CHIRAL HPLC APPLICATION GUIDE IV

Transcription

1 CIRAL PLC APPLICATI GUIDE IV

2 DURABLE, IG EFFICIECY CIRAL CLUM Regis Technologies, Inc. is proud to be a leader in chiral separations and serve the analytical and preparative needs of chromatographers and researchers worldwide. Regis offers three different types of Chiral tationary Phases: Pirkle-Concept Davankov Ligand Exchange Protein-based Regis manufactures a complete line of Pirkle Chiral tationary Phases and Davankov Ligand Exchange columns on-site at its quality manufacturing facility. Columns range from analytical to custom preparative in size. A line of protein-based chiral stationary phases is also available. All products must meet rigorous manufacturing and quality control specifications before release. As evidence of our leadership in, and our commitment to chiral separations, Regis is pleased to present its Chiral PLC Application Guide IV. This guide contains over 500 specific chiral applications using a variety of chiral column types. For those applications not listed in this guide, Regis maintains a dedicated chiral applications laboratory offering a free chiral screening service where technical staff are able to assist in the selection of appropriate chiral stationary phases and resolution techniques (ee page 88). For applications or specific column types not listed in this guide, additional references, information about our free chiral screening service, or further product informationplease contact Regis directly. All Regis Chiral eparations products must meet rigorous manufacturing and quality control specifications before release. Regis Technologies, Inc. 1

3 CIRAL PLC APPLICATI GUIDE IV C T E T Introduction to Regis Chiral tationary Phases 4 Pirkle Chiral tationary Phases Davankov Ligand Exchange Chiral tationary Phase Protein-based Chiral tationary Phases Regis PLC Chiral Column Product List 7 Aryl Propionic Acid on teroidal Anti-Inflammatory Drugs (AID) 8 ther Carboxylic Acids 11 Esters, Lactones, Ketones, Anhydrides 15 Basic itrogen 19 Amides, Imides, Carbamates, etc. 20 Epoxides 25 Alcohols 26 Diols, ydroxyketones, etc. 32 ulfoxides 33 Phosphorous Compounds 37 Atropisomers 41 ptical witches, Liquid Crystal Components 49 Allenes 50 rganometallic Compounds 55 Agricultural Compounds 58 Miscellaneous Pharmaceuticals 64 atural Products 76 Barbiturates 78 Ureas 78 Diol Epoxides 78 β-blockers 79 Amino Acids 82 Amino Acid Derivatives 83 References 84 Index Applications by Compound 85 Applications by Column Type 86 Customer ervice Form (Fax) 87 Chiral creening Forms 88 Regis Technologies, Inc. 3

4 REGI Introduction ITRDUCTI T REGI CIRAL TATIARY PAE Pirkle Chiral tationary Phases Regis Technologies, Inc., along with Dr. William Pirkle, of the University of Illinois, introduced the Pirkle Chiral tationary Phases (CPs) starting in Regis Pirkle Chiral tationary Phases offer the following advantages: eparation of the enantiomers of a wide variety of compound groups Column durability due to covalent phase bonding Ability to invert elution order Excellent chromatographic efficiency Available in analytical to preparative size columns Accurate determination of enantiomeric excess, especially in trace analyses Universal solvent compatibility Regis manufactures a full line of Pirkle Columns. The Pirkle-Concept Chiral tationary Phases generally fall into three classes: π-electron acceptor/π-electron donors, the π- electron acceptors and the π-electron donors. With Pirkle Phases, chiral recognition occurs at binding sites. Major binding sites are classified as π-basic or π-acidic aromatic rings, acidic sites, basic sites, and steric interaction sites. Aromatic rings are potential sites for π-π interactions. Acidic sites supply hydrogens for potential intermolecular hydrogen bonds-the hydrogen is often an amido proton (-) from an amide, carbamate, urea, or amine. Basic sites, such as π-electrons, sulfinyl or phosphinyl oxygens, and hydroxy or ether oxygens, may also be involved in hydrogen bond formation. teric interactions may also occur between large groups. Whelk- 1 () (,) more stable adsorbate (R) (,) less stable adsorbate π-electron Acceptor/π-Electron Donor Phases WELK- 1 WELK- 2 ULM DAC-DB The latest and most revolutionary addition to the Pirkle-Concept series is the π-electron acceptor/π-electron donor phase. This concept is an innovative incorporation of both π-acceptor and π-donor characteristics, resulting in a phase that can be used for a wide variety of compound groups. WELK- 1 The Whelk- 1 Chiral tationary Phase is based on 1-(3,5-Dinitrobenzamido)-1,2,3,4,- tetrahydrophenanthrene. This phase allows separation of underivatized racemates from a number of families including amides, epoxides, esters, ureas, carbamates, ethers, aziridines, phosphonates, aldehydes, ketones, carboxylic acids, and alcohols. The Whelk- 1 was originally designed for the separation of underivatized non-steroidal anti-inflammatory drugs (AIDs). This π-electron acceptor/π-electron donor phase exhibits an extraordinary degree of generality, allowing resolution of a wide variety of underivatized racemates. This broad versatility observed on the Whelk- 1 column, compares favorably with polysaccharide-derived chiral stationary phases. In addition, because of the Whelk- 1 s covalent nature, this chiral phase is compatible with all commonly used mobile phases, including aqueous systems a distinct advantage over polysaccharide-derived chiral stationary phases. ther advantages include column a) chematic diagram showing key functional groups of the Whelk- 1 involved in chiral recognition. b) chematic diagram showing generalized structure of analytes which are resolved on the Whelk- 1. a) b) D B -bond donor Tetrahydrophenanthrene π system Dinitrobenzamide π-system -bond acceptor Conjugated π-system 4 REGI TECLGIE CIRAL APPLICATI GUIDE IV

5 Introduction REGI durability, excellent efficiency, elution order inversion allowing availability of both enantiomeric forms, and excellent preparative capacity. WELK- 2 The Whelk- 2 is the covalent trifunctional version of the Whelk- 1. The Whelk- 2 retains the same chiral selector but modifies the support to silica from a monofunctional linkage to a trifunctional. In most cases, the enantioselectivity remains the same allowing the separation of the analogous family of racemates as does the Whelk- 1. Whelk- 2 was designed to enhance the stability of the stationary phase due to hydrolysis while using strong organic modifiers such as trifluoroacetic acid. The Whelk- 2 is ideal for preparative separations since the material is bonded on 10 µm 100 Å spherical Kromasil silica. This allows the preparative chromatographer to perform method development on their analytical column and immediately scale-up to larger diameter columns. Whelk- 2 ULM The ULM chiral stationary phase was developed by Austrian Researchers, Uray, Lindner, and Maier. This CP has a general ability to separate the enantiomers of many racemate classes, and is particularly good at separating the enantiomers of aryl carbinols. The ULM CP is based on a 3,5-dinitrobenzoyl derivative of diphenylethylene-diamine. DAC-DB The innovative DAC-DB CP was designed by Italian chemists Dr. Francesco Gasparrini, Misiti and Villani at Rome University La apienza. The DAC-DB CP; which contains the 3,5-dinitrobenzoyl derivative of 1,2-diaminocyclohexane, has been found to resolve a broad range of racemate classes including amides, alcohols, esters, ketones, acids, sulfoxides, phosphine oxides, selenoxides, phosphonates, thiophosphineoxides, phosphineselenides, phosphine-boranes, β-lactams, organometallics, atropisomers and heterocycles. ULM DAC-DB π-electron Acceptor Phases Pirkle 1-J α-burke 2 β-gem 1 Leucine Phenylglycine The π-electron acceptor Pirkle Chiral tationary Phases can be used to separate a wide range of enantiomers without derivatization, as demonstrated for the following classes of solutes: secondary benzyl alcohols, mandelic acid analogs, α-hydroxy-α-aryl phosphates, α-tetralol analogs, propranolol analogs, β-hydroxy-aryl sulfoxides, alkylaryl sulfoxides, diaryl sulfoxides, aryl-substituted cyclic phthalides, aryl-substituted lactams, aryl-substituted succinimides, aryl-substituted hydantoins, bi-β-naphthol and its analogs, and α-aryl acetamides. Pirkle 1-J PIRKLE 1-J The Pirkle 1-J CP is based on 3-(3,5-dinitrobenzamido)-4-pheny-β-lactam. This unusual β-lactam structure significantly alters its molecular recognition properties. The Pirkle 1J is useful for the direct separation of underivatized β-blocker enantiomers. It can also be used for the separation of the enantiomers of aryl propionic acid AID s as well as other drugs. α-burke 2 The α-burke 2 phase, first prepared by J. A. Burke III, a graduate student of Dr. Pirkle, is derived from dimethyl -3,5-dinitro-benzoyl-α-amino-2,2-dimethyl-4-pentenyl phosphonate The α-burke 2 has been specifically designed to directly separate the enantiomers of β-blockers without chemical derivatization, but this chiral phase is not limited solely to the separation of β-blocker enantiomers. It also resolves the enantiomers of many compounds separated on π-acceptor Pirkle type chiral stationary phases. α-burke 2 REGI TECLGIE CIRAL APPLICATI GUIDE IV 5

