CLS_HALO_Catalogue

HPLC/UHPLC Columns SMALL MOLECULE HPLC/UHPLC Columns 90 Å 2 micron particle 90 Å 2.7 micron particle 90 Å 5 micron particle BIOCLASS 160 Å 2 micron particle 160 Å 2.7 micron particle 160 Å 5 micron particle PEPTIDE 1000 Å 2.7 micron particle 400 Å 3.4 micron particle 90 Å 2.7 micron particle PROTEIN GLYCAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-1 LIFE SCIENCE

Colonnes HPLC/UHPLCColonnes HPLC/UHPLC Superior Performance of HALO® Fused-Core® Columns • HALO 2 µm columns will deliver reliable high speed and high resolution separations at pressures lower than most standard sub-2 µm columns. • HALO 2.7 µm columns can meet or exceed the performance of most standard sub-2 µm columns at pressures one-third to one-half the back pressure under the same conditions. • HALO 5 µm columns will match the performance of totally porous 3 µm columns at roughly half the back pressure under the same conditions. Some of the early and current explanations for the excellent performance of Fused-Core columns are described below. Early Explanations for Superior Performance • Faster Mass Transfer due to a thin porous bonded-phase layer exterior to particle’s solid silica core • More Uniform and Stable Column beds due to very narrow particle size distribution (~4–6% RSD vs. ~20% RSD for standard particles) Current Understanding of SPP Performance The superior performance of Fused-Core SPP columns is now believed to be due to: • Much smaller reduced plate heights and high efficiencies for SPP columns due to smaller van Deemter A and B terms for SPP particles • Reduction in eddy diffusion - 40% smaller van Deemter “A term” due to more uniform analyte flow paths through the column bed • Much lower longitudinal broadening, flat van Deemter plot and higher optimum linear velocity (flow rate) - Due to the presence of the particle’s solid core (25–30% smaller van Deemter “B term”) SEM image of first commercial HALO SEM image of a focused-ion beam cleaved SEM image of a focused-ion beam cleaved particle with 2.7 μm total size consisting HALO 1000 Å 2.7 μm silica particle. HALO Protein 3.4 μm silica particle. This of a 1.7 μm solid silica core and a 0.5 μm This “cut-away” view shows the solid “cut-away” view shows the solid core with shell. core and shell with large pores allowing its very thin 0.2 μm outer porous layer. unrestricted access of macromolecules to the bonded phase. D-2 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns Van Deemter Plot of Plate Height vs. Linear Velocity (flow rate) HPLC/UHPLC Columns Effect of Particle Size and Type Columns: 50 x 4.6 mm, non-core C18, 5 µm; non-core C18, 3.5 µm; Non-core C18, 1.8 µm; HALO C18, 2.7 µm Solute: naphthalene Mobile phase: 60% ACN/40% water Temperature: 24°C m, Plate Height, µm 20 5 µm FPP CN/40% water 18 16 3.5 µm FPP 14 1.8 µm FPP 12 10 2.7 µm Fused-Core 8 1 2 3 4 5 6 7 8 9 10 6 Mobile Phase Velocity, mm/s 4 2 0 0 H = A + —Bµ + Cµ The shorter diffusion path of HALO reduces axial dispersion Van Deemter Equation 3 µm totally porous 1.8 µm totally porous HALO H = height equivalent to theoretical plate 0.5 µm Diffusion Path A = eddy diffusion term 0.9 µm B = longitudinal diffusion term 1.5 µm Diffusion C = resistance to mass transfer term Path µ = mobile phase linear velocity (L/t0) The shorter diffusion path of HALO particles reduces axial dispersion of solutes and minimizes *G.J. Kennedy, J.H. Knox, peak broadening. Because of the shorter diffusion path, the performance advantages of HALO J. Chromatogr. Sci 10 (1972) 549 become even more apparent when separating peptides and operating at faster mobile phase flow rates. American Laboratory, April 2007 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-3 LIFE SCIENCE

Colonnes HPLC/UHPLC Colonnes HPLC/UHPLC Key advantages of HALO® Fused-Core® Columns High Speed Separations • Smaller reduced plate heights lead to high efficiencies; narrower and taller peaks, for improved resolution and lower detection limits (LODs and LOQs) • Flat van Deemter plot and higher linear velocity optimum allow higher flow rates with minimal column efficiency loss High Resolution Separations • High efficiency with longer geometries (100, 150, 250 mm) provides greater resolving power for challenging applications • Lower back pressure permits columns to be used in series for the most demanding UHPLC and HPLC separations Excellent Ruggedness and Reproducibility • Less plugging, longer usable column lifetime and greater uptime due to larger porosity frits (vs. sub-2 µm totally porous (non-core) columns) - 2 µm frits for HALO 2.7 and HALO 5 - 1 μm frits for HALO 2 vs. 0.2–0.5 μm frits for sub-2 μm non-core columns • Excellent column-to-column and lot-to-lot reproducibility • Robust pores in multiple sizes for a tailored application solution (90 Å, 160 Å, 400 Å and 1000 Å) HALO BIOCLASS - Solutions for Proteins, Peptides and Glycans • Application specific columns for bioseparations that outperform non-core columns • Up to 1/2 the back pressure • Offer better peak shape and peak capacity • Breakthrough 1000 Å pore particles for large molecules Separation of 14 Cannabinoids on HALO C18 A HALO C18 column is used to separate a mixture of fourteen cannabinoids, showing fast results and high resolution within critical pairs. Cannabinoids are a class of chemical compounds primarily found in the marijuana plant. Many of these compounds have been found to provide medicinal benefits such as reduction in pain and inflammation. Testing Conditions: Temperature: 30°C PEAK IDENTITIES 9. delta-9- Tetrahydrocannabinol (Δ9-THC) Column: HALO 90Å C18, 2.7 μm, 3.0 x Detection: UV 220 nm, PDA 1. Cannabidivarinic acid (CBDVA) 150mm 10. delta-8-Tetrahydrocannabinol Injection Volume: 0.6 μL 2. Cannabidvarin (CBDV) (Δ8-THC) Part Number: HAL92813-702 Dwell Volume: 0.471 mL 3. Cannabidiolic acid (CBDA) 11. Cannabicyclol (CBL) AppMlicoabtiiloenPhNaostee:: 162-CN 12. Cannabichromene (CBC) A= Water/ 0.1% formic acid Sample Solvent: 75/25 methanol/water 4. Cannabigerolic acid (CBGA) 13. delta-9-Tetrahydrocannabinolic Response Time: 0.025 sec. 5. Cannabigerol (CBG) acid A (THCA) B= Acetonitrile/ 0.085% foSrmeicpacaidrationDatoa Rfat1e:4100CHaz nnabinoids on6H. ACLanOnabCidi1ol 8(CBD) 14. Cannabichromenic acid (CBCA) Gradient: 70-88%B in 6 min. LC System: Shimadzu Nexera X2 7. Tetrahydrocannabivarin (THCV) Flow Rate: 1.0 mL/min. Flow Cell: 1 μL 8. Cannabinol (CBN) Initial Pressure: 350 bar 1 G0154 Absorbance @ 220nm 2 3 4 8 5 6 7 9 11 12 13 10 0 14 0 1 2 3 4 5 6 7 Time, min. D-4TEST CONDITIONSC: ANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca Column: HALO 90ÅLCIF1E8, S2.C7 IµEmN,C3E.0 x 150mm

HPLC/UHPLC Columns HALO: Dependable quality and reproducibility HPLC/UHPLC Columns Consistent reproducible performance from column to column and lot to lot is ensured because of well-designed processes and practices in the manufacture of HALO Fused-Core particles, HALO phases and HALO columns. Representative chromatograms of QA and QC tests are shown below, along with a historical plot of selec­tivity between a neutral and basic probe. Advanced Materials Technology incorporates the know-how from its principal scientists, each with over 40 years of experience in liquid chromatography, particle synthesis and column manufacturing to bring high quality, innovat­ive products to our customers. QA TEST 220 QC TEST 200 180 180 1-Cl-4-NO2-benzene 160 160 butyl paraben 140 120 140 HALO 90 Å C18, 2.7 µm, 4.6 x 50 mm 100 phenol HALO 90 Å C18, 2.7 µm, 4.6 x 50 mm 75:25 CH3OH/20 mM phosphate (pH 7) uracil 60:40 CH3CN/water 80 1.8 mL/min., 30 ˚C 120 1.5 mL/min., 25 ˚C 60 40 naphthalene Absorbance (mAU) 20 Absorbance (mAU) 100 acenaphthene amitriptyline 0 uracil propranolol -20 0 80 naphthalene 60 40 20 0 0 0.5 1 1.5 2 2.5 3 0.2 0.4 0.6 0.8 1 1.2 -20 Time (min.) Time (min.) Reproducible performance over time HALO Reproducibility Data for 10 Years (QA RetentiHoAnLO FReapcrotdourci)bility Data for Ten Years Advanced Materials Technology (AMT) is one of only a few HPLC column manufacturers that (QA Retention Factor) completes the entire column manufacturing process in-house. The scientists and engineers 4.0 at AMT have expertise in every aspect of the column development process. Every step that 3.8 comprises the creation of a HALO column is monitored and controlled. From the solid silica 3.6 cores to the bonded Fused-Core particles to the final loaded and QC-tested column, customers 3.4 can be confident that the HALO products they receive are reliable and reproducible. The graph Retention Factor 3.2 RSD <1% demonstrates the superior reproducibility of the retention of HALO 90 Å C18, 2.7 μm columns 3.0 over a 10- year period. 2.8 2.6 2.4 2.2 2.0 10 20 30 40 50 60 70 80 90 100 0 Chronological Lot Data 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-5 LIFE SCIENCE

Colonnes HPLC/UHPLC Colonnes HPLC/UHPLC Selecting the appropriate pore size Guidance for Pore Size Selection • Match the column pore size according to the range of MW Range Pore Application Particle Sizes Column molecular weights (MWs) of the analytes in your sample Size (Å) (μm) Family • S mall molecules (< 5000 Da) are usually analyzed using Small 90 Small Mol- 2, 2.7, 5 HALO® HALO 90 Å columns < 5000 Da ecules - Packing materials with smaller pores have greater surface HALO® area, which allows improved retention and loading Small 90 Glycan 2.7 BioClass capacity for lower MW analytes < 20 kDa* - When an analyte is too large for the pores, restricted diffusion can occur, which can lead to peak broadening Medium 160 Peptides 2, 2.7, 5 and reduced retention 100 Da < MW < 15 kDa • For macrocyclic antibiotics and biomolecules such as Large 400 Proteins 3.4 peptides and proteins, use larger pore sizes such as 2 kDa < MW < 500 kDa 1000 2.7 160 Å HALO and 400 Å HALO columns Large • F or mAbs and intact proteins of molecular sizes > 50 kDa > 50 kDa, consider the HALO 1000 Å products * for glycans, glycopeptides and glycoproteins Restricted diffusion for higher MW Polypeptides on HALO 90 Å In this exa­ mple, broad peaks are observed in the upper chromatogram for bovine and human insulin, cytochrome C and lysozyme (MWs > ~5700 Da) chromat­ographed using a HALO 90 Ångstrom column. This broaden­ing is due to restricted diffusion using the smaller pore column. However, those same four peaks are much narrower and taller in the lower chromato­gram using the 160 Ångstrom HALO Peptide ES-C18 column. HALO C18, Cyt C Lyso TEST CONDITIONS: 100 x 4.6 mm, 2.7μm, 90 Å Column: 4.6 x 100 mm Part Number: H ALO 90 Å C18, 2.7 μm: HAL92814-602 Lyso HALO 160 Å ES-C18, 2.7 μm: HAL92124-602 Cyt C Mobile Phase A: 90/10 water/acetonitrile/0.1% TFA Mobile Phase B: 30/70 water/acetonitrile/0.1% TFA HALO ES-C18, Gradient: 15% B to 50% B in 15 min. 100 x 4.6 mm, 2.7μm, 160 Å Flow Rate: 1.5 mL/min. Temperature: 30 ˚C 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Detection: UV 220nm, VWD Injection Volume: 5.0 µL Time (min.) LC System: Agilent 1100 Peptides (in RT order): Leu-Enk, Bovine Insulin, Human Insulin, Cytochrome C, Lysozyme D-6 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns HALO Columns for small molecule analyses HPLC/UHPLC Columns Of the three variables in the general resolution equation, including efficiency (N) and retention (k), selectivity (α) is the most powerful parameter for adjusting and improving resolution between peaks in a chromatographic separation. EFFICIENCY SELECTIVITY RETENTION Parameters That Affect HPLC Selectivity in Order of Increasing Effectiveness HPLC Parameter Effectiveness for Changing Selectivity Mobile phase pH Most Effective (ionizable analytes only) Organic modifier choice Percent organic modifier or gradient steepness Column stationary phase Column temperature Buffer choice Buffer concentration Least Effective Moreover, column phase selectivity is one of the four most powerful and useful parameters for adjusting HPLC separation selectivity. For ionizable analytes, mobile phase pH is, by far, the most effective parameter. However, column stationary phase is comparable to organic modifier choice (acetonitrile vs. methanol) and percent organic modifier/gradient steepness in its ability to change relative retention for UHPLC and HPLC separations. For example, if you were looking for a column with a different selectivity to a HALO C18 column at low pH, you might consider and select a HALO PFP column as one most likely to be orthogonal to C18. However, the other available HALO phases (Phenyl-Hexyl, RP-Amide, ES-CN) also retain and separate analytes via retention mechanisms different from HALO C18 and HALO C8, so it might be prudent to consider one or more of the former phases as part of a comprehensive column screening or method development strategy. Example Strategy for Comprehensive Method Development Using Multiple HALO Stationary Phases and Column/Condition Screening, Followed by Optimization of Gradient Time, Temperature and pH Reversed-Phase Screening Conditions HALO C18 Screening Select phase, Optimize t G, T, pH HALO Phenyl-Hexyl gradients pH, solvent Optimize t G, T, pH Select phase, HALO Biphenyl pH, solvent HALO RP-Amide 5-95% CCCCHHHH³³³³OCOCNNHH,,,, low pH HALO PFP 5-95% low pH 5-95% mid pH 5-95% mid pH HILIC Mode Screening Conditions HALO HILIC Screening HALO PFP gradients HALO Penta-HILIC 95-50% CH3CN, NH4HCO2, low pH 95-50% CH3CN, NH4HCO2, mid pH 95-50% CH3CN, NH4CH3CO2, low pH 95-50% CH3CN, NH4CH3CO2, mid pH 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-7 LIFE SCIENCE

