ISO 20369:2012 : ( ) ( ) () () ( ) : (ISO/TC126) () ( ) () KT&G () KT&G () () ( ) KT&G : (http:/www.standard.go.kr) ： ：2012 9 19 2012-0452 ： ：(ISO/TC126) ( 02-509-7266) (http:/www.kats.g
2、o.kr). 10 5 , . KS H ISO 20369:2012 i ii . iii 1 1 2 1 3 .1 4 2 5 2 5.1 2 5.2 2 5.3 (citric acid monohydrate) .3 6 .3 7 3 7.1 3 7.2 4 8 .4 9 5 10 .5 7 KS H ISO 20369:2012 ii ISO(the International Organization for Standardization) (ISO ) . ISO . . ISO . ISO (IEC) . ISO/IEC , 2 . . . 75 % . . ISO . IS
3、O/TC 126, . KS H ISO 20369:2012 iii 2009 1 ISO 20369, Material used for producing wrappings for cigarette filters, cigarettes and other tobacco products Determination of citrate content . KS H ISO 20369:2012 , Material used for producing wrappings for cigarette filters, cigarettes and other tobacco
4、productsDetermination of citrate content , . . . 1 , (citrate) . 2 . . ( ) . KS M ISO 187, , KS M ISO 287, KS M ISO 3696, 3 . 3.1 (citrate content) , , , . KS H ISO 20369:2012 2 4 (citrate lyase, CL) (oxaloacetate) (acetate) . CL citrate oxaloacetate acetate (1) (pyruvate) L- (L-malate dehydrogenase
5、 L-MDH) L- (L-lactate dehydro-genase, L-LDH) , (reduced nicotinamide adenine dinucleotide, NADH) L- L- . L-MDH oxaloacetate NADH H L-malate NAD (2) L-LDH pyruvate NADH H L-lactate NAD(3) NADH . NADH 340 nm . 5 5.1 . KS M ISO 3696 3 . 5.2 5.2.1 (kit)Roche-Biopharm 10.139.076.035 .1) . . , . 5.2.2 1
6、1 1 . (glycylglycine) pH 7.8 1 . 1) Roche-Biopharm 10.139.076.035 . , ISO . KS H ISO 20369:2012 3 L- (L-MDH), 12 IU2)/mL L- (L-LDH), 23 IU/mL (NADH), 0.5 mg/mL (stabilizers). 1 4 2 20 4 . 5.2.3 2 2 2 . 2 40 IU/mL . 2 4 2 20 4 . (1005) % . 5.3 (citric acid monohydrate) 6 . 6.1 , 250 mL . 6.2 , 80 mm
7、 6.3 , 125 mmWhatman No. 40 3). 6.4 , 1 mL, 2 mL, 5 mL 10 mL . 2. 6.5 , 20 L . 6.6 (double-beam spectrophotometer), 340 nm . 6.7 (cuvet), 10 mm , 5 mL . 6.8 (ultrasonic bath) (magnetic stirrer) 6.9 , 0.001 g . 7 7.1 KS M ISO 187 1.0 g 0.001 g 2) IU(international unit) 1 1 mol ( ) . 3) Whatman No.40
8、 . , ISO . KS H ISO 20369:2012 4 250 mL (6.1) 100 mL (6.8) 30 . (6.3) . 7.2 20 25 . ( 1 ) ( ) (7.1 ) . (6.6) 340 nm , ( ) . V 3.02 mL . , 50 mg/L, 25 mg/L 12.5 mg/L (5.3) . 1 mL mL 1(5.2.2) 1.00 2.00 1.00 1.80 0.20 5 (A1) . . 2(5.2.3) 0.02 0.02 ( 5 ) (A2) . 8 NADH , A (4. ) . . blank21sample21)()( A
9、AAAA = (4) (A1 A2)blank . (4) , |(A1 A2)blank| (A1 A2)sample . 0.1 . 0.850 , . 80 g . L g C . AVFMV =0001PC(5) , V ： (mL)( 3.02 mL) M ： ( ) F ： ： 340 nm NADH ： 6.30 L mmol 1 cm 1 ： (cm) KS H ISO 20369:2012 5 VP： (mL) A ： 7.2 , L g C (M 192.1 g/mol) . A = 461.0C(6) C . %100PCC= (7) P L g . 1 g 100 mL
10、 , P 10 g/L , C . %10CC= (8) ( ) . 9 2 sr sR . 9 2005 , 0.6 % 2.5 % 3 . 2 mNsrsR , n (%) C1 8 0.68 0.025 0.038 C2 6 2.44 0.046 0.060 C3 8 0.60 0.012 0.022 10 . a) ( , ) b) c) d) e) f) g) KS M ISO 287 h) KS H ISO 20369:2012 6 i) j) , . KS H ISO 20369:2012 7 1 CORESTA Recommended Method No 34, Determi
11、nation of Citrate in Cigarette Paper, January 1933 2 DIN 12699, Laboratory glassware Graduated pipettes for enzymatic analyses, short delivery time, waiting time 15 s, class AS , 153787 1 92 3(13) (02)26240114 (02)262401489 http:/ Korean Agency for Technology and Standards http:/www.kats.go.kr KS H ISO 20369:2012 KSKSKS SKSKS KSKS SKS KS SKS KSKS SKSKS KSKSKS Material used for producing wrappings for cigarette filters, cigarettes and other tobacco products Determination of citrate content ICS 65.160