6 REGI Introduction β-gem 1 Leucine Phenylglycine β-gem 1 β-gem 1 is a π-acceptor chiral stationary phase and is derived from -3,5- dinitrobenzoyl-3-amino-3-phenyl-2-(1,1-dimethylethyl)-propanoate. For a great many analytes, this chiral phase considerably outperforms its widely used analog, phenylglycine. It can separate anilide derivatives of a wide variety of chiral carboxylic acids, including nonsteroidal anti-inflammatory agents. LEUCIE The leucine CP is based on the 3,5-dinitrobenzoyl derivative of leucine. This π-acceptor phase demonstrates enhanced enantioselectivities for several classes of compounds, including benzodiazapines. PEYLGLYCIE Phenylglycine is based on the 3,5-dinitrobenzoyl derivative of phenylglycine. This CP resolves a wide variety of compounds which contain π-basic groups. These include: aryl-substituted cyclic sulfoxides, bi-β-naphthol and its analogs, α-indanol and α-tetralol analogs, and aryl-substituted hydantoins. π-electron Donor Phases APTYLLEUCIE The naphthylleucine phase is based on the -(1-naphthyl) derivative of leucine. These columns resolve essentially the same class of compounds as does the naphthylalanine column, but typically with enhanced enantioselectivities for amino acids. This π-donor chiral phase resolves DB derivatives of amino acids as the free acid when using the reversed-phase mode. In the classic normal-phase, this CP can resolve the amides and esters of DB amino acids and will display typical alphas between 10 and 40. Davankov Ligand Exchange Chiral tationary Phase DAVAKV The Davankov chiral stationary phase is useful for the separation of underivatized amino acid enantiomers. This phase operates according to the principles of ligand-exchange chromatography (LEC), a technique pioneered by professor V. Davankov. The Davankov column typically requires a mobile phase of aqueous methanol containing copper (II) acetate. Regis provides either an PLC column, or a kit which allows the user to convert a standard D column into a Davankov Chiral tationary Phase. aphthylleucine Protein-Based Chiral tationary Phases Regis Technologies carries a line of protein-based chiral columns manufactured by ChromTech AB, weden. For additional product information and a Protein-Based Chiral tationary Phase application guide, please contact Regis directly. Chiral AGP (α glycoprotein) Chiral CB (cellobiohydrolase) Chiral A (human serum albumin) As described above, Regis Chiral columns can be used to separate a wide variety of enantiomers in numerous compound groups. Please refer to the Product List on page 7 for particular column types, sizes, configurations and product numbers. Consult the application separation data section that begins on page 8 for information regarding specific chiral separations on a wide variety of compounds. ee Application Indexes on pages 85 and 86 for particular compounds or column types of interest. For your convenience, a customer service form can be found on page 87. For additional chromatography product information or catalog requests, please fill out the form and fax or mail directly to Regis. 6 REGI TECLGIE CIRAL APPLICATI GUIDE IV

7 Introduction REGI REGI PLC CIRAL CLUM PRDUCT LIT PRDUCT PARTICLE IZE CLUM DIMEI PRDUCT # (R,R)-Whelk- 1 5 µm, 100 Å (R,R)-Whelk- 1 5 µm, 100 Å 25 cm x 10.0 mm (,)-Whelk- 1 5 µm, 100 Å (,)-Whelk- 1 5 µm, 100 Å 25 cm x 10.0 mm (R,R)-Whelk µm, 100 Å (R,R)-Whelk µm, 100 Å 25 cm x 10.0 mm (R,R)-Whelk µm, 100 Å 25 cm x 21.1 mm (R,R)-Whelk µm, 100 Å 50 cm x 21.1 mm (,)-Whelk µm, 100 Å (,)-Whelk µm, 100 Å 25 cm x 10.0 mm (,)-Whelk µm, 100 Å 25 cm x 21.1mm (,)-Whelk µm, 100 Å 50 cm x 21.1 mm (R,R)-Whelk µm, 100 Å (R,R)-Whelk µm, 100 Å 25 cm x 10.0 mm (R,R)-Whelk µm, 100 Å 25 cm x 21.1 mm (R,R)-Whelk µm, 100 Å 50 cm x 21.1 mm (,)-Whelk µm, 100 Å (,)-Whelk µm, 100 Å 25 cm x 10.0 mm (,)-Whelk µm, 100 Å 25 cm x 21.1mm (,)-Whelk µm, 100 Å 50 cm x 21.1 mm (R,R)-ULM 5 µm, 100 Å (R,R)-ULM 5 µm, 100 Å 25 cm x 10.0 mm (,)-ULM 5 µm, 100 Å (,)-ULM 5 µm, 100 Å 25 cm x 10.0 mm (R,R)-DAC-DB 5 µm, 100 Å (R,R)-DAC-DB 5 µm, 100 Å 25 cm x 10.0 mm (,)-DAC-DB 5 µm, 100 Å (,)-DAC-DB 5 µm, 100 Å 25 cm x 10.0 mm (3R,4)-Pirkle 1-J 5 µm, 100 Å (3R,4)-Pirkle 1-J 5 µm, 100 Å 25 cm x 10.0 mm (3,4R)-Pirkle 1-J 5 µm, 100 Å (3,4R)-Pirkle 1-J 5 µm, 100 Å 25 cm x 10.0 mm (R,R)-α-Burke 2 5 µm, 100 Å (R,R)-α-Burke 2 5 µm, 100 Å 25 cm x 10.0 mm (,)-α-burke 2 5 µm, 100 Å (,)-α-burke 2 5 µm, 100 Å 25 cm x 10.0 mm (R,R)-β-Gem 1 5 µm, 100 Å (R,R)-β-Gem 1 5 µm, 100 Å 25 cm x 10.0 mm (,)-β-gem 1 5 µm, 100 Å (,)-β-gem 1 5 µm, 100 Å 25 cm x 10.0 mm D-Leucine 5 µm, 100 Å D-Leucine 5 µm, 100 Å 25 cm x 10.0 mm L-Leucine 5 µm, 100 Å L-Leucine 5 µm, 100 Å 25 cm x 10.0 mm D-Phenylglycine 5 µm, 100 Å D-Phenylglycine 5 µm, 100 Å 25 cm x 10.0 mm L-Phenylglycine 5 µm, 100 Å L-Phenylglycine 5 µm, 100 Å 25 cm x 10.0 mm L-aphthylleucine 5 µm, 100 Å L-aphthylleucine 5 µm, 100 Å 25 cm x 10.0mm Davankov 5 µm, 100 Å 15 cm x 4.6 mm Chiral AGP 5 µm, 300 Å 10 cm x 4.0 mm Chiral AGP 5 µm, 300 Å 15 cm x 4.0 mm Chiral CB 5 µm, 300 Å 10 cm x 4.0 mm Chiral CB 5 µm, 300 Å 15 cm x 4.0 mm Chiral A 5 µm, 300 Å 10 cm x 4.0 mm Chiral A 5 µm, 300 Å 15 cm x 4.0 mm Bulk material available. FC columns available. TE: All columns listed contain chiral stationary phases that are covalently bound on 5 µm or 10 µm 100 Å spherical silica. A large variety of column dimensions and/or particle sizes are available upon request. REGI TECLGIE CIRAL APPLICATI GUIDE IV 7