Colonnes HPLC/UHPLC HALO Columns for Small Molecule Separations Colonnes HPLC/UHPLC HALO Small Molecule Column Specification USP Particle Carbon Surface Low pH High pH Max. Temp. Max. Temp. Phase Designation Size Load (%) Area Limit Limit Lower pH Upper pH Endcapped (µm) (m2/g) Limit Limit C18 L1 2 7.5 120 2 9 60 40 Yes 2.7 7.7 135 5 6.4 90 2 AQ-C18 L1 2.7 6.5 120 2 9 60 40 Yes 5 6.7 135 2 5.6 90 2.7 C8 L7 5 4.8 120 2 9 60 40 Yes 2 5.4 135 2.7 3.7 90 5 Phenyl-Hexyl L11 2 6.3 120 2 9 60 40 Yes 2.7 7.1 135 5 5.2 90 2 Biphenyl L11 2.7 6.7 120 2 9 60 40 Yes 5 7.0 135 2 5.5 90 2.7 PFP L43 5 5.3 120 2 8 60 40 Yes 2 5.5 135 2.7 3.9 90 5 ES-CN L10 2 3.4 120 1 8 80 40 Yes 2.7 3.5 135 5 2.5 90 2 RP-Amide L60 2.7 7.3 120 2 9 60 40 Yes 5 8.2 135 5.1 90 2.7 HILIC L3 Unbonded 120 1 8 60 40 N.A. 135 90 Penta-HILIC L95 2.8 120 2 9 60 40 No 3.2 135 2.1 90 C30 L62 4.5 160 2 9 60 40 Yes Relative Reversed-Phase Orthogonality of HALO Phases vs. HALO C18 at Low and Mid pH Most Similar pH 2.8 pH 7 Most Orthogonal HALO C18 HALO C18 HALO C8 HALO C8 HALO AQ-C18 HALO AQ-C18 HALO Phenyl-Hexyl HALO PFP HALO ES-CN HALO Phenyl-Hexyl HALO Biphenyl HALO Biphenyl HALO RP-Amide HALO ES-CN HALO PFP HALO RP-Amide D-8 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns HALO Columns for Small Molecule Separations Phase Features & Benefits Target Analytes Typical Applications HPLC/UHPLC Columns C18 • Excellent performance for broad • Diverse analytes ranging from polar • Pharmaceutical (octadecyldimethylsilane) range of analyte polarities to non-polar • Environmental • Uncharged acids and bases, • Cannabinoid AQ-C18 • Resistant to dewetting, making uncharged ion pairs • General purpose (polar modified) it 100% aqueous mobile phase compatible • Acids, bases, polar analytes • Pesticides C8 • Nucleobases (octyldimethylsilane) • Enhanced retention for polar • Neurotransmitters molecules • Polar acids • Excellent performance for broad • Diverse analytes ranging from polar • Pharmaceutical range of analyte polarities to non-polar • Environmental • Uncharged acids and bases, • Higher hydrophobic uncharged ion pairs compounds Phenyl-Hexyl • Complementary selectivity to alkyl • Electron-poor molecules, aromatic • Benzodiazepines (phenylhexyldimethylsilane) • phases or unsaturated compounds • Aromatics Enhanced selectivity for aromatic (ketones, nitriles, alkenes) • Drugs of abuse Biphenyl compounds (dimethylbiphenyl) • Complementary selectivity to • Aromatic PFP alkyl phases • Electron-poor molecules, aromatic • Heterocycles (pentafluorophenylpropylsilane) or unsaturated compounds • Drugs of abuse • Enhanced selectivity for aromatic (ketones, nitriles, alkenes) • Pain management drugs ES-CN compounds • Highly aqueous conditions (diisopropylcyanopropylsilane) • Complementary selectivity to alkyl RP-Amide phases (C16 Amide) • Enhanced selectivity • Electron-rich compounds, • Steroids aromatics, unsaturated compounds • Isomeric compounds for stereoisomers with double and/or triple bonds • Substituted aromatics • Can be used in RPLC and HILIC modes • Complementary selectivity • Polar and very polar bases, acids to alkyl phases and neutrals • Explosives • More retention for polar analytes • Aromatics and much less reten­tion for non- • Very hydrophobic analytes • Polar compounds polar analytes retained too strongly on C18 phase • Complementary selectivity to alkyl • Alcohols, Acids, Phenols, Catechins • Phenols • phases • Alcohols Enhanced stability for mini­mum • Catechins bleed and long life HILIC • Can be used in HILIC and normal- • Polar and very polar bases, acids • Polar compounds (Bare Silica) phase modes and neutrals, especially with log P<0.5 Penta-HILIC • Ideal for separation of highly • Polar analytes with log P values • Polar basic compounds (proprietary penta-hydroxy ligand) polar compounds that are poorly near or less than 0 • Fat/Water Soluble Vitamins retained in RPLC • Carotenoids • Lipids • Highly reproducible bonded phase • Steroids coverage resulting in fast, highly • Hydrophobic, long-chain C30 efficient, rugged separations • structurally related isomers • 100 % Aqueous Compatibility 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-9 LIFE SCIENCE

Colonnes HPLC/UHPLC Reversed-Phase Separations with HALO To illustrate the selectivity differences among the various HALO RPLC phases, the following examples are provided. Colonnes HPLC/UHPLC Benzodiazepines on HALO Fused-Core bonded phases Absorbance (mAU) TEST CONDITIONS: PEAK IDENTITIES: 5. Clonazepam Column: HALO 90 Å, 50 x 4.6 mm 1. Oxazepam 6. Temazepam Part Numbers: HAL92814-406, HAL92814-402, 2. Lorazepam 7. Flunitrazepam HAL92814-407, HAL92814-409 3. Nitrazepam 8. Diazepam Mobile Phase A: 25 mM Ammonium Acetate 4. Alprazolam Mobile Phase B: Acetonitrile Gradient: 34 - 63% B in 3.5 min. 1 3 67 10 Flow Rate: 1.5 mL/min; 2 45 HALO Phenyl-Hexyl Temperature: 35 ˚C Detection: UV 254 nm Injection volume: 1 µL Pressure: 200 bar 1 3 76 10 2 45 10 HALO Phenyl-Hexyl 1 2, 3 5,6 7 HALO C18 is the most retentive 4 HALO RP-Amide phase for these anti- anxiety drugs due to 3 4,6 its propensity for π-π 5 interactions. 1 7 8 2 1.5 2 2.5 3 HALO PFP Time (min.) 3.5 4 Aromatic and Nitroaromatic Compounds TEST CONDITIONS: Temperature: 30 ˚C PEAK IDENTITIES: 7. 2,4-DNT Column: HALO ES-CN 90 Å, 100 x 4.6 mm Detection: UV 254 mn, VWD 1. Resorcinol 8. Anisole Injection Volume: 1.0 μL 2. Benzyl alcohol 9. 1-Chloro-4-nitrobenzene HALO C18 90 Å, 100 x 4.6 mm Sample Solvent: water/methanol 1:1 3. Phenylacetonitrile 10. Toluene Part Numbers: C 18: HAL92814-602 (v/v) Response Time: 0.02 sec. 4. 1-Indanol DNT = dinitrotoluene Flow Cell: 2.5 μL semi-micro 5. 3,4-DNT i = impurity ES-CN: HAL92814-604 LC System: Shimadzu Prominence 6. 2,3-DNT Mobile Phase: 40/60-A/B for C18, UFLC XR 50/50-A/B for ES-CN Extra Column Volume: about 14 μL 6 A= water B= methanol Flow Rate: 1.25 mL/min. Pressure: About 300 bar 1 5 3 10 9 2 7 HALO C18 and HALO ES-CN columns may be used as Absorbance (mAU) 4 HALO C18 orthogonal confirmatory columns for explosives analysis. ii 10 1 3 10 HALO ES-CN 6 2 75 9 4 i 10 0 1 2 3 4 5 6 Time (min.) D-10 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns HALO C18 vs RP-Amide for Phenolics HPLC/UHPLC Columns HALO RP-Amide provides greater retention and resolution compared to HALO C18 for this phenol mixture. TEST CONDITIONS: HALO RP-AMIDE 2 6 Column: HALO RP-Amide 90 Å 1 7 & HALO C18 90 Å, 50 x 4.6 mm, 2.7 µm Part Number: HAL92814-407 & HAL92814-402 4 Mobile Phase: 60/40: A/B A= 0.02 M Potassium phosphate, pH = 7.0 3 5 B= Acetonitrile Flow Rate: 2.0 mL/min. 6 Temperature: ambient (25 ˚C) Absorbance (mAU) HALO C18 4 PEAK IDENTITIES: 1 5 1. Uracil 2 2. Catechol 3 7 3. 2,4,6-Trinitrophenol 4. 2,4,6-Trichlorophenol 5. 2,2’-Biphenol 6. 2’,6’-Dihydroxyacetophenone 7. 4-Chlorophenol 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 Time (min.) Separation of Structurally similar steroids on HALO C18 and PFP HALO PFP delivers improved resolution and different elution order compared to HALO C18 for this mixture of steroids. TEST CONDITIONS: PEAK IDENTITIES: HALO C18 3,4 Column: HALO C18 & HALO PFP, 1. Prednisone 1 100 x 4.6 mm, 2.7 µm, 90 Å 2. Cortisone Part Number: 3. Prednisolone 2 C18: HAL92814-602 4. Hydrocortisone PFP: HAL92814-609 Mobile Phase: 50/50: water/methanol Absorbance (mAU) 41 Flow Rate: 1.0 mL/min. 3 Pressure: about 230 bar HALO PFP 2 Temperature: 35 ˚C Detection: UV 240 nm, VWD 0 1 2 3 4 5 6 7 Injection Volume: 0.5 μL Time (min.) Sample Solvent: 80% methanol in water Response Time: 0.02 sec. Flow Cell: 2.5 µL semi-micro LC System: Shimadzu Prominence UFLC XR ECV: about 14 μL 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-11 LIFE SCIENCE

Colonnes HPLC/UHPLC Colonnes HPLC/UHPLC HILIC Separations with HALO Hydrophilic interaction liquid chromatography (HILIC) is a useful UHPLC and HPLC mode for the following situations: • Polar analytes that are poorly or not retained in RPLC • Basic analytes that have poor peak shape (overloading) and/or poor retention at low pH in RPLC • Analytes that have log P values near or less than zero • When conditions orthogonal to RPLC mode are needed (elution order change) HALO columns are currently available in two different phases for HILIC separations: • HALO HILIC A Fused-Core silica phase that can be used either in HILIC mode or in normal-phase mode with water-immiscible solvents (NPLC). • HALO Penta-HILIC A bonded silica phase, which has a highly polar ligand with 5 hydroxyl groups tethered via novel proprietary linkage chemistry to Fused-Core silica particles. Some Typical Analytes for HILIC Separations: • Basic pharmaceuticals • Peptides • Polar organic acids • Catecholamines and other neurotransmitters • Nucleosides and nucleobases • Drug glycoside and glycuronide metabolites • Mono-, di-, tri- and other oligosaccharides • Opiates • Glycosylceramides • Polar triazines and pyrimidines • Analytes from metabolomic profiling Retention order reversal and improved retention with HILIC You can often obtain adenosine + TEST CONDITIONS: a complete reversal cytosine Column: HALO C18 90 Å, 2.7 µm, 50 x 4.6 mm in elution order and acenaphthene Part Number: HAL92814-402 different selectivity Mobile Phase A: 90/10 ACN/0.1 M Ammonium Formate using HILIC mode cytosine Flow Rate: 1.8 mL/min. compared to pH: 3.0 reversed-phase adenosine mode under the TEST CONDITIONS: same or appropriate acenaphthene Column: HALO HILIC 90 Å, 2.7 µm, 50 x 4.6 mm conditions. Part Number: HAL92814-401 Mobile Phase A: 90/10 ACN/0.1 M Ammonium Formate Flow Rate: 1.8 mL/min pH: 3.0 0 0.5 1 1.5 Time (min.) D-12 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns Nucleosides and Nucleobases on HALO Penta-HILIC HPLC/UHPLC Columns These 15 nucleosides and nucleobases are separated in under 10 minutes using a HALO 2.7 Penta-HILIC column. 90 TEST CONDITIONS: PEAK IDENTITIES: 70 7 Column: HALO Penta-HILIC 90 Å, 1. Thymine 1 6 2.7 µm, 100 x 4.6 mm 2. Uracil Part Number: HAL92814-605 3. Thymidine Absorbance (260 mm) 50 2 45 Mobile Phase A: 10 mM 4. 2’-Deoxyadenosine 3 Ammonium formate in 85% 5. Adenine 13 Acetoni­trile/15% water 6. Uridine 30 8 14 Mobile Phase B: 10mM 7. Adenosine Ammonium Formate in 93% 8. Hypoxanthine 10 11 15 Acetonitrile/7% water 10 13 12 Gradient: Time %B 9. Xanthine 0-3 min. 94 10. Cytosine -10 3-9 min. 94-0 11. 2’-Deoxycytidine 0 1 2 3 4 5 6 7 8 9 Flow Rate: 1.5 mL/min. 12. Guanine Time (min.) 13. 2’-Deoxyguanosine Temperature: 35 ˚C Detection: UV 260 nm, VWD 14. Cytidine Injection volume: 3.0 µL 15. Guanosine Pressure: about 230 bar Cephalosporins on HALO Penta-HILIC and HALO HILIC HALO Penta-HILIC shows increased retention and different selectivity vs. HALO HILIC for these 10 cephalosporins. 1 HALO Penta-HILIC TEST CONDITIONS: Gradient: HALO Penta-HILIC: PEAK IDENTITIES: 5 67 8 85-65% B in 10 min. 2 Column:  1. Cephalothin 3 HALO HILIC: 85-70% B in 10 min. 2. Cefoxitin 4 13 10 HALO HILIC & Penta-HILIC, 90 Å, Absorbance @ 254 nm 9 2.7 µm, 150 x 2.1 mm Flow Rate: 0.5 mL/min. 3. Cefotaxime 8 9 3 HALO HILIC Part Number: Pressure: 195 bar with HALO 4. Cefazolin 1 2,3 5 68 7 HALO Penta-HILIC: HAL92812-705 Penta-HILIC 5. Cefaclor HALO HILIC: HAL92812-701 163 bar with HALO HILIC 6. Cephalexin 4 Mobile Phase: A= 95/5 ACN/H20 Temperature: 30 ˚C 7. Cephradine with 5 mM Detection: UV 254 nm, VWD 8. Cefadroxil 10 Ammonium formate, pH=3.0 Injection Volume: 0.5 µL 9. Ceftazidime (adj.) Sample Solvent: 50/50 ACN/water 10. Cephalosporin C 0 1 2 3 4 5 6 7 10 Mobile Phase: B= 50/50 ACN/H20 Flow Cell: 5 µL semi-micro with 5 mM Ammonium formate, LC System: Agilent 1100 Time (min.) pH=3.0 (adj.) Reversed-Phase separation of cephalosporins using HALO ES-CN HALO HILIC and Penta-HILIC columns often offer an orthogonal separation relative to reversed-phase separations, as shown here for HALO ES-CN for a subset of the same cephalosporins shown. 6 TEST CONDITIONS: Sample Solvent: PEAK IDENTITIES: 1 Column: HALO ES-CN 90 Å, 2.7 µm 70/30: Water/Methanol 1. Cefadroxil 100 x 4.6 mm Response Time: 0.02 sec. 2. Ceftazidime 4 5 Part Number: HAL92814-604 Flow Cell: 2.5 µL semi-micro 3. Cefaclor 3 Mobile Phase A: 0.02 M LC System: Shimadzu 4. Cephalexin Absorbance @ 254 nm Phosphate buffer, pH 2.7 (adj.) Prominence UFLC XR 5. Cephradine 2 7 9 Mobile Phase B: Methanol ECV: about 14 µL 6. Cefotaxime 8 Gradient: 20% B to 40% B Pressure: 225 bar at start of 7. Cefoxitin in 2.5 min. gradient 8. Cefazolin Flow Rate: 2.0 mL/min. Temperature: 40 ˚C 0.0 0.5 Detection: UV 254 nm, VWD 1 1.5 2.0 2.5 Injection Volume: 1.0 µL Time (min.) 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-13 LIFE SCIENCE

