ISO

ISO

ISO

ISO(International Standardization Organization) 135 ISO

CD(Committee Draft) DIS(Draft for International Standard) FDIS(Final DIS) ISO ISO23500 2004.01.13 ANSI/AAMI RD52 2004.04.13 6 2004.08.09 ANSI/AAMI RD52 2004.09.08 2004.11.15 ISO/CD23500 2005.03.14 45 2005.05.03 8 4 0 162 2005.10.03 2005.10.10 12 ISO 2006.03.22 ISO/DIS23500 2006.07.30 ISO/CD13958 13959 26722 2006.08.22 ISO/DIS23500 2006.09.11 15

ISO/TC150/SC2/WG5 ISO/DIS 23500 CD 2nd 23500 13958, 13959, 26722 4 ISO23500 NWI 2 ISO23500 2009

ISO/TC150/SC2/WG5 ISO/DIS 23500 CD 2nd 23500 13958, 13959, 26722 4 ISO23500 NWI 2

ISO/CD DIS ISO/DIS 23500 : Fluids for hemodialysis and related therapies ISO/CD 13958 : Concentrates for hemodialysis and related therapies ISO/CD 13959 : Water for hemodialysis and related therapies ISO/CD 26722 : Water equipment for hemodialysis application NWI ; RD62

13958 13959 26722

CD13959 26722 RO 0.1mg/L ISO/CD 13959 0.1mg/L ISO/CD 26722 0.1mg/L 1

ISO/TC150/SC2/WG5 ISO/DIS 23500 CD 2nd 23500 13958, 13959, 26722 4 ISO23500 NWI 2

Water for hemodialysis ET cfu/ml EU/L 100 (Action Level 50) 250 ISO 1 Dialysis fluid Ultrapure dialysis fluid Online prepared substitution fluid 100 (Action Level 50) 500 0.1 30 10 6 30 ( ) 1 2 1 1 2 1 TGEA, R2A, MEA 17 23 7

ISO 23500 Dr.Ward 07 5 CD NWI 07 8 07. 9.10-14 ISO ( )

ISO/CD23500 Ver. 1.05

ET

1 1 100 /ml 26 0.1mg/L 2 27 0.2mg/L 3 0.01mg/L 28 0.03mg/L 4 0.0005mg/L 29 0.09mg/L 5 0.01mg/L 30 0.08mg/L 6 0.01mg/L 31 1.0mg/L 7 0.01mg/L 32 0.2mg/L 8 0.05mg/L 33 0.3mg/L 9 0.01mg/L 34 1.0mg/L 10 10mg/L 35 200mg/L 11 0.8mg/L 36 0.05mg/L 12 1.0mg/L 37 200mg/L 13 0.002mg/L 38 ( ) 300mg/L 14 1,4-0.05mg/L 39 500mg/L 15 1,1-0.02mg/L 40 0.2mg/L 16-1,2-0.04mg/L 41 0.00001mg/L 17 0.02mg/L 42 2-0.00001mg/L 18 0.01mg/L 43 0.02mg/L 19 0.03mg/L 44 0.005mg/L 20 0.01mg/L 45 (TOC) 5mg/L 21 0.02mg/L 46 ph 5.8 8.6 22 0.06mg/L 47 23 0.04mg/L 48 24 0.1mg/L 49 5 25 0.01mg/L 50 2

ISO 13959 ISO/CD 23500 1 (mg/l) 1. 2(0.1mEq/L) 2. 4(0.3mEq/L) 3. 8(0.2mEq/L) 4. 70(3.0mEq/L) 5. 0.006 6. 0.005 7. 0.10 8. 0.0004 9. 0.001 10. 0.014 11. 0.005 12. 0.0002 13. 0.09 14. 0.005 15. 0.01 16. 0.10 17. 0.50 18. 0.10 19. 0.20 20. 2.0 21. 100 22. 0.002 23. 0.10

Water for hemodialysis Dialysis fluid Ultrapure dialysis fluid Online prepared substitution fluid ISO ET ET cfu/ml EU/L cfu/ml EU/L 100 (Action Level 50) 100 (Action Level 50) 250 500 100 50 1) ( 10) ( 0.1 30 1 1 10 6 30 ( ) TGEA, R2A, MEA 17 23 R2A or equivalent (agar or broth or sheet) 15 25 7 7

ET 68 ET ET ET 1 (1 15) 10 4 EU/ 250EU/L (1.5 10 2 ) (2.5 10 3 ) cfu/ml Ex). 15. PDA Journal of GMP and Validation in Japan,6,38-45,2004 Eschrichia coli (dried cell) 1 1.4 10 4 EU/ 250EU/L E. coli 1800 /ml

Imidazoquinolines Flagellin Bcterial DNA (CpG DNA) Mycoplasmal Lipopeptide Peptidoglycan LPS LBP TLR 7 TLR 5 TLR 9 TLR 2 TLR 6 CD14 CD14 MD-2 TLR 4 MyD88 TRAF 6 IRAK Cytokine NF- B DNA

0.3 1.0 m 10nm 2000 6000 MDP 400 -D glcan 6800 200 DNA fragment 1300 6000

MF

30 300 CFU MF 10 100 CFU

Culture and Non-culture Methods Culture method : Non-culturable cells (Not colony forming VBNC or dead cells) Nucleic acid staining Non-culture method 4,6-diamidino-2-phenylindole (DAPI) Propidium iodide (PI) Intracellular enzyme activity 6-carboxyfluorescein diacetate (6CFDA) 6CF 6CFDA : Culturable cells (Colony-forming ability) Colony Microcolony CTC CTC-formazan 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) Respiratory activity

Culture and Non-culture Methods Culture Method Fluorescent Method 5 Fig. Culture for colony Fig. Non-culture Method - Direct Epifluorescent Filtration Technique (Escherichia coli ) Microbial cells stained with DAPI

Bacterial number and endotoxin level in dialysis fluids Yamaguchi, N., et al : Rapid monitoring of bacteria in dialysis fluids by fluorescent vital staining

Rapid and Automated Bacterial Detection Systems asu, M. (2006): JP s standpoint for chapters on rapid microbiological methods presented in New microbiology chapters of the European armacopoeia - Session II: Alternative methods for control of microbiological quality (Part 1). Strasbourg, France. 2-3 Oct. 2006.

Water for hemodialysis ET cfu/ml EU/L 100 (Action Level 50) 250 ISO 1 Dialysis fluid Ultrapure dialysis fluid Online prepared substitution fluid 100 (Action Level 50) 500 0.1 30 10 6 30 ( ) 1 2 1 1 2 1 TGEA, R2A, MEA 17 23 7

(SAL:Sterility assurance level) D 10 1 10 6 10 5 10 4 10 3 10 2 10 1 1 10 1 10 2 10 3 10 4 10 5 10 6 *D SAL *D 10 1 1 2 3 4 5 6 7 8 9 10 11 12 SAL 10 6

ISO

68 1. 2. 1 3. 2 4. (DQ Design Qualification) 5. (IQ:Installation Qualification) 6. (OQ:Operational Qualification) 7. (PQ:Performance Qualification)

(PQ:Performance Qualification) (a) (b) (c) Phase 1 / RO 3 1 1 Phase 2 1 Phase 2,

Ver. 1.05

RO RO RO ETCF Ca

25 7000 8000h 50 75%

ET 2 70% 50%