8 REGI Aryl Propionic Acid on-teroidal Anti-Inflammatory Drugs (AIDs) Ibuprofen Ketoprofen aproxen (normal phase) 3 C ibuprofen k' 1 = 1.13 α = :2:0.5 hexane/ipa/ac 0.9 ml/min; 254 nm run time = 8 min 4.6 mm x 25 cm Whelk- 1 reference 26 ketoprofen k' 1 = 3.63 α = 1.35 Column = (R,R)-Whelk- 1 Mobile Phase = (47/47/6) C 2 2 /exane/ethanol M Ammonium Acetate Flow Rate = 1.5 ml/min Run Time = 11.0 min naproxen (normal phase) k' 1 = 2.09 α = :20:0.5 hexane/ipa/ac run time = 16 min 4.6 mm x 25 cm Whelk- 1 reference 26 aproxen (reversed phase) aproxen (semi prep) Fenoprofen 3 C 3 C naproxen (reversed phase) k' 1 = 2.24 α = /40 / % Acetic Acid run time = 16 min 4.6 mm x 25 cm Whelk- 1 reference 26 naproxen (semi prep on analytical column) 80:20:0.5 hexane/ipa/ac 1 ml/min; 300 nm run time = 18 min inject mg/ml = 12.6 mg 4.6 mm x 25 cm Whelk- 1 reference 6 fenoprofen k' 1 = 2.62 α = 1.66 Column = (R,R)-Whelk- 1 Mobile Phase = (98/2) exane/ipa + 0.1% Acetic Acid Run Time = 14.5 min 8 REGI TECLGIE CIRAL APPLICATI GUIDE IV

9 Aryl Propionic Acid on-teroidal Anti-Inflammatory Drugs (AIDs) REGI Etodolac Cicloprofen etodolac k' 1 = 2.43 α = 1.50 Column = (,)-ULM Mobile Phase = (98/2) exane/ipa + 0.1% TFA Run Time = 14.5 min reference 48 cicloprofen k' 1 = 1.16 α = % IPA/hex, 1g/L 4 Ac 4.6 mm x 25 cm Whelk- 1 reference 4 Tiaprofenic Acid Pirprofen tiaprofenic acid k' 1 = 2.02 α = % IPA/hex, 1g/L 4 Ac 4.6 mm x 25 cm Whelk- 1 reference 4 pirprofen k' 1 = 0.85 α = % IPA/hex, 1g/L 4 Ac 4.6 mm x 25 cm Whelk- 1 reference 4 Ketoprofen as 1-naphthylamide aproxen (R:=30:70) Column: (,)-ULM Mobile Phase = (70/30) eptane/ipa Detection = UV 230 nm Run Time = 13 min k' 1 = 1.51 α = 1.25 reference 48 Flurbiprofen Column = (,)-ULM Mobile Phase (90/10) eptane/ipa + 0.1% TFA Detection = UV 230 nm Run Time = 8.5 min k' 1 = 1.54 α = 1.34 reference 48 Cicloprofen flurbiprofen k' 1 = 2.27 α = :5:0.5 hexane/et/ac 4.6 mm x 25 cm Whelk- 1 reference 26 F cicloprofen k' 1 = 0.48 α = :30:0.5 hexane/ipa/ac 4.6 mm x 25 cm Whelk- 1 reference 26 REGI TECLGIE CIRAL APPLICATI GUIDE IV 9

10 REGI Aryl Propionic Acid on-teroidal Anti-Inflammatory Drugs (AIDs) aproxen Diisopropyl Amide 3,5-Dimethylanilide-R,-Ibuprofen naproxen diisopropyl amide k' 1 = 2.23 α = %Et/hex 3 C 4.6 mm x 25 cm Whelk- 1 reference 26 Column = (3R,4)-Pirkle 1-J Mobile Phase = (85/15) exane/ipa Run Time = 13.0 min k' 1 = 2.91 α = 1.36 aproxen Methyl Ester aproxen Methyl Amide naproxen methyl ester k' 1 = 3.42 α = % IPA/hex, 1g/L 4 Ac 4.6 mm x 25 cm Whelk- 1 reference 14 3 C naproxen methyl amide k' 1 = α = % IPA/hex, 1g/L 4 Ac 4.6 mm x 25 cm Whelk- 1 reference 14 3 C aproxen aproxen Dimethyl Amide naproxen Extract from Aleve tablet (99.4%ee) 80:20:0.5 hexane/ipa/ac run time = 10 min 3C 4.6 mm x 25 cm (,) Whelk- 1 ample prep: 1/2 tablet partitioned between 1M C1 (2 ml) and C 2 C1 2 (5 ml) with sonication. C 2 C1 2 layer filtered through glass wool and injected naproxen dimethyl amide k' 1 = 5.24 α = % IPA/hex, 1g/L 4 Ac 4.6 mm x 25 cm Whelk- 1 reference 14 3 C Indoprofen indoprofen k' 1 = 8.93 α = 1.32 Column = (,)-Whelk- 1 10/100 (FEC) Mobile Phase = (80/20) exane/ethanol M Ammonium Acetate Flow Rate = 2.0 ml/min Run Time 17.0 min α-trityl-2-naphthalene Propionic Acid α-trityl-2-naphthalene propionic acid k' 1 = 1.57 α = 1.79 Column = (R,R)-ULM Mobile Phase = (97/3)exane/IPA Run Time = 10.0 min 10 REGI TECLGIE CIRAL APPLICATI GUIDE IV

11 ther Carboxylic Acids REGI Tetrahydropyrimindine Carboxylic Acid Column: (,)-ULM Mobile Phase: (90/10) eptane/ipa + 0.1% TFA Flow Rate: 1.0 ml/min Detection: UV 215 nm Run Time: 14 min k' 1 = 3.38 α = 1.21 reference 48 Column = (,)-ULM Mobile Phase = (99/1) eptane/ipa + 0.1% TFA Flow Rate = 2.0 ml/min Detection = UV 215 nm Run Time = 6.5 min k' 1 = 3.19 α = 1.16 reference 48 Phenylbutyric acid k' 1 = 4.06 α = :1:0.1 hexane/ipa/ac run time = 17 min 4.6 mm x 25 cm Whelk- 1 k' 1 = 3.45 α = :1:0.1 hexane/ipa/ac run time = 16 min 4.6 mm x 25 cm Whelk- 1 Ph Ph Ph 2-Phenylcyclopropane Carboxylate Trolox 2-phenylcyclopropane carboxylate k' 1 = 4.19 α = :1 hexane/ipa 1 ml/min; 220 nm run time = 18 min 4.6 mm x 25 cm Whelk- 1 trolox k' 1 = 5.09 α = C 95:5:0.1 hexane/ipa/ac run time = 19 min 4.6 mm x 25 cm Whelk- 1 Mandelic Acid 1,1'-binaphthyl-2,2'-diylhydrogen phosphate mandelic acid k' 1 = 3.08 α = % Ac in water run time = 13 min 4.6 mm x 25 cm Whelk- 1 k' 1 = 4.46 α = :44 2 /Me, 0.1% Ac run time = 18 min 4.6 mm x 25 cm Whelk- 1 P REGI TECLGIE CIRAL APPLICATI GUIDE IV 11