Colonnes HPLC/UHPLC Colonnes HPLC/UHPLC HALO® 90 Å 2 μm (UHPLC) HALO 2 µm 1.2 µm 2 µm Highest UHPLC performance possible without the disadvantages of sub-2 µm columns 0.4 µm • Use when the highest efficiency is needed Shell with 90 Å pores • Excellent for fast method development and column/condition screening • Best performance obtained with instrumentation having extra column volume (IBW< 10 µL) • Ruggedness for R&D • 1 µm inlet frit • Pressure limit, 1000 bar/14,500 psi Note: IBW is instrumental band width, and is a measure of extracolumn dispersion. Application Note: 113 Higher Efficiency of HALO Fused-Core® C18, 2 µm compared to a 1.7 μm Totally Porous C18 Column pWoiHrtohiugashH1eA.r7LOμmE2ffiCC11c88ieccnooclluuymmn1n.,.o7ofneHµcAmaLnOaTc2ohtieavllCey1m8oPreo(2rsoeµpumasraFtiuCos1ne8edffiCcioe-lnuCcmyoanrtele®s)s pcroemsspuraerethdantowaith a competitor’s totally 3 HALO 2 µm C18 TEST CONDITIONS: Injection Volume: 0.2 μL 2 N (peak 4) = 15500 Column 1: HALO 90Å C18, 2 μm Sample Solvent: 20/80: 4 Water/Acetonitrile Pressure = 190 bar CHMAo=aolluWobmiPlaeanterP2trh:Na2H2su..Ae11m:LxxbO1e55541P32Cr/00....:18EmmH85ADUDP,:AymmoerK9ArLacde09/caIenBÅ1iDnlce8,oaE11npN2.oh7-Tpe4μInh0mTo2eInnEoeSn:e Response Time: 0.16 sec. 1 Flow Cell: 1 μL B= Acetonitrile LC System: Shimadzu Nexera Aborbance @ 254 nm Totally Porous 1.7 µm C18 Flow Rate: 0.5 mL/min ECV: ~ 7 μL Pressure: See chart PNr(epsesaku4r)e=S=9T620R80U0CbTaUr R Temperature: 25°C PEAK IDENTITIES: Detection: UV 254 nm, PDA 1. Uracil 0.0 0.5 1.0 1.5 2.0 2. Pyrene Time, min 3. Decanophenone 4. Dodecanophenone CToESlSutTmenrCoO1i:NdD2s.eI1TpxIOa50rNamSt:imo,nHAuLsOin2 gµmHCA1L8O 2 PFP STRUCTURES: Halo Part Number: 91812 -402 CoHluAmLnO2 2: 2µ.1mx 5c0omlumm, TnostaollyftPeonroussh1o.7w exµcmeCll1e8nt selectivity for steroids. HALO 2 µm is able to readily separate a mixture of M9obsilteePrhoaside s: i1n5/l8e5s:sAt/hBan 8 minutes in gradient mode. A= Water B= Acetonitrile Flow Rate: 0.5 mL/min 6 Uracil TEST CONDITIONS:D ecanophenone Injection volume: 2 μL Pressure: See chart Column: H ALO PFP 90 Å, 2 μm Sample Solvent: methanol 3 Response Time: 0.02 sec. 50 x 3.0 mm Flow Cell: 2.5 μL semi-micro Temperature: 25 °C Part Number: HAL91813-409 ECV: about 14 μL Detection: UV 254 nm, PDA Mobile Phase A: water LC System: Injection Volume: 0.2 μL 8 Mobile Phase B: methanol Shimadzu Prominence UFLC XR Gradient: Time %B Absorbance @ 245 mm (mAU)SRaemsppolensSeoTlvimen1et::20/80: Water/Acetonit5rile7 0 min. D od4e7canophenone 0.16 sec. 3 min. 47 Flow Cell: 1 μL 8 min. 88 LC System: S himadzu Nexera Flow Rate: 0.4 mL/min. ECV: ~ 7 μL Pyrene Temperature: 35 ˚C Pressure: 180 bar initial PEAK IDENTITIES: 5. Prednisone With a HALO 2 C18 colum2 n, one can 9 1. Uracil 6. Dexamethasone achieve more separationeffic4 iency at less 2. Hydrocortisone 7. ß-Estradiol pressurethan with a competitor’s totally 3. Prednisolone 8. Estrone porous 1.7 µm C18 column. 4. Cortisone 9. Halcinonide 0 2 4 6 8 Detection: UV 280 nm, VWD Time (min.) D-14 CANADIAN 1 88PF8OLA-R2CM2E6OA-NR2E7OI7RND5FOERR,:M:COATNiInTOfANoC@OT:RliTfeOscience.ca :: www.lifescience.ca www.advanced -materials -tecLh.cIFomE SCIENCE

HPLC/UHPLC Columns HALO® 90 Å 2 μm (UHPLC) Columns Ordering Information HPLC/UHPLC Columns The part numbers for HALO 90 Å 2 μm columns are presented below and available in 2.1 and 3.0 mm internal diameters. Guard columns are also available for these IDs for UHPLC to provide additional protection when necessary. Dimensions 2 μm, 90 Å ID x Length (mm) C18 AQ-C18 C8 Phenyl- Biphenyl RP-Amide PFP ES-CN Penta- HILIC Hexyl HILIC 2.1 x 20 HAL91812-202 HAL91812-222 HAL91812-208 HAL91812-211 HAL91812-207 HAL91812-209 HAL91812-204 HAL91812-201 2.1 x 30 HAL91812-302 HAL91812-322 HAL91812-308 HAL91812-206 HAL91812-311 HAL91812-307 HAL91812-309 HAL91812-304 HAL91812-205 HAL91812-301 2.1 x 50 HAL91812-402 HAL91812-422 HAL91812-408 HAL91812-306 HAL91812-411 HAL91812-407 HAL91812-409 HAL91812-404 HAL91812-305 HAL91812-401 2.1 x 75 HAL91812-502 HAL91812-522 HAL91812-508 HAL91812-406 HAL91812-511 HAL91812-507 HAL91812-509 HAL91812-504 HAL91812-405 HAL91812-501 2.1 x 100 HAL91812-602 HAL91812-622 HAL91812-608 HAL91812-506 HAL91812-611 HAL91812-607 HAL91812-609 HAL91812-604 HAL91812-505 HAL91812-601 2.1 x 150 HAL91812-702 HAL91812-722 HAL91812-708 HAL91812-606 HAL91812-711 HAL91812-707 HAL91812-709 HAL91812-704 HAL91812-605 HAL91812-701 2.1 x 250 HAL91812-902 HAL91812-922 HAL91812-908 HAL91812-706 HAL91812-911 HAL91812-907 HAL91812-909 HAL91812-904 HAL91812-705 HAL91812-901 3.0 x 20 HAL91813-202 HAL91813-222 HAL91813-208 HAL91812-906 HAL91813-211 HAL91813-207 HAL91813-209 HAL91813-204 HAL91812-905 HAL91813-201 3.0 x 30 HAL91813-302 HAL91813-322 HAL91813-308 HAL91813-206 HAL91813-311 HAL91813-307 HAL91813-309 HAL91813-304 HAL91813-205 HAL91813-301 3.0 x 50 HAL91813-402 HAL91813-422 HAL91813-408 HAL91813-306 HAL91813-411 HAL91813-407 HAL91813-409 HAL91813-404 HAL91813-305 HAL91813-401 3.0 x 75 HAL91813-502 HAL91813-522 HAL91813-508 HAL91813-406 HAL91813-511 HAL91813-507 HAL91813-509 HAL91813-504 HAL91813-405 HAL91813-501 3.0 x 100 HAL91813-602 HAL91813-622 HAL91813-608 HAL91813-506 HAL91813-611 HAL91813-607 HAL91813-609 HAL91813-604 HAL91813-505 HAL91813-601 3.0 x 150 HAL91813-702 HAL91813-722 HAL91813-708 HAL91813-606 HAL91813-711 HAL91813-707 HAL91813-709 HAL91813-704 HAL91813-605 HAL91813-701 3.0 x 250 HAL91813-902 HAL91813-922 HAL91813-908 HAL91813-706 HAL91813-911 HAL91812-907 HAL91813-909 HAL91813-904 HAL91813-705 HAL91813-901 HAL91813-906 HAL91813-905 HALO guard cartridge and guard holder For more details please go to page D-36 Dimensions 2 μm, 90 Å ID x Length (mm) C18 AQ-C18 C8 Phenyl- Biphenyl RP-Amide PFP ES-CN Penta- HILIC Hexyl HILIC 2.1 x 5 HAL91812-102 HAL91812-122 HAL91812-109 HAL91812-104 HAL91812-101 3.0 x 5 HAL91813-102 HAL91813-122 2 µm, 90 Å Guard Cartridges, 3-pack HAL91813-109 HAL91813-104 HAL91812-105 HAL91813-101 HAL91813-105 HAL91812-108 HAL91812-106 HAL91812-111 HAL91812-107 HAL91813-108 HAL91813-106 HAL91813-111 HAL91813-107 Guard Column Holder - HAL94900-001 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-15 LIFE SCIENCE

Colonnes HPLC/UHPLC Colonnes HPLC/UHPLC HALO 2.7 µm HALO® 90 Å 2.7 μm (UHPLC and HPLC) Reliable, efficient performance with lower back pressure compared to all sub-2 µm totally porous columns. • High speed or high resolution UHPLC or HPLC applications 1.7 µm 2.7 µm • Excellent for R&D and routine analyses 0.5 µm • 2 µm inlet frit • Pressure limit, 600 bar/9000 psi Shell with 90 Å pores LC-MS Analysis of Multiple Mycotoxins on HALO® 90 Å Biphenyl ThAepspelicbaatisoincNtortiec:y1c7l6i-cMantidepressants are separated in less than two minutes, with excellent peak shape, using a HALO Biphenyl column. LC-MS Analysis of Multiple Mycotoxins on HALOTE®ST C9O0NDÅITIOBNipS:henyl Detection: LC-MS Column: HALO 90 Å Biphenyl, 2.7 μm, 2.1 Injection Volume: 2.0 μL x 100mm MS System: Thermo Fisher Orbitrap EIC 6 G0170 Part Number: HAL92812-611 VelosPro ETD ESI: +4 PEMAoKbiIlDe EPhNaTseITAI:EwSa:ter with 0.1% formic Heat Block: 350°C 1. Fumonisin Bac1id(m/5/mz:M72a2m.8m) onium Sheath Gas Flow: 34.88 4 2. Aflatoxin Gf2o(rmm/azt:e331.3) Aux Gas Flow: 10.00 8 M3o.bilAefPlahtaosxeinBB: 2ac(emto/zn:it3r1ile5.w3)ith 0.1% PEAK IDENTITIES: 4. Aflatoxin Gf1o(rmm/icz:a3c2id9/.53m) M 1. Fumonisin B1 (m/z: 722.8) 5. Fumonisin aBm2 m(mo/nzi:u7m06fo.8r)mate 2. Aflatoxin G2 (m/z: 331.3) 1 23 G6ra. dieAnflta:tToimxine B1 (m%B/z: 313.3) 3. Aflatoxin B2 (m/z: 315.3) 4. Aflatoxin G1 (m/z: 329.3) 5 7. Zearal0e.n0one (m32/z: 319.4) 8. Ochrat5o.0xin A (3m4/z: 404.8) 7 10.0 60 5. Fumonisin B2 (m/z: 706.8) Flow Rate: 0.4 mL/min 6. Aflatoxin B1 (m/z: 313.3) Initial Pressure: 182 bar 7. Zearalenone (m/z: 319.4) Temperature: 40°C 8. Ochratoxin A (m/z: 404.8) TEST CONDITIONS: STRUCTURES: HiCgohlurmesno: HluAtLiOon90seÅpBaiprahetinoynl, 2o.f7 eµxmp,l2o.s1ivx e1s00mm InMthoibsileexPahPmaasrpetlNeA:,uwamabHteeArr:Lw9O2it8Ch11028.-16%9101foÅrm, 2ic.7acµidm/ , 150 x 4.6 mm column is used to resolve 17 explosives in 20 minutes. This separation is quite sensitive to5mteMmapmemraotnuiurem, faonrmd awteas optimized using gradient time x temperature (tG x T) computer modeling and simulation usMinogbDileryPLhabse®Bs:oafctewtoanrietr.ile with 0.1% formic acid/ TEST CONDITIONS: Injection Volume: 40 µL PEAK IDENTITIES: 5mM ammonium formate Gradient: Time %B Column: HALO C18 90 Å, Sample Solvent: 50/50: 1. HMX 2.7 µm, 150 x 4.6 mm Water/methanol Response 0.0 32 Part Number: Time: 0.02 sec. 2. RDX 5.0 34 HAL92814-702 Data rate: 25 Hz 3. 1,3,5-Trinitrotoluene 10.0 60 Pressure: 366 bar to start, 4. 1,3-Dinitrotoluene Flow Rate: 0.4 mL/min Mobile Phase A: Water max. 405 bar 5. 3,5-Dinitroaniline Initial Pressure: 182 bar Mobile Phase B: Methanol Flow Cell: 2.5 µL semi- 6. Nitrobenzene Temperature: 40°C 3 15 16 Gradient: Time %B micro 7. Nitroglycerin Detection: LC-MS 0.0 25 LC System: Shimadzu 8. Tetryl Prominence UFLC XR ECV: 9. 2,4,6-Trinitrotoluene Injection Volume: 2.0 μL 8 14.0 35 about 14 µL 10. 2-Amino-4,6-Dinitrotoluene Absorbance @ 220 nm MS System: Thermo Fisher Orbitrap VelosPro ETD 14 20.0 62 ESI: +4 1 4 13 Flow Rate: 1.5 mL/min. 11. 4-Amino-2,6-Dinitrotoluene Heat Block: 3502°C 12 Sheath Gas Flow: 34.88 9 Temperature: 43 ˚C 12. 2,4-Dinitrotoluene 10 6 Detection: UV 220 nm, 17 VWD Aux Gas Flow: 10.00 5 11 13. 2,6-Dinitrotoluene 7 14. 2-Nitrotoluene -5 15. 4-Nitrotoluene 0 5 10 15 20 FOR MORE INFORMATION OR TO 16. 3-Nitrotoluene PLACE AN ORDER, CONTACT: www.advanced-materials-tech.com Time (min.) 17. PETN (pentaerythritol tetranitrate) ® HALO and Fused-Core are registered trademarks of Advanced Materials Technology, Inc. D-16 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns HALO® 90 Å 2.7 μm (UHPLC and HPLC) Column Ordering Information HALO 90 Å 2.7 μm columns are available in nano, capillary and analytical diameters, as well as in a 10 mm semi-preparative diameter. HPLC/UHPLC Columns Guard columns are available for analytical diameters of 2.1, 3.0 and 4.6 mm to provide additional protection when necessary. Dimensions 2.7 μm, 90 Å ID x Length (mm) C18 AQ-C18 C8 Phenyl- Biphenyl RP-Amide PFP ES-CN Penta- HILIC Hexyl HILIC .075 x 50 HAL98219-402 HAL98219-422 HAL98219-408 HAL98219-411 HAL98219-407 HAL98219-409 HAL98219-404 HAL98219-401 .075 x 100 HAL98219-602 HAL98219-622 HAL98219-608 HAL98219-406 HAL98219-611 HAL98219-607 HAL98219-609 HAL98219-604 HAL98219-405 HAL98219-601 .075 x 150 HAL98219-702 HAL98219-722 HAL98219-708 HAL98219-606 HAL98219-711 HAL98219-707 HAL98219-709 HAL98219-704 HAL98219-605 HAL98219-701 0.1 x 50 HAL98218-402 HAL98218-422 HAL98218-408 HAL98219-706 HAL98218-411 HAL98218-407 HAL98218-409 HAL98218-404 HAL98219-705 HAL98218-401 0.1 x 100 HAL98218-602 HAL98218-622 HAL98218-608 HAL98218-406 HAL98218-611 HAL98218-607 HAL98218-609 HAL98218-604 HAL98218-405 HAL98218-601 0.1 x 150 HAL98218-702 HAL98218-722 HAL98218-708 HAL98218-606 HAL98218-711 HAL98218-707 HAL98218-709 HAL98218-704 HAL98218-605 HAL98218-701 0.2 x 50 HAL98217-402 HAL98217-422 HAL98217-408 HAL98218-706 HAL98217-411 HAL98217-407 HAL98217-409 HAL98217-404 HAL98218-705 HAL98217-401 0.2 x 100 HAL98217-602 HAL98217-622 HAL98217-608 HAL98217-406 HAL98217-611 HAL98217-607 HAL98217-609 HAL98217-604 HAL98217-405 HAL98217-601 0.2 x 150 HAL98217-702 HAL98217-722 HAL98217-708 HAL98217-606 HAL98217-711 HAL98217-707 HAL98217-709 HAL98217-704 HAL98217-605 HAL98217-701 0.3 x 50 HAL98216-402 HAL98216-422 HAL98216-408 HAL98217-706 HAL98216-411 HAL98216-407 HAL98216-409 HAL98216-404 HAL98217-705 HAL98216-401 0.3 x 100 HAL98216-602 HAL98216-622 HAL98216-608 HAL98216-406 HAL98216-611 HAL98216-607 HAL98216-609 HAL98216-604 HAL98216-405 HAL98216-601 0.3 x 150 HAL98216-702 HAL98216-722 HAL98216-708 HAL98216-606 HAL98216-711 HAL98216-707 HAL98216-709 HAL98216-704 HAL98216-605 HAL98216-701 0.5 x 50 HAL98215-402 HAL98215-422 HAL98215-408 HAL98216-706 HAL98215-411 HAL98215-407 HAL98215-409 HAL98215-404 HAL98216-705 HAL98215-401 0.5 x 100 HAL98215-602 HAL98215-622 HAL98215-608 HAL98215-406 HAL98215-611 HAL98215-607 HAL98215-609 HAL98215-604 HAL98215-405 HAL98215-601 0.5 x 150 HAL98215-702 HAL98215-722 HAL98215-708 HAL98215-606 HAL98215-711 HAL98215-707 HAL98215-709 HAL98215-704 HAL98215-605 HAL98215-701 1.0 x 30 HAL92811-302 HAL92811-322 HAL92811-308 HAL98215-706 HAL92811-311 HAL92811-307 HAL92811-309 HAL92811-304 HAL98215-705 HAL92811-301 1.0 x 50 HAL92811-402 HAL92811-422 HAL92811-408 HAL92811-306 HAL92811-411 HAL92811-407 HAL92811-409 HAL92811-404 HAL92811-305 HAL92811-401 1.0 x 75 HAL92811-502 HAL92811-522 HAL92811-508 HAL92811-406 HAL92811-511 HAL92811-507 HAL92811-509 HAL92811-504 HAL92811-405 HAL92811-501 1.0 x 100 HAL92811-602 HAL92811-622 HAL92811-608 HAL92811-506 HAL92811-611 HAL92811-607 HAL92811-609 HAL92811-604 HAL92811-505 HAL92811-601 1.0 x 150 HAL92811-702 HAL92811-722 HAL92811-708 HAL92811-606 HAL92811-711 HAL92811-707 HAL92811-709 HAL92811-704 HAL92811-605 HAL92811-701 2.1 x 20 HAL92812-202 HAL92812-222 HAL92812-208 HAL92811-706 HAL92812-211 HAL92812-207 HAL92812-209 HAL92812-204 HAL92811-705 HAL92812-201 2.1 x 30 HAL92812-302 HAL92812-322 HAL92812-308 HAL92812-206 HAL92812-311 HAL92812-307 HAL92812-309 HAL92812-304 HAL92812-205 HAL92812-301 2.1 x 50 HAL92812-402 HAL92812-422 HAL92812-408 HAL92812-306 HAL92812-411 HAL92812-407 HAL92812-409 HAL92812-404 HAL92812-305 HAL92812-401 2.1 x 75 HAL92812-502 HAL92812-522 HAL92812-508 HAL92812-406 HAL92812-511 HAL92812-507 HAL92812-509 HAL92812-504 HAL92812-405 HAL92812-501 2.1 x 100 HAL92812-602 HAL92812-622 HAL92812-608 HAL92812-506 HAL92812-611 HAL92812-607 HAL92812-609 HAL92812-604 HAL92812-505 HAL92812-601 2.1 x 150 HAL92812-702 HAL92812-722 HAL92812-708 HAL92812-606 HAL92812-711 HAL92812-707 HAL92812-709 HAL92812-704 HAL92812-605 HAL92812-701 2.1 x 250 HAL92812-902 HAL92812-922 HAL92812-908 HAL92812-706 HAL92812-911 HAL92812-907 HAL92812-909 HAL92812-904 HAL92812-705 HAL92812-901 3.0 x 20 HAL92813-202 HAL92813-222 HAL92813-208 HAL92812-906 HAL92813-211 HAL92813-207 HAL92813-209 HAL92813-204 HAL92812-905 HAL92813-201 3.0 x 30 HAL92813-302 HAL92813-322 HAL92813-308 HAL92813-206 HAL92813-311 HAL92813-307 HAL92813-309 HAL92813-304 HAL92813-205 HAL92813-301 3.0 x 50 HAL92813-402 HAL92813-422 HAL92813-408 HAL92813-306 HAL92813-411 HAL92813-407 HAL92813-409 HAL92813-404 HAL92813-305 HAL92813-401 3.0 x 75 HAL92813-502 HAL92813-522 HAL92813-508 HAL92813-406 HAL92813-511 HAL92813-507 HAL92813-509 HAL92813-504 HAL92813-405 HAL92813-501 3.0 x 100 HAL92813-602 HAL92813-622 HAL92813-608 HAL92813-506 HAL92813-611 HAL92813-607 HAL92813-609 HAL92813-604 HAL92813-505 HAL92813-601 3.0 x 150 HAL92813-702 HAL92813-722 HAL92813-708 HAL92813-606 HAL92813-711 HAL92813-707 HAL92813-709 HAL92813-704 HAL92813-605 HAL92813-701 3.0 x 250 HAL92813-902 HAL92813-922 HAL92813-908 HAL92813-706 HAL92813-911 HAL92813-907 HAL92813-909 HAL92813-904 HAL92813-705 HAL92813-901 HAL92813-906 HAL92813-905 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-17 LIFE SCIENCE

