β−Blockers REGIS

Alprenolol Betaxolol 90:5:5 CH2C12/EtOH/MeOH 85:10:5 CH2C12/EtOH/MeOH 10 mM NH4OAc 10 mM NH4OAc O 1 ml/min; 254 nm 1 ml/min; 254 nm run time = 10 min run time = 11 min 4.6 mm x 25 cm 4.6 mm x 25 cm α-Burke 2 O N α-Burke 2 k' = 1.44 k' = 2.36 1 1 O N α = 1.44 OH H α = 1.25 reference 33 reference 33 OH H

Atenolol Bupranolol

Atenolol H2N Bupranolol 85:10:5 Column: (3R,4S)-Pirkle 1-J

CH2C12/EtOH/MeOH O 25 cm x 4.6 mm 15 mM NH4OAc O N Mobile Phase: (85/15) 1 ml/min; 254 nm CH2Cl2/Ethanol OH H run time = 16 min + 0.015M 4.6 mm x 25 cm Ammonium Acetate α-Burke 2 Flow Rate: 1.0 mL/min k'1 = 4.41 Detection: UV 254 nm α = 1.13 Run Time: 8.5 min reference 33 k’1: 1.44 α: 1.47 reference 46 Bufuralol

Bufuralol metoprolol Column: (3R,4S)-Pirkle 1-J 85:10:5 CH2C12/EtOH/MeOH 25 cm x 4.6 mm 10 mM NH OAc 4 O

Mobile Phase: (90/10) 1 ml/min; 254 nm CH3 CH2Cl2/Ethanol run time = 13 min + 0.02 M Ammonium 4.6 mm x 25 cm Acetate α-Burke 2 Flow Rate: 1.0 mL/min k'1 = 2.66 Detection: UV 254 nm α = 1.28 O N Run Time: 7.0 min reference 33 OH H k’1: 0.91 α: 2.01 reference 46 HCI β-Blocker

Tulobuterol HCI β-Blocker Column: (S)-α-Burke 2 Column: (S,S)- 25 cm x 4.6 mm DACH-DNB Mobile Phase: (91/9) 25 cm x 4.6 mm CH2Cl2/Ethanol Mobile Phase: + 0.01 M (90/10) Ammonium Acetate CH2Cl2/IPA Flow Rate: 1.5 mL/min Flow Rate: 1.0 mL/min Detection: UV 254 nm Detection: UV 254 nm Run Time: 15.0 min Run Time: 18.0 min k’1: 6.38 k'1: 4.52 α: 1.13 α: 1.29 reference 46 reference 59 REGIS TECHNOLOGIES CHIRAL APPLICATION GUIDE VI • WWW.REGISTECH.COM/chiral/ 77 REGIS β−Blockers Maleate

Nadolol Timolol Maleate Column = (S,S)-Whelk-O 1 Column = (3R,4S)-Pirkle 1-J 10/100 (FEC) 25 cm x 4.6 mm 25 cm x 4.6 mm Mobile Phase = (78/22) Mobile Phase = Hexane/Ethanol + (94/3/3) CH2Cl2/ 0.01 M Ammonium Ethanol/IPA Acetate + 0.01M Ammonium Acetate Flow Rate = 1.5 mL/min Flow Rate = 1.0 mL/min Detection = UV 270 nm Detection = UV 294 nm Run Time = 9.5 min Run Time = 16.0 min k'1 = 3.05 k'1 = 3.72 α = 1.43 α = 1.33 reference 46 reference 46 Carazolol

Carazolol Pindolol Column = (R)-α-Burke 2 Column: (3R,4S)-Pirkle 1-J 25 cm x 4.6 mm 25 cm x 4.6 mm Mobile Phase = Mobile Phase: (80/20) (46/46/8) CH2Cl2/ CH2Cl2/Ethanol + 0.04M Methanol/Ethanol + 0.01 M Ammonium Acetate Ammonium Acetate Flow Rate: 1.0 mL/min Flow Rate = 1.5 mL/min Detection: UV 254 nm Detection = UV 254 nm Run Time: 11.0 min Run Time = 15.0 min k’1: 1.56 k'1 = 6.73 α: 2.06 α = 1.10 reference 46 reference 46

Practolol Oxprenolol 85:10:5 CH2C12/ Column: (3R,4S)-Pirkle 1-J EtOH/MeOH H3C O 25 cm x 4.6 mm 15 mM NH OAc Mobile Phase: (90/10) 4 N 1 ml/min; 254 nm H CH2Cl2/Ethanol + 0.015M run time = 19 min Ammonium Acetate 4.6 mm x 25 cm O N Flow Rate: 1.0 mL/min

α-Burke 2 OH H practolol Detection: UV 254 nm k'1 = 4.78 Run Time: 13.5 min α = 1.14 k’1: 3.55 reference 33 α: 1.15 reference 46

Pronethalol

Pronethalol Propranolol 90:10 CH2C12/EtOH Column: (3R,4S)-Pirkle 1-J 15 mM NH4OAc 25 cm x 4.6 mm 1 ml/min; 254 nm Mobile Phase: (80/20) run time = 15 min CH2Cl2/Ethanol + 0.04M 4.6 mm x 25 cm Ammonium Acetate α -Burke 2 N Flow Rate: 1.0 mL/min k'1 = 3.26 Detection: UV 254 nm α = 1.31 OH H Run Time: 6.5 min reference 33 k’1: 0.80 α: 1.80 reference 46

78 REGIS TECHNOLOGIES CHIRAL APPLICATION GUIDE VI • WWW.REGISTECH.COM/chiral/