12 REGI ther Carboxylic Acids Column = (,)-Whelk- 1 Mobile Phase = (95/5) exane/ipa + 0.1% Trifluoroacetic Acid Flow Rate = 2.0 ml/min Run Time = 8.5 min k' 1 = 2.03 α = 2.10 reference 49 Column = (,)-Whelk- 1 Mobile Phase = (95/5) exane/ipa + 0.1% Trifluoroacetic Acid Flow Rate = 2.0 ml/min Run Time = 8.5 min k' 1 = 4.20 α = 1.11 reference 50 Column = (,)-Whelk- 1 Mobile Phase = (95/5) exane/ipa + 0.1% Trifluoroacetic Acid Flow Rate = 2.0 ml/min Run Time = 10.0 min k' 1 = 2.07 α = 2.62 reference 49 Column = (,)-Whelk- 1 Mobile Phase = (95/5) exane/ipa + 0.1% Trifluoroacetic Acid Flow Rate = 2.0 ml/min Run Time = 14.5 min k' 1 = 7.24 α = 1.22 reference 50 Trolox Column = (R,R)-ULM Mobile Phase = (95/5) exane/ipa + 0.1% Acetic acid Flow Rate = 1.5 ml/min Detection = UV 280 nm Run Time = 12.5 min k' 1 = 2.18 α = C Column = (,)-Whelk- 1 Mobile Phase = (95/5) exane/ipa + 0.1% Trifluoroacetic Acid Flow Rate = 2.0 ml/min Run Time = 11.5 min k' 1 = 5.44 α = 1.34 reference 50 Calcium Channel Blocker Column = (,)-Whelk- 1 Mobile Phase: (95/5) exane/ipa + 0.1% Trifluoroacetic Acid Flow Rate = 2.0 ml/min Run Time = 3.5 min k' 1 = 0.84 α = 1.36 reference 49 Column = (,)-ULM Mobile Phase = (99/1) eptane/ipa + 0.1% TFA Detection = UV 230 nm Run Time = 6 min k' 1 = 0.55 α = 2.06 reference REGI TECLGIE CIRAL APPLICATI GUIDE IV

13 ther Carboxylic Acids REGI Phenylsuccinic Acid 4-Chloromandelic Acid α-methoxyphenyl Acetic Acid phenylsuccinic acid Column = (,)-ULM Mobile Phase = (95/5) exane/ipa + 0.1% TFA Run Time = 8.5 min k' 1 = 1.71 α = 1.22 reference 48 4-chloromandelic acid Column = (R,R)-Whelk- 2 Mobile Phase = (70/30) 2 / + 0.1% Acetic Acid Run Time = 10.0 min k' 1 = 1.95 α = 1.43 α-methoxyphenyl Acetic Acid Column = (,)-Whelk- 1 10/100 (FEC) Mobile Phase = (90/10) exane/ethanol M Ammonium Acetate Flow Rate = 1.5 ml/min Detection = UV 220 nm Run Time = 10.0 min k' 1 = 2.96 α = 1.61 Ketorolac uprofen Ditoluoyltartaric Acid ketorolac Column = (R,R)-Whelk- 1 Mobile Phase = (98/2) exane/ipa + 0.1% TFA Flow Rate = 1.5 ml/min Run Time = 20.0 min k' 1 = 8.87 α = 1.15 suprofen Column = (,)-Whelk- 1 10/100 (FEC) 25 cm x 4.6 mm Mobile Phase = (80/20) exane/ipa M Ammonium Acetate Flow Rate = 2.0 ml/min Run Time = 18.0 min k' 1 = 9.76 α = 1.27 ditoluoyltartaric acid k' 1 = 2.47 α = 1.19 column: (,)-ULM mobile phase: (90/10) exane/ipa + 0.1% TFA flow rate: 1.0 ml/min detection: UV 254 nm run time = 12.0 min reference 48 REGI TECLGIE CIRAL APPLICATI GUIDE IV 13

14 REGI ther Carboxylic Acids Trolox-methylether 1-Cyclopentyl-1-phenylacetic Acid trolox-methylether k' 1 = 0.32 α = 2.50 column: (,)-ULM 25 cm x 4.6 mm mobile phase: (90/10) exane/ipa + 0.1% TFA flow rate: 1.0 ml/min detection: UV 254 nm run time = 6.0 min reference 48 1-cyclopentyl-1-phenylacetic acid k' 1 = 2.46 α = 1.19 column: (,)-ULM mobile phase: (99/1) exane/ipa + 0.1% TFA flow rate: 1.0 ml/min detection: UV 254 nm run time = 12.0 min reference 48 1-Cyclohexyl-1-phenylacetic Acid 2-(2-Chloro-4-methylphenoxy)propionic acid 1-cyclohexyl-1-phenylacetic acid k' 1 = 2.53 α = 1.18 column: (,)-ULM mobile phase: (99/1) exane/ipa + 0.1% TFA flow rate: 1.0 ml/min detection: UV 254 nm run time = 13.0 min reference 48 2-(2-chloro- 4-methylphenoxy)propionic acid k' 1 = 2.22 α = 1.11 column: (,)-ULM mobile phase: (99/1) exane/ipa + 0.1% TFA flow rate: 1.0 ml/min detection: UV 254 nm run time = 11.0 min reference REGI TECLGIE CIRAL APPLICATI GUIDE IV

15 Esters, Lactones, Ketones, Anhydrides REGI Et Et 3 C k' 1 = 1.27 α = :5:0.5 hexane/ipa/ac 1 ml/min; 280 nm 4.6 mm x 25 cm Whelk- 1 reference 26 k' 1 = 2.85 α = :5:0.5 hexane/ipa/ac 1 ml/min; 280 nm 4.6 mm x 25 cm Whelk- 1 reference 26 k' 1 = 2.75 α = :5:0.5 hexane/ipa/ac 1 ml/min; 280 nm 4.6 mm x 25 cm Whelk- 1 reference 26 Methyl Mandelate 2-Methyl-1-Indanone k' 1 = 8.10 α = :7:0.1 hexane/ipa/ac run time = 30 min 4.6 mm x 25 cm Whelk- 1 methyl mandelate k' 1 = 5.27 α = :27:0.1 2 / C/Ac run time = 20 min 4.6 mm x 25 cm Whelk- 1 2-methyl-1-indanone k' 1 = 4.00 α = :1 hexane/ipa run time = 15 min 4.6 mm x 25 cm Whelk- 1 REGI TECLGIE CIRAL APPLICATI GUIDE IV 15

16 REGI Esters, Lactones, Ketones, Anhydrides k' 1 = 3.41 α = :10 hexane/ipa 4.6 mm x 25 cm Whelk- 1 reference 7 k' 1 = 5.66 α = % IPA/hex 4.6 mm x 25 cm Whelk- 1 reference 7 3 C k' 1 = 1.17 α = % IPA/hex 4.6 mm x 25 cm Whelk- 1 reference 7 3 C k' 1 = 7.82 α = :2 hexane/ipa 4.6 mm x 25 cm Whelk- 1 reference 7 3-Methyl-1-Indanone 3-methyl-1-indanone k' 1 = 6.11 α = :1 hexane/ipa run time = 20 min 4.6 mm x 25 cm Whelk- 1 k' 1 = α = 1.16 Me/IPA/hexane run time = 17 min 4.6 mm x 25 cm Whelk- 1 reference 19 Et Et DPB DPB 6% Et/hexane run time = 41 min 4.6 mm x 25 cm Whelk- 1 reference 29 Column = L-Leucine Mobile Phase = (99.5/0.5) exane/ipa Run Time = 11.5 min k' 1 = 2.42 α = 1.21 reference REGI TECLGIE CIRAL APPLICATI GUIDE IV