Colonnes HPLC/UHPLC HALO® 90 Å 2.7 μm (UHPLC and HPLC) Column Ordering Information Colonnes HPLC/UHPLC Dimensions 2.7 μm, 90 Å ID x Length (mm) C18 AQ-C18 C8 Phenyl- Biphenyl RP-Amide PFP ES-CN Penta- HILIC Hexyl HILIC 4.6 x 20 HAL92814-202 HAL92814-222 HAL92814-208 HAL92814-211 HAL92814-207 HAL92814-209 HAL92814-204 HAL92814-201 4.6 x 30 HAL92814-302 HAL92814-322 HAL92814-308 HAL92814-206 HAL92814-311 HAL92814-307 HAL92814-309 HAL92814-304 HAL92814-205 HAL92814-301 4.6 x 50 HAL92814-402 HAL92814-422 HAL92814-408 HAL92814-306 HAL92814-411 HAL92814-407 HAL92814-409 HAL92814-404 HAL92814-305 HAL92814-401 4.6 x 75 HAL92814-502 HAL92814-522 HAL92814-508 HAL92814-406 HAL92814-511 HAL92814-507 HAL92814-509 HAL92814-504 HAL92814-405 HAL92814-501 4.6 x 100 HAL92814-602 HAL92814-622 HAL92814-608 HAL92814-506 HAL92814-611 HAL92814-607 HAL92814-609 HAL92814-604 HAL92814-505 HAL92814-601 4.6 x 150 HAL92814-702 HAL92814-722 HAL92814-708 HAL92814-606 HAL92814-711 HAL92814-707 HAL92814-709 HAL92814-704 HAL92814-605 HAL92814-701 4.6 x 250 HAL92814-902 HAL92814-922 HAL92814-908 HAL92814-706 HAL92814-911 HAL92814-907 HAL92814-909 HAL92814-904 HAL92814-705 HAL92814-901 10.0 x 50 HAL92810-402 HAL92810-422 HAL92810-408 HAL92814-906 HAL92810-411 HAL92810-407 HAL92810-409 HAL92810-404 HAL92814-905 HAL92810-401 10.0 x 75 HAL92810-502 HAL92810-522 HAL92810-508 HAL92810-406 HAL92810-511 HAL92810-507 HAL92810-509 HAL92810-504 HAL92810-405 HAL92810-501 10.0 x 100 HAL92810-602 HAL92810-622 HAL92810-608 HAL92810-506 HAL92810-611 HAL92810-607 HAL92810-609 HAL92810-604 HAL92810-505 HAL92810-601 10.0 x 150 HAL92810-702 HAL92810-722 HAL92810-708 HAL92810-606 HAL92810-711 HAL92810-707 HAL92810-709 HAL92810-704 HAL92810-605 HAL92810-701 HAL92810-706 HAL92810-705 HALO guard cartridge and guard holder For more details please go to page D-36 Dimensions 2.7 μm, 90 Å ID x Length (mm) C18 AQ-C18 C8 Phenyl- Biphenyl RP-Amide PFP ES-CN Penta- HILIC Hexyl HILIC 2.1 x 5 HAL92812-102 HAL92812-122 HAL92812-109 HAL92812-104 HAL92812-101 3.0 x 5 HAL92813-102 HAL92813-122 2.7 µm, 90 Å Guard Cartridges, 3-pack HAL92813-109 HAL92813-104 HAL92812-105 HAL92813-101 4.6 x 5 HAL92814-102 HAL92814-122 HAL92814-109 HAL92814-104 HAL92813-105 HAL92814-101 HAL92812-108 HAL92812-106 HAL92812-111 HAL92812-107 HAL92814-105 HAL92813-108 HAL92813-106 HAL92813-111 HAL92813-107 HAL92814-108 HAL92814-106 HAL92814-111 HAL92814-107 Guard Column Holder - HAL94900-001 Did you know? We sell a wide range of HPLC maintenance products such as Pump Seals, Check Valves, Piston Seals and Lamps. For more information please ask your Sales Representative today. D-18 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns HALO 5 µm HPLC/UHPLC Columns HALO® 90 Å 5 μm (HPLC) Performance of 3 µm non-core column at 5 µm column pressures. Ideal for: 4.6 µm • QC laboratories 3.3 µm • Dirty samples • High throughput, ballistic gradient and isocratic applications 0.6 µm • High resolution at HPLC back pressures (using columns in series) • 2 µm inlet frit • Pressure limit, 600 bar/9000 psi Benzodiazepine separation using HALO 5 µm PFP Shell with 90 Å pores These six benzodiazepine drugs are separated in 5 minutes with better performance than a 3 µm non-core column at ½ the pressure. 0.0461 0.0510 TEST CONDITIONS: PEAK IDENTITIES: Column: HALO PFP 90 Å, 5 μm 1. Oxazepam 0.0467 100 x 4.6 mm 2. Lorazepam Part Number: HAL95814-609 3. Nitrazepam Absorbance (mAU) HALO 5 PFP (5µm) 0.0601 0.0686 Mobile Phase A: 25 mM Ammonium 4. Clonazeparm P=46 bar Acetate, pH 5.5 5. Flunitrazepam 3 µm PFP 0.0625 Mobile Phase B: ACN, 36-65% B in 7 min. 6. Diazepam P=93 bar Temperature: 35 ˚C Flow: 0.75mL/min. NOTE: Detector: UV at 254 nm Peak widths at half height are labeled Injection: 1 μL for comparable peaks on both columns. 0 1 2 3 4 5 6 7 Time (min.) Comparative results presented here may not be representative for all applications. Polar aromatic compounds on HALO 5 µm RP-Amide HALO 5 µm RP-Amide shows excellent resolution and peak shape for this mixture of polar aromatic compounds. TEST CONDITIONS: Pressure: 308 bar 145 1 Column: HALO RP-Amide 90 Å, 5 μm Temperature: 26 °C 100 x 4.6 mm Detection: UV 254 nm, VWD Part Number: HAL95814-607 Injection Volume: 5.0 μL Mobile Phase: 70/30: A/B Sample Solvent: 50/50: Water/Acetonitrile A: 20 mM Potassium Phosphate, pH 7 Response Time: 0.12 sec. Absorbance @ 254nm (mAU) 115 B: Acetonitrile Flow Cell: 5 μL semi-micro 3 Flow Rate: 4.0 mL/min LC System: Optimized Agilent 1100 2 85 PEAK IDENTITIES: 5. Dimethyl Phthalate 4 5 6 7 8 1. Uracil 6. 2-Nitroaniline 2. Benzamide 7. 4’-Bromoacetanilide 55 3. Aniline 8. 2,2’-Biphenol 9 25 4. Cinnamyl Alcohol 9. 4,4’-Biphenol -5 0.5 1.0 1.5 2.0 2.5 3.0 0.0 Time (min.) 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-19 LIFE SCIENCE

