GPC/SEC

GPC/SEC

Agilent GPC/SEC...3... 5...5...7...9...11...12... 13...13...14... 15...15...16... 18...18...19 /... 21...21... 23 GPC/SEC...24 Q Q Q Q Q 35 GPC/SEC 24 365 GPC/SEC GPC/SEC GPC 2

80 % 2006 - Polyplasdone Ultra (ISP) Lµtrol micro 68 127 Kollidon CL-F & CL-SF ( BASF) Swelstar MX 1 ( ) GalenIQ 721 (Palatinit) - Spectrablend HS (Sensient) Prosolv ODT (JRS) Ludiflash (BASF) Aquarius (Ashland) Avicel DG (FMC) Sepitrap (Seppic) Starcap 1500 (Colorcon) - Solutol HS 15 Soluplus Kollicoat Smartseal 30 D ( BASF) Spress B818 Pregelatinized Corn Starch NF (Grain Processing Corporation) ICH M4Q (Quality) 1 () ( ) 1 The Common Technical Document for the Registration of Pharmaceuticals for Human Use:Quality - M4Q(R1) EU 3

Quality by Design QbD (Quality by Design) 20 J. M. Juran Juran QbD 2002 2 2008 2009 300 % QbD QbD (GPC SEC ) GPC/SEC HPLC 2 Pharmaceutical Quality for the 21st Century - A Risk-Based Approach.USFDA 4

(PVP ) 100 3 : PLgel 10 µm MIXED-B 7.5 x 300 mm x 2 ( PL1110-6100) : DMF + 0.1 % LiBr : 70 ºC : 1260 Infinity GPC/SEC (RI) (DMF) RI DMF Agilent PLgel 10 µm MIXED-B ( 1) 10 min 17 1. Agilent PLgel 10 µm MIXED-B 5

PVP (DMAc) PLgel 10 µm MIXED-B ( 2) : PLgel 10 µm MIXED-B 7.5 x 300 mm x 3 ( PL1110-6100) : DMAc + 0.5 % LiBr : 0.2 % w/v 100 µl : 60 ºC : 1260 Infinity GPC/SEC (RI) 5 3 : Agilent PL aquagel-oh 60 8 µm 7.5 x 300 mm ( PL1149-6860) PL aquagel-oh 50 8 µm 7.5 x 300 mm ( PL1149-6850) : 0.2 M NaNO 3 + 0.01 M NaH 2 PO 4 ph 3 : 50 ºC : RI 15 24 2. Agilent PLgel 10 µm MIXED-B 3 10 30 3. Agilent PL aquagel-oh 6

- (Liquid chromatography under critical conditions: LCCC) PEG PEG MW SEC LCCC PEG PEG 4 PEG PEG GPC/SEC PEG 5 SEC ACN/ PEG 22,800 PEG 8,500 PEG 960 SEC 70 % ACN 49 % ACN 0.0 0.6 1.2 1.8 2.4 3.0 40 % ACN 5. SEC 4. ( ) ( ) 7

6 PEG : Agilent PLRP-S 100Å 5 µm 4.6 x 150 mm ( PL1111-3500) : 49 % : 20 µl : Agilent 1260 Infinity 1 ( PEG ) 2 ( ) 2 PEG 8 % PEG 45 % 47 % SEC (PEG ) PEG HCI PEG- SEC PEG- PEG (8 %) (45 %) (47 %) 0 2 4 6 8 10 6. ( ) ( ) Agilent 1260 Infinity ( ) 1290 Infinity ( ) 8

Agilent PL aquagel-oh MIXED-H 8 µm (PEO/PEG 1000 ) (>35,000 /m) PL aquagel-oh MIXED-H 8 µm 2 mg/ml 0.45 µm 7 Mp 1,600,000 Mp 788,000 Mp 212,000 8 : PL aquagel-oh MIXED-H 8 µm 7.5 x 300 mm x 2 ( PL1149-6800) : 0.2 M NaNO 3 + 0.01 M NaH 2 PO 4 ph 7 : 50 ºC : RI D C Log M Mp 47,300 Mp 11,800 B Mp 667 A 11.7 17.6 5 20 7. Agilent PL aquagel-oh MIXED-H 8 µm 8. Agilent PL aquagel-oh MIXED 4 Agilent GPC/SEC GPC/SEC Agilent EasiVial Agilent EasiCal Q Q Q Q Q GPC Calibrating GPC Columns - A Guide to Best Practice (5991-2720EN : ) 9

9 : PL aquagel-oh MIXED-H 8 µm 7.5 x 300 mm x 2 ( PL1149-6800) : 0.2 M NaNO 3 + 0.01 M NaH 2 PO 4 ph 7 : 50 ºC : RI A B C D D 1 1. 4 (Mw/Mn) Mn Mw A 6,520 17,560 2.7 B 21,720 88,480 4.1 C 67,980 243,120 3.6 D 128,360 459,990 3.6 dw/dlogm (Mw/Mn) 2 log M 8 PL aquagel-oh MIXED-H 8 µm 9. 4 10