17 Esters, Lactones, Ketones, Anhydrides REGI 2-Methyl-1-Tetralone 3 C 2-methyl-1-tetralone k' 1 = 2.76 α = :1 hexane/ipa run time = 12 min 4.6 mm x 25 cm Whelk- 1 k' 1 = 8.46 α = :3 hexane/ipa run time = 27 min 4.6 mm x 25 cm Whelk- 1 k' 1 = 2.27 α = % IPA/hexane 4.6 mm x 25 cm Whelk- 1 reference 26 Buckminsterfullerene-Enone [2+2] Photoadducts k' 1 = 0.67 α = % IPA/hexane 4.6 mm x 25 cm Whelk- 1 reference 26 semi-prep separation on analytical column 2:1 toluene/hexane 1 ml/min; 400 nm run time = 22 min sample: 100µ1 of 5 mg/ml solution (0.5 mg) 4.6 mm x 25 cm Whelk- 1 reference 8 REGI TECLGIE CIRAL APPLICATI GUIDE IV 17

18 REGI Esters, Lactones, Ketones, Anhydrides Column = (,)-Whelk- 1 Mobile Phase = (98/2) exane/ipa Run Time = 34.0 min k' 1 = 8.00 α = 1.44 reference 51 Column = (,)-Whelk- 1 Mobile Phase = (98/2) exane/ipa Detection = UV 254 nm Run Time = 20.5 min k' 1 = 1.62 α = 4.18 reference 51 Column = D-Phenylglycine Mobile Phase = 99/1) exane/ipa Run Time = 13.5 min k' 1 = 3.10 α = 1.18 reference 57 Diperodon 1,3,5-Triphenylpent-4-yn-1-one Column = (,)-β-gem 1 Mobile Phase = (99.5/0.5) exane/ipa Run Time = 14.5 min k' 1 = 2.67 α = 1.43 reference 57 Diperodon Column = (R)-α-Burke 2 Mobile Phase = (48/48/4) C 2 2 /exane/ethanol mm Ammonium Acetate Run Time = 9.0 min k' 1 = 1.7 α = ,3,5-Triphenylpent-4-yn-1-one Column = (,)-ULM Mobile Phase = exane + 0.5% IPA Run Time = 6.5 min k' 1 = 1.19 α = REGI TECLGIE CIRAL APPLICATI GUIDE IV

19 Basic itrogen REGI Troger s Base Bz 3 C Troger's base k' 1 = 4.65 α = :3 hexane/ipa run time = 20 min 4.6 mm x 25 cm Whelk- 1 k' 1 = 2.46 α = % Et/hexane run time = 18 min 4.6 mm x 25 cm Whelk- 1 REGI TECLGIE CIRAL APPLICATI GUIDE IV 19

20 REGI Amides, Imides, Carbamates, etc. k' 1 = 0.50 α = 1.56 ethyl acetate 4.6 mm x 25 cm Whelk- 1 reference 7 k' 1 = 0.64 α = 2.06 ethyl acetate 4.6 mm x 25 cm Whelk- 1 reference 26 k' 1 = 0.16 α = 5.56 ethyl acetate 4.6 mm x 25 cm Whelk- 1 reference 7 0. k' 1 = 0.14 α = 3.21 ethyl acetate 4.6 mm x 25 cm Whelk- 1 reference 26 k' 1 = α = :5:0.5 hexane/ipa/ac 4.6 mm x 25 cm Whelk- 1 reference 7 Me Et k' 1 = 5.13 α = %Et/hex 4.6 mm x 25 cm Whelk- 1 reference 26 p-ts p-ts k' 1 = 3.64 α = % IPA/hexane 4.6 mm x 25 cm Whelk- 1 reference 26 k' 1 = 4.45 α = % IPA/hexane 4.6 mm x 25 cm Whelk- 1 reference REGI TECLGIE CIRAL APPLICATI GUIDE IV

21 Amides, Imides, Carbamates, etc. REGI k' 1 = 3.72 α = % IPA/hexane run time = 4 min (,) Whelk- 1 reference 38 k' 1 = 1.48 α = % IPA/hexane run time = 4 min (,) Whelk- 1 reference 38 k' 1 = 4.10 α = % IPA/hexane run time = 4 min (,) Whelk- 1 reference 38 I k' 1 = 1.61 α = % IPA/hexane run time = 4 min (,) Whelk- 1 reference 38 Br k' 1 = 1.39 α = % IPA/hexane run time = 4 min (,) Whelk- 1 reference 38 k' 1 = 1.75 α = % IPA/hexane run time = 4 min (,) Whelk- 1 reference 38 Br k' 1 = 1.17 α = % IPA/hexane run time = 4 min (,) Whelk- 1 reference 38 F k' 1 = α = % IPA/hexane run time = 4 min (,) Whelk- 1 reference 38 3 C Et P Et REGI TECLGIE CIRAL APPLICATI GUIDE IV 21

22 REGI Amides, Imides, Carbamates, etc. k' 1 = 0.83 α = % IPA/hexane run time = 4 min (,) Whelk- 1 reference 38 3 C Et P Et k' 1 = 0.84 α = % IPA/hexane run time = 4 min (,) Whelk- 1 reference 38 3 C Et P Et F k' 1 = 0.86 α = % IPA/hexane run time = 4 min (,) Whelk- 1 reference 38 3 C Et P Et k' 1 = 8.5 α = % acetonitrile in C 2 (,) Whelk- 1 reference 39 D D Br D D D TM k' 1 = 25 α = % acetonitrile in C 2 (,) Whelk- 1 reference 39 D D k' 1 = 19.7 α = % acetonitrile in C 2 (,) Whelk- 1 reference 39 Br D D D D D D TM Br Column = (,)-Whelk- 1 Mobile Phase = (90/10) exane/ipa Run Time = 46.0 min k' 1 = 2.70 α = 6.02 reference 51 Column = (R,R)-Whelk- 1 Mobile Phase = (80/20) exane/ipa Flow Rate = 2.0 ml/min Run Time = 19.0 min k' 1 = 7.53 α = 1.77 reference REGI TECLGIE CIRAL APPLICATI GUIDE IV

23 Amides, Imides, Carbamates, etc. REGI EEDQ CBZ nornicotine k' 1 = 1.75 EEDQ k' 1 = 1.53 α = :10 hexane/ipa run time = 25 min 4.6 mm x 25 cm Whelk- 1 CBZ nornicotine k' 1 = 0.37 α = :3 Me/dichloromethane run time = 5 min 4.6 mm x 25 cm Whelk- 1 reference 7 k' 1 = 1.75 α = 1.20 methanol run time = 6 min 4.6 mm x 25 cm Whelk- 1 reference 7 CBZ-Val BC-Ala CBZ-Phe CBZ-Val k' 1 = 5.49 α = :5:0.1 hexane/ipa/ac run time = 19 min 4.6 mm x 25 cm Whelk- 1 BC-Ala k' 1 = 4.43 α = :2:0.2 hexane/ipa/ac 1 ml/min; 220 nm run time = 17 min 4.6 mm x 25 cm Whelk- 1 CBZ-Phe k' 1 = 10.2 α = :5:0.1 hexane/ipa/ac run time = 40 min 4.6 mm x 25 cm Whelk- 1 REGI TECLGIE CIRAL APPLICATI GUIDE IV 23