Colonnes HPLC/UHPLC Colonnes HPLC/UHPLC HALO® 90 Å 5 μm (HPLC) Column Ordering Information HALO 90 Å 5 μm columns are available in nano, capillary and analytical diameters, and in a 10 mm semi-preparative diameter. Guard columns are available for analytical diameters of 2.1, 3.0 and 4.6 mm. Dimensions 5 μm, 90 Å ID x Length (mm) C18 AQ-C18 C8 Phenyl- Biphenyl RP-Amide PFP ES-CN Penta- HILIC Hexyl HILIC .075 x 50 HAL98519-402 HAL98519-422 HAL98519-408 Enquire HAL98519-407 HAL98519-409 HAL98519-404 HAL98519-401 .075 x 100 HAL98519-602 HAL98519-622 HAL98519-608 HAL98519-406 HAL98519-609 HAL98519-604 HAL98519-405 HAL98519-601 .075 x 150 HAL98519-702 HAL98519-722 HAL98519-708 HAL98519-606 Enquire HAL98519-607 HAL98519-709 HAL98519-704 HAL98519-605 HAL98519-701 0.1 x 50 HAL98518-402 HAL98518-422 HAL98518-408 HAL98519-706 HAL98518-409 HAL98518-404 HAL98519-705 HAL98518-401 0.1 x 100 HAL98518-602 HAL98518-622 HAL98518-608 HAL98518-406 Enquire HAL98519-707 HAL98518-609 HAL98518-604 HAL98518-405 HAL98518-601 0.1 x 150 HAL98518-702 HAL98518-722 HAL98518-708 HAL98518-606 HAL98518-709 HAL98518-704 HAL98518-605 HAL98518-701 0.2 x 50 HAL98517-402 HAL98517-422 HAL98517-408 HAL98518-706 Enquire HAL98518-407 HAL98517-409 HAL98517-404 HAL98518-705 HAL98517-401 0.2 x 100 HAL98517-602 HAL98517-622 HAL98517-608 HAL98517-406 HAL98517-609 HAL98517-604 HAL98517-405 HAL98517-601 0.2 x 150 HAL98517-702 HAL98517-722 HAL98517-708 HAL98517-606 Enquire HAL98518-607 HAL98517-709 HAL98517-704 HAL98517-605 HAL98517-701 0.3 x 50 HAL98516-402 HAL98516-422 HAL98516-408 HAL98517-706 HAL98516-409 HAL98516-404 HAL98517-705 HAL98516-401 0.3 x 100 HAL98516-602 HAL98516-622 HAL98516-608 HAL98516-406 Enquire HAL98518-707 HAL98516-609 HAL98516-604 HAL98516-405 HAL98516-601 0.3 x 150 HAL98516-702 HAL98516-722 HAL98516-708 HAL98516-606 HAL98516-709 HAL98516-704 HAL98516-605 HAL98516-701 0.5 x 50 HAL98515-402 HAL98515-422 HAL98515-408 HAL98516-706 Enquire HAL98517-407 HAL98515-409 HAL98515-404 HAL98516-705 HAL98515-401 0.5 x 100 HAL98515-602 HAL98515-622 HAL98515-608 HAL98515-406 HAL98515-609 HAL98515-604 HAL98515-405 HAL98515-601 0.5 x 150 HAL98515-702 HAL98515-722 HAL98515-708 HAL98515-606 Enquire HAL98517-607 HAL98515-709 HAL98515-704 HAL98515-605 HAL98515-701 1.0 x 30 HAL95811-302 HAL95811-322 HAL95811-308 HAL98515-706 HAL95811-309 HAL95811-304 HAL98515-705 HAL95811-301 1.0 x 50 HAL95811-402 HAL95811-422 HAL95811-408 HAL95811-306 Enquire HAL98517-707 HAL95811-409 HAL95811-404 HAL95811-305 HAL95811-401 1.0 x 75 HAL95811-502 HAL95811-522 HAL95811-508 HAL95811-406 HAL95811-509 HAL95811-504 HAL95811-405 HAL95811-501 1.0 x 100 HAL95811-602 HAL95811-622 HAL95811-608 HAL95811-506 Enquire HAL98516-407 HAL95811-609 HAL95811-604 HAL95811-505 HAL95811-601 1.0 x 150 HAL95811-702 HAL95811-722 HAL95811-708 HAL95811-606 HAL95811-709 HAL95811-704 HAL95811-605 HAL95811-701 2.1 x 20 HAL95812-202 HAL95812-222 HAL95812-208 HAL95811-706 Enquire HAL98516-607 HAL95812-209 HAL95812-204 HAL95811-705 HAL95812-201 2.1 x 30 HAL95812-302 HAL95812-322 HAL95812-308 HAL95812-206 HAL95812-309 HAL95812-304 HAL95812-205 HAL95812-301 2.1 x 50 HAL95812-402 HAL95812-422 HAL95812-408 HAL95812-306 Enquire HAL98516-707 HAL95812-409 HAL95812-404 HAL95812-305 HAL95812-401 2.1 x 75 HAL95812-502 HAL95812-522 HAL95812-508 HAL95812-406 HAL95812-509 HAL95812-504 HAL95812-405 HAL95812-501 2.1 x 100 HAL95812-602 HAL95812-622 HAL95812-608 HAL95812-506 Enquire HAL98515-407 HAL95812-609 HAL95812-604 HAL95812-505 HAL95812-601 2.1 x 150 HAL95812-702 HAL95812-722 HAL95812-708 HAL95812-606 HAL95812-709 HAL95812-704 HAL95812-605 HAL95812-701 2.1 x 250 HAL95812-902 HAL95812-922 HAL95812-908 HAL95812-706 Enquire HAL98515-607 HAL95812-909 HAL95812-904 HAL95812-705 HAL95812-901 3.0 x 20 HAL95813-202 HAL95813-222 HAL95813-208 HAL95812-906 HAL95813-209 HAL95813-204 HAL95812-905 HAL95813-201 3.0 x 30 HAL95813-302 HAL95813-322 HAL95813-308 HAL95813-206 Enquire HAL98515-707 HAL95813-309 HAL95813-304 HAL95813-205 HAL95813-301 3.0 x 50 HAL95813-402 HAL95813-422 HAL95813-408 HAL95813-306 HAL95813-409 HAL95813-404 HAL95813-305 HAL95813-401 3.0 x 75 HAL95813-502 HAL95813-522 HAL95813-508 HAL95813-406 Enquire HAL95811-307 HAL95813-509 HAL95813-504 HAL95813-405 HAL95813-501 3.0 x 100 HAL95813-602 HAL95813-622 HAL95813-608 HAL95813-506 HAL95813-609 HAL95813-604 HAL95813-505 HAL95813-601 3.0 x 150 HAL95813-702 HAL95813-722 HAL95813-708 HAL95813-606 Enquire HAL95811-407 HAL95813-709 HAL95813-704 HAL95813-605 HAL95813-701 3.0 x 250 HAL95813-902 HAL95813-922 HAL95813-908 HAL95813-706 HAL95813-909 HAL95813-904 HAL95813-705 HAL95813-901 HAL95813-906 Enquire HAL95811-507 HAL95813-905 Enquire HAL95811-607 Enquire HAL95811-707 HAL95812-211 HAL95812-207 HAL95812-311 HAL95812-307 HAL95812-411 HAL95812-407 HAL95812-511 HAL95812-507 HAL95812-611 HAL95812-607 HAL95812-711 HAL95812-707 HAL95812-911 HAL95812-907 HAL95813-211 HAL95813-207 HAL95813-311 HAL95813-307 HAL95813-411 HAL95813-407 HAL95813-511 HAL95813-507 HAL95813-611 HAL95813-607 HAL95813-711 HAL95813-707 HAL95813-911 HAL95813-907 D-20 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns HALO® 90 Å 5 μm (HPLC) Column Ordering Information (continued) Dimensions 5 μm, 90 Å HPLC/UHPLC Columns ID x Length (mm) C18 AQ-C18 C8 Phenyl- Biphenyl RP-Amide PFP ES-CN Penta- HILIC Hexyl HILIC 4.6 x 20 HAL95814-202 HAL95814-222 HAL95814-208 HAL95814-211 HAL95814-207 HAL95814-209 HAL95814-204 HAL95814-201 4.6 x 30 HAL95814-302 HAL95814-322 HAL95814-308 HAL95814-206 HAL95814-311 HAL95814-307 HAL95814-309 HAL95814-304 HAL95814-205 HAL95814-301 4.6 x 50 HAL95814-402 HAL95814-422 HAL95814-408 HAL95814-306 HAL95814-411 HAL95814-407 HAL95814-409 HAL95814-404 HAL95814-305 HAL95814-401 4.6 x 75 HAL95814-502 HAL95814-522 HAL95814-508 HAL95814-406 HAL95814-511 HAL95814-507 HAL95814-509 HAL95814-504 HAL95814-405 HAL95814-501 4.6 x 100 HAL95814-602 HAL95814-622 HAL95814-608 HAL95814-506 HAL95814-611 HAL95814-607 HAL95814-609 HAL95814-604 HAL95814-505 HAL95814-601 4.6 x 150 HAL95814-702 HAL95814-722 HAL95814-708 HAL95814-606 HAL95814-711 HAL95814-707 HAL95814-709 HAL95814-704 HAL95814-605 HAL95814-701 4.6 x 250 HAL95814-902 HAL95814-922 HAL95814-908 HAL95814-706 HAL95814-911 HAL95814-907 HAL95814-909 HAL95814-904 HAL95814-705 HAL95814-901 10.0 x 50 HAL95810-402 HAL95810-422 HAL95810-408 HAL95814-906 HAL95810-407 HAL95810-409 HAL95810-404 HAL95814-905 HAL95810-401 10.0 x 75 HAL95810-502 HAL95810-522 HAL95810-508 HAL95810-406 Enquire HAL95810-507 HAL95810-509 HAL95810-504 HAL95810-405 HAL95810-501 10.0 x 100 HAL95810-602 HAL95810-622 HAL95810-608 HAL95810-506 Enquire HAL95810-607 HAL95810-609 HAL95810-604 HAL95810-505 HAL95810-601 10.0 x 150 HAL95810-702 HAL95810-722 HAL95810-708 HAL95810-606 Enquire HAL95810-707 HAL95810-709 HAL95810-704 HAL95810-605 HAL95810-701 10.0 x 250 HAL95810-902 HAL95810-922 HAL95810-908 HAL95810-706 Enquire HAL95810-907 HAL95810-909 HAL95810-904 HAL95810-705 HAL95810-901 HAL95810-906 Enquire HAL95810-905 HALO guard cartridge and guard holder For more details please go to page D-36 Dimensions 5 μm, 90 Å ID x Length (mm) C18 AQ-C18 C8 Phenyl- Biphenyl RP-Amide PFP ES-CN Penta- HILIC Hexyl HILIC 2.1 x 5 HAL95812-102 HAL95812-122 HAL95812-109 3.0 x 5 HAL95813-102 HAL95813-122 5 µm, 90 Å Guard Cartridges, 3-pack HAL95813-109 4.6 x 5 HAL95814-102 HAL95814-122 HAL95814-109 HAL95812-108 HAL95812-106 HAL95812-111 HAL95812-107 HAL95812-104 HAL95812-105 HAL95812-101 HAL95813-104 HAL95813-105 HAL95813-101 HAL95813-108 HAL95813-106 HAL95813-111 HAL95813-107 HAL95814-104 HAL95814-105 HAL95814-101 HAL95814-108 HAL95814-106 HAL95814-111 HAL95814-107 Guard Column Holder - HAL94900-001 Did you know? Canadian Life Science is an Authorized Distributor for Honeywell Research Chemicals? For more information please ask your Sales Representative today. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-21 LIFE SCIENCE

Colonnes HPLC/UHPLC PEAK IDENTITIES • Colonnes HPLC/UHPLC HSAMALLLOMO®LECCUL3E 0 160 Å 2.7 μm 1. Retinyl acetate (A) • 2. Delta tocopherol (E) B • Built on proven Fused-Core® particile technology, the HALO® C30 is designed 3. Ergocalciferol (D2) to deliver fast separations ideal for lipids and isomers compared to your C18 4. Cholecalciferol (D3) • 5. Alpha tocopherol (E) • 6. DL-alpha-tocopherol acetate (E) 7. 2,3-trans-phylloquinone (K) 8. Retinyl palmitate (A) • 90 High shapeÅ 2 micron particle selectivity90 Å 2.7 micron particle for hydrophobic,90 Å 5 micron particle long-chain structurally related isomers COLUMN • BIOCHLAiSgShly reproducible bonded phase coverage resulting in fast, highly efficient, rugged separatioHnAsLO 160 Å C30, 2.7 µm, 4.6 x 150 mm • 100 % Aqueous Compatibility *For method conditions please see full application at advanced-materials-tech.com A HALO® C30 column shows increased retention and baseline resolution of vitamin K1 trans and cis isomers compared to a SPP C30 column. Since160 Å 2 micron particle the cis isomer160 Å 2.7 micron particle of K1 is biologically160 Å 5 micron particle inactive, it is important to know how much of each isomer is present in vitamin PEPTIDE enriched products. 1 2 *For method conditions PEAK IDENTITIES please see full application at 1. Menadione (K3) Absorbance @ 280nm1000 Å 2.7 micron particle 400 Å 3.4 micron particle 90 Å 2.7 micron particle advanced-materials-tech.com 2. Menaquinone 4 (K2) 3. 2,3-trans-phylloquinone (K1) PROTEIN GLYCAN 3 4. cis-phylloquinone (K1) 1 2 Fully resolved HALO® C30 COLUMNS SPP C30 Unresolved 3,4 HALO 160 Å C30, 2.7 µm, 4.6 x 150 mm SPP C30, 2.6 μm, 4.6 x 150 mm Part Number: HAL9211P4E-A7K3I0DENTITIES 0 2 4 6 8 10 12 14 *For method conditions please see full Time, min. HALO ® C30 PEAK IDENTITIES application at advanced-materials-tech.com *For metho please see 1 4 PEAPKEAIDKPEEIANDKTEIDINETNTITEIITTSIEIES S 2 TEST CONDITIONS advanced HALO ® C30 HALO ® C18 Absorbance @ 280nm HALO ® C30 Columns: HALO 160 Å C30 and Tocopherols, a form of vitamin E, known for their antioxidant properties contain many isomers which are easily baseline separatedHALO ® C184 1 on the HALO® C30 compared to a C18 due to the shape selectivity property of the phase.STRUCTURE HAHLOAL®OSCT®3R0UCC3T0URE HALO ® C30 HAHLOAL®OC®18C18 HATLOES®TC1C8ONDITIONS 4 2 90 Å C18 , 2.7 µm, 4.6 x 150mm PartTNESuTmCbOeNr:DHIATILO9N21S14-730 4CCRHHHH2S₃₃TRTUAoCclTpoUhpRaEh(eαr9PMCP9IIFTD)onnel05ahoeojsomlr%ealttwÅutbercspmuNietcBClReietmou1nBraionam8Pset:net:hnV4,ubM::at2oreUH:s1ee.lreA7uVA.::t5hgmLAHµCH12RiamO:lmA09eAHen1°L:0LW1nLo,C/9₃196t4lnm2a.T20I.1G5ms6t18i2AoeonÅD1μPT9CPB,a1x0rcc4lPL4C0aoe0mreEpT1EM-GoD-a3r7l50l7SAtÅohSuAttDA0top3mBa0La0iacmacTblN0ma2CehmKep:iCo(anlu1R(nlm(eethβCdtδHpmαmI8as2(raa)DO:)hγ)o₃,b(a()(eEβ2lNeδHαT(.rGNr)7)γADA)oo:DBa)LTHcµllHemIepOomITAlAthtTpm4aLIa1aL,hOI996(a4E(2(eβ2SHδ20α.N(r4S61)CC8)γP)oRÅA1HH)1SHHxlP144CLPCMP9T9P1₃₃-CC-3C057ahRo5OE7EHHoT0HHGr%3al0031AÅoSuADtb®Bas0ma₃₃2cl0emmepTNieBoCKnllthCtepmmau1anBadhC(a3CCmI8P(s(:eβRδαDHHO:(HHr0h))γ1,)oCCbMR)a₃₃El2NHHeHHHs4e2.Nre7AD₃₃:tCChRLTAHµHHHHHIaO2:mITAAnT₃₃LCCIW1LRo,HHOIHH9CC961R4lEH₃₃H2aHH20I.2NSs6t18₃₃eoÅ11Sxrc44Cr1M--a37CC57RtHH0oHH3020ic₃₃bM9TPCFTD9IIP0m6a2nn:eli05ahoneEolsjomer%ealtdtwÅSutberTM9CPcPITID9FspnnmuTeNietcBl05ahCoelEReoisjeotmmMPT9IDIFor%eual1ntBtrainnwÅSCuetlb5oherencsam8oPsjset:pomTm%ueTaNiettncBOtet:wChlRnbeeiVrics,MP9CIFDPT9IubetmMpm:nn:oueu1naileBtc05ahBotraieo2ClsjRoNeroemioUr%eHaelt:mnatmt8Psw1oÅsuet:etebeBe.raricslrpnOeomeAtu:7NientcuBahVA.PC,nDet:l:R:teVi,uetm5bMnou:e1t:nBhhraigmmanLtAVHoµ2unaMm8PoNHsret:12e::UIHa:intamOset:1honereV:l.,mUTubAMi:lr09e::s1AeaeAe7tennu2VA.olreD:U8:tHe:As1uV.5eL°:0eL.I:tWl1rnL.ehAo5g7,muVA.LA::tHChµgmO/59HA112hg9m6IHLt1A2Hiµ4amOlnH12imamia:lmOmT2Aa:l.09:elmA029e0A9e0eAAeIne.1nn5mN°L:0sL6t1W1nL°L°::00LLo,IW8W1innCLLo/o29,1e9C6ontÅC4/lnO/1m91μ912at,.916S2l0ntxI.41m0l5mnrmcas6t1.282i4aI.2PL1e52m4onÅ0CI1.1μ0st,r5m1N1xM08irsc6t12-4eDoPLn8-a4iC302μr17,Me1o5nÅ-0D7-aSr13μc7,t51S7SA0PLxo04t3rA0o00cr3L00iML0i4PL4cDCc-ba00bmra021mM:a27S-iD:tn-aAUl3oim7e5L0in7dSlctbmA0eo32n0dL0:iicblr0ma2e:e1inlsm0eodlved 3,4 HALO ® CS3T0RSUTRCUTUCRTUE RE Tocopherol R1 12 HALO ® C18 Alpha (α) CH₃ Beta (β) CH₃ Gamma (γ) H Delta (δ) H HALO ® C18 STRUCTURE STRUCTURE CThaeroseteTGsnAoBaeclomepovthpimaeadh(an(eβα(Sr)γ)oce)lapraortaetnCCiRGHHHoo1DBa₃₃neimedlttmaa(sL(a(βδae())γnCCfR)HHHtd2)₃₃ CH₃ MobileHPhase A: Water FMloowbiRleate: 1.5 mL/min H PhaseCBH: ₃Methanol IsocrTaetmicp: erature: 10°C H 95% B H Injection Volume: 1.5 μL related Flow Rate: 1.5 mL/min Instrument: Agilent 1200 SL Temperature: 10°C Detection: UV 290 nm, PDA isomersDeflrtao(mδ) a commH ercialHly availablIenjection Volume: 1.5 μL vitamin formulation are easily resolvedInstrument: Agilent 1200 SL Detection: UV 290 nm, PDA under simple mobile phase conditions.TEST CONDITIONS Column: HALO 160 Å C30, 2.7 µm, 3.0 x 150 mm Part Number: HAL92113-730 Mobile Phase A: Methanol PEAK IDENTITIES TEST CONDITIONS PEAK IDENTITIES Mobile Phase B: Ethanol Column: HALO 160 Å C30, 2.7 µm, 4.6 x 150 mm 1. Prednisone Gradient: 100% A with gradient to 40% B at 20 min. 1. Lutein Flow Rate: 0.65 mL/min 2. cis-carotenoid 1 Part Number: HAL92114-730 2. Cortisone Temperature: 38°C 3. cis-carotenoid 2 Mobile Phase A: Water 3. Prednisolone Steroids Separations (Right)TEST CONDITIONS Mobile Phase B: 50/50 Acetonitrile/ Methanol 4. Hydrocortisone Data acquisition rate: 2.5 Hz -carotene Isocratic: 50% B 5. Dexamethasone CANADIAN aiGasgsmnthlnluenfiuadlnrcuacpouopmmoeitcrdce.bmoeoshersratcaortitirncriocioombdofnieidcodd.dosnesIn1naemaadsr8trboeihea8tonies8oatmseh-ent2pprexsna2oaaacta6rwnmeaat-seu2utpeertr7sxlofadee7culdel5cwblyolfebliopentlyohr:nrt:morwitnodploustienhifcxafreoked@e lifescience.ca :: www.lifescience.caGTCMMPIFTDIDDnnelaoaearEojsoomarelttttwSutbbaaedrcpmuTNiitciacllReeieetmouoncrainConqamuPP:ettnOetu:hhrnvubt::HiaatoesNre:1ssAielsrtueeU00yD::LiAom.0VOAB6oHIng%::e53Tf4i1Alr:8N7eIAm6aLO°n1Ea09tMCwLettt2Nn0/Åuh:ei11m.mtra6tS1h1Cehni0,033n’agsoP0-0µnr27lSDT,aLo.3uA2d5lE0n.iH7esSnhLzµTtinmteoIC,F34PO.0r0oE%NxdBu1Dc5aSt0ItsT.2mIC0mOmNIiEn.SNCGMPCMTTFelaor761P23EooomEarl.....wESutbbdLccLcPCTMGMFTpAymuiiiTe-Nii-liasssollrERcceoecteeoo---mKarauoleanlccranwSCuytbbrdraaipamPP:tpnoctoImTrrNiieOiet:DohhllRtooeetubeene:epHunaattranECeNrneeene1amPPss:AtetNennernOeeet00:hhD::Looeub:T.0HaaOBAii6NreddHI1%IssAe::53TTree100A12D::L8IIAm6.0LOEABO6°HIE09%MCS::w53LTt12AN/Å8heIAim61mLtO°atSE0h91CMChwLnit233nN/Åaghoei1-0mtnra7ltSh1C,aIMCDMPIGFTDDhon3nnie3l2d3aoneaaroagljso0oomare-.l0ittttwnrut7ebb7laaedrcp,amuoNiintci3acllReµe2dieetmlotuo0cnrain.mionMPTCMFBITIIDqamuPP7e:etttnsnenletauo:aohhreEnono,vsjuboomtµ::recHilaatttcowSetsu3trebb4ke:rc1mssrApielsmurT.tNiittacueeU000PllyR0e::LiioeetmtAomo,.0un%VrOBAraii6CooHcengnx%a3mPP:et4::e53f:4nOiset1Ahh.nlBr:V018ubsN7e0:A:m6HaaaLtu°5No%r5en1E:a09tMssaCAewrxL0lret0ee1ttU2uetn0D:/:LÅuh%:eSBi.111mm:VO.mABtH52mrah6tI1h1C5e:6hB:TnAa0eii02m,0133nm’0mags3o:tL2PI06-0µmnr°LO9207l0Sa0D502mWC,a/Lo2.93.ud0mA2d550NinÅl10nnzam/b.imH7eu1iμ5stmS.Cnaenh4L0z,µrN3r-tiiPnm0A7netDe,3xco.,2Aee034P.tr7.oa0r0oIDIDn%µnnXxP213deaijsm2ettttBu...1aerrcEu,itcc5cLccliatmoe4tAouiii012P3tosnssue/.n.t6r...2--mnKvEt:Mecctoe0Lccx:mlaaisAuuUiienyIm1ssAmrrVtDto--oo5gKheiefcc4in0l:ttaN7eEaai.eenn1naImrrtNtnnDn0ooou1m.mrool61tteTE0,0eeii’sPd0dµINnnSDLTuA12oonT6721P3IsiiE.....ddhEIicLcLcTnSAyP53461212uiii-e-sssccIc......t---EKaPEoealccPDCCHPyrrAraaiporrooSneyocoIeerrdeDrrodKxtootuddttneaepriicttEnnoecsntmeenIesooiiDcNenn.ensseonsooooeEttTernliiehtoddNIieranTso12soTeInonEIenTeSeIMPTCMEaoE67Soorl..SutbbLcMCPTyi-msTNiiccao-llEaoleeoyrruplnocSCutbeotmPP:nempTNiOenhhlebeenHunaaCeNemPss:ArOeehCMPTMIIIDFBTD:LsnnbelHaoaaOeEoABHosjNooemIreclsttcAwS:u:tTbbAkerrcer1pDmu:TLNiitacLIPllRe6iOeetmAtHoOu9nrraiIi5C0WocenamP:P:T2Aet01:nOsetNhhÅn1VLubIsa/:6:HaatuO915Nore5tS:0CssWAer0elr24ee10UueD3::LN%ÅrS1-.m:VaOABH50A7hI1BTDIIFt:6SnnB:TACe,eil32mac1eeom4sjm0e3c:Lt22It3we06kerrc-°LO9p00a0u.5tc00tW7CP/Rei729tm.doo0mrrai,5Nn3Åoe1nazeta/nbµ2nmset0u1niμ5VtuSsC:na:it.emu4orL0t,:7rN3erlrr-1UueP:0A7Se,.im:Vµ5Dlh,3xce46ei2mm2Amee003:2/.°L.t0r60a0C7/o,a9.Mdm5nznx4µbmXuiμnaie.m2Lt1,rN6rtPe5,ihDlxxe40Aea/.r16anmM5Xox20mel1t5mh0amnmoml 6. Corticosterone PEAK IDENTITIES 1. Prednisone 2. Cortisone 3. Prednisolone 4. Hydrocortisone 5. Dexamethasone 6. Corticosterone PEAPKEAIDKEINDTEINTTIEITSIES D-22 1. PHCPrr213oyee...rdddtPCPrinnosrrooeeiicrssddontooinnresnltooiiiessnsnoooeelnnoeene 2. 3. 4.