N- 2 GPC/SEC Mw 2 Agilent PL aquagel-oh MIXED-H 8 µm 3 ph 0.5 M PL aquagel-oh MIXED 8 µm 7 (9 ) 3 (Mw) 10 ( /) RI : PL aquagel-oh MIXED-H 8 µm 7.5 x 300 mm x 2 ( PL1149-6800) : 0.5 M NaNO 3 + 0.01 M NaH 2 PO 4 ph 2 : 50 ºC : RI A (Mw 367,600) B (Mw 385,800) C (Mw 449,350) 5 20 RI 10. 3 HPLC GC www.agilent.com/chem/jp 11

2 PL aquagel-oh SEC 11 2 2 : PL aquagel-oh 60 8 µm 7.5 x 300 mm ( PL1149-6860) PL aquagel-oh 40 8 µm 7.5 x 300 mm ( PL1149-6840) : 0.05 M NaH 2 PO 4 + 0.25 M NaCl ph 7 : 50 ºC : RI 0.5 2. 2 dw/dlogm A B (cps) 85 115 4,000 6,000 Mn 131,000 484,000 Mw 369,000 1,023,000 Mz 691,000 1,884,000 0.0 4 log M 6 11. Agilent PL aquagel-oh 2 12

1 PL aquagel-oh MIXED-H 8 µm 0.1 M NaOH ph 1.0 mg/ml Mp 186,000 1.0 mg/ml Mp dn/dc dn/dc dn/dc 90º 15º Mw RI LS SEC Mw 174,000 (90º) 189,850 (15º) 184,800 (SEC) 90º 15º RI RI : PL aquagel-oh MIXED-H 8 µm 7.5 x 300 mm x 2 ( PL1149-6800) : + 0.2 M NaNO 3 + 0.01 M NaH 2 PO 4 ph 7 : 1260 Infinity GPC/SEC RI 18 (mv) 12 RI 90º 15º -2 0 22 12. Agilent PL aquagel-oh MIXED-H 2 90º 15º 13

: PLgel 10 µm MIXED-B 7.5 x 300 mm x 3 ( PL1110-6100) : DMAc + 0.5 % LiBr : 0.2 % w/v 100 µl : 60 ºC : 1260 Infinity GPC/SEC (RI) 15 min 25 15 25 13. Agilent PLgel 10 µm MIXED-B Agilent 1260 Infinity GPC/SEC 14

(CMC) Agilent PL aquagel-oh 40 8 µm PL aquagel-oh 60 8 µm CMC (>35,000 /m) 10 4 10 7 2 PL aquagel-oh 14 3 : PL aquagel-oh 60 8 µm 7.5 x 300 mm ( PL1149-6860) PL aquagel-oh 40 8 µm 1.5 x 300 mm ( PL1149-6840) : 0.5 M Na 2 SO 4 : RI 0.3 dw/dlogm 0.0 5 log M 7 14. 3 Mw Agilent PL aquagel-oh 15

- HPLC (CD) 6 7 8 - - - CD HPLC UV RI (ELSD) HPLC ELSD 1260 Infinity 1 Agilent ELSD : C18 5 µm 4.6 x 150 mm A : B : : 20 µl : 5 50 95 % B : 1260 Infinity ( = 30 ºC = 50 ºC = 1.0 SLM) 1. - 2. - 3. 1 2 0 12 3 Agilent ELSD UV 221 nm 15. Agilent ELS () 15 ELSD 16

Agilent ELSD 16 30 ºC 50 ºC Agilent ELSD 1 Agilent ELSD UV UV HPLC ELSD ESLD HPLC : C18 5 µm 4.6 x 150 mm A : B : : 20 µl : 5 50 95 % B : 1260 Infinity ( = 30 ºC = 50 ºC = 1.0 SLM) 1. - 2. - 3. 1 2 3 50 ºC 30 ºC 0 12 16. Agilent ELSD 17

in vitro 17 2 Agilent PLgel 5 µm MIXED-C THF 80,000 2.5 : PLgel 5 µm MIXED-C 7.5 x 300 mm x 2 ( PL1110-6500) : THF ( ) : 200 µl : RI ~ 0 22 17. Agilent PLgel MIXED-C ( ) 18

() - (GPEC) GPEC GPEC GPEC LC HPLC (GPEC ) ( ) / ( HPLC ) / 2 18 18. 19

99 % 2 10 1 % 99 % 19 20 : Agilent PLRP-S 100Å 5 µm 4.6 x 150 mm ( PL1111-3500) : THF : 20 µl : : Agilent 1260 Infinity : 1260 Infinity (%) (%) 50 (%) log Mn 5.0 log Mn 0 % R 2 = 0.9927 0 4.7 15 % 7.5 Rt/min 12.5 25 % 20. 35 % 1.0 16.2 19. 50 % GPEC GPEC 20

/ (HEC) HEC GPC (DMF) PLgel 5 µm MIXED-C LiBr ( 21) DMF PEO/PEG 3 3 3. 3 Mn Mw Mp (Mw/Mn) A 27,000 140,000 80,000 5.2 B 30,000 159,000 102,000 5.2 C 39,000 345,000 190,000 8.9 : PLgel 5 µm MIXED-C 7.5 x 300 mm ( PL1110-6500) : DMF + 0.1 % LiBr : 50 ºC : RI B A dw/dlogm 1K 10M 21. Agilent PLgel 5 µm MIXED-C 3 3 C 21

3 PL aquagel-oh 22 4 4. 3 Mn Mw Mz (cps) A 60,300 179,000 139,000 75 112 B 413,000 849,000 1,552,000 250 324 C 914,000 2,016,000 3,422,000 1,500 2,500 : PL aquagel-oh 60 8 µm 7.5 x 300 mm ( PL1149-6860) PL aquagel-oh 40 8 µm 7.5 x 300 mm ( PL1149-6840) : 0.05 M NaH 2 PO 4 + 0.25 M NaCl ph 7 : 50 ºC : RI 0.5 A B C HEC PL aquagel-oh 40 PL aquagel-oh 60 8 µm 10 4 10 7 2 PL aquagel-oh 23 HEC A HEC B A : : PL aquagel-oh 60 8 µm 7.5 x 300 mm ( PL1149-6860) PL aquagel-oh 40 8 µm 7.5 x 300 mm ( PL1149-6840) : 0.05 M NaH 2 PO 4 + 0.25 M NaCl ph 7 : 50 ºC : RI 0.4 B A dw/dlogm dw/dlogm 0.0 4.0 log M 7.0 22. Agilent PL aquagel-oh 0.0 23. 22

NMR FTIR HPLC GC/MS SEC GC/MS UV-Vis GPC-FTIR (GPC) (FTIR) 2 1 GC/MS (GC) / (GC/MS) GC GC/MS LC LC/MS ( ) Agilent 6100 Agilent 6200 Accurate-Mass TOF LC/MS Easy Access Agilent 6500 Accurate-Mass Q-TOF LC/MS MassHunter Agilent Poroshell 120 LC 2 µm LC Poroshell LC LC/MS ICP-MS Agilent 7700x ICP-MS (HMI) ORS3 () 9 7700x Apparatus 1 2 5 6 Agilent 708-DS Apparatus 3 Agilent BIO-DIS Apparatus 7 Agilent 400-DS Apparatus 7 Agilent 400-DS Agilent Poroshell 120 23

GPC/SEC (5 7) Agilent 1260 Infinity GPC/SEC 5. Agilent 1260 Infinity GPC/SEC ( ) G1310B G1322A G1316A G1329B Agilent 1260 Infinity 1260 Infinity 1260 Infinity 1260 Infinity G1328C 1260 Infinity GPC/SEC GPC 80 ºC 2 7.5 300 mm GPC GPC/SEC 20 50 100 200 µl (PEG) (PEG) (HPMC) (PVAP) (CMC) * * * (HEC) * 24

6. Agilent 1260 Infinity GPC/SEC ( ) * G1362A G1314F G7850AA G7854AA 1260 Agilent 1260 Infinity Agilent GPC/SEC Agilent GPC/SEC G1365D Agilent 1260 Infinity 8 µl LAN GPC (PEG) (PEG) (HPMC) (PVAP) (CMC) (HEC) * 1 (G1362A G1314F G1365D ) HPLC LC LC/MS HPLC GC CrossLab CrossLab Bruker/Varian CTC PerkinElmerThermo/Dionex Waters CrossLab 90 www.agilent.com/chem/jp 25

Agilent 1260 Infinity GPC/SEC 7. Agilent 1260 Infinity GPC/SEC ( ) G1310B G1322A G1316A G1329B G7800A Agilent 1260 Infinity 1260 Infinity * 1 1260 Infinity 1260 Infinity 1260 Infinity G1328C 1260 Infinity GPC/SEC GPC 80 ºC 2 7.5 x 300 mm GPC GPC/SEC 20 50 100 µl (PEG) (PEG) (HPMC) (PVAP) (CMC) * 2 * 2 * 2 (HEC) * 1 * 2 * 3 1 (G7801A G7802A G7803A ) * 4 1260 Infinity MDS (G7801A) * 5 1260 Infinity MDS /1260 Infinity MDS 26

G7801A* 3 G7802A* 3 G7803A* 3 G1314F* 4 G7850AA G7852AA G7854AA 1260 Infinity MDS 1260 Infinity MDS 1260 Infinity MDS Agilent 1260 Infinity G1365D* 4 Agilent 1260 Infinity Agilent GPC/SEC Agilent GPC/SEC * 5 Agilent GPC/SEC GPC 15º 90º Mw 1 GPC GPC Agilent GPC/SEC 27

agilent.com/chem/navigator LC 4 Q Q Q Q - LC - - USP : www.agilent.com/chem/jp : Agilent EasiVial Agilent EasiCal GPC Calibrating GPC Columns - A Guide to Best Practice (5991-2720EN ) 0120-477-111 email_japan@agilent.com Agilent Technologies, Inc. 2014 Published in Japan, March 14, 2014 5991-2519JAJP