24 REGI Amides, Imides, Carbamates, etc. Ph Ph k' 1 = 5.97 α = :20:0.1 hexane/ipa/ac run time = 25 min 4.6 mm x 25 cm Whelk- 1 k' 1 = 3.79 α = % Et/hexane run time = 16 min 4.6 mm x 25 cm Whelk- 1 β-lactam β-lactam Column: (,)-DAC-DB Mobile Phase: (48/48/2) ex/c 2 2 /IPA Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 14.0 min k 1 : 3.40 α: 1.33 reference REGI TECLGIE CIRAL APPLICATI GUIDE IV

25 Epoxides REGI tyrene xide tilbene xide styrene oxide k' 1 = 1.37 α = % IPA/hexane 4.6 mm x 25 cm Whelk- 1 stilbene oxide k' 1 = 0.45 α = % IPA/hexane 4.6 mm x 25 cm Whelk- 1 m- tyrene xide ene k' 1 = 5.92 α = % Ac in hexane run time = 20 min 4.6 mm x 25 cm Whelk- 1 m- styrene oxide hexane 1 ml/min; 220 nm 4.6 mm x 25 cm Whelk- 1 reference 30 REGI TECLGIE CIRAL APPLICATI GUIDE IV 25

26 REGI Alcohols Ibuprofenol 3 C Tert Butyl Phenyl Carbinol ibuprofenol k' 1 = 3.38 α = :1 hexane/ipa run time = 14 min 4.6 mm x 25 cm Whelk- 1 reference 26 k' 1 = 1.22 α = :20 hexane/ipa run time = 10 min 4.6 mm x 25 cm Whelk- 1 reference 26 Column = (,)-ULM Mobile Phase = (99/1) eptane/ipa Detection = UV 215 nm Run Time = 6.0 min k' 1 = 4.60 α = ,2,3,4-Tetrahydro-1-aphtol α-aphthyl Methyl Carbinol 9-Anthryl Trifluoromethyl Carbinol CF 3 Column = (R,R)-ULM Mobile Phase = (99/1) exane/ipa Run Time = 10.5 min k' 1 = 2.17 α = 1.30 Column = (R,R)-ULM Mobile Phase = (99/1) exane/ipa Run Time = 14.5 min k' 1 = 3.49 α = 1.25 Column = (R,R)-ULM Mobile Phase = (95/5) exane/ipa Run Time = 10 min k' 1 = 1.36 α = REGI TECLGIE CIRAL APPLICATI GUIDE IV

27 Acenaphthenol Alcohols REGI Beta aphthyl Methyl Carbinol k' 1 = 1.76 α = % IPA/hexane 1 ml/min; 220 nm 4.6 mm x 25 cm Whelk- 1 Column: (R,R)-ULM Mobile Phase: (95/5) exane/ipa Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 10 min k 1 : 1.68 α: 1.46 Column: (R,R)-ULM Mobile Phase: (97/3) exane/ipa Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 9 min k 1 : 1.64 α: ,1 -Bi-2-aphthol Tetrahydrobenzopyrene-7-ol 1,1 -bi-2-naphthol k' 1 = 4.84 α = 1.24 Column = (,)-ULM Mobile Phase = (98/2) exane/ipa + 0.1% TFA Run Time = 18.0 min reference 48 tetrahydrobenzopyrene-7-ol k' 1 = 6.10 α = :20 hexane/ipa run time = 22 min 4.6 mm x 25 cm Whelk- 1 k' 1 = 3.38 α = % IPA/hexane 4.6 mm x 25 cm Whelk- 1 reference 7 REGI TECLGIE CIRAL APPLICATI GUIDE IV 27

28 REGI Alcohols 9-Anthrylethanol 1-aphthyl-2-butanol Column = (,)-Whelk- 1 Mobile Phase = (99/1) exane/ipa Run Time = 18.5 min k' 1 = 5.59 α = 1.09 reference 55 Column = (,)-ULM Mobile Phase = (95/5) eptane/ipa Detection = UV 215 nm Run Time = 12 min k' 1 = 1.82 α = 1.74 reference 48 Column = (,)-ULM Mobile Phase = (95/5) eptane/ipa Detection = UV 215 nm Run Time = 6 min k' 1 = 0.80 α = 1.35 reference 48 2-aphthyl-2-butanol 1-Phenylpentanol Column = (,)-ULM Mobile Phase = (95/5) eptane/ipa Detection = UV 215 nm Run Time = 8 min k' 1 = 1.00 α = 1.93 reference 48 Column = (,)-ULM Mobile Phase = (99/1) eptane/ipa Detection = UV 215 nm Run Time = 7 min k' 1 = 1.18 α = 1.30 reference 48 Column = (,)-Whelk- 1 Mobile Phase = (80/20) exane/ipa Flow Rate = 2.0 ml/min Run Time = 24.0 min k' 1 = α = 1.11 reference REGI TECLGIE CIRAL APPLICATI GUIDE IV

29 Phenyl Tribromomethyl Carbinol Phenyl Ethyl Carbinol Alcohols REGI Phenyl Propyl Carbinol CBr 3 Column = (R,R)-ULM Mobile Phase = (99/1) exane/ipa Run Time = 9 min k' 1 = 1.87 α = 1.25 Column = (R,R)-ULM Mobile Phase = (99/1) exane/ipa Run Time = 6.5 min k' 1 = 1.06 α = 1.30 Column = (R,R)-ULM Mobile Phase = 100% exane Run Time = 12 min k' 1 = 2.25 α = 1.56 Phenyl Methyl Carbinol 1,1,2,-Triphenyl-1,2-Ethanediol Column = (R,R)-ULM Mobile Phase = 100% exane Run Time = 14 min k' 1 = 3.11 α = 1.30 Column = (R,R)-ULM Mobile Phase = (99/1) exane/ipa Run Time = 10 min k' 1 = 1.97 α = 1.37 reference 48 Column = (,)-ULM Mobile Phase = (99/1) eptane/ipa Detection = UV 215 nm Run Time = 13 min k' 1 = 2.59 α = 1.14 reference 48 REGI TECLGIE CIRAL APPLICATI GUIDE IV 29

30 REGI Alcohols 1,1 -Binaphthol Monomethylether Terfenadine 1,1'-Binaphthol Monomethylether k' 1 = 2.23 α = 1.28 column: (,)-ULM 25 cm x 4.6 mm mobile phase: (98/2) exane/ipa + 0.1% TFA flow rate: 1.0 ml/min detection: UV 254 nm run time = 11.0 min reference 48 Terfenadine Column = (R,R)-Whelk- 1 Mobile Phase = (97/3) exane/ethanol M Ammonium Acetate Flow Rate = 1.5 ml/min Run Time = 15.0 min k' 1 = 5.91 α = 1.20 Propafenone Trans Phenyl Cyclohexanol Analytical vs. Preparative Run propafenone Column = (R,R)-Whelk- 1 Mobile Phase = (47/47/6) C 2 2 /exane/ethanol M Ammonium Acetate Flow Rate = 1.5 ml/min Run Time = 11.0 min k' 1 = 3.99 α = 1.25 Column = (,)-ULM Mobile Phase = (99/1) eptane/ipa Detection = UV 270 nm Run Time = 7.0 min reference MG 1.4 MG 0.14 MG 30 REGI TECLGIE CIRAL APPLICATI GUIDE IV