HPLC/UHPLC Columns HALO® C30 160 Å 2.7 μm Ordering Information HPLC/UHPLC Columns HALO® C30 160 Å 2.7 μm columns are available in analytical diameters of 2.1, 3.0 and 4.6 mm. Guard columns are available for analytical diameters of 2.1, 3.0 and 4.6 mm to provide additional protection when necessary. Dimensions 2.7 μm, 160 Å ID x Length (mm) HAL92112-230 2.1 x 20 HAL92112-330 2.1 x 30 HAL92112-430 2.1 x 50 HAL92112-530 2.1 x 75 HAL92112-630 2.1 x 100 HAL92112-730 2.1 x 150 HAL92112-930 2.1 x 250 HAL92113-230 3.0 x 20 HAL92113-330 3.0 x 30 HAL92113-430 3.0 x 50 HAl92113-530 3.0 x 75 HAL92113-630 3.0 x 100 HAL92113-730 3.0 x 150 HAL92113-930 3.0 x 250 HAL92114-230 4.6 x 20 HAL92114-330 4.6 x 30 HAL92114-430 4.6 x 50 HAL92114-530 4.6 x 75 HAL92114-630 4.6 x 100 HAL92114-730 4.6 x 150 HAL92114-930 4.6 x 250 HALO guard cartridge and guard holder For more details please go to page D-36 2.1 x 5 Guard Cartridges, 3-pack 3.0 x 5 4.6 x 5 HAL92112-130 HAL92113-130 HAL92114-130 Guard Column Holder - HAL94900-001 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-23 LIFE SCIENCE

Colonnes HPLC/UHPLC Colonnes HPLC/UHPLC HALO® Protein, Peptide and Glycan Today, researchers are interested in both fast and high resolution separations of numerous biomolecules to support the development of novel therapeutic proteins and peptides in pharmaceutical drug development, to advance understanding in modern university laboratories, to characterize protein post-translation modifications, and to fully assess subtle differences in biosimilars and other products of bioengineering and manufacturing. HALO BioClass columns have been developed to make such tasks easier and more effective, and are aimed at the following areas of bioseparations: • Intact proteins, monoclonal antibodies (mAbs), biosimilars, and other large biomolecules such as pegylated proteins, antibody drug conjugates (ADCs), etc. • Peptide mapping (analysis of enzyme digests) for characterization and monitoring of synthetic protein drugs • Analysis of therapeutic peptides and peptide biomarkers (protein surrogates) • High resolution separations of complex mixtures of glycans released from N- and O-linked glycoproteins Currently, the HALO BioClass column family is comprised of the following products. HALO BioClass Column Specification Bonded USP Particle Pore Size Carbon Surface Low pH/T High pH/T Endcapped Phase Designation Size (Å) Load (%) Area Limit Limit (µm) (m2/g) L26 C4 2.7 1000 0.6 22 2/90 °C 9/40 °C Yes ES-C18 L1 2.7 1000 1.4 22 1/90 °C 8/40 °C Yes Protein C4 L26 3.4 400 0.4 15 2/90 °C 9/40 °C Yes ES-C18 L1 3.4 400 1.0 15 1/90 °C 8/40 °C Yes Diphenyl L11 2.7 1000 1.0 22 2/90 °C 9/40 °C Yes 2 4 65 ES-C18 L1 2.7 160 4.6 90 1/90 °C 8/40 °C No ES-CN 5 4.0 60 Phenyl-Hexyl Peptide Proprietary 2.7 2.2 90 1/90 °C 8/40 °C Yes Glycan L10 160 Ligand Yes 5 1.5 60 No L11 2.7 160 4.7 90 2/90 °C 9/40 °C 2/65 °C 9/40 °C L95 2.7 90 3.2 135 D-24 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns High resolution of light and heavy chain variants of IgG1 Very high resolution is obtained Masses deconvoluted using TEST CONDITIONS: HPLC/UHPLC Columns bet­ ween variants of light and heavy MagTran software* Column: HALO C4 400 Å, 3.4 μm, 100 x 2.1 mm chains of a reduced and alkylated Part Number: HAL93412-614 monoclonal antibody (IgG1) sample 3.5 Mobile Phase A: 0.5% formic acid with 20 mM using a HALO Protein C4 column. 1366 .0 Ammonium Formate 3 1290 .0 Mobile Phase B: 45% ACN/45% IPA/10% A solvent Gradient: 29–32% B in 20 min. 2.5 1221 .9 Temperature: 80 ˚C 1161 .4 Detection: 280 nm and MS using 2pps scan rate 2 1105 .9 1451 .6 from 500 to 2000 m/z Injection Volume: 2 μL of 2 μg/μL reduced 1.5 and alkylated lgG1 Sample Solvent: 0.25% (v/v) formic acid in water 1 1546 .7 1655 .6 MS Parameters: Positive ion mode, ESI at +4.5 kV, 0.5 400 ˚C heatblock, 225 ˚C capillary 1010 .1 1783 .1 LC-MS System: Shimadzu Nexera and LCMS-2020 0 (single quadrupole MS) 7 LC1: 23,204 500 930 .2 1604 .6 1931 .5 6 773 .6 859 .4 1859 .1 20 5 4 750 1000 1250 1500 1750 m/z 3 Absorbance @ 280 nm 2 LC= light chain HC= HC2: 50,424 1 heavy chain HC3: 50,668 0 HC1: 50,539 -5 LC3: 23,203 HC4: 50,680 LC2: 23,192 HC5: 28,862 0 2.5 5 7.5 10 12.5 15 17.5 Time (min.) Ultrafast protein separation using HALO Protein ES-C18 TEST CONDITIONS: PEAK IDENTITIES Column: HALO ES-C18 400 Å, 1. Ribonuclease A An ultrafast protein HALO Protein ES-C18, 3.4 µm, 400 Å 0.0065 3.4 μm, 100 x 4.6 mm 2. Cytochrome c separa­tion is achieved 75 Pressure: 234 bar 0.0071 Part Number: HAL93414-602 3. Lysozyme on a HALO Protein ES-C18 Instrument: Agilent 1200 SL 4. α-Lactalbumin 400 Å column in less 55 0.0065 Injection Volume: 5 μL 5. Catalase than 1 minute. Detection: 215 nm NOTE: 35 0.0069 0.0082 Temperature: 60 ˚C Peak widths at half height Flow Rate: 3 mL/min. are shown above respective Absorbance (mAU) 15 Mobile Phase A: water/0.1% peaks. TFA -5 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 Mobile Phase B: acetonitrile/0.1% TFA Non-core C18, 3 µm, 300 Å 0.0102 Gradient: 23–85% B in 1 min. 75 Pressure: 244 bar 0.0114 Using 3 and 6 µL heat 55 exchangers in column 35 0.0104 0.0107 0.0150 compartment 15 -5 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 Time (min.) Comparative results presented here may not be representative for all applications. HALO protein C4 provides narrower and taller peaks than totally porous column HALO Protein HALO Protein C4, 3.4 µm, 400 Å Pressure: C4 dramatically 55 103 bar outperforms a 45 1 2 TEST CONDITIONS: 2. Cytochrome c conventional non-core 35 3 Column: HALO C4 400 Å, 3.4 μm, 3. Lysozyme C4 column, deliv­ 5 100 x 2.1 mm 4. Holotransterrin ering narrower and 25 5 7 Part Number: HAL93412-614 5. Apomyoglobin taller peaks with 15 Instrument: Agilent 1200 SL 6. Catalase improved recovery 4 Injection Volume: 1 μL 7. Enolase of holotransferrin, Detection: 215 nm apomyoglobin, 5 Temperature: 60 ˚C catalase and enolase. Flow Rate: 0.5 mL/min. Absorbance (mAU) -5 6 7 8 9 10 Mobile Phase A: water/0.1% TFA 0 1 2 3 4 5 Mobile Phase B: acetonitrile/0.1% TFA Gradient: Time (min.) 25–52% B in 10 min. PEAK IDENTITIES Non-core C4, 5 µm, 300 Å 1. Ribonuclease A 55 Pressure: 55 bar 45 35 1 23 57 25 15 -5 10 0 1 2 3 4 5 6 7 8 9 Time (min.) Comparative results presented here may not be representative for all applications. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-25 LIFE SCIENCE