31 2-Methoxyphenyl Phenyl Carbinol Phenyl cyclohexyl carbinol Alcohols REGI Phenyl isopropyl carbinol Column = (,)-ULM Mobile Phase = (99/1) eptane/ipa Detection = UV 215 nm Run Time = 12.0 min k' 1 = 2.92 α = 1.13 reference 48 Column = (,)-ULM Mobile Phase: (99/1) eptane/ipa Detection = UV 215 nm Run Time = 6.5 min k' 1 = 0.97 α = 1.39 reference 48 Column = (,)-ULM Mobile Phase = (99/1) eptane/ipa Detection = UV 215 nm Run Time: 6 min k' 1 = 0.86 α = 1.38 reference 48 Phenyl phenylethyl carbinol Methyl 3-phenyl-3azido- 2hydroxypropanoate (Erythro-diastereomer) Column = (,)-ULM Mobile Phase = (99/1) eptane/ipa Detection = UV 215 nm Run Time = 9.5 min k' 1 = 1.81 α = 1.30 reference 48 Column = (,)-ULM Mobile Phase = (97/3) eptane/glyme Detection = UV 215 nm Run Tim = 10.5 min k' 1 = 2.34 α = 1.16 reference 48 REGI TECLGIE CIRAL APPLICATI GUIDE IV 31

32 REGI Diols, ydroxyketones, etc. ydrobenzoin hydrobenzoin k' 1 = 1.14 α = :5 hexane/ipa run time = 18 min 4.6 mm x 25 cm Whelk- 1 k' 1 = 7.54 α = :2:0.5 hexane/ipa/ac 4.6 mm x 25 cm Whelk- 1 reference 7 k' 1 = 4.20 α = :2:0.5 hexane/et/ac 1 ml/min; 240 nm 4.6 mm x 25 cm Whelk- 1 reference 26 k' 1 = 2.03 α = :40 hexane/et 1 ml/min; 240 nm 4.6 mm x 25 cm Whelk- 1 reference 26 3 C irvanol Benzoin Benzoin benzoin k' 1 = 0.86 α = :20:0.5 hexane/ipa/ac 4.6 mm x 25 cm Whelk- 1 reference 7 benzoin k' 1 = 0.24 α = :1 dichloromethane/methanol 4.6 mm x 25 cm Whelk- 1 reference 26 Anisoin Ipsdienol anisoin k' 1 = 3.07 α = :20:0.5 hexane/ipa/ac 4.6 mm x 25 cm Whelk- 1 reference 26 3 C ipsdienol k' 1 = 0.95 α = % IPA/hexane run time = 8 min 4.6 mm x 25 cm Whelk REGI TECLGIE CIRAL APPLICATI GUIDE IV

33 ulfoxides REGI 3 C C3 C3 C3 Br k' 1 = 3.83 α = :2:1 hexane/ipa/c 2 C mm x 25 cm Whelk- 1 reference 7 k' 1 = 4.10 α = :2:1 hexane/ipa/c 2 C1 2 run time = 9 min 4.6 mm x 25 cm Whelk- 1 k' 1 = 3.75 α = :2:1 hexane/ipa/c 2 C1 2 run time = 8 min 4.6 mm x 25 cm Whelk- 1 C3 Br 3 C k' 1 = 3.75 α = :2:1 hexane/ipa/c 2 C1 2 run time = 8 min 4.6 mm x 25 cm Whelk- 1 k' 1 = 1.90 α = :2:1 hexane/ipa/c 2 C1 2 run time = 6 min 4.6 mm x 25 cm Whelk- 1 k' 1 = 2.11 α = :20 hexane/ipa 1.5 ml/min; 254 nm run time = 14 min 4.6 mm x 25 cm Whelk- 1 reference 31 REGI TECLGIE CIRAL APPLICATI GUIDE IV 33

34 REGI ulfoxides C3 k' 1 = 3.29 α = :2:1 hexane/ipa/c 2 C mm x 25 cm Whelk- 1 reference 7 k' 1 = 5.02 α = :2:1 hexane/ipa/c 2 C1 2 run time = 11 min 4.6 mm x 25 cm Whelk- 1 C3 k' 1 = 5.04 α = :2:1 hexane/ipa/c 2 C1 2 run time = 11 min 4.6 mm x 25 cm Whelk- 1 k' 1 = α = :2:1 hexane/ipa/c 2 C1 2 run time = 6 min 4.6 mm x 25 cm Whelk REGI TECLGIE CIRAL APPLICATI GUIDE IV

35 ulfoxides REGI ulfinpyrazone meprazole sulfinpyrazone k' 1 = 3.74 α = 1.35 Column = (R,R)-Whelk- 1 Mobile Phase = (75/25) exane/ethanol + 15 mm Ammonium Acetate Flow Rate = 1.5 ml/min Run Time = 11.0 min omeprazole k' 1 = 0.64 α = 3.04 Column = ()-α-burke 2 Mobile Phase = (95/5) C 2 2 / Detection = UV 302 nm Run Time = 8.0 min Column: (,)-DAC-DB Mobile Phase: (27.5/27.5/45) C 2 2 /Dioxane/ex Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 35.0 min k 1 : α: 1.15 reference 59 Column: (,)-DAC-DB Mobile Phase: (98/2) C 2 2 /IPA Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 13.0 min k 1 : 3.08 α: 1.26 reference 59 Column: (,)-DAC-DB Mobile Phase: (98/2) C 2 2 /IPA Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 17.0 min k 1 : 3.33 α: 1.63 reference 59 Column: (R,R)-DAC- DB Mobile Phase: (98/2) C 2 2 /IPA Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 16.0 min k 1 : 2.34 α: 2.07 reference 59 REGI TECLGIE CIRAL APPLICATI GUIDE IV 35

36 REGI ulfoxides Column: (,)-DAC-DB Mobile Phase: (40/40/20) C 2 2 /Dioxane/ex Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 27.0 min k 1 : 7.51 α: 1.21 reference 59 Column: (R,R)-DAC-DB Mobile Phase: (27.5/27.5/45) C 2 2 /Dioxane/ex Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 20.0 min k 1 : 5.16 α: 1.26 reference 59 Column: (R,R)-DAC-DB Mobile Phase: (27.5/27.5/45) C 2 2 /Dioxane/ex Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 18.0 min k 1 : 4.77 α: 1.18 reference 59 Column: (R,R)-DAC-DB Mobile Phase: (95/5) C 2 2 /IPA Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 15.0 min k 1 : 2.15 α: 2.05 reference REGI TECLGIE CIRAL APPLICATI GUIDE IV

37 Phosphorous Compounds REGI F 3 C 3 C Et P Et Et P Et Et P Et k' 1 = 2.26 α = % 2-propanol in hexane (,) Whelk- 1 reference 40 k' 1 = 4.72 α = % 2-propanol in hexane (,) Whelk- 1 reference 40 k' 1 = 4.26 α = % 2-propanol in hexane (,) Whelk- 1 reference 40 CF 3 Et P Et F F Et P Et CF 3 Et P Et k' 1 = 4.10 α = % 2-propanol in hexane (,) Whelk- 1 reference 40 k' 1 = 4.09 α = % 2-propanol in hexane (,) Whelk- 1 reference 40 k' 1 = 6.05 α = % 2-propanol in hexane (,) Whelk- 1 reference 40 REGI TECLGIE CIRAL APPLICATI GUIDE IV 37

38 REGI Phosphorous Compounds Et P Et Et P Et Et P Et k' 1 = 9.61 α = % 2-propanol in hexane (,) Whelk- 1 reference 40 k' 1 = 4.58 α = % 2-propanol in hexane (,) Whelk- 1 reference 40 k' 1 = 7.35 α = % 2-propanol in hexane (,) Whelk- 1 reference 40 Et P Et Et P Et 3 C P 2 k' 1 = 5.87 α = % 2-propanol in hexane (,) Whelk- 1 reference 40 k' 1 = α = % 2-propanol in hexane (,) Whelk- 1 reference 40 k' 1 = 1.11 α = :4:1 hexane/c 2 C1 2 / C 4.6 mm x 25 cm Whelk- 1 reference 7 38 REGI TECLGIE CIRAL APPLICATI GUIDE IV