Colonnes HPLC/UHPLC Colonnes HPLC/UHPLC Protein Solutions 1.7 µm 2.7 µm • A s the first manufacturer of the 1000 Å fused-core particle, AMT recognizes the benefit of unrestricted pore access and offers both 400 Å and 1000 Å products to tailor the perfect large molecule solution. Benefits of HALO protein solutions include: 0.5 µm • Provides narrower peaks and better recoveries for large biomolecules Shell with 1000 Å pores (vs. smaller pore sizes and standard particles) 3.0 µm 3.4 µm • Allows HALO Protein columns to be used with both UHPLC and HPLC 0.2 µm instrumentation for fast bioseparations at moderate back pressures • C 4, Diphenyl and sterically- protected ES-C18 phases - Excellent high temperature stability (up to 90 ˚C) for improved peak shape and recovery • 2 μm inlet frit • Pressure limit, 600 bar/9000 psi Large protein separation using HALO Protein C4, 400 Å Shell with 400 Å pores High resolution separation of light and heavy chains of a denatured contractile protein (whole myosin from purified rabbit skeletal muscle) using HALO Protein C4 at 80 ˚C. TEST CONDITIONS: 9 Light chain subunits ~17- Expanded view Column: HALO C4 400 Å, 3.4 µm, 100 x 2.1 mm 8 25 kDa 8 Part Number: HAL93412-614 7 7 Instrument: Agilent 1200 SL Absorbance (mAU) 6 Heavy chain subunits 6 Injection Volume: 1 µL 5 ~220 kDa 5 Sample: denatured myosin 4 4 Detection: 215 nm 3 5 10 15 20 25 3 Flow Rate: 0.45 mL/min. 2 Time (min.) 2 Mobile Phase A: water/0.1% TFA 1 1 Mobile Phase B: acetonitrile/0.09% TFA 0 0 Gradient: 35–55% B in 30 min. 30 6.5 7.5 8.5 9.5 Temperature: 80 ˚C -1 0 High Resolution Separation of a Monoclonal Antibody using HALO C4 Protein, 1000Å TEST CONDITIONS: Flow Rate: 0.4 mL/min Column: HALO C4 1000 Å, Pressure: 210 bar 110 2.7 µm, 100 x 2.1 mm Temperature: 80 °C Part Number: HAL92712-614 90 Injection Volume: 2 μL A= Water, 0.1% TFA Absorbance, mAU Sample Solvent: 70/30 Water/ 70 B= 80/20 ACN/Water, ACN 0.085% TFA Detection: UV 280 nm, PDA 50 Gradient: Data Rate: 12.5 Hz 30 Time (min.) %B Response Time: 0.05 sec. 0.00 40 Flow Cell: 1 μL 10 12.00 47.5 LC System: Shimadzu Nexera X2 -10 0 2 4 6 8 10 12 Time, min D-26 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns HALO 1000 Å Diphenyl HPLC/UHPLC Columns Advantages of the HALO 1000 Å Diphenyl: • Extremely stable bonded phase using proven Fused-Core® technology • Alternate selectivity for large biomolecules • High resolution High Temperature Comparison of Comparison of HALO 1000 Å Diphenyl to HALO 1000 Å Phases Competitor PolyPhenyl at 40 °C Figure 1. Three bonded phases (C4, ES-C18, and Diphenyl) are Figure 2. HALO 1000 Å Diphenyl shows improved resolution, now available on HALO 1000 Å superficially porous particles. retention, and peak area compared to a competitor column Each phase offers unique selectivity for mAbs as demonstrated of 450 Å SPPs with PolyPhenyl phase. The increased retention by this high temperature separation of trastuzumab. clearly demonstrates the benefit of unrestricted large pore access to the bonded phase. The HALO Diphenyl phase can also be operated at 40 °C without showing peak area loss. 135 45 40 135 115 45 35 115 95 40 35 30 SPP 45 , mAU 25 95 , mAU 75 SPP 450 Å P2o0 lyPhenyl Absorbance, mAU Absorbance, mAU HALO 130000 Å Diphenyl 75 55 25 15 55 10 HALO 1000 Å Diphenyl HALO 120000 Å ES-C18 35 35 HALO151000 Å ES-C18 15 5 HALO 15 -5 HALO 1000 Å C4 10 HALO 1000 0Å Diphenyl 5 HALO 1000 Å C4 6 7 8 9 10 11 Time, min. 0 -5 -5 7.5 8.0 8.5 9.0 6 7 8 9 10 11 -5 Time, min. Time, min. 7.5 8.0 8.5 9.0 9.5 TEST CONDITIONS Time, min. TEST CONDITIONS Column: TEST CONDITIONS HALO 1000 Å C4, 2.7 µm, 2.1 x 150 mm HAL92712-714 Column: HALO 1000 Å ES-C18, 2.7 µm, 2.1 x 150 mm HAL92712-7T0E2 ST CONDITIONS HALO 1000 Å Diphenyl, 2.7 µm, 2.1 x 150 mm - HAL Column: HAHL9A2L7O121-701040 Å Diphenyl, 2.7 µm, 2.1 x 150 mm SPP 450 Å PolyPhenyl, 2.7 µm, 2.1 x 150 mm HALO 1000 Å C4, 2.7 µm, 2.1 x 150 mm HAL92712-726 Column: HALO 1000 Å ES-C18, 2.7 µm, 2.1 x 150 mm HAL92712-702 HALO 1000 Å Diphenyl, 2.7 µm, 2.1 x 150 mm - HAL92712-726 HALO 1000 Å Diphenyl, 2.7 µm, 2.1 x 150 mm HAFL9lo27w12R-7a2t6e: 0.4 mL/min Temperature: 80 °C MobSiPleP P45h0aÅsePoAly:Phweantyel,r/20.7.1µ%m,T2F.1Ax 150 mm Mobile Phase B: ACN/0.1% TFA Flow Rate: 0.4 mL/min Injection Volume: 2 µL of 2 mg/mL Temperature: 40 °C ItnrassMMttruuoozmbbuiiemlleenatPPb:hhiaaSnssheewimaBAat::edrAwz/u0Ca.tNN1e%/er/0x0.Te1.F1r%a%ATTFFAA MItnroajbestciluetizouPnmhaVasobeluiAnm:wewa:at2teerµr/L/00.o.11f%%2 TmTFFgAA/mL Flow Rate: 0.4 mL/min Temperature: 80 °C G3322r--a34dItFT80nriel%%aeojmesnwtcBBtput:Rieziionnuranamt11teV62ua:obrmme0lui:ii.nnn4m4w0m((eDCa:°L4itC2p/emahrµn/eiL0ndn.o1yEf%l)S2-TCmF1gA8/)mL Mobile Phase B: ACN/0.1% TFA Injection Volume: 2 µL of 2 mg/mL Gradient: trastuzumab in water/0.1% TFA Instrument: Shimadzu Nexera Gradient: 32-40% B in 16 min (C4 and ES-C18) 30-45% B in 15 min 32-38% B in 12 min (Diphenyl) 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-27 LIFE SCIENCE

Colonnes HPLC/UHPLC HALO® 1000 Å and 400 Å Column Ordering Information Colonnes HPLC/UHPLC Part numbers for nano, capillary, analytical and semi-preparative HALO 1000 Å and 400 Å phases in 2.7 and 3.4 µm particle sizes are provided below. Guard columns are available in 2.1, 3.0 and 4.6 mm IDs for UHPLC and HPLC applications to provide additional column protection when desired. Dimensions 400 Å, 3.4 μm 1000 Å, 2.7 μm ES-C18 ID x Length (mm) C4 ES-C18 C4 Diphenyl HAL97219-402 0.075 x 50 HAL94319-414 HAL94319-402 HAL97219-414 HAL97219-602 HAL97219-426 0.075 x 100 HAL97219-614 HAL97219-702 HAL97219-626 0.075 x 150 HAL94319-614 HAL94319-602 HAL97219-714 HAL97218-402 HAL97219-726 0.1 x 50 HAL97218-414 HAL97218-602 HAL97218-426 0.1 x 100 HAL94319-714 HAL94319-702 HAL97218-614 HAL97218-702 HAL97218-626 0.1 x 150 HAL97218-714 HAL97217-402 HAL97218-726 0.2 x 50 HAL94318-414 HAL94318-402 HAL97217-414 HAL97217-602 HAL97217-426 0.2 x 100 HAL97217-614 HAL97217-702 HAL97217-626 0.2 x 150 HAL94318-614 HAL94318-602 HAL97217-714 HAL97216-402 HAL97217-726 0.3 x 50 HAL97216-414 HAL97216-602 HAL97216-426 0.3 x 100 HAL94318-714 HAL94318-702 HAL97216-614 HAL97216-702 HAL97216-626 0.3 x 150 HAL97216-714 HAL97215-402 HAL97216-726 0.5 x 50 HAL94317-414 HAL94317-402 HAL97215-414 HAL97215-602 HAL97215-426 0.5 x 100 HAL97215-614 HAL97215-702 HAL97215-626 0.5 x 150 HAL94317-614 HAL94317-602 HAL97215-714 HAL92711-302 HAL97215-726 1.0 x 30 HAL92711-314 HAL92711-402 HAL92711-326 1.0 x 50 HAL94317-714 HAL94317-702 HAL92711-414 HAL92711-502 HAL92711-426 1.0 x 75 HAL92711-514 HAL92711-602 HAL92711-526 1.0 x 100 HAL94316-414 HAL94316-402 HAL92711-614 HAL92711-702 HAL92711-626 1.0 x 150 HAL92711-714 HAL92712-202 HAL92711-726 2.1 x 20 HAL94316-614 HAL94316-602 HAL92712-214 HAL92712-302 HAL92712-226 2.1 x 30 HAL92712-314 HAL92712-402 HAL92712-326 2.1 x 50 HAL94316-714 HAL94316-702 HAL92712-414 HAL92712-502 HAL92712-426 2.1 x 75 HAL92712-514 HAL92712-602 HAL92712-526 2.1 x 100 HAL94315-414 HAL94315-402 HAL92712-614 HAL92712-702 HAL92712-626 2.1 x 150 HAL92712-714 HAL92712-902 HAL92712-726 2.1 x 250 HAL94315-614 HAL94315-602 HAL92712-914 HAL92712-926 HAL94315-714 HAL94315-702 HAL93411-314 HAL93411-302 HAL93411-414 HAL93411-402 HAL93411-514 HAL93411-502 HAL93411-614 HAL93411-602 HAL93411-714 HAL93411-702 HAL93412-214 HAL93412-202 HAL93412-314 HAL93412-302 HAL93412-414 HAL93412-402 HAL93412-514 HAL93412-502 HAL93412-614 HAL93412-602 HAL93412-714 HAL93412-702 HAL93412-914 HAL93412-902 Did you know? We also sell a wide selection of solid phase extraction, syringe filters, flash cartridges and a lot more. For more information please ask your Sales Representative today. NEXT DAY DELIVERY! D-28 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns HALO® 1000 Å and 400 Å Column Ordering Information (continued) Dimensions 400 Å, 3.4 μm C4 1000 Å, 2.7 μm Diphenyl HPLC/UHPLC Columns C4 ES-C18 ES-C18 ID x Length (mm) HAL92713-214 HAL92713-226 HAL93413-214 HAL93413-202 HAL92713-314 HAL92713-202 HAL92713-326 3.0 x 20 HAL92713-414 HAL92713-302 HAl92713-426 3.0 x 30 HAL93413-314 HAL93413-302 HAL92713-514 HAL92713-402 HAL92713-526 3.0 x 50 HAL92713-614 HAL92713-502 HAL92713-626 3.0 x 75 HAL93413-414 HAL93413-402 HAL92713-714 HAL92713-602 HAL92713-726 3.0 x 100 HAL92713-914 HAL92713-702 HAL92713-926 3.0 x 150 HAL93413-514 HAL93413-502 HAL92714-214 HAL92713-902 HAL92714-226 3.0 x 250 HAL92714-314 HAL92714-202 HAL92714-326 4.6 x 20 HAL93413-614 HAL93413-602 HAL92714-414 HAL92714-302 HAL92714-426 4.6 x 30 HAL92714-514 HAL92714-402 HAL92714-526 4.6 x 50 HAL93413-714 HAL93413-702 HAL92714-614 HAL92714-502 HAL92714-626 4.6 x 75 HAL92714-714 HAL92714-602 HAL92714-726 4.6 x 100 HAL93413-914 HAL93413-902 HAL92714-914 HAL92714-702 HAL92714-926 4.6 x 150 HAL92710-414 HAL92714-902 HAL92710-426 4.6 x 250 HAL93414-214 HAL93414-202 HAL92710-514 HAL92710-402 HAL92710-526 10.0 x 50 HAL92710-614 HAL92710-502 HAL92710-626 10.0 x 75 HAL93414-314 HAL93414-302 HAL92710-714 HAL92710-602 HAL92710-726 10.0 x 100 HAL92710-702 10.0 x 150 HAL93414-414 HAL93414-402 HAL93414-514 HAL93414-502 HAL93414-614 HAL93414-602 HAL93414-714 HAL93414-702 HAL93414-914 HAL93414-902 HAL93410-414 HAL93410-402 HAL93410-514 HAL93410-502 HAL93410-614 HAL93410-602 HAL93410-714 HAL93410-702 HALO guard cartridge and guard holder For more details please go to page D-36 Dimensions 400 Å, 3.4 μm 1000 Å, 2.7 μm ES-C18 ID x Length (mm) C4 ES-C18 C4 Diphenyl HAL92712-102 2.1 x 5 Guard Cartridges, 3-pack HAL92713-102 HAL92712-126 3.0 x 5 HAL92714-102 HAL92713-126 4.6 x 5 HAL93412-114 HAL93412-102 HAL92712-114 HAL92714-126 HAL93413-114 HAL93413-102 HAL92713-114 HAL93414-114 HAL93414-102 HAL92714-114 Guard Column Holder - HAL94900-001 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-29 LIFE SCIENCE

Colonnes HPLC/UHPLC Colonnes HPLC/UHPLC Peptide Solutions 2 µm 1.2 µm HALO 160 Å columns currently available in three particle sizes (2, 2.7 and 5 µm) 0.4 µm and in two sterically protected phases: Shell with 160 Å pores ES-C18 (all particle sizes) and ES-CN (2.7 and 5 µm) 2.7 µm • Extremely stable at high temperatures and low pH 1.7 µm • Ideal for both ultrafast and ultrahigh resolution separations of peptides and 0.5 µm Shell with 160 Å pores polypeptides up to 20 kDa • Outperforms non-core 3 µm, 300 Å columns in terms of peak width, peak capacity 3.3 µm 4.6 µm and peak height • Offers comparable peak capacity to sub-2 µm non-core columns at 40–50% back 0.6 µm pressure (2.7 µm) Shell with 160 Å pores • ~ 20% higher peak capacity than sub-2 µm non-core columns at comparable back pressure (2 µm) • Columns (Peptide 2.7 and 5 µm) can be used in series to increase peak capacity for UHPLC and HPLC analyses of complex tryptic digest samples. • HALO Peptide ES-CN (2.7 and 5 µm) offers different selectivity and improved retention for polar peptides. • 2 µm inlet frit (2.7 and 5 µm); 1 µm inlet frit (2 µm) provides extra protection from plugging • Pressure limit, 600 bar/9000 psi (2.7 and 5 µm); Pressure limit, 1000 bar/14500 psi (2 µm) High Temperature and Low pH Stability with HALO 160 Å ES-C18 The sterically-protected C18 phase on the HALO 2 µm Peptide column enables high temperature stability with low pH mobile phases. The replicate injections were stopped at injection 480 (15,500 column volumes). The column is expected to have a lifetime of ~ 1000 injections, depending on the type of sample and conditions used. TEST CONDITIONS: Gradient: 6% B to 54% B in 10 min Flow Cell: 1 μL Column: Initial pressure: 395 bar PEAK IDENTITIES MW (g/mol) HALO ES-C18 160 Å, 2 μm Maximum pressure: 417 bar 1. Gly-Tyr 238 100 x 2.1 mm Temperature: 60 °C 2. Val-Tyr-Val 380 Part Number: HAL91122-602 Detection: UV 215 nm, PDA 3. Met-enkephalin 574 Mobile Phase: Injection Volume: 0.5 μL 4. Angiotensin II 1046 A = 0.1% Trifluoroacetic acid in water Sample Solvent: mobile phase A 5. Leu-enkephalin 556 B = 0 .1% Trifluoroacetic acid Response Time: 0.025 sec. 6. Ribonuclease A 13,700 Data Rate: 40 Hz 7. Bovine Insulin 5733 in 80/20 acetonitrile/water LC System: Shimadzu Nexera X2 Flow Rate: 0.5 mL/min. 7 1 2 3 45 G0 122 Injection 480 6 Injection 400 Injection 300 7 8 9 10 Injection 200 Injection 100 Injection 1 0123456 Time, min. D-30 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns Comparison of Fused-Core with non-core columns for peptide separations HPLC/UHPLC Columns HALO 160 Å ES-C18, 2.7 µm column produces significantly taller peaks and higher peak capacity than a non-core 5 µm column. 150 0.0136 0.0121 0.0123 0.0125 0.0128 TEST CONDITIONS: PEAK IDENTITIES: 125 0.0116 0.0121 0.0128 Columns: HALO ES-C18 160Å, 1. Gly-Tyr 100 0.0125 2.7 µm, 100 x 4.6 mm 2. Angiotensin 1/2 (1-7) amide Non-core C18 5 µm, 300 Å and Non-core C18 300 Å, 5 µm, 3. Val-Tyr-Val 75 Pmax = 174 bar 100 x 4.6 mm 4. Met-Enk 50 5. Angiotensin 1/2 (1-8) amide Absorbance (mAU) 25 Part Number: HAL92124-602 6. Angiotensin II 7. Leu-Enk 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 Mobile Phase A: 90% water/10% 8. Angiotensin (1-12) human -25 9. Angiotensin (1-12) mouse Time (min.) ACN/0.1% TFA 0 NOTE: 0.0102 0.0083 0.0081 0.0084 0.0085 Mobile Phase B: 30% water/70% Peak widths at half height are 150 0.0076 ACN/0.1% TFA shown above respective peaks. 125 0.0078 Gradient: 0-87.5% B in 2 min. 100 Flow Rate: 2.5 mL/min. 0.0082 Temperature: 60 ˚C 75 Injection Volume: 5 μL 50 HALO 160Å ES-C18, 0.0086 25 2.7 µm, 100 x 4.6 mm Pmax = 260 bar 0 -25 LC System: Agilent 1100 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 Time (min.) Comparative results presented here may not be representative for all applications. Coupled HALO Peptide columns for maximum peak capacity Three HALO ES-C18 150 mm columns (450 mm total length) were connected in series to achieve a peak capacity of 560 for this mixture of tryptic digests of α-1-glycoprotein and apotransferrin. Absorbance (mAU) 5.0 TEST CONDITIONS: 4.5 Columns: HALO ES-C18, 2.7 µm, 150 x 2.1 mm 4.0 10 20 30 40 50 60 70 80 90 100 110 120 110 Part Number: 3 of HAL92122-702 3.5 Time (min.) Mobile Phase A: water/0.1% formic acid/20 mM ammonium formate 3.0 Mobile Phase B: A with 80% acetonitrile 2.5 Gradient: 5–55% B in 150 min. 2.0 Flow Rate: 0.5 mL/min. 1.5 Temperature: 70 ˚C 1.0 Detection: 220 nm 0.5 Injection Volume: 50 μL [25 μg each] of α-1-glycoprotein tryptic digest and apotransferrin tryptic digest 0 0 Alternate selectivity using HALO 160 Å ES-CN, 5 µm HALO 160 Å ES-CN offers alternative selectivity to HALO 160 Å ES-C18 for this mixture of 10 peptides and polypeptides. 245 56 7 8 9 10 TEST CONDITIONS: PEAK IDENTITIES: HALO Peptide ES-CN 4 Column: HALO 160Å ES-CN, 5 µm and 1. Asp-Phe HALO 160Å ES-C18, 5 µm, 150 x 4.6 mm 2. Angiotensin (1-7) amide Absorbance (mAU) 195 Part Numbers: 3. Tyr-Tyr-Tyr 3 HALO ES-CN: HAL95124-704 4. Bradykinin HALO ES-C18: HAL95124-702 5. Leu-Enk 145 Instrument: Optimized Agilent 1100 6. Angiotensin II Injection Volume: 10 μL 7. Neurotensin 95 2 Detection: 215 nm 8. ß–endorphin 1 Temperature: 40 ˚C 9. Sauvagine Flow Rate: 1.0 mL/min 10. Mellitin 45 Mobile Phase A: water/0.1% TFA Mobile Phase B: ACN/0.1% TFA -5 5 10 31 20 25 30 Gradient: 5–50% B in 30 min. 0 10 Time (min.) 30 245 HALO 160 Å ES-C18 3 6 195 2 5 145 4 7 95 1 89 45 -5 5 10 31 20 25 0 Time (min.) 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-31 LIFE SCIENCE