39 Phosphorous Compounds REGI Cyclophosphamide Tertiary Phosphine xide Phosphine elenium xide cyclophosphamide k' 1 = 6.31 α = 1.27 Column = (,)-Whelk- 1 10/100 (FEC) Mobile Phase = (95/5) exane/ethanol Flow Rate = 1.5 ml/min Detection = UV 195 nm Run Time = 16.0 min tertiary phosphine oxide Column: (R,R)-DAC-DB Mobile Phase: (37.5/37.5/25) ex/dioxane/ipa Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 14.0 min k 1 : 2.19 α: 1.48 reference 59 phosphine selenium oxide Column: (,)-DAC-DB Mobile Phase: (70/30) ex/c 2 2 Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 13.0 min k 1 : 2.49 α: 1.48 reference 59 econdary Phosphine xide econdary Phosphine xide econdary Phosphine xide secondary phosphine oxide Column: (,)-DAC-DB Mobile Phase: (75/25) C 2 2 /IPA Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 19.0 min k 1 : 1.49 α: 4.11 reference 59 secondary phosphine oxide Column: (,)-DAC-DB Mobile Phase: (90/10) C 2 2 /IPA Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 8.0 min k 1 : 1.23 α: 1.81 reference 59 secondary phosphine oxide Column: (,)-DAC-DB Mobile Phase: (90/10) C 2 2 /IPA Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 14.5 min k 1 : 2.20 α: 1.97 reference 59 REGI TECLGIE CIRAL APPLICATI GUIDE IV 39

40 REGI Phosphorous Compounds Et Et P Tertiary Phosphine xide P k' 1 = 1.35 α = % IPA/hex 4.6 mm x 25 cm Whelk- 1 reference 7 k' 1 = 3.07 α = % Et/hexane run time = 13 min 4.6 mm x 25 cm Whelk- 1 tertiary phosphine oxide Column: (R,R)-DAC-DB Mobile Phase: (42.5/42.5/15) ex/dioxane/ipa Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 28.0 min k 1 : 8.11 α: 1.17 reference 59 Tertiary Phosphine xide P k' 1 = 3.75 α = % Et/hexane run time = 18 min 4.6 mm x 25 cm Whelk- 1 tertiary phosphine oxide Column: (R,R)-DAC-DB Mobile Phase: (40/40/20) ex/dioxane/ipa Flow Rate: 1.0 ml/min Detection: UV 254 nm Run Time: 14.0 min k 1 : 4.19 α: 1.25 reference REGI TECLGIE CIRAL APPLICATI GUIDE IV

41 Atropisomers REGI k' 1 = 0.46 α = 2.17 EtAc 4.6 mm x 25 cm Whelk- 1 reference 7 Et Et k' 1 = 2.26 α = % IPA/hexane 4.6 mm x 25 cm Whelk- 1 reference 7 k' 1 = 1.73 α = % IPA/hexane 4.6 mm x 25 cm Whelk- 1 reference 7 k' 1 = 1.91 α = % IPA/hexane 4.6 mm x 25 cm Whelk- 1 reference 7 k' 1 = 3.29 α = :20:0.1% hexane/ipa/ac 4.6 mm x 25 cm Whelk- 1 reference 10 k' 1 = 3.23 α = :20:0.1% hexane/ipa/ac 4.6 mm x 25 cm Whelk--1 reference 10 k' 1 = 6.24 α = :20:0.1% hexane/ipa/ac 4.6 mm x 25 cm Whelk--1 reference 10 k' 1 = 4.46 α = :20:0.1% hexane/ipa/ac 4.6 mm x 25 cm Whelk--1 reference 10 REGI TECLGIE CIRAL APPLICATI GUIDE IV 41

42 REGI Atropisomers k' 1 = α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 4.06 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 6.29 α = % 2-propanol in hexane (,) Whelk- 1 reference REGI TECLGIE CIRAL APPLICATI GUIDE IV

43 Atropisomers REGI k' 1 = 7.95 α = :20:0.1% hexane/ipa/ac 4.6 mm x 25 cm Whelk- 1 reference 10 mixture of stereoisomers 80:20:0.1% hexane/ipa/ac 4.6 mm x 25 cm Whelk- 1 reference 10 k' 1 = 2.05 α = :20:0.1% hexane/ipa/ac 4.6 mm x 25 cm Whelk- 1 reference 10 k' 1 = 1.71 α = :20:0.1% hexane/ipa/ac 4.6 mm x 25 cm Whelk- 1 ()(-) retained on (,) CP reference 10 sulfoxide atropisomer k' 1 = 5.11 α = :20:0.1% hexane/ipa/ac 4.6 mm x 25 cm Whelk- 1 reference 10 sulfoxide atropisomer Z diastereomer k' 1 = 1.32 α = % Me in C ml/min; 300 nm, -40 o C 4.6 mm x 25 cm Whelk- 1 reference 21 sulfone atropisomer sulfone atropisomer sulfone atropisomer k' 1 = 0.54 α = % Me in C ml/min; 300 nm, -80 C 4.6 mm x 25 cm Whelk- 1 reference 21 sulfone atropisomer -80 C reference 22 3 C REGI TECLGIE CIRAL APPLICATI GUIDE IV 43

44 REGI Atropisomers k' 1 = 3.06 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 3.00 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 3.25 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 Br k' 1 = 2.06 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 1.94 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 5.65 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 Me k' 1 = 2.41 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 2.78 α = % 2-propanol in hexane (,) Whelk- 1 reference REGI TECLGIE CIRAL APPLICATI GUIDE IV

45 Atropisomers REGI k' 1 = 7.12 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 6.94 α = % 2-propanol in hexane (,) Whelk- CP reference 41 k' 1 = 6.65 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 7.41 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 7.12 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 6.65 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 5.65 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 3.06 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 REGI TECLGIE CIRAL APPLICATI GUIDE IV 45

46 REGI Atropisomers k' 1 = 1.94 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 2.24 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 2.59 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 3.12 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 4.12 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 3.82 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 2.14 α = % IPA/hexane (,) Whelk- 1 reference 42 k' 1 = 2.86 α = % IPA/hexane (,) Whelk- 1 reference REGI TECLGIE CIRAL APPLICATI GUIDE IV

47 Atropisomers REGI k' 1 = 3.12 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 2.41 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 1.82 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 2.41 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 2.24 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 Br k' 1 = 1.53 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 2.18 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 k' 1 = 2.29 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 REGI TECLGIE CIRAL APPLICATI GUIDE IV 47

48 REGI Atropisomers Amlodipine Vapol Ph Ph amlodipine k' 1 = 5.13 α = 1.22 Column = (R,R)-Whelk- 1 Mobile Phase = (46/46/8) C 2 2 /exane/ethanol M Ammonium Acetate Flow Rate = 1.5 ml/min Run Time = 13.0 min Column = (R,R)-ULM Mobile Phase = 100% Methanol Flow Rate = 1.5 ml/min Run Time = 13 min k' 1 = 1.74 α = 3.37 reference 48 k' 1 = 4.00 α = % 2-propanol in hexane (,) Whelk- 1 reference 41 Adam s Acid Diethylamide F 2 Column = (3R,4)-Pirkle 1-J Mobile Phase = (70/30) exane/ipa Run Time = 17.0 min k' 1 = 4.11 α = 1.27 k' 1 = 11.8 α = % IPA/hexane (,) Whelk- 1 reference REGI TECLGIE CIRAL APPLICATI GUIDE IV