Colonnes HPLC/UHPLC HALO 160 Å Peptide Ordering Information Colonnes HPLC/UHPLC The part numbers are provided below for the nano, capillary, analytical and semi-preparative HALO 160 Å 2, 2.7 and 5 μm phases. Guard columns are available for 2.1, 3.0 and 4.6 mm internal diameters for UHPLC and HPLC applications, if additional protection is desired. Dimensions 160 Å, 2 μm 160 Å, 2.7 μm 160 Å, 5 μm ES-C18 ES-CN ID x Length (mm) ES-C18 Phenyl-Hexyl ES-C18 ES-CN - HAL91229-404 .075 x 50 - HAL91229-402 HAL91229-604 HAL91219-406 HAL91529-402 HAL91529-404 .075 x 100 - HAL91229-602 HAL91229-704 HAL91219-606 .075 x 150 - HAL91229-702 HAL91228-404 HAL91219-706 HAL91529-602 HAL91529-604 0.1 x 50 - HAL91228-402 HAL91228-604 HAL91218-406 0.1 x 100 - HAL91228-602 HAL91228-704 HAL91218-606 HAL91529-702 HAL91529-704 0.1 x 150 - HAL91228-702 HAL91227-404 HAL91218-706 0.2 x 50 - HAL91227-402 HAL91227-604 HAL91217-406 HAL91528-402 HAL91528-404 0.2 x 100 - HAL91227-602 HAL91227-704 HAL91217-606 0.2 x 150 - HAL91227-702 HAL91226-404 HAL91217-706 HAL91528-602 HAL91528-604 0.3 x 50 - HAL91226-402 HAL91226-604 HAL91216-406 0.3 x 100 - HAL91226-602 HAL91226-704 HAL91216-606 HAL91528-702 HAL91528-704 0.3 x 150 - HAL91226-702 HAL91225-404 HAL91216-706 0.5 x 50 - HAL91225-402 HAL91225-604 HAL91215-406 HAL91527-402 HAL91527-404 0.5 x 100 - HAL91225-602 HAL91225-704 HAL91215-606 0.5 x 150 - HAL91225-702 HAL92121-304 HAL91215-706 HAL91527-602 HAL91527-604 1.0 x 30 - HAL92121-302 HAL92121-404 HAL92111-306 1.0 x 50 - HAL92121-402 HAL92121-504 HAL92111-406 HAL91527-702 HAL91527-704 1.0 x 75 - HAL92121-502 HAL92121-604 HAL92111-506 1.0 x 100 - HAL92121-602 HAL92121-704 HAL92111-606 HAL91526-402 HAL91526-404 1.0 x 150 HAL91122-202 HAL92121-702 HAL92122-204 HAL92111-706 2.1 x 20 HAL91122-302 HAL92122-202 HAL92122-304 HAL92112-206 HAL91526-602 HAL91526-604 2.1 x 30 HAL91122-402 HAL92122-302 HAL92122-404 HAL92112-306 2.1 x 50 HAL91122-502 HAL92122-402 HAL92122-504 HAL92112-406 HAL91526-702 HAL91526-704 2.1 x 75 HAL91122-602 HAL92122-502 HAL92122-604 HAL92112-506 2.1 x 100 HAL91122-702 HAL92122-602 HAL92122-704 HAL92112-606 HAL91525-402 HAL91525-404 2.1 x 150 HAL91122-902 HAL92122-702 HAL92122-904 HAL92112-706 2.1 x 250 HAL91123-202 HAL92122-902 HAL92123-204 HAL92112-906 HAL91525-602 HAL91525-604 3.0 x 20 HAL91123-302 HAL92123-202 HAL92123-304 HAL92113-206 3.0 x 30 HAL91123-402 HAL92123-302 HAL92123-404 HAL92113-306 HAL91525-702 HAL91525-704 3.0 x 50 HAL91123-502 HAL92123-402 HAL92123-504 HAL92113-406 3.0 x 75 HAL91123-602 HAL92123-502 HAL92123-604 HAL92113-506 HAL95121-302 HAL95121-304 3.0 x 100 HAL91123-702 HAL92123-602 HAL92123-704 HAL92113-606 3.0 x 150 HAL91123-902 HAL92123-702 HAL92123-904 HAL92113-706 HAL95121-402 HAL95121-404 3.0 x 250 HAL92123-902 HAL92113-906 HAL95121-502 HAL95121-504 HAL95121-602 HAL95121-604 HAL95121-702 HAL95121-704 HAL95122-202 HAL95122-204 HAL95122-302 HAL95122-304 HAL95122-402 HAL95122-404 HAL95122-502 HAL95122-504 HAL95122-602 HAL95122-604 HAL95122-702 HAL95122-704 HAL95122-902 HAL95122-904 HAL95123-202 HAL95123-204 HAL95123-302 HAL95123-304 HAL95123-402 HAL95123-404 HAL95123-502 HAL95123-504 HAL95123-602 HAL95123-604 HAL95123-702 HAL95123-704 HAL95123-902 HAL95123-904 D-32 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns HALO 160 Å Peptide Ordering Information (continued) Dimensions 160 Å, 2 μm 160 Å, 2.7 μm 160 Å, 5 μm HPLC/UHPLC Columns ES-C18 ES-CN ID x Length (mm) ES-C18 Phenyl-Hexyl ES-C18 ES-CN - HAL92124-204 4.6 x 20 - HAL92124-202 HAL92124-304 HAL92114-206 HAL95124-202 HAL95124-204 4.6 x 30 - HAL92124-302 HAL92124-404 HAL92114-306 4.6 x 50 - HAL92124-402 HAL92124-504 HAL92114-406 HAL95124-302 HAL95124-304 4.6 x 75 - HAL92124-502 HAL92124-604 HAL92114-506 4.6 x 100 - HAL92124-602 HAL92124-704 HAL92114-606 HAL95124-402 HAL95124-404 4.6 x 150 - HAL92124-702 HAL92124-904 HAL92114-706 4.6 x 250 - HAL92124-902 HAL92120-404 HAL92114-906 HAL95124-502 HAL95124-504 10.0 x 50 - HAL92120-402 HAL92120-504 HAL92110-406 10.0 x 75 - HAL92120-502 HAL92120-604 HAL92110-506 HAL95124-602 HAL95124-604 10.0 x 100 - HAL92120-602 HAL92120-704 HAL92110-606 10.0 x 150 - HAL92120-702 HAL92110-706 HAL95124-702 HAL95124-704 10.0 x 250 - - - HAL95124-902 HAL95124-904 HAL95120-402 HAL95120-404 HAL95120-502 HAL95120-504 HAL95120-602 HAL95120-604 HAL95120-702 HAL95120-704 HAL95120-902 HAL95120-904 HALO guard cartridge and guard holder For more details please go to page D-36 Dimensions 160 Å, 2 μm 160 Å, 2.7 μm 160 Å, 5 μm ES-C18 ID x Length (mm) ES-C18 ES-CN Phenyl-Hexyl ES-C18 ES-CN HAL91122-102 2.1 x 5 HAL91123-102 Guard Cartridges, 3-pack 3.0 x 5 4.6 x 5 - HAL92122-102 HAL92122-104 HAL92112-106 HAL95122-102 HAL95122-104 HAL95123-102 HAL95123-104 HAL92123-102 HAL92123-104 HAL92113-106 HAL95124-102 HAL95124-104 HAL92124-102 HAL92124-104 HAL92114-106 Guard Column Holder - HAL94900-001 Did you know? We sell Soft Grip One-Piece High Pressure Fingertight PEEK Fittings? For more information please ask your Sales Representative today. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-33 LIFE SCIENCE

Colonnes HPLC/UHPLC Colonnes HPLC/UHPLC Glycan Solutions HALO Glycan 2.7 µm 1.7 µm • Incorporates a highly polar ligand that contains 5 hydroxyl Shell with 90 Å pores 0.5 µm groups tethered to 2.7 µm Fused-Core silica particles via novel, proprietary linkage chemistry • Ideal for hydrophilic interaction liquid chromatography (HILIC) separations of oligosaccharides, and particularly, of released and labeled glycans from glycoproteins and proteoglycans • Mobile phases typically consist of acetonitrile and aqueous ammonium formate buffer (50 mM, pH 4.4) used to form a gradient of increasing water content during elution • Each lot of HALO Glycan material is tested for quality assurance by separation of a procainamide-reducing-end- labeled glycan ladder of oligosaccharides having 2–25 glucose units (GU). - Peaks for oligosaccharides composed of 5 and 10 GU must meet tight specifications for retention and peak width before lot is approved for glycan analysis • 2 µm inlet frit • Pressure limit, 600 bar/9000 psi Separation of N-Linked Glycans from Human IgG Released and procainamide-labeled glycans from human IgG were separated using a 2.1 x 150 mm HALO Glycan column and detected using UV and selected-ion-monitoring MS detection. 3.0 10..584130 17.595 21.817 21.516 TEST CONDITIONS: 4.871 25.586 Column: HALO Glycan 90 Å, 2.7 µm, 2.5 G0F G1F G2F 150 x 2.1 mm 11.432 13.924 18.357 22.2432.0254 23.529 33.788 Part Number: HAL92922-705 UV2.0 15.001 18.669 24.289 Mobile Phase A: 50 mM Ammonium 26.365 Formate, pH 4.4 1.5 0.247 10.961 12.518 14.490 19.527 27.342 30.008 Mobile Phase B: Acetonitrile 15.481 20.190 30.870 30.366 Gradient: 1.0 11.875 16.318 77.5 – 60% B in 60 min 31.273 35.834 39.537 Flow Rate: 0.5 mL/min. 0.5 0.418 31.937 34.740 40.024 Temperature: 60 °C Detection: UV 300 nm 0 32.616 41.617 Injection Volume: 4 μL 42.916 LC System: Shimadzu Nexera -0.5 MS: Shimadzu LCMS 2020 (single 0 5 10 15 20 25 30 35 40 24 50 quadrupole) ESI: +4.7 kV 300,000 2:1536.50(+) G1F G2F Scan range: 500-2000 m/z 250,000 2:1479.50(+) Scan rate: 2 pps 200,000 2:1682.60(+) MS 150,000 2:1454.50(+) 100,000 2:922.80(+) G0F G1FB G2F+NeuAc 2:1024.20(+) G0F-N G0 50,000 2:1003.80(+) G2 G2FB G2+NeuAc G2FB+NeuAc G2F+NeuAc2 0 2:1105.20(+) 10 15 20 G2FB+NeuAc2 0 2:1076.30(+) 2:1149.30(+) 25 30 35 40 24 50 2:1250.70(+) 2:1396.80(+) 2:1295.40(+) 2:1084.30(+) 2:1157.30(+) 2:1258.90(+) 2:930.80(+) 5 D-34 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns HALO 90 Å Glycan Ordering Information HPLC/UHPLC Columns HALO Glycan columns are available in 2.1 and 4.6 mm diameters in the following lengths as a 2.7 μm particle size. Guard columns are available for UHPLC and HPLC applications if additional protection is desired. Dimensions HALO Glycan Column ID x Length (mm) HAL92922-405 2.1 x 50 HAL92922-605 2.1 x 100 HAL92922-705 2.1 x 150 HAL92924-405 4.6 x 50 HAL92924-605 4.6 x 100 HAL92924-705 4.6 x 150 HALO guard cartridge and guard holder For more details please go to page D-36 Guard Cartridges, 3-pack 2.1 x 5 HAL92922-105 4.6 x 5 HAL92924-105 Guard Column Holder - HAL94900-001 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN D-35 LIFE SCIENCE

Colonnes HPLC/UHPLC HALO UHPLC and HPLC Guard Columns Colonnes HPLC/UHPLC • Collect strongly retained • Finger-tight, direct-connect units • Available for all HALO analytical compounds from the sample and that auto-adjust to any column geometries (2.1, 3.0 and 4.6 mm minimizes column fouling with a 10–32 inlet port ID) and phases • Ultra-low dispersion, easy to use, • Easily replace guard cartridge operate at pressures up to 1000 without removing guard holder bar from the flow path HALO Guard Columns: Protection + Performance HALO guard columns provide optimum protection for your HALO HPLC and UHPLC column without sacrificing column efficiency. Absorbance @ 254 nm 30 TEST CONDITIONS: Column: HALO 90 Å C18, 2.7 µm, 4.6 x 50 mm 26 with guard column Part Number: HAL92814-402 Mobile Phase: 60/40 ACN/water 22 Flow Rate: 1.8 mL/min. 18 Temperature: 30 ˚C 14 Detection: 254 nm 10 Injection Volume: 1 µL Pressure: 1 58 bar with guard column 6 2 146 bar without guard column -2 Instrument: Optimized Agilent 1100 bypassed semi-micro flow cell 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0.005” ID tubing Time (min.) 14 Hz data rate 34 30 no guard column 26 22 18 14 10 6 2 -2 0.0 0.2 0.4 0.6 0.8 1.0 1.2 Time (min.) See below for an exploded view of the HALO guard cartridge and guard holder. Finger-tight guard Finger-tight guard Titanium hybrid cartridge holder, Inlet cartridge holder, Outlet ferrule Provides easy replacement of Allows for replacement of guard cartridge without Adds durability for guard cartridge removing the guard holder multiple connections from the flow path The Optimize Technologies EXP® Direct HALO guard cartridge Auto-adjusting zero dead Connect Holder: U.S. Patent No. 8,201,854 All HALO phases available volume (ZDV) end fitting & 8,696,902 and Foreign Patents Pending. Ensures optimum, ZDV connection to the analytical column D-36 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE