CLS_ACE_Catalogue

HPLC/UHPLC Columns ACE® Ultra-Inert, Base Deactivated HPLC and ACE® Excel UHPLC columns give you the choices HPLC/UHPLC Columns you need to achieve successful separations. 17 Phases C18 | C18-AR | C18-PFP | C18-Amide | SuperC18 | CN-ES | AQ | C8 | C4 | CN | Phenyl | Silica | C18-HL | NH2 | HILIC-A | HILIC-B| HILIC-N • Leverage 5 different mechanisms of separation offered by these bonded phases to achieve optimum selectivity for all peak pairs in your sample. 3 Pore Sizes 90 Å | 100 Å | 300 Å • Match the stationary phase support to the molecular size of your analytes. For peptides and proteins, choose a 300 Å pore. For molecules smaller than 5,000 daltons, choose 100 Å pore. Plus, optimize your column’s hydrophobic strength and loading capacity for your particular separation needs. The 90 Å pore C18-HL stationary phase offers greater surface area for higher loading capacity. 6 Particle Sizes 1.7 μm | 2 μm | 3 μm | 5 μm | 10 μm | 15 μm • Choose the right particle size for UHPLC, HPLC or preparative HPLC. 11 Standard Column Lengths 300 mm |250 mm | 200 mm | 150 mm | 125 mm |100 mm | 75 mm | 50 mm | 35 mm | 30 mm | 20 mm • Find the right balance between analysis time, resolution and peak capacity. Enquire for lengths not listed. 12 Standard Column IDs 0.075 mm | 0.10 mm | 0.30 mm | 0.50 mm | 1.0 mm | 2.1 mm | 3.0 mm | 4.0 mm | 4.6 mm | 10.0 mm | 21.2 mm | 30.0 mm • ACE: from capillary to preparative applications. ACE® Column Configurations • ACE 100 Å HPLC Columns • ACE Excel UHPLC Columns • ACE 300 Å Bioanalytical Columns • Capillary & Nano Columns • Preparative HPLC Columns • Guard Columns 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-1 LIFE SCIENCE

Colonnes HPLC/UHPLC Leverage the power of stationary phase selectivity with ACE® bonded phases. Colonnes HPLC/UHPLC Rs = N α-1 k 4 α 1+k Rs The resolution equation identifies the parameters that contribute to resolution: efficiency (N), retention (k) and selectivity (α). There has been a great deal of emphasis in the past several years on column efficiency and the use of ultra-efficient columns, so called “UHPLC” columns, as a means of achieving separation goals. UHPLC has proved its value by contributing to improvements in laboratory productivity through faster separations and faster method development. However, selectivity is often overlooked and its importance overshadowed by the emphasis on column efficiency. This is unfortunate. Of the three parameters that affect resolution, selectivity is the most powerful. By leveraging both efficiency and selectivity, better and faster separations often can be achieved. The effect of N, α and k on resolution (Rs) α N k Increasing N, α or k increases Resolution (Rs). However, as can be seen from these plots, increasing either N or k suffers from quickly diminishing returns. Increasing selectivity (α), on the other hand, does not have this problem and, therefore, becomes the most powerful of these three variables to optimize when developing a separation. In reversed-phase chromatography, there are multiple mechanisms of interaction between the stationary phase and analytes that can be leveraged to achieve a separation. These mechanisms of interaction include: hydrophobic binding, π–π, hydrogen bonding, dipole-dipole, and shape selectivity. Different types of bonded phases will offer one or more of these mechanisms of interaction. ACE reversed bonded phases are listed below and the principal mechanisms of interaction that are possible with each, depending on the analyte and the mobile phase conditions. Comparison of separation mechanisms/interactions offered by different bonded phases ACE Hydrophobic π–π Hydrogen Dipole- Shape Bonded Binding Bonding Dipole Selectivity Phase C18 Strong None None None Weak C18-HL Very strong None None None Weak C18-AR Strong Strong Moderate Moderate Moderate C18-PFP Strong Strong Strong Strong Strong AQ Moderate None Moderate Weak None C8 Moderate None None None None C4 Weak None None None None Phenyl Moderate Strong Weak Moderate Weak CN Weak None Weak Strong None Amino Weak None Strong Weak None A-2 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns Leverage bonded phase selectivity to achieve better separations. HPLC/UHPLC Columns ACE bonded phases provide a wide variety of powerful retention mechanisms to choose from when developing separations. By leveraging both efficiency and selectivity, much better separations can be achieved. Below illustrates the selectivity difference between two ACE Here is another illustration of the power of bonded-phase bonded phases, the C18-AR and the Phenyl. Although both selectivity. One separation was done on a UHPLC column bonded phases offer the possibility of moderate-strong π–π packed with a C18 bonded phase and the other separation and dipole-dipole interactions, they differ significantly in was done on a UHPLC column packed with a C18-PFP bonded the strength of other possible mechanisms of interaction, phase. The additional mechanisms of separation offered by the particularly hydrophobic binding. The additional, or different, C18-PFP bonded phase generates a better overall separation. mechanisms of separation that are possible with the C18-AR yield a better overall separation for this complex mixture. Comparison of Selectivity Differences Between C18-AR ACE Excel delivers excellent resolution and peak shape: and Phenyl Phases Pharmaceuticals and Related Compounds by UHPLC Column Dimensions: 50 x 2.1 mm, 3 μm 1. Metronidazole Conditions Sample Mobile vP/hva)pBpseHH=:222A0..=77m2iiMnn0mMHK2MHeOO2;PKHOH/42,HPO2O4, 2. 3-Hydroxybenzoic acid Column Dimensions: 50 x 2.1mm 1. Paracetamol 3. Phenol Mobile Phase: A = 5 mM formic acid in H2O 2. Hydrochlorothiazide 4. Benzyl alcohol B = 5 mM formic acid in MeOH 3. Methylphenylsulphoxide 5. Caffeine Gradient: 3 to 100% B in 5 minutes 4. Methylphenylsulphone (65:35, 6. Salicylic acid Flow Rate: 0.6 mL/min Flow rate: 0.6 mL/min 7. Quinoxaline Temperature: 40 °C Temperature: 60 °C 8. Benzoic acid Detection: UV 254 nm Detection: UV 214 nm 9. Quinine Gradient: 3 to 100% B in 5 min. 10. Phenacetin and hold for 1 min. 11. 1,4-Dinitrobenzene 12. 1,3,5-Trinitrobenzene 13. Furosemide 14. 1,3,5-Trimethoxybenzene 15. Piroxicam 16. Carvedilol 17. Ethyl benzoate 18. Nortriptyline ACE 3 C18-AR 56 Waters ACQUITY 2 Pmax = 581 bar 1.7 BEH C18 1 3 15 4 4 14 18 2 17 13 78 910 11 12 13 16 0 1 2 Time (sec) ACE 3 Phenyl 65 ACE Excel 2 C18-PFP 1 2 Pmax = 365 bar 15,17 3 4 24 12,13,14 18 3 87 10 9 11 16 1 The greater hydrophobicity of the C18-AR phase provides more retention 0 1 2 and better selectivity for the peak pairs (13, 14) and (15, 17). Also note the Time (sec) numerous elution order changes on the C18-AR versus the Phenyl phase. The chromatograms generated on the C18 UHPLC column and the C18-PFP column are equally fast. However, the C18-PFP column provides better selectivity for peak pairs (1, 2) and (3, 4) and, therefore, is able to provide a superior overall separation. Note: Comparative separations may not be representative of all applications. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-3 LIFE SCIENCE

Colonnes HPLC/UHPLC Colonnes HPLC/UHPLC ACE® Excel brings renowned ACE® HPLC performance to UHPLC It is important to recognize that column selectivity and column efficiency are independent variables and you do not have to choose to use one rather than the other when developing a separation. In fact, for the best overall separation you should optimize both, as well as retention. This is especially true when there is a need for a high speed separation. ACE Excel UHPLC columns are engineered to take full advantage of low dispersion, ultra-high pressure UHPLC instruments (up to 1000 bar) and are designed to be compatible with all systems. ACE Excel UHPLC columns deliver better peak shape for basic compounds, improved column-to-column reproducibility, additional stationary phases that leverage multiple mechanisms of separations, and improved column ruggedness. The availability of ACE Excel UHPLC columns means chromatographers now have more choices to achieve better outcomes with their UHPLC instruments. Utilizing both column efficiency and bonded phase selectivity to develop a ACE Excel columns provide high-speed, high-resolution separations faster separation Sample: Conditions Conditions: Compound: 1. Aspirin Mobile Phase: A = 5mM Formic acid Columns: ACE Excel C18, 2 µm, 3.0 x 50 mm 1. Acetanilide 2. Phenacetin 5. Nimesulide (0.189)ml/L); Mobile Phase: 30% B to 100% B in 60 seconds 2. Acetophenone 3. 1,3-dinitrobenzene 6. Ibuprofen B = 5mM Formic acid in MeOH A = Water + 0.1% TFA B = Acetonitrile 3. Propiophenone 4. Ethyl benzoate 7. Indomethacin Column Temperature: 40°C Flow Rate: 2.5 mL/min 4. Butyrophenone Detection: PDA, 254nm Column Temperature: 40 ºC 5. Benzophenone Column: ACE C18, 5μm, 4.6 x 100 mm Pressure: 703 bar (10,335 psi) 6. Valerophenone Flow rate: 1.0 mL/min 7 Detection: PDA, 254 nm 7. Hexanophenone tG = 30.0 mins Instrument: Agilent 1290 UHPLC 8. Heptanophenone Injection Volume: 3µL 9. Octanophenone 5 2 3 5 2 1 46 1 25 minutes 3 4 67 89 Column: ACE Excel C18, 2μm, 2.1 x 50 mm 7 0 12 24 36 48 60 Flow rate: 0.6 mL/min 2 5 Time (sec) 46 tG = 5.0 mins Chromatographic data supplied courtesy of a west coast U.S. pharmaceutical company. Injection Volume: 1µL 3 An ACE Excel C18 column provides baseline separation of these 9 analytes in less than 60 seconds. 1 4.5 minutes Column: ACE Excel C18-PFP, 2 μm, 2.1 x 30 mm 7 Flow rate: 2.5 mL/min 2 tG = 0.7 mins Injection Volume: 1µL 3 5 1 46 45 seconds By utilizing the higher efficiency of a UHPLC column, a shorter length column run at higher mobile phase linear velocity can be used to reduce the separation time of this mixture by 80% compared to the HPLC column. However, by also optimizing bonded phase selectivity, in this case using the enhanced selectivity of a C18-PFP bonded phase, separation time can be reduced even further. The ACE Excel C18-PFP UHPLC column reduced separation time of this mixture by over 80% compared to the C18 UHPLC column and by 96% compared to the C18 HPLC column, while maintaining excellent resolution of all peaks. A-4 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns ACE® Excel™ UHPLC columns HPLC/UHPLC Columns ACE Excel UHPLC columns: • Fully compatible with all commercial UHPLC instruments • Brings renowned ACE HPLC columns’ advantages to UHPLC columns • Provides users of UHPLC with additional choices in stationary phases, including the powerful C18-AR and C18-PFP phases • Delivers a high level of reliability and ruggedness • Offers easy scalability from UHPLC to HPLC to preparative LC • Available in 1.7, 2, 3 and 5 µm particle sizes for UHPLC Manufacturing UHPLC columns is tough and there is a general perception that they are not as rugged and reliable as HPLC Columns. By drawing on their vast experience in manufacturing the finest HPLC columns, Advanced Chromatography Technologies is able to produce very reliable UHPLC columns. Now, chromatographers will get even more from their UHPLC instruments. Excellent peak shape for basic compounds, improved column-to-column reproducibility, additional stationary phases that leverage multiple mechanisms of separations, and improved column ruggedness are now available to UHPLC users with ACE Excel UHPLC columns. ACE Excel delivers excellent resolution and peak shape Conditions: Analytes: 6. Phenacetin 11. Ibuprofen Mobile Phase: A = 5 mM formic acid in H2O and B = 5 mM 1. Paracetamol 7. 1,3-Dinitrobenzene 12. Indomethacin formic acid in MeOH 2. Hydrochlorothiazide 8. 1,2,4-Trimethoxybenzene 13. Mefenamic acid Gradient: 3 to 100% B in 5 minutes 3. Methylphenylsulphoxide 9. Ethyl benzoate Flow Rate: 0.6 ml/min 4. Methylphenylsulphone 10. Nimesulide Temperature: 40 °C 5. Aspirin Detection: UV, 254 nm Column Dimensions: 50 x 2.1mm Waters ACQUITY 1.7 BEH C18 Phenomenex Kinetex 1.7 C18 1,2 21 7 12 Pmax = 581 bar Pmax = 540 bar 36 13 7 12 4 58 10 13 5 6 9 11 3 6 10 4 58 9 11 4 0 1 2 3 5 6 0 1 2 3 4 ACE Excel 2 C18-PFP ZORBAX Eclipse 1.8 XDB C18 7 12 21 3 7 12 Pmax = 365 bar 12 Pmax = 540 bar 4 10 13 10 13 6 8 9 11 36 5 45 9 11 8 0 1 2 3 4 5 6 0 1 2 3 4 5 6 Time (min) Time (min) The C18 bonded phases shown in these comparison chromatograms are not able to fully separate all 13 analytes. The ACE Excel C18-PFP, due to its extra separation mechanisms, is able to baseline separate all analytes. Note: Comparative separations may not be representative of all applications. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-5 LIFE SCIENCE

Colonnes HPLC/UHPLCColonnes HPLC/UHPLC ACE® Excel columns are very forgiving and provide the same durability and lifetime that you would expect from HPLC columns. ACE Excel will change your perception about the durability of UHPLC columns. Not only are the ACE Excel columns packed better, they incorporate a proprietary inlet frit design that is much less prone to plugging than other UHPLC columns. It is still recommended that you filter samples and mobile phases as you would with any UHPLC column, but ACE Excel columns are much more forgiving than other UHPLC columns and provide the same durability and lifetime that you would expect from HPLC columns. ACE® columns have a well-deserved reputation for reproducibility. Column-to-column reproducibility is affected by the production of the silica stationary phase support, the bonding of the stationary phase to the stationary phase support, and the packing of the stationary phase into the column. The better each stage of the column manufacturing process is controlled, the better will be the reproducibility and the quality of the column. ACE columns are subjected to extensive controls at each stage of the column manufacturing process. These controls ensure that ACE columns produce reliable, predictable performance from column-to-column and lot-to-lot. To the right provides an example of a typical Batch Validation test. To request a batch validation certificate please contact your sales representative. Subtle changes in silanol activity are one of the primary causes of column-to-column selectivity changes. ACE and ACE Excel columns have excellent reproducibility due to the use of ultra pure reagents and strict control of the manufacturing process that results in a high purity silica stationary phase support with uniform surface characteristics. Combining this high purity silica with advanced bonding techniques results in a family of highly inert phases that provide an outstanding level of column-to-column reproducibility by virtually eliminating silanol interference in the chromatography. ACE Excel columns have a well deserved reputation for durability Initial injection at 1000 bar 0 1 2 3 4 5 6 7 After 2000 runs 0 1 2 3 4 5 6 7 Time (min) A 2.1 x 100 mm ACE Excel C18 UHPLC column was subjected to over 2,000 gradient runs at an average pressure of 1,000 bar (14,500 psi). The column efficiency (plate number) and retention times were essentially unchanged at the end of this stability study. A-6 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns HPLC/UHPLC Columns ACE® Guard Columns (Not suitable for ACE Excel UHPLC columns) ACE® Integral Guard Columns • Guard column is incorporated into the analytical column’s inlet end-fitting • Ultra low dead volume design provides protection without degrading performance • Easy to change cartridge design ACE analytical columns are manufactured with state-of-the-art column fittings that incorporate the guard column as an integral part of the analytical column. To install a guard column on an ACE analytical column, simply replace the standard column inlet end- fitting with the end-fitting designed as a guard column holder. Then, insert a guard cartridge packed with the desired stationary phase into the guard column holder (See photograph). Don’t worry about disturbing the packing bed when installing the guard column holder. It is well protected by a PEEK frit cap, even with the column end-fitting removed. The integral guard column system is available for columns with internal diameters of 2.1, 3.0, and 4.6 mm. For preparative columns (> 10 mm), a typical stand-alone style of guard column is available, packed with the appropriate stationary phase. Stand-alone style guard column holders are also available for situations where they are preferred or required. A column coupler (p/n ACE-C0001), or connection tubing and fittings, will be required to use this style of guard column. Using a guard column to protect your analytical column can substantially increase the column lifetime and improve the quality of your chromatography. But, to be effective, the guard column must be replaced often enough to prevent contaminants from saturating the guard column and bleeding through to the analytical column. The best way to determine the optimum time to replace a guard column for a specific set of sample and mobile phase conditions is through experience. However, it is helpful to have some quantitative measure to help make the replacement decision. By monitoring plate number (N), pressure (P), and resolution (Rs), the performance of the guard and analytical column can be closely watched to determine when a guard column should be replaced. We suggest replacing the guard column when any one of these parameters changes by more than 10%. Column with no guard Integral guard column systems 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-7 LIFE SCIENCE

Mobile Phase: A: 0.1M CH 3 COONH 4 3. Simazine B: MeCN 4. Cyanazine Colonnes HPLC/UHPLC 5. Atrazine Flow Rate: 0.3 mL/min 6. Internal standard 7. Sebuthylazine Gradient: T(mins) %A %B 8. Propazine 9. Terbuthylazine 0 90 10 10. Prometryn 11. Terbutryn 40 20 80 12. Alachlor 13. Pendimethalin 47 10 90 49 90 10 Temperature: 40° C ACE® and ACE Excel™ C18 Detection: UV, 220nm (pendimethalin at 245nm) Injection Volume: 25µl Colonnes HPLC/UHPLC Sample: 0.05 µg/l standards in 10:90 MeCN/H2O Physical Properties 35 Pesticides in Water Silica : Ultra-Inert Base Deactivated Particle Size : 1.7 µm, 2 µm, 3 µm, 5 µm 30 Surface Area : 300 m²/g 25 6 20 Pore Size : 100 Å 15 Bonded Phase : Octadecyl groups 10 1 End-Capping : Yes 2 3 5 45 0 78 9 10 11 Carbon Load : 15.5% -5 USP Code : L1 12 13 pH Range : 2.0-8.0 Mechanisms of Separation Strength of Interaction 0 10 15 20 25 30 35 40 Hydrophobic binding Strong Time (min) interactions Weak Shape selectivity Reproduced with kind permission of Amt der Tiroler Landesregierung, Chemisch-technische Umweltschutzanstalt, Innsbruck, Austria. Conditions: Compounds: Column: ACE 3 C18, 150 x 2.1mm 1. Desisopropylatrazine Part Number: ACE-111-1502 2. Desethylatrazine Mobile Phase: A: CH3COONH4 in H2O 3. Simazine B: MeCN 4. Cyanazine Target Analytes Flow Rate: 0.3 mL/min 5. Atrazine • Analytes differing in hydrophobicity Gradient: T(mins) %A %B 6. Internal standard • Polar, moderately polar, and non-polar analytes 0 90 10 7. Sebuthylazine • Uncharged acids and bases 40 20 80 8. Propazine • Ionized acids or bases using ion-pairing 47 10 90 9. Terbuthylazine 49 90 10 10. Prometryn Recommended Applications Temperature: 40° C 11. Terbutryn • Analytes differing in hydrophobicity Detection: UV, 220nm (pendimethalin at 245nm) 12. Alachlor • Homologous compounds differing by –CH2 Injection Volume: 25μl 13. Pendimethalin Sample: 0.05 μg/l standards in 10:90 MeCN/H2O Antihistamines and Expectorants Conditions: Compounds: Column: ACE 5 C18, 150 x 4.6mm 1. Maleic acid Part Number: ACE-121-1546 2. Salicylamide Mobile Phase: 45:55 MeOH/20mM KH2PO4 (pH 3.0) 3. Guaifenesin Flow Rate: 1.0 mL/min 4. Guaiacol Temperature: Ambient 5. Chlorpheniramine maleate Detection: UV, 220nm 6. Dextromethorphan Injection Volume: 0.4µl 240 220 200 180 2 Response–MilliVolts 160 140 1 3 120 4 100 80 60 56 40 20 0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Time (min) A-8 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE & ACE Excel C18) Dimensions ACE® & ACE® Excel™ 100Å C18 Analytical HPLC Columns HPLC/UHPLC Columns ID x Length (mm) 5 µm 3 µm 2 µm 1.7 µm 4.6 x 250 ACE-121-2546 -- - 4.6 x 150 ACE-121-1546 - 4.6 x 100 ACE-121-1046 ACE-111-1546 EXL-101-1546U - 4.6 x 75 ACE-121-7546 - 4.6 x 50 ACE-121-0546 ACE-111-1046 EXL-101-1046U - 4.6 x 30 ACE-121-0346 - 4.6 x 20 ACE-121-0246 ACE-111-7546 EXL-101-7546U - 3.0 x 250 ACE-121-2503 - 3.0 x 150 ACE-121-1503 ACE-111-0546 EXL-101-0546U - 3.0 x 100 ACE-121-1003 EXL-171-1003U 3.0 x 75 ACE-121-7503 ACE-111-0346 EXL-101-0346U EXL-171-7503U 3.0 x 50 ACE-121-0503 EXL-171-0503U 3.0 x 30 ACE-121-0303 ACE-111-0246 EXL-101-0246U EXL-171-0303U 3.0 x 20 ACE-121-0203 EXL-171-0203U 2.1 x 250 ACE-121-2502 -- - 2.1 x 150 ACE-121-1502 - 2.1 x 100 ACE-121-1002 ACE-111-1503 EXL-101-1503U EXL-171-1002U 2.1 x 75 ACE-121-7502 EXL-171-7502U 2.1 x 50 ACE-121-0502 ACE-111-1003 EXL-101-1003U EXL-171-0502U 2.1 x 30 ACE-121-0302 EXL-171-0302U 2.1 x 20 ACE-121-0202 ACE-111-7503 EXL-101-7503U EXL-171-0202U 1.0 x 250 ACE-121-2501 - 1.0 x 150 ACE-121-1501 ACE-111-0503 EXL-101-0503U - 1.0 x 100 ACE-121-1001 - 1.0 x 75 ACE-121-7501 ACE-111-0303 EXL-101-0303U - 1.0 x 50 ACE-121-0501 - ACE-111-0203 EXL-101-0203U -- ACE-111-1502 EXL-101-1502U ACE-111-1002 EXL-101-1002U ACE-111-7502 EXL-101-7502U ACE-111-0502 EXL-101-0502U ACE-111-0302 EXL-101-0302U ACE-111-0202 EXL-101-0202U -- ACE-111-1501 - ACE-111-1001 - ACE-111-7501 - ACE-111-0501 - Semi-Prep and 10 µm also available, please visit lifescience.ca ACE® Analytical HPLC Guard Cartridge Ordering Information Dimensions ACE® 100Å C18 ID x Length (mm) 5 µm 3 µm Cartridges for 10 x 2.1mm ID Columns (pkg of 5) ACE-121-0102GD ACE-111-0102GD Cartridges for 10 x 3.0 - 4.6mm ID Columns (pkg of 5) ACE-121-0103GD ACE-111-0103GD Integral guard cartridge holder for 2.1mm ID Columns ACE-H0004 Integral guard cartridge holder for 3.0 - 4.6mm ID Columns ACE-H0005 Stand-alone universal guard cartridge holder. Requires Column coupler item # ACE-C0001 ACE-H0001 Other dimensions are available. Please enquire for details. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-9 LIFE SCIENCE

Colonnes HPLC/UHPLC ACE® & ACE® Excel™ C18-AR ACE UHPLC and HPLC Columns Colonnes HPLC/UHPLC Physical Properties Leveraging ACE Excel C18-AR Selectivity Silica : Ultra-Inert Base Deactivated Conditions: Analytes: Particle SizAe nalge: s1i.c7 Sµmep, a2rµamti,o3nµm, 5 µm Part Number: EXL-109-0502U 1. Pyridoxine Surface Area : 300 m²/g 1A. An4pa-pAlylcicteaetMTFGBtaeslirmo=oomawnbdi2dpii0#olReeeApnarmaPhttNetM:ehu2:2nar400osKe9.elH:3t0:o412AP07mO=0°4lC%/2, mp0BHimnin2M.57K/mMHi2nePuOOtHe4,s(p5H0:25.07va/nvd) 2. p-Aminobenzoic acid PBoornedSeidzeP hCaosen diti::o1On0cs0taÅdecyl with phenyl groups 2. 4-AminDoebteenctziooinc:aUciVd, 205 nm 3. Pantothenic acid CEnardb-CoanpLpoianPDCdgaoimr ltueNmnusnmi:o::bn1Yese:5r:s.5A1A%5CC0EE-x3114C9.61-18m5-4Am6R 3. 4-HydrCooxlyubmennzDoiicmaecnidsions: 50 x 2.1mm 4. Folic acid USP Code Mobile Ph: aLs1e: A: 0.1% formic acid in H2O 5. D-Biotin pH Range : 2.0-8B.:00.1% formic acid in MeCN 4. Caffeine 6. Cyanocobalamin Gradient: Time (mins) %B 5. 2-Acetamidophenol 7. Riboflavin 6. 3-Hydroxybenzoic acid 1 05 7. Salicylamide 4 6 9 35 8. Acetanilide 3 2 ACE Excel 2 C18-AR 5 Pmax = 209 bar Mechanisms of Separa14tion Stre3n5gth of Interact9i.onPhenol π-π interaFTceltomiwopneRsraatteu:re: 1 mL/min 10. Acetylsalicylic acid 40 °C Strong 11. Benzoic acid For additional7 12. Sorbic acid Dipole-dipDoelteecitniotne:ractiUoVn, s240 nmModerate 13. Salicylic acid 0 column2 3 dim4 ensio5ns 111 14. Phenacetin Minutes Hydrophobic binding 15. Salicylaldehyde 1 6 interactions Strong 1144 Please enquire 113 email: [email protected] Shape selectivity Modera88te 77 11 33 1100 1122 Recommended Applications 66 • Analytes differing in hydrophobicity, homologous Target Analytes • Analytes with π bonding55 compounds differing by –CH2 • Stereoisomers • Awnitahldytreaswwinigthgeroleucptsro, snu4d4cehloascahliazlaotgioennsa,nnditerolegcrtorounp-s, 1155 • Steroids ketones, este22rs, and acids 99 • Analytes with different dipole moments • Taxanes • Analytes differing in hydrophobicity 12 min • Substituted aromatics • Polar, m4 odera5 tely p6 olar, a7 nd no8T nim-ep-oMl9ian urteasna10ly tes 11 • Highly aqueous conditions • Uncharged acids and bases • Particularly recommended for applications where a typical C18 does not provide an adequate separation Analgesics / Cough & Cold Medicine Ingredients Analgesics / Cough & Cold Medicine Ingredients Application #AN1930 Conditions Analytes mAU 11 PCADCMBGaoo::irmoar0nl0utbd..de1m1Niiilne%%teunisnomiP:tffono:ohAAnbsrraCCs:emms:EEreT0is1iMCDPGc33c::i5amo iraAAaoCCm0ralcctCCebu11dxiieEEdN88imdi(lne4--me-ui11iAsnn.nn6mP11ii:RtonM:hH19msbn--a2)ees11Oms:rC55es44N::%566AA1AAA90TBB5iCCCC::m000EEEE..e--x11331111%%(4mCC19.6-11-1ffi188oon5m5-rrs44Ammm)66Riicc HM2eOCN869141A12537...1..0....nA3CP2S44P..aaa----ABhacl32569711841HHAAyl.ref.e1e0s.......itfacmc.yyptnne.eceayddiioz2P4PCA3ASB4sierlnntrrno:i----ialhaacesatcooAHAHneioiarllepefmncixxiafcamcyybmdtinezyycreaddcyaioeeioibbdienintolncrrninidlaooeeoctcieilodalzmxxnnipbaadomyyzzhccieeibbdoocoieinddeeoiilnzaccnnpooczzaahiiloocdcceiiiinccaddocaaildcciidd 22 77 88 1111 9 3514 1133 1144 11,,22 33 acid in 55 66 1100 1122 1155 acid in 99 %B 44 5 ACE C18-AR 35 33,,44 35 55 66 1133,,1144 1111,,1122 14 3515 5 12. S1o2r.bSicorabcicidacid 1F5low Rate: 51 mL/min 13. S1a3l.icSyalilcicyalcicidacid Flow Rate: 1 mTeLm/mpeinrature: 40 °C 14. P1h4e. nPahceentaicnetin 88 Temperature:D4e0te°cCtion: UV, 240 nm 15. S1a5l.icSyalalicldyelahldyedheyde 77 Detection: UV, 240 nm 1100 99 1155 ACE C18 4 5 6 7 8 9 10 11 12 min Time - Minutes A-10 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE & ACE Excel C18-AR) Dimensions ACE® & ACE® Excel™ 100Å C18-AR Analytical HPLC Columns* HPLC/UHPLC Columns ID x Length (mm) 5 µm 3 µm 2 µm 1.7 µm 4.6 x 250 ACE-129-2546 ACE-119-2546 - - 4.6 x 150 ACE-129-1546 - 4.6 x 100 ACE-129-1046 ACE-119-1546 EXL-109-1546U - 4.6 x 75 ACE-129-7546 - 4.6 x 50 ACE-129-0546 ACE-119-1046 EXL-109-1046U - 4.6 x 30 ACE-129-0346 - 4.6 x 20 ACE-129-0246 ACE-119-7546 EXL-109-7546U - 3.0 x 250 ACE-129-2503 - 3.0 x 150 ACE-129-1503 ACE-119-0546 EXL-109-0546U - 3.0 x 100 ACE-129-1003 EXL-179-1003U 3.0 x 75 ACE-129-7503 ACE-119-0346 EXL-109-0346U EXL-179-7503U 3.0 x 50 ACE-129-0503 EXL-179-0503U 3.0 x 30 ACE-129-0303 ACE-119-0246 EXL-109-0246U EXL-179-0303U 3.0 x 20 ACE-129-0203 EXL-179-0203U 2.1 x 250 ACE-129-2502 ACE-119-2503 - - 2.1 x 150 ACE-129-1502 - 2.1 x 100 ACE-129-1002 ACE-119-1503 EXL-109-1503U EXL-179-1002U 2.1 x 75 ACE-129-7502 EXL-179-7502U 2.1 x 50 ACE-129-0502 ACE-119-1003 EXL-109-1003U EXL-179-0502U 2.1 x 30 ACE-129-0302 EXL-179-0302U 2.1 x 20 ACE-129-0202 ACE-119-7503 EXL-109-7503U EXL-179-0202U 1.0 x 250 ACE-129-2501 - 1.0 x 150 ACE-129-1501 ACE-119-0503 EXL-109-0503U - 1.0 x 100 ACE-129-1001 - 1.0 x 75 ACE-129-7501 ACE-119-0303 EXL-109-0303U - 1.0 x 50 ACE-129-0501 - ACE-119-0203 EXL-109-0203U ACE-119-2502 - ACE-119-1502 EXL-109-1502U ACE-119-1002 EXL-109-1002U ACE-119-7502 EXL-109-7502U ACE-119-0502 EXL-109-0502U ACE-119-0302 EXL-109-0302U ACE-119-0202 EXL-109-0202U ACE-119-2501 - ACE-119-1501 - ACE-119-1001 - ACE-119-7501 - ACE-119-0501 - Semi-Prep and 10 µm also available, please visit lifescience.ca ACE® Analytical HPLC Guard Cartridge Ordering Information Dimensions ACE® 100Å C18-AR ID x Length (mm) 5 µm 3 µm Cartridges for 10 x 2.1mm ID Columns (pkg of 5) ACE-129-0102GD ACE-119-0102GD Cartridges for 10 x 3.0 - 4.6mm ID Columns (pkg of 5) ACE-129-0103GD ACE-119-0103GD Integral guard cartridge holder for 2.1mm ID Columns ACE-H0004 Integral guard cartridge holder for 3.0 - 4.6mm ID Columns ACE-H0005 Stand-alone universal guard cartridge holder. Requires Column coupler item # ACE-C0001 ACE-H0001 Other dimensions are available. Please enquire for details. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-11 LIFE SCIENCE

Colonnes HPLC/UHPLC ACE UHPLC and HPLC Columns Catecholamines and their Metabolites in Urine by LC-MS/MS ACE® & ACE® Excel™ C18-PFP Conditions Analytes Colonnes HPLC/UHPLC Column: ACE UltraCore 2.5 1. Norepinephrine Physical Properties LeveragDinimgetnhseionusn: iqS1u0ue0peAxrPC2h.E1enmEyxmlHceexlyCl 18-PFP selectivity2. (m/z 170 107) Epinephrine Silica : Ultra-Inert Base Deactivated Particle Size : 1.7 µm, 2 µm, 3 µm, 5 µm CCoonludmitnio:nAPMsCa:oErtbENilxecuemPlh2baeCsre1::8-CAPO:F2PRmE-M25Bam-10m0o2nUium formA1.na1tae,l2y+,t3e0-sT.:0ri5m%ethox3y. bNeonrmz(meen/ztea1n8e4phri1n6e6) Surface Area : 300 m²/g Dimensions: 50 x 2.1mm formic acid in H2O 2. 1,2-Dimethoxybenzen(me/z 184 166) Part Number: EXL-1010-0B5:022UmM ammonium form3.a1te,2+,40-T.0ri5m%ethox4y. bDeonpzaemniene Pore Size : 100 Å Mobile Phase: MeOH/H2Ofor(m50ic:5a0civd/ivn) MeOH 4. Methoxybenzene (m/z 154 91) Bonded Phase : O ctadecyl with pentaflurophenyl groups Flow RateG: r0a.2d1iemntl:/min Time (mins) %B 5. 1,3-Dimethoxyb5e. nMzeetnaenephrine Temperature: 40 °C 0.00 0 6. 1,3,5-Trimethoxybenz(men/ze198 180) End-Capping : Yes 1.00 70 7. Toluene 6. 3-Methoxytyramine Carbon Load : 14.3% USP Code : L1 1.10 70 (m/z 181 91) pH Range : 2.0-8.0 1.11 0 ACE Excel 2FlCo1w8-PRFaPte(f:ully 4.50 0 37.5e5 porous ultIrnajiencetritosnili:ca) 35.0e5 0.3 mL/min 32.5e5 Temperature: 30.0e5 Detection: 10 µL 6 27.5e5 Sample: 310 °C 7 25.0e5 Shima3dzu LCMS-8040 22.5e5 20.0e5 Mechanisms of Separation Strength of Interaction ESI in positive ion mode π-π interactions Strong Dipole-dipole interactions Strong Standard 100 ng/mL in urine 5 Hydrophobic binding interactions Strong (aft2er SPE purification4) Shape selectivity Strong 17.5e5 15.0e5 12.5e5 0 1 2 3 4 5 6 710.0e5 Minutes 7.5e5 5.0e5 2.5e5 Target Analytes Recommended Applications 0.0 • Analytes with π bonding • Analytes with electron donating groups, such as • RepSrotedurceeodiswoitmh peerrsmission of Shimadzu, France phenols, aromatic ethers and amines • Steroids • Analytes with proton donor groups • CaTtaexcahnoeslamines and Metanephrines Separation (Gradient) • Analytes with different dipole moments • Analytes differing in hydrophobicity • Substituted aromatics • Polar, moderately polar, and non-polar analytes • Uncharged acids and bases • CoAnndaitlyiotenssdiffering in hydrophobicity, homAonlaolgyotues Colcuommn:poundAsCEdi5ffCe1r8in-PgFPby –CH2 1. Norepinephrine • PDaimrHteNingushmiolybnesa:r:quA1e5Co0Eu-x1s241.m60-mo15mb4i6le phase conditions32.. Epinephrine Normetanephrine • MoPbialertPichuaslea:rlAy:r2e0cmoMmmamemnodneiudmffoorrmaapteplicatio4n.sDwophamerinee a typical C18 does npHot3.p0rionvHi2dOe an adequate separ5a.tMioentanephrine No B: 20 mM ammonium formate Nor Catecholamines and Metanephrines Separation (Gradient) Gradient: pH 3.0 in MeOH/H2O (90:10 v/v) Time (mins) %B 00 5 40 44 Conditions: Analytes: 6 40 33 Column: ACE 5 C18-PFP 1. Norepinephrine 70 2. Epinephrine Dimensions: 150 x 4.6 mm 3. Normetanephrine 17 0 5 4. Dopamine 1 mL/min Part Number: ACE-1210-1546 5. Metanephrine Flow Rate: Mobile Phase:  Injection: 5 µL A: 20 mM ammonium formate Temperature: 25 °C pH 3.0 in H2O Detection: UV, 260 nm B: 20 mM ammonium formate 22 pH 3.0 in MeOH/H2O (90:10 v/v) Gradient: Time (mins) %B 11 00 5 40 6 40 M 70 17 0 Flow Rate: 1 mL/min Injection: 5 μL Temperature: 25 °C Detection: UV, 260 nm 1 2 3 4 5 6 min Time - Minutes A-12 CANADIAN 1 888-2265-42775 w:w: [email protected]|.c+a44::(0)w1w22w4.l7if0e4sc5ie5n4ce|[email protected] LIFE SCIENCE

HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE® & ACE® Excel™ C18-PFP) Dimensions ACE® & ACE® Excel™ 100Å C18-PFP Analytical HPLC Columns HPLC/UHPLC Columns ID x Length (mm) 5 µm 3 µm 2 µm 1.7 µm 4.6 x 250 ACE-1210-2546 -- - 4.6 x 150 ACE-1210-1546 - 4.6 x 100 ACE-1210-1046 ACE-1110-1546 EXL-1010-1546U - 4.6 x 75 ACE-1210-7546 - 4.6 x 50 ACE-1210-0546 ACE-1110-1046 EXL-1010-1046U - 4.6 x 30 ACE-1210-0346 - 4.6 x 20 ACE-1210-0246 ACE-1110-7546 EXL-1010-7546U - 3.0 x 250 ACE-1210-2503 - 3.0 x 150 ACE-1210-1503 ACE-1110-0546 EXL-1010-0546U - 3.0 x 100 ACE-1210-1003 EXL-1710-1003U 3.0 x 75 ACE-1210-7503 ACE-1110-0346 EXL-1010-0346U EXL-1710-7503U 3.0 x 50 ACE-1210-0503 EXL-1710-0503U 3.0 x 30 ACE-1210-0303 ACE-1110-0246 EXL-1010-0246U EXL-1710-0303U 3.0 x 20 ACE-1210-0203 EXL-1710-0203U 2.1 x 250 ACE-1210-2502 -- - 2.1 x 150 ACE-1210-1502 - 2.1 x 100 ACE-1210-1002 ACE-1110-1503 EXL-1010-1503U EXL-1710-1002U 2.1 x 75 ACE-1210-7502 EXL-1710-7502U 2.1 x 50 ACE-1210-0502 ACE-1110-1003 EXL-1010-1003U EXL-1710-0502U 2.1 x 30 ACE-1210-0302 EXL-1710-0302U 2.1 x 20 ACE-1210-0202 ACE-1110-7503 EXL-1010-7503U EXL-1710-0202U 1.0 x 250 ACE-1210-2501 - 1.0 x 150 ACE-1210-1501 ACE-1110-0503 EXL-1010-0503U - 1.0 x 100 ACE-1210-1001 - 1.0 x 75 ACE-1210-7501 ACE-1110-0303 EXL-1010-0303U - 1.0 x 50 ACE-1210-0501 - ACE-1110-0203 EXL-1010-0203U -- ACE-1110-1502 EXL-1010-1502U ACE-1110-1002 EXL-1010-1002U ACE-1110-7502 EXL-1010-7502U ACE-1110-0502 EXL-1010-0502U ACE-1110-0302 EXL-1010-0302U ACE-1110-0202 EXL-1010-0202U -- ACE-1110-1501 - ACE-1110-1001 - ACE-1110-7501 - ACE-1110-0501 - Semi-Prep and 10 µm also available, please visit lifescience.ca ACE® Analytical HPLC Guard Cartridge Ordering Information Dimensions ACE® 100Å C18-PFP ID x Length (mm) 5 µm 3 µm Cartridges for 10 x 2.1mm ID Columns (pkg of 5) ACE-1210-0102GD ACE-1110-0102GD Cartridges for 10 x 3.0 - 4.6mm ID Columns (pkg of 5) ACE-1210-0103GD ACE-1110-0103GD Integral guard cartridge holder for 2.1mm ID Columns ACE-H0004 Integral guard cartridge holder for 3.0 - 4.6mm ID Columns ACE-H0005 Stand-alone universal guard cartridge holder. Requires Column coupler item # ACE-C0001 ACE-H0001 Other dimensions are available. Please enquire for details. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-13 LIFE SCIENCE

Reproduced with permission of Nufarm UK Ltd Phenolic Compounds in Ground Water & Landfill Leachates ColonnePsheHAnCoPlEiLcCUAc/HiUdPsLHCPaLnCd HPLC Columns Application #AN3070 Application #AN3030 Conditions Analytes Conditions Analytes ACE® ECxolcuemOnl™:rgaCn1iAc8CAE-Ac5iCdm1s8ide 1. Caffeic acid Column: ACE Excel 3 C18-Amide 1. Pyrocatechol Dimensions: 150 x 4.6 mm 2. p-CouAmpaprilcicaactiiodn #AN2780 Dimensions: 150 x 4.6 mm 2. Resorcinol Colonnes HPLC/UHPLC Physical PPMraoortbpNileeuCrmPtohibneaesdsre:i:tioAMCneEsC-N12/01-.115%46formic acid in 3. Ferulic acid PMPhaoretbnNileoulmPihcbaeCsreo::mE0.pX1Lo%-u11fno1r2dm-1si5ci4na6cUGidrvo/vuinnHd2OWate34..r &Pmh-LeCnareonsl dolfill Leachates Response - MilliVolts Analytes PSSPBuiaoolirrrnctfeadiac Sceleeidz eASP DTIFirnhzeleeojemaetaw ecsp cteeRito CMDPFiraoanlaoitoonmtr:elwutbu::e:::::rNimlenRe1U1O3u:nsamP0.0i:ltco7the00HUA11tbnr:aaµesam2VµmsÅmO:dr,-emL:bL2I:e²(in/5e/2,mcA25A1H4eng02y05CC2ti:mrnnO80lEEmtµm0L/w-xM51BM/mvm24iCe/a1t.viK6,On1-hs)2H83eHm52a4Pmµ(Dm67Om0e4:i3adp,0Hce5tv5i/gµvv.7r)amointuepd51234s..... CDPCMDTIFnaooeilemoojrlnmeuttbwdeecmNipicltnteeRiutnisooiramiP:oanonthAnt:nbesuaC:s::ers:Eree1::E:E5x0X0c.13M1UL1ex00V%lm-e,41µ3°CL2.C1fL6CNo/71m41rm2(m86in-nm-15imAc5:3m4a5c6idivUde/vv)/v in H2O 638A57412........n9615171183mPoRP32.1a023.....,,,hye-l...445-yCrseC2233223o1---toronDDD-e-r,,-,,,,ecNN4C44543reosiiisacmmm-,,,--saa:rolDD656Diteopnpel---eeeiisiTTTlhhommmctttorrrtthlhhhiiihhlmmmeeeoyyyootttlllleeehhhllpppyyyttthhhhhhlllpppeeeyyyhhhlllnnnpppeeeooohhhnnnllleeeooonnnlll ooolll L -Ascorbic acid MeCN (65:35 v/v) Maleic acid Acetylsalicylic acid Flow Rate: 1 mL/min Benzoic acid Injection: 10 μL Salicylic acid Temperature: 30 °C End-Capping Temp:eYreatsure: 22 °C Carbon Load Dete:ct1io6n.:4% UV, 220 nm USP Code : L1 & L60 Detection: UV, 274 nm 9. 1-Naphthol pH Range : 2.0-8.0 For further applications10. 3,4,5-Trimethylphenol 1 11. 2,3,6-Trimethylphenol Mechanisms of SLe-pACsaa2cfrfoearibctiicao3canicdid p-CMouamleaicriaccaidcid FAecreutlyiclsaacliicdylic acid mmAAUU 11 visit: www.ace-h1123p.. 22lc,-4N.,a6cp-oThrtimhmoelthylphenol St1rength of Interaction 22 or 14 π-π interactions Strong 12 email: [email protected] Dipole-dipole interBaecntziooicnascid StrongSalicylic acid22 3 4 10 Hydrophobic binding Strong interactions 3 2 8 6 44 5 1100 4 99 11111122 Shape selectivity Strong1 35 2 66 77 88 133 0 Target Analytes Recommended Applications -2 • Organic acids •T ime -SMminautlelswater soluble 5 10 15 20 25 min Time - Minutes • Phenolic compounds analytes Reproduced with permission of Environmental Scientific Group Ltd, UK • Stero1i3d6s 0.0ww0w.5 .ac1e.0-hp1.l5c.co2.m0 |aH2T.n5+i-mbd4eo4-p3nM.o(0di0nlua)dtre1o3sc.25no2om4r4sp7.,0o0au4cni45d.d55ss,4 5|.0 [email protected] • • bases Vitamin D2/D3 Organic Acids – Fast Saenpdaprhaetinoonlic compounds • Small peptides Application #AN2200 Conditions Analytes Column: ACE Excel 3 C18-Amide 1. Sinapic acid ACE Excel 1.7 C18-Amide 2. Ferulic acid Organic Acids PD–aimrFtaeNnsutsmioSbneesp:r:ar22a55t00ioxxn22..11 mm, 50 x 3.0 mm 3. 3,4-Dimethoxycinnamic acid mmAAUU 11 22 33 ACE 3 Excel C18-Amide mm (EXL -1112-2502U), 4. Cinnamic acid 220000 44 50 x 3 mm (EXL -1712-0503U) 5. 4-Methoxycinnamic acid 55 Conditions: Mobile Phase: 20 mM H3PO4 in Analytes: 117755 2200 25 3300 3355 4400 min Column: ACE Excel 3 C18-Amide MeOH/H2O (40:60 v/v) 1. Sinapic acid ACE ExceFl l1o.w7 CR1a8te-A: mide0.21 mL/min (250 x 2.1 mm)2. Ferulic acid 115500 Dimensions: 250 x 2.1 mm, 50 x 30..08mmmL/min (50 x 3.0 mm) 3. 3,4-Dimethoxycinnamic acid 112255 Part Numbers: 2 50Inxje2c.t1iomn:m (EXL5-µ1L11(225-025x022.U1 )m, m) 4. Cinnamic acid 110000 50 x 3 mm (EXL -1721µ2L-0(5500x3U3.)0 mm) 5. 4-Methoxycinnamic acid 7755 Mobile Phase: 20 TmeMmpHe3raPtOu4rei:n M20eO°CH/H2O (40:60 v/v) Flow Rate: 0 .21 mDL/emteicnti(o2n5:0 x 2.1UVm, 2m1)0 nm 5500 0.8 mL/min (50 x 3.0 mm) 2255 ACE 3 C18-Amide Injection: 5 μL (250 x 2.1 mm) Sinapic acid 00 2 μL (50 x 3.0 mm) Temperature: 20 °C I 0 55 1100 1155 Detection: UV, 210 nm mmAAUU 44 ACE Excel 1.7 C18-Amide 117755 ACE Excel 1.7 C18-Amide 1100 1155 2200 2255 330 3355 4400 min Ferulic acid 3,4-Dimethoxycinnamic 115500 acid Time - Minutes 112255 22 33 110000 11 7755 55 5500 2255 Cinnamic acid 4-Methoxycinnamic 00 55 acid I 0 A-14 112CANwwAwD.aIAceN-hplc.com | +44 (0) 1224 704 5158488| -i2n2fo6@-27a7ce5-hp::[email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE® Excel™ C18-Amide) Dimensions ACE® Excel™ 100Å C18-Amide Analytical HPLC Columns HPLC/UHPLC Columns ID x Length (mm) 5 µm 3 µm 2 µm 1.7 µm 4.6 x 250 EXL-1212-2546U -- - 4.6 x 150 EXL-1212-1546U - 4.6 x 100 EXL-1212-1046U EXL-1112-1546U EXL-1012-1546U - 4.6 x 75 EXL-1212-7546U - 4.6 x 50 EXL-1212-0546U EXL-1112-1046U EXL-1012-1046U - 4.6 x 30 EXL-1212-0346U - 4.6 x 20 EXL-1212-0246U EXL-1112-7546U EXL-1012-7546U - 3.0 x 250 EXL-1212-2503U - 3.0 x 150 EXL-1212-1503U EXL-1112-0546U EXL-1012-0546U - 3.0 x 100 EXL-1212-1003U EXL-1712-1003U 3.0 x 75 EXL-1212-7503U EXL-1112-0346U EXL-1012-0346U EXL-1712-7503U 3.0 x 50 EXL-1212-0503U EXL-1712-0503U 3.0 x 30 EXL-1212-0303U EXL-1112-0246U EXL-1012-0246U EXL-1712-0303U 3.0 x 20 EXL-1212-0203U EXL-1712-0203U 2.1 x 250 EXL-1212-2502U -- - 2.1 x 150 EXL-1212-1502U - 2.1 x 100 EXL-1212-1002U EXL-1112-1503U EXL-1012-1503U EXL-1712-1002U 2.1 x 75 EXL-1212-7502U EXL-1712-7502U 2.1 x 50 EXL-1212-0502U EXL-1112-1003U EXL-1012-1003U EXL-1712-0502U 2.1 x 30 EXL-1212-0302U EXL-1712-0302U 2.1 x 20 EXL-1212-0202U EXL-1112-7503U EXL-1012-7503U EXL-1712-0202U 1.0 x 250 ACE-1212-2501 - 1.0 x 150 ACE-1212-1501 EXL-1112-0503U EXL-1012-0503U - 1.0 x 100 ACE-1212-1001 - 1.0 x 75 ACE-1212-7501 EXL-1112-0303U EXL-1012-0303U - 1.0 x 50 ACE-1212-0501 - EXL-1112-0203U EXL-1012-0203U -- EXL-1112-1502U EXL-1012-1502U EXL-1112-1002U EXL-1012-1002U EXL-1112-7502U EXL-1012-7502U EXL-1112-0502U EXL-1012-0502U EXL-1112-0302U EXL-1012-0302U EXL-1112-0202U EXL-1012-0202U ACE-1112-2501 ACE-1012-2501 ACE-1112-1501 ACE-1012-1501 ACE-1112-1001 ACE-1012-1001 ACE-1112-7501 ACE-1012-7501 ACE-1112-0501 ACE-1012-0501 Semi-Prep and 10 µm also available, please visit lifescience.ca ACE® Analytical HPLC Guard Cartridge Ordering Information Dimensions ACE® 100Å C18-Amide ID x Length (mm) 5 µm 3 µm Cartridges for 10 x 2.1mm ID Columns (pkg of 5) ACE-1212-0102GD ACE-1112-0102GD Cartridges for 10 x 3.0 - 4.6mm ID Columns (pkg of 5) ACE-1212-0103GD ACE-1112-0103GD Integral guard cartridge holder for 2.1mm ID Columns ACE-H0004 Integral guard cartridge holder for 3.0 - 4.6mm ID Columns ACE-H0005 Stand-alone universal guard cartridge holder. Requires Column coupler item # ACE-C0001 ACE-H0001 Other dimensions are available. Please enquire for details. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-15 LIFE SCIENCE

Conditions Analytes Colonnes HPLC/UHPLC Column: ACE 5 SuperC18 1. Omeprazole Dimensions: 150 x 4.6 mm 2. Lansoprazole Part Number: EXL -1211-1546U Mobile Phase: A: 0.1% ammonia in H2O B: 0.1% ammonia in MeCN/H2O (90:10 v/v) ACE® Excel™ SuperC18 Anti-UlcGerardDiernut:gs in B0Tiamseic(mMinosb) ile1%0PBhase Conditions Colonnes HPLC/UHPLC 5 90 Physical Properties ConditionFsl:ow Rate: 1 mL/min Analytes: Column: AInCjEecEtxiocenl: 5 Supe5rCµ1L8 1. Omeprazole Silica : Ultra-Inert Base Deactivated DimensioTnesm: 1p5e0raxt4u.r6e:mm30 °C 2. Lansoprazole Particle Size : 1.7 µm, 2 µm, 3 µm, 5 µm Part NumDbeert:ecEtXiLo-n1:211-1U5V4,62U80 nm Mobile Phase: 2 Surface Area : 400 m²/g A: 0.1% ammonia in MH2eOCN/H2O 1 Pore Size : 90 Å B: 0.1% ammonia in Bonded Phase : Octadecyl encapsulated (90:10 v/v) Gradient: Time (mins) %B End-Capping : Yes 0 10 Carbon Load : 14.8% 5 Omepra9z0ole USP Code : L1 Flow Rate: 1 mL/min Injection: 5 μL pH Range : 1.5-11.5 Temperature: 30 °C Detection: UV, 280 nm Mechanisms of Separation Strength of Interaction Lansoprazole Hydrophobic binding Strong interactions Weak 12345 Shape selectivity Time - Minutes www.ace-hplc.com Target Analytes Recommended Applications • Analytes differing in hydrophobicity • Analytes differing in hydrophobicity • Polar, moderately polar, and non-polar analytes • Homologous compounds differing by –CH2 • Uncharged acids and bases • Ionized acids or bases using ion-pairing Encapsulated Bonding Technology (EBT™) for Improved Chromatography and Stability Traditional C18 Bonding ACE SuperC18 with Encapsulated Bonding Technology (EBT™) C18 Endcapping group C18 Unbonded silanol group A-16 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE





HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE® Excel™ CN-ES) Dimensions ACE® Excel™ 100Å CN-ES Analytical HPLC Columns HPLC/UHPLC Columns ID x Length (mm) 5 µm 3 µm 2 µm 1.7 µm 4.6 x 250 EXL-1213-2546U EXL-1113-2546U - - 4.6 x 150 EXL-1213-1546U - 4.6 x 100 EXL-1213-1046U EXL-1113-1546U EXL-1013-1546U - 4.6 x 75 EXL-1213-7546U - 4.6 x 50 EXL-1213-0546U EXL-1113-1046U EXL-1013-1046U - 4.6 x 30 EXL-1213-0346U - 4.6 x 20 EXL-1213-0246U EXL-1113-7546U EXL-1013-7546U - 3.0 x 250 EXL-1213-2503U - 3.0 x 150 EXL-1213-1503U EXL-1113-0546U EXL-1013-0546U - 3.0 x 100 EXL-1213-1003U EXL-1713-1003U 3.0 x 75 EXL-1213-7503U EXL-1113-0346U EXL-1013-0346U EXL-1713-7503U 3.0 x 50 EXL-1213-0503U EXL-1713-0503U 3.0 x 30 EXL-1213-0303U EXL-1113-0246U EXL-1013-0246U EXL-1713-0303U 3.0 x 20 EXL-1213-0203U EXL-1713-0203U 2.1 x 250 EXL-1213-2502U EXL-1113-2503U - - 2.1 x 150 EXL-1213-1502U - 2.1 x 100 EXL-1213-1002U EXL-1113-1503U EXL-1013-1503U EXL-1713-1002U 2.1 x 75 EXL-1213-7502U EXL-1713-7502U 2.1 x 50 EXL-1213-0502U EXL-1113-1003U EXL-1013-1003U EXL-1713-0502U 2.1 x 30 EXL-1213-0302U EXL-1713-0302U 2.1 x 20 EXL-1213-0202U EXL-1113-7503U EXL-1013-7503U EXL-1713-0202U EXL-1113-0503U EXL-1013-0503U EXL-1113-0303U EXL-1013-0303U EXL-1113-0203U EXL-1013-0203U EXL-1113-2502U - EXL-1113-1502U EXL-1013-1502U EXL-1113-1002U EXL-1013-1002U EXL-1113-7502U EXL-1013-7502U EXL-1113-0502U EXL-1013-0502U EXL-1113-0302U EXL-1013-0302U EXL-1113-0202U EXL-1013-0202U Semi-Prep and 10 µm also available, please visit lifescience.ca Did you know? We sell Gas Chromatography Columns & Accessories? For more information please ask your Sales Representative today. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-19 LIFE SCIENCE

Colonnes HPLC/UHPLC ACE® & ACE® Excel™ C8 Colonnes HPLC/UHPLCPhysical Properties Vitamins–Water Soluble (Gradient) Response–MilliVoltsSilica : Ultra-Inert Base DeactivatedConditions: Compounds: Particle Size : 2 µm, 3 µm, 5 µm 1. Pyridoxamine Column: ACE 5 C8, 250 x 4.6mm 2. Thiamine (Vitamin B1) Part Number: ACE-122-2546 3. L-Ascorbic acid (Vitamin C) Surface Area : 300 m²/g Mobile Phase: A. 50mM KH2PO4 (pH 2.5) 4. Niacinamide (Vitamin B3) Pore Size : 100 Å B. MeOH 5. Nicotinic acid Bonded Phase : Octyl groups 19.5 6. Pyridoxal Flow Rate: 1.0ml/min 16.5 20 7. Pyridoxine Gradient: T(mins) 0 3 55 80 8. p-Aminobenzoic acid End-Capping : Yes %A 100 100 45 9. Pantothenic acid (Vitamin B5) Carbon Load : 9.0% 10. Folic acid (Vitamin M) USP Code : L7 %B 0 0 11. Cyanocobalamin (Vitamin B12) Temperature: Ambient 12. Riboflavin (Vitamin B2) Detection: UV, 205nm 13. d-Biotin (Vitamin H) pH Range : 2.0-8.0 14. Thioctic acid Mechanisms of Separation Strength of Interaction 1200 Hydrophobic binding Strong/Moderate 100 interactions 800 4 Target Analytes • Analytes differing in hydrophobicity • Polar, moderately polar, and non-polar analytes 600 25 • Uncharged acids and bases 400 1 67 12 • Ionized acids or bases using ion-pairing 8 11 14 Recommended Applications 200 3 ACE UHPLACC9aEnUd10HHPPLL1CC3 aCnodluHmPnLsC Columns • Analytes differing in hydrophobicity, 0 • Homologous compounds differing by –CH2 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 Time (min) •U SPMMixtounreosgcUroaSnptPhaiMn–inoMgneoptogrloraanrpiadhnad–zvoBeluerydheysdornoipdheobic USP MonogUraSpPhM– oNnaopgroraxpehn – Doxepin analytes Application #AN1A80p0plication #AN1720 Application #AN1A8p1p0lication #AN1730 •C onGodfoitpoiordonstsetainrCtsionangnddpihtlaiaorsgneesfpoor ldyepveepltoidpeinsAg. (ns3ea0ply0atÅreationAnalytes Conditions Conditions Analytes Analytes DCoimluermnesnci:oonms:meA7n5CDEodixmle43ue.md6Cn)8snmi:omns: ACE 5 C18 1. Metronidazole1. Budesonide B Column: AUCSEoPlUumlMtrnaoC: onroe g2.r5aASCpuEhpe3–rCD818oxep1i.nNaproxen 1. Doxepin isomer Dimensions: 150 x 4.6 mm 2. Budesonide A 50Dixm3e.0nsmiomns: 50 x 4.6 mm 2. Butyrophenone2. Doxepin isomer Part Number: CCPOoanRrdtEi-Nt2iu5oAmn-s0b:5e0r:3UACE-112-0546 Compounds: Part Number: ACPaEr-t11N2u-7m54b6er: ACE-121-1546 0(HPDC4.MF2ao59iOmlr1:loo1utwwebm)mN/nMiilLutRnseh/emi:amoPCgtAnbheilNnaCse:ac:Er(si55a:e30lA0:a:CC5xc00p80Ee..H425-tv1.i6c62/M1v.ma)5m2cm-Liim0nd/om5Hn4io2n6Oba/MsiceONHaH(32P0O:740 v/v) 1. Doxepin isomer 2. Doxepin isomer UMSoPbiMle oPhnaoseg:raHpM2Oho/Mb–ieleMOPHhe(at4sr:e1o:vn/1vi8d) mazMolmeonobasic sodium Mobile Phase: Flow Rate: 1 mL/min phosphate pH 3.2 in Flow Rate: CIonnjedcittiioonn:s: 15 µL H2O/MeCN (68:32 vC/ov)mpounds: A2UMF.DITmlV8noeo,ejbwmµ2ebtiLe5eciplRct4neeitatonirPto(anme2hnt::2au:0sr°e.eC5::)60pm.H5U42.0V5L2M,/°.µ2Cm5L5mii4nnonHnmo2Ob/aMsiceONHaNH(a23pP0rOo:7x4e0nv/v) CToelummpner:aAtCurEe3: C380Fl°oCw Rate: 1.5 mL/min 1. Metronidazole Injection: Temperature: DDimeteencstiioonns: : 75 xU4IVn.,6j3e1mc9timonnm: 20 µL Detection: Part Number: ACET-e1m12p-e7r5a4tu6re: Ambient (22 °C) Injection: 4.5 μL MFloowbilReaPteh:a1sem: HL/2mOD/ienMteecOtiHon(:4:1 v/Uv)V, 254 nm Temperature: 50 °C mAU Detection: U22V, 254 nm Injection: 15 μL °CmmAAUU Temperature: 30 Detection: UV, 319 1n16m600 225 mAU 114400 11 1 mAU 11 112200 2 Butyrophenone 707000 110000 2200 mAU 606000 1155 22 505000 8800 1100 11171.75.5 404000 6600 1155 303000 4400 5 202000 2200 121.25.5 101000 1100 77.5.5 55 22.5.5 00 00 00 00 00..55 11 11..55 Ti5me 22 221..550 33 15 33..55 min 11 2 55 11030 1155 4 2200 min 2255 min - Minutes Time Minutes - 20 Time - Minutes Time - Minutes Translated methodTraacncsolardteindgmtoetUhSoPd a<c6c2o1r>dingguitdoanUcSeP. A<l6w2a1y>s gchueidcakntchee. Always chTercaknsthlaeted method aTcrcaonrsdlaintegdtomUeSthPod<6a2c1co>rdgiunigdatnocUeS. APlw<6a2ys1>chgeucikdathnece. Always check the latest and official mlateetshtoadnidnfofrfmicaiatliomneftrhoomdtihneforremleavtaionnt fprhoamrmthaecorpeoleevianptrpiohrarmacloapteosetiaanpdrioorfficial melattheosdt ainnfdoromffaictioanl mfreotmhotdheinrfeolremvantitonphfarormatchoeproeeleiavpanritorpharmacopoeia prior to analysis. to analysis. to analysis. to analysis. A-20 USP MonogUraSCpPhAM–NoPAnaorDagcIrAeatpNahm–oEl/sAtrsapdiriionl/Caffeine 1 888-226-2775US::P Minfoon@olgiferascpiehn–ceG.claime::pirwidwew.lifescience.ca LIFE SCIENCE Application #AN1A8p2p0lication #AN1740 Application #AN1760

HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE & ACE Excel C8) Dimensions 5 µm ACE® & ACE® Excel™ 100Å C8 Analytical HPLC Columns 2 µm HPLC/UHPLC Columns 3 µm ID x Length (mm) ACE-122-2546 - ACE-122-1546 - EXL-102-1546U 4.6 x 250 ACE-122-1046 ACE-112-1546 EXL-102-1046U 4.6 x 150 ACE-122-7546 ACE-112-1046 EXL-102-7546U 4.6 x 100 ACE-122-0546 ACE-112-7546 EXL-102-0546U 4.6 x 75 ACE-122-0346 ACE-112-0546 EXL-102-0346U 4.6 x 50 ACE-122-0246 ACE-112-0346 EXL-102-0246U 4.6 x 30 ACE-122-2503 ACE-112-0246 4.6 x 20 ACE-122-1503 - 3.0 x 250 ACE-122-1003 - EXL-102-1503U 3.0 x 150 ACE-122-7503 ACE-112-1503 EXL-102-1003U 3.0 x 100 ACE-122-0503 ACE-112-1003 EXL-102-7503U 3.0 x 75 ACE-122-0303 ACE-112-7503 EXL-102-0503U 3.0 x 50 ACE-122-0203 ACE-112-0503 EXL-102-0303U 3.0 x 30 ACE-122-2502 ACE-112-0303 EXL-102-0203U 3.0 x 20 ACE-122-1502 ACE-112-0203 2.1 x 250 ACE-122-1002 - 2.1 x 150 ACE-122-7502 - EXL-102-1502U 2.1 x 100 ACE-122-0502 ACE-112-1502 EXL-102-1002U 2.1 x 75 ACE-122-0302 ACE-112-1002 EXL-102-7502U 2.1 x 50 ACE-122-0202 ACE-112-7502 EXL-102-0502U 2.1 x 30 ACE-122-2501 ACE-112-0502 EXL-102-0302U 2.1 x 20 ACE-122-1501 ACE-112-0302 EXL-102-0202U 1.0 x 250 ACE-122-1001 ACE-112-0202 1.0 x 150 ACE-122-7501 - 1.0 x 100 ACE-122-0501 - - 1.0 x 75 ACE-112-1501 - 1.0 x 50 ACE-112-1001 - ACE-112-7501 - ACE-112-0501 Semi-Prep and 10 µm also available, please visit lifescience.ca ACE® Analytical HPLC Guard Cartridge Ordering Information Dimensions 5 µm ACE® 100Å C8 3 µm ID x Length (mm) ACE-H0004 ACE-112-0102GD ACE-H0005 ACE-112-0103GD Cartridges for 10 x 2.1mm ID Columns (pkg of 5) ACE-122-0102GD ACE-H0001 Cartridges for 10 x 3.0 - 4.6mm ID Columns (pkg of 5) ACE-122-0103GD Integral guard cartridge holder for 2.1mm ID Columns Integral guard cartridge holder for 3.0 - 4.6mm ID Columns Stand-alone universal guard cartridge holder. Requires Column coupler item # ACE-C0001 Other dimensions are available. Please enquire for details. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-21 LIFE SCIENCE

Colonnes HPLC/UHPLC ACE® & ACE® Excel™ C4 Colonnes HPLC/UHPLC Physical Properties Silica : Ultra-Inert Base Deactivated Particle Size : 2 µm, 3 µm, 5 µm Surface Area : 300 m²/g Pore Size : 100 Å Bonded Phase : Butyl groups End-Capping : Yes Carbon Load : 5.5% USP Code : L26 pH Range : 2.0-8.0 Mechanisms of Separation Strength of Interaction Hydrophobic binding Moderate/Weak interactions Target Analytes Recommended Applications • Analytes differing in hydrophobicity • Analytes differing in hydrophobicity • Polar, moderately polar, and non-polar analytes • Homologous compounds differing by –CH2 • Uncharged acids and bases • Larger molecules, such as polypeptides and • IonizAedCaEcidUs HorPbLasCesaunsidngHioPnL-pCairCinoglumns proteins Phosphatidylethanol Biomarker Analysis by UHPLC-MS/MS Application #AN3400 Vitamins–Water Soluble (Gradient) Phosphatidylethanol Conditions: Conditions CompoundAs:nalytes 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 Column: ACE Excel 2 C4 1. R1/R2 =11(.m8R:/11z//R17280:1=1.418:1/182:181.2) Time - Minutes Dimensions: C1o00luxm2n.1: mm ACE 2 C4 Part NumberD: EimXLen-1s0io3n-1s0: 021U00 x 2.1 mm (m/z 701.4 281.2) Mobile PhaseP:aArt: N2 ummMbearm: mEoXnLiu-1m03-1002U 2. R1/R2 =21.6R:11//R162:=1 16:1/16:1 MoacbeilteatPeh/aMsee:CNA:(220m:8M0 va/mv)monium (m/z 7(m0/1z.7401.4 225555..11)) B: IPA acetate/MeCN (20:80 v/v) Gradient: Time (mins) B:%IPBA 1 G0ra.0d0ient: Ti1m0e (mins) %B 2 1.00 0.100 10 3.2e5 3.00 1.060 10 3.0e5 3.01 3.1000 60 2.8e5 5.00 3.10100 5.10 5.1000 100 100 2.6e5 Flow Rate: 0.4 mL/min 5.10 10 2.4e5 Injection: 5 μFLlow Rate: 0.4 mL/min TemperatureI:n4je0c°tCion: 2.2e5 5 µL 2.0e5 Detection: ABTeSmCIpEeXratrtiuprlee:qu4a0d°5C500 1.8e5 TTIoeunmrbSppoerIaroayntVuSoprelrtDa:ay6eg5tene0:ec-g°t4Caio5ti0nv0:eVmodITAoeuBnrEbSSSopCIrIaIoEynXVSotprlritpaaylgeenq:eu-g4aa5dt0iv50e5V0m0ode ESI 1.6e5 Intensity (cps) Phosphatidylethanol (PEth) mTeemaspuerreamtueren:t6i5n0 °C 1.4e5 busloeo/adbius sues.edPaashsoasabpbihoiamotmiadrayklreekrtehorafoncofhlcr(ohPnrEoictnhai)clcmaolehcaooslhuuorselem/aebnutsien. blood is used 1.2e5 1.0e5 8.0e4 6.0e4 4.0e4 2.0e4 0.0 1.5 2.0 2.5 0.0 0.5 1.0 Reproduced with permission of Biotage GB Ltd, UK A-22 PhytoCesAtrNogAeDnsIAfrNom Hop Extract by LC-MS1/M88S8-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE Application #AN1160

HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE & ACE Excel C4) Dimensions 5 µm ACE® & ACE® Excel™ 100Å C4 Analytical HPLC Columns 2 µm HPLC/UHPLC Columns 3 µm ID x Length (mm) ACE-123-2546 - ACE-123-1546 - EXL-103-1546U 4.6 x 250 ACE-123-1046 ACE-113-1546 EXL-103-1046U 4.6 x 150 ACE-123-7546 ACE-113-1046 EXL-103-7546U 4.6 x 100 ACE-123-0546 ACE-113-7546 EXL-103-0546U 4.6 x 75 ACE-123-0346 ACE-113-0546 EXL-103-0346U 4.6 x 50 ACE-123-0246 ACE-113-0346 EXL-103-0246U 4.6 x 30 ACE-123-2503 ACE-113-0246 4.6 x 20 ACE-123-1503 - 3.0 x 250 ACE-123-1003 - EXL-103-1503U 3.0 x 150 ACE-123-7503 ACE-113-1503 EXL-103-1003U 3.0 x 100 ACE-123-0503 ACE-113-1003 EXL-103-7503U 3.0 x 75 ACE-123-0303 ACE-113-7503 EXL-103-0503U 3.0 x 50 ACE-123-0203 ACE-113-0503 EXL-103-0303U 3.0 x 30 ACE-123-2502 ACE-113-0303 EXL-103-0203U 3.0 x 20 ACE-123-1502 ACE-113-0203 2.1 x 250 ACE-123-1002 - 2.1 x 150 ACE-123-7502 - EXL-103-1502U 2.1 x 100 ACE-123-0502 ACE-113-1502 EXL-103-1002U 2.1 x 75 ACE-123-0302 ACE-113-1002 EXL-103-7502U 2.1 x 50 ACE-123-0202 ACE-113-7502 EXL-103-0502U 2.1 x 30 ACE-123-2501 ACE-113-0502 EXL-103-0302U 2.1 x 20 ACE-123-1501 ACE-113-0302 EXL-103-0202U 1.0 x 250 ACE-123-1001 ACE-113-0202 1.0 x 150 ACE-123-7501 - 1.0 x 100 ACE-123-0501 - - 1.0 x 75 ACE-113-1501 - 1.0 x 50 ACE-113-1001 - ACE-113-7501 - ACE-113-0501 Semi-Prep and 10 µm also available, please visit lifescience.ca ACE® Analytical HPLC Guard Cartridge Ordering Information Dimensions 5 µm ACE® 100Å C4 3 µm ID x Length (mm) ACE-H0004 ACE-113-0102GD ACE-H0005 ACE-113-0103GD Cartridges for 10 x 2.1mm ID Columns (pkg of 5) ACE-123-0102GD ACE-H0001 Cartridges for 10 x 3.0 - 4.6mm ID Columns (pkg of 5) ACE-123-0103GD Integral guard cartridge holder for 2.1mm ID Columns Integral guard cartridge holder for 3.0 - 4.6mm ID Columns Stand-alone universal guard cartridge holder. Requires Column coupler item # ACE-C0001 Other dimensions are available. Please enquire for details. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-23 LIFE SCIENCE

Colonnes HPLC/UHPLC Catechins Cat ACE® & ACE® Excel™ CN Application #AN3950 Con Colonnes HPLC/UHPLC Physical Properties Conditions Analytes Colu Silica : Ultra-Inert Base Deactivated ColCuamten:chins ACE 5 C18, ACE 5 CN Dime Particle Size : 2 µm, 3 µm, 5 µm 1. Epigallocatechin Part Surface Area : 300 m²/g Dimensions: 150 x 4.6 mm 2. (+)-Epicatechin Mob Pore Size : 100 Å ParCtoNnudimtiobnesr:: ACE-121-1546, ACE-124-1546 Co3m. Eppoiugnadllso:catechin gallate MIFnlojoePCMHDwbcaoi2imotrllORiuteboemaNi(nlnP2etuns:he5miP:o:a:7hAnbs5aCeses:Ev:re1:/5:vMH21A5M)C0C2mµeO1EexLO8-OL14(,H/2H2.mA65/1/C0i0:m-n7E.1.115m55%%4vC6/fNvf,oo)ArrmCmEiic-c1aa2cc4idi-d1i5nin46 1. Epigallocatechin Flow TemFlpoweraRtautere: :1 mALm/mbiinent 2. (+)-Epicatechin Temp DetInejcetcitoionn: : 2 μLUV, 280 nm 3. Epigallocatechin gallate Dete Bonded Phase : Cyano Temperature: Ambient End-Capping : Yes Detection: UV, 280 nm Carbon Load : 5.5% USP Code : L10 pH Range : 2-7 RespoRenspsonsee --MilliVolMtsilliVolts 240 3 220 Mechanisms of Separation Strength of Interaction 200 Dipole-dipole interactions Strong Hydrophobic binding 180 2 interactions Weak 160 140 120 100 1 80 60 40 ACE C18 20 0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Time - Minutes Target Analytes 400 • Polar analytes • Analytes with double and triple bonds RespoResnposnsee- M-illiVoltsMilliVolts 350 3 • Non-polar analytes having too much 300 retention on alkyl phases 250 Recommended Applications 200 • Mixtures of very polar and polar analytes • Antihistamines 150 2 • Anaesthetics • As an orthogonal phase in RPLC method 100 1 50 development ACE CN 0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Time - Minutes Time - Minutes Catecholamine Analysis (II) Cat Application #AN3910 Con Conditions Analytes Colu Dime Column: ACE 5 AQ 1. Noradrenaline (Norepinephrine) Part Mob Oxymetazoline in Nasal Spray Formulation Dimensions: 150 x 4.6 mm 2. Adrenaline (Epinephrine) Part Number: ACE-126-1546 3. L(O-DH)OPA Flow Injec Conditions: Compounds: Mobile Phase: 50 mM KH2PO4 pH 3.0 in(HH3C2)O3C 4. DopamineCH3 Temp Column: ACE 5 CN, 150 x 4.6mm 1. Oxymetazoline Flow Rate: 1 mL/min 5. L -Tyrosine N Dete Part Number: ACE-124-1546 2. Benzalkonium chloride Mobile Phase: 50:50 MeCN/aqueous 24 Injection: 2 µL 6. VMA (Vanillylmandelic acid) cps 3.0e4 Na2HPO4, pH 7.0 22 Temperature: Ambient N 2.8e4 Flow Rate: 1.5 mL/min 20 Detection: UV1, 210 nm H 2.6e4 Temperature: 30° C CH3 2.4e4 Detection: UV, 214nm Oxymetazoline 2.2e4 2.0e4 18 1.8e4 1.6e4 Response -MiliVolts 16 1.4e4 1.2e4 14 H3C 1.0e4 R 8000 6000 12 N CI 4000 2000 10 CH3 8 6 Benzalkonium chloride 4 ReRsesppoonnses -eMi-lliVolMtilsliVolts 2 8 10 12 2 350 46 Time (Mins) 14 16 2 14 300 0 0 250 2 200 150 A-24 CANADIAN 1 888-2261-002775 :: [email protected] :: www.lifescience.ca 50 5 6 LIFE SCIENCE

HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE & ACE Excel CN) Dimensions 5 µm ACE® & ACE® Excel™ 100Å CN Analytical HPLC Columns 2 µm HPLC/UHPLC Columns 3 µm ID x Length (mm) ACE-124-2546 - ACE-124-1546 - EXL-104-1546U 4.6 x 250 ACE-124-1046 ACE-114-1546 EXL-104-1046U 4.6 x 150 ACE-124-7546 ACE-114-1046 EXL-104-7546U 4.6 x 100 ACE-124-0546 ACE-114-7546 EXL-104-0546U 4.6 x 75 ACE-124-0346 ACE-114-0546 EXL-104-0346U 4.6 x 50 ACE-124-0246 ACE-114-0346 EXL-104-0246U 4.6 x 30 ACE-124-2503 ACE-114-0246 4.6 x 20 ACE-124-1503 ACE-114-2503 - 3.0 x 250 ACE-124-1003 ACE-114-1503 EXL-104-1503U 3.0 x 150 ACE-124-7503 ACE-114-1003 EXL-104-1003U 3.0 x 100 ACE-124-0503 ACE-114-7503 EXL-104-7503U 3.0 x 75 ACE-124-0303 ACE-114-0503 EXL-104-0503U 3.0 x 50 ACE-124-0203 ACE-114-0303 EXL-104-0303U 3.0 x 30 ACE-124-2502 ACE-114-0203 EXL-104-0203U 3.0 x 20 ACE-124-1502 2.1 x 250 ACE-124-1002 - - 2.1 x 150 ACE-124-7502 ACE-114-1502 EXL-104-1502U 2.1 x 100 ACE-124-0502 ACE-114-1002 EXL-104-1002U 2.1 x 75 ACE-124-0302 ACE-114-7502 EXL-104-7502U 2.1 x 50 ACE-124-0202 ACE-114-0502 EXL-104-0502U 2.1 x 30 ACE-124-2501 ACE-114-0302 EXL-104-0302U 2.1 x 20 ACE-124-1501 ACE-114-0202 EXL-104-0202U 1.0 x 250 ACE-124-1001 ACE-114-2501 1.0 x 150 ACE-124-7501 ACE-114-1501 - 1.0 x 100 ACE-124-0501 ACE-114-1001 - 1.0 x 75 ACE-114-7501 - 1.0 x 50 ACE-114-0501 - - Semi-Prep and 10 µm also available, please visit lifescience.ca ACE® Analytical HPLC Guard Cartridge Ordering Information Dimensions 5 µm ACE® 100Å CN 3 µm ID x Length (mm) ACE-H0004 ACE-114-0102GD ACE-H0005 ACE-114-0103GD Cartridges for 10 x 2.1mm ID Columns (pkg of 5) ACE-124-0102GD ACE-H0001 Cartridges for 10 x 3.0 - 4.6mm ID Columns (pkg of 5) ACE-124-0103GD Integral guard cartridge holder for 2.1mm ID Columns Integral guard cartridge holder for 3.0 - 4.6mm ID Columns Stand-alone universal guard cartridge holder. Requires Column coupler item # ACE-C0001 Other dimensions are available. Please enquire for details. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-25 LIFE SCIENCE

Colonnes HPLC/UHPLC ACE® & ACE® Excel™ Phenyl Colonnes HPLC/UHPLC Physical Properties Comparison of Selectivity differences between C18 and Phenyl phases Silica : Ultra-Inert Base Deactivated Particle Size : 2 µm, 3 µm, 5 µm Column: 4.6 x 250 mm, 5.0 μm Surface Area : 300 m²/g Part Numbers: ACE C18 - ACE-121-2546 1. Norephedrine Pore Size : 100 Å 2. Nortriptyline ACE Phenyl - ACE-125-2546 3. Toluene 4. Imipramine Bonded Phase : Phenyl groups Mobile Phase: 20% 0.025M KH2PO4, pH 6.0 5. Amitriptyline 80% MeOH End-Capping : Yes Carbon Load : 9.5% Flow Rate: 1.0 mL/min Temperature: 22 °C USP Code : L11 pH Range : 2.0-8.0 Mechanisms of Separation Strength of Interaction ACE C18 23 4 5 1 π-π interactions Strong Dipole-dipole interactions Moderate Hydrophobic binding Moderate interactions 0 1 2 3 4 5 6 7 8 9 10 11 12 13 Target Analytes Time (min) • Analytes with π bonding • Analytes with electron delocalization and ACE Phenyl 45 ehlaelcotgreAonnCs-,wEnitithUrdoHrgaPrwoLiunCpgsg,arkoneutdopnsH,essPu, cLehsCtaesrCs,olumns 1 2 3 and acids • Analytes with proton donor groups 0 1 2 3 4 5 6 7 8 9 10 • AnalytePsawraitbhedniffPerreenstedrvipaotlievemsoments Time (min) •R ecoSmtemreeoniCCdsooeolmdunmdeAnripts:ipolnicsatAioCEns3 Phenyl Analytes These comparison chromatograms show the substantial differences in seleAcptipvliitcyabteiotwn e#eAnN1250 • SteroidDsimensions: 150 x 2.1 mm a C18 and a phenyl phase. Peaks 2 and 3 have reversed elution order on the phenyl phase 1. Methyl paraben compared to the C18 phase 2. Ethyl paraben • TaxanesMPaortbNileumPhbaesre:: ACE-115-1502 3. n-Propyl paraben 25 mM ammonium 4. i-Butyl paraben • Substituted aromatiaccse tate pH 6.8 in 5. n-Butyl paraben • Highly aFqlouweRoautes: condH0.i22tOimo/MLne/smOiHn (50:50 v/v) 6. Benzyl paraben Methyl paraben Ethyl paraben n-Propyl paraben Injection: 2 µL Temperature: 40 °C Intens. [m AU] Paraben PreDseetercvtaiotniv: es UV, 240 nm 11 Conditions: Analytes: -10 22 Column: ACE 3 Phenyl 1. Methyl paraben -15 33 Dimensions: 150 x 2.1 mm 2. Ethyl paraben -20 Part Number: ACE-115-1502 3. n-Propyl paraben -25 Mobile Phase: 25 mM ammonium 4. i-Butyl paraben 5. n-nB-Buutytyl lppaarraabbeenn acetatie-BpuHty6l .p8arinaben 6. Benzyl paraben H2O/MeOH (50:50 v/v) Flow Rate: 0.2 mL/min 44 55 Injection: 2 μL Temperature: 40 °C 66 Detection: UV, 240 nm Benzyl paraben -30 0 2 4 6 8 10 12 14 16 18 Time [min] 2 4 6 8 10 12 14 16 18 mins Time - Minutes Reproduced with permission of The Chemical Analysis Facility, University of Reading A-26 ParaLCceIFAtEaNmSAoClDIEaINnACdNERelated Compounds 1 888-226-2775Par::[email protected]:: Suwbwswta.linfecsecsience.ca

HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE & ACE Excel Phenyl) Dimensions 5 µm ACE® & ACE® Excel™ 100Å Phenyl Analytical HPLC Columns 2 µm HPLC/UHPLC Columns 3 µm ID x Length (mm) ACE-125-2546 - ACE-125-1546 - EXL-105-1546U 4.6 x 250 ACE-125-1046 ACE-115-1546 EXL-105-1046U 4.6 x 150 ACE-125-7546 ACE-115-1046 EXL-105-7546U 4.6 x 100 ACE-125-0546 ACE-115-7546 EXL-105-0546U 4.6 x 75 ACE-125-0346 ACE-115-0546 EXL-105-0346U 4.6 x 50 ACE-125-0246 ACE-115-0346 EXL-105-0246U 4.6 x 30 ACE-125-2503 ACE-115-0246 4.6 x 20 ACE-125-1503 - 3.0 x 250 ACE-125-1003 - EXL-105-1503U 3.0 x 150 ACE-125-7503 ACE-115-1503 EXL-105-1003U 3.0 x 100 ACE-125-0503 ACE-115-1003 EXL-105-7503U 3.0 x 75 ACE-125-0303 ACE-115-7503 EXL-105-0503U 3.0 x 50 ACE-125-0203 ACE-115-0503 EXL-105-0303U 3.0 x 30 ACE-125-2502 ACE-115-0303 EXL-105-0203U 3.0 x 20 ACE-125-1502 ACE-115-0203 2.1 x 250 ACE-125-1002 - 2.1 x 150 ACE-125-7502 - EXL-105-1502U 2.1 x 100 ACE-125-0502 ACE-115-1502 EXL-105-1002U 2.1 x 75 ACE-125-0302 ACE-115-1002 EXL-105-7502U 2.1 x 50 ACE-125-0202 ACE-115-7502 EXL-105-0502U 2.1 x 30 ACE-125-2501 ACE-115-0502 EXL-105-0302U 2.1 x 20 ACE-125-1501 ACE-115-0302 EXL-105-0202U 1.0 x 250 ACE-125-1001 ACE-115-0202 1.0 x 150 ACE-125-7501 - 1.0 x 100 ACE-125-0501 - - 1.0 x 75 ACE-115-1501 - 1.0 x 50 ACE-115-1001 - ACE-115-7501 - ACE-115-0501 Semi-Prep and 10 µm also available, please visit lifescience.ca ACE® Analytical HPLC Guard Cartridge Ordering Information Dimensions ACE® 100Å Phenyl ID x Length (mm) 5 µm 3 µm Cartridges for 10 x 2.1mm ID Columns (pkg of 5) ACE-125-0102GD ACE-115-0102GD Cartridges for 10 x 3.0 - 4.6mm ID Columns (pkg of 5) ACE-125-0103GD ACE-115-0103GD Integral guard cartridge holder for 2.1mm ID Columns ACE-H0004 Integral guard cartridge holder for 3.0 - 4.6mm ID Columns ACE-H0005 Stand-alone universal guard cartridge holder. Requires Column coupler item # ACE-C0001 ACE-H0001 Other dimensions are available. Please enquire for details. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-27 LIFE SCIENCE

Colonnes HPLC/UHPLC ACE® & ACE® Excel™ Silica Colonnes HPLC/UHPLC Physical Properties Silica : Ultra-Inert Base Deactivated Particle Size : 2 µm, 3 µm, 5 µm Surface Area : 300 m²/g Pore Size : 100 Å Bonded Phase : N.A. End-Capping : N.A. Carbon Load : N.A. USP Code : L3 pH Range : 2.0-8.0 Mechanisms of Separation Strength of Interaction ACE UHPLC and HPLC Column Dipole-dipole interactions Weak Weak cation exchange (HILIC) Moderate Normal phase Moderate Target Analytes Recommended for HILIC Applications •L ipPidolaCrlaansaslyetess Separation f•r omVeDryrpooslaorpahndilabaMsicealnaanlyotegsaster Application #AN1530 • Antihistamines TIC +ve ion Conditions • AnaestheticsAnalytes TIC -ve ion Column: ACE 3 SIL 1. Triglyceride (TG) 24 26 28 2. Phosphoserine (PS) Dimensions: 150 x 3.0 mm 3i. Phosphoethanolamine (PE) Part Number: ACE-117-1503 Mobile Phase: A: IPA/MeCN (20:80 v/v) 3ii. Lyso phosphoethanolamine (Lyso PE) Lipid Classes SeBp:aIPraAt/i0o.0n2fMroammDmroonsiuomphfiolramMateelanoga4s.tSeprhingomyelin phosphoethanolamine (SMPE) (20:80 v/v) 5i. Phosphatidylcholine (PC) CGornadditieonst: Time (mins) %B 5ii. Lyso phosphatidylcholine (Lyso PC) Column: ACE 3 SIL 0.0 6. Glycerophosphoglycerol (GPG) 8 7. Phosphoinositol (PI) PMDaimortbeNilneusmiPohnbases:re1::A5BA0C::ExIIP-P15113AA.01..0//00.70M0m-.10em52C0NM3(a2m0:m80ovn298/i0uv)m formate 100 55ii (20:1860.v0/v) 25 Gradient: Time2(3m.0ins) %B 35 0.0 26.0 88 Flow Rate: 1.0 0.3 mL/min8 Detection: 5.0 LTQ Orbitra9p MS 11 1106..00Positive and2205negative ion mode Relative abundance 33ii 23.0 35 100 22 44 33iii 26.0 8 55iiii 77 33ii Flow Rate: 0.3 mL/min Detection: LTQ Orbitrap MS Positive and negative ion mode Analytes: 1. Triglyceride (TG) 66 5ii 2. Phosphoserine (PS) 22 5iii 3i. Phosphoethanolamine (PE) 3ii. Lyso phosphoethanolamine (Lyso PE) 2 4 6 8 10 12 14 16 18 20 22 4. Sphingomyelin phosphoethanolamine (SMPE) Time - Minutes 55iii..PLhyosospphhaotisdpyhlachtDiodrlyionlcsehoo(pPlihCni)ela(LMyseolaPnCo) gaster 6. Glycerophosphoglycerol (GPG) 7. Phosphoinositol (PI) Reproduced with permission of University of Strathclyde, Glasgow, UK Liquorice Extracts Fingerprint A-28 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescienceA.pcaplication #AN2090 LIFE SCIENCE

HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE® & ACE® Excel Silica) Dimensions 5 µm ACE® & ACE® Excel™ 100Å Silica Analytical HPLC Columns 2 µm HPLC/UHPLC Columns 3 µm ID x Length (mm) ACE-127-2546 - ACE-127-1546 - EXL-107-1546U 4.6 x 250 ACE-127-1046 ACE-117-1546 EXL-107-1046U 4.6 x 150 ACE-127-7546 ACE-117-1046 EXL-107-7546U 4.6 x 100 ACE-127-0546 ACE-117-7546 EXL-107-0546U 4.6 x 75 ACE-127-0346 ACE-117-0546 EXL-107-0346U 4.6 x 50 ACE-127-0246 ACE-117-0346 EXL-107-0246U 4.6 x 30 ACE-127-2503 ACE-117-0246 4.6 x 20 ACE-127-1503 - 3.0 x 250 ACE-127-1003 - EXL-107-1503U 3.0 x 150 ACE-127-7503 ACE-117-1503 EXL-107-1003U 3.0 x 100 ACE-127-0503 ACE-117-1003 EXL-107-7503U 3.0 x 75 ACE-127-0303 ACE-117-7503 EXL-107-0503U 3.0 x 50 ACE-127-0203 ACE-117-0503 EXL-107-0303U 3.0 x 30 ACE-127-2502 ACE-117-0303 EXL-107-0203U 3.0 x 20 ACE-127-1502 ACE-117-0203 2.1 x 250 ACE-127-1002 - 2.1 x 150 ACE-127-7502 - EXL-107-1502U 2.1 x 100 ACE-127-0502 ACE-117-1502 EXL-107-1002U 2.1 x 75 ACE-127-0302 ACE-117-1002 EXL-107-7502U 2.1 x 50 ACE-127-0202 ACE-117-7502 EXL-107-0502U 2.1 x 30 ACE-127-2501 ACE-117-0502 EXL-107-0302U 2.1 x 20 ACE-127-1501 ACE-117-0302 EXL-107-0202U 1.0 x 250 ACE-127-1001 ACE-117-0202 1.0 x 150 ACE-127-7501 ACE-117-2501 - 1.0 x 100 ACE-127-0501 ACE-117-1501 - 1.0 x 75 ACE-117-1001 - 1.0 x 50 ACE-117-7501 - ACE-117-0501 - Semi-Prep and 10 µm also available, please visit lifescience.ca ACE® Analytical HPLC Guard Cartridge Ordering Information Dimensions ACE® 100Å Silica ID x Length (mm) 5 µm 3 µm Cartridges for 10 x 2.1mm ID Columns (pkg of 5) ACE-127-0102GD ACE-117-0102GD Cartridges for 10 x 3.0 - 4.6mm ID Columns (pkg of 5) ACE-127-0103GD ACE-117-0103GD Integral guard cartridge holder for 2.1mm ID Columns ACE-H0004 Integral guard cartridge holder for 3.0 - 4.6mm ID Columns ACE-H0005 Stand-alone universal guard cartridge holder. Requires Column coupler item # ACE-C0001 ACE-H0001 Other dimensions are available. Please enquire for details. 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-29 LIFE SCIENCE

Colonnes HPLC/UHPLC ACE® Excel™ 100Å HILIC-A Colonnes HPLC/UHPLC Physical Properties Silica : Ultra-Inert Base Deactivated Particle Size : 1.7 µm, 3 µm, 5 µm Surface Area : 300 m²/g Pore Size : 100 Å Bonded Phase : Proprietaryl SIL End-Capping : No Carbon Load : N.A. USP Code : L3 pH Range : 2.0-7.0 ACE HILIC-A Creatine and Metabolite Creatinine using HILIC Mode• An acidic character phase with an ionisable negative surface charge depending upon mobile phase pH. β-blockers (ACE AN#4530) Appli mmAUAU 2 Conditions 2. Sotalol ACE 5 HILIC-A 600 1 1. PropranDCoiolmolulemnsni:ons: 150 x 4.6 mm 550000 440000 Part Number: HILA-5-1546U Mobile Phase: 330000 A: 2 mM ammonium formate pH 3.0 in MeCN/H2O ( 220000 3 B: 2 mM ammonium formate pH 3.0 in MeCN/H2O ( Time ( mins ) %B 05 110000 4 5 10 5 00 22..55 55 77..55 mmiinn 3. Acebutalol 4. Salbutamol 11 55 12 5. A5te5nolol ComlVumn: ACE HILIC-A, 150 x 4.6 mm, 5 µm Part Number: ACE-HILA-5-1546U Mobile Phase: 12 mM ammonium formate pH 4.7 in MeCN/H2O (8381:12 v/v) 5 Flow Rate: 1.5 mL/min Injection: 2 µL Temperature: 25 ºC Detection: 230 nm Flow Rate: 1.5 mL/min 120 Injection: 5 µL ACE HILIC-B 25 °C Temperature: UV, 230 nm Cr•e1A0a0 tbianseicacnhdarMacetetrapbhoalisteewCirtheaatninioinneisaubslienpgoHsiItLivICe sMurofadcee charge dependDientgecutpioonn: mobile phase pH. Succinic acid and methylmalonic acid (ACE AN#4520) 80 1 DCCooimmln6u6Vde00mintnisoi:onAnsCs: :E15S5oH0lveIxLnIt4Cf.-r6Aonmt m 200 Part Number: ACE-HILA-5-1546U MB:o24b400mileMPhamasme:oAn:iu2mmfMorammamteopnHiu3m.0foinrmMaetCeNp/HH23O.0(5in0M:50e1CvN/v/)H2O2 (95:5 v/v) 150 1 2 Tim2e2000(mins) %B 1. Succinic acid 2. Methylmalonic acid 5 10 5 100 2211 550044 66 88 1100 1122 1144 1166 1188 mmin in 12 5500 Col3u1mn: ACE HIL5IC-B, 150 x 4.6 mm, 5 µm FlFolwowRRaattee:: 1..55mmLL/m/mininTemperature: 25 ºC Part Number: ACE-HILB-5-1546U Mobile Phase:5100 mM ammonium formate pH 3.0 in MeCN/H2O (90:10 v/v) Injection: 5 µL Detection: ELSD (Evaporator temp: 30 ºC, Nebuliser temp: 30 ºC, Gas speed: 1 SLM) Injection: 5 μL 2 Temperature: 25 °C 0 DeAteCctEionH: UIVL,I2C30-Nnm An•aAlytpeos:lar neutral character phase 1. Creatinine with the capability of H-bonding amo-n5g0st other mechanisms of interaction. 2.NCurecalteinoebases and nucleosides (ACE AN#4550) 0 2 4 6 8 mAU 12 1.0E+1.01E+1 3 8.0E+8.00E+0 6.0E+6.00E+0Int ensity (mAU) A-30 4.0E+4.00E+0 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca 2.0E+2.00E+0 LIFE SCIENCE 1. Deoxyadenosine 2. Adenine 3. Adenosine

HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE® HILIC-A) Dimensions 5 µm ACE® Excel™ 100Å HILIC-A Analytical HPLC Columns 1.7 µm HPLC/UHPLC Columns 3 µm ID x Length (mm) ACE-HILA-5-2546U - ACE-HILA-5-1546U ACE-HILA-3-2546U - 4.6 x 250 ACE-HILA-5-1046U ACE-HILA-3-1546U - 4.6 x 150 ACE-HILA-5-7546U ACE-HILA-3-1046U - 4.6 x 100 ACE-HILA-5-0546U ACE-HILA-3-7546U - 4.6 x 75 ACE-HILA-5-0346U ACE-HILA-3-0546U - 4.6 x 50 ACE-HILA-5-0246U ACE-HILA-3-0346U - 4.6 x 30 ACE-HILA-5-2503U ACE-HILA-3-0246U - 4.6 x 20 ACE-HILA-5-1503U ACE-HILA-3-2503U - 3.0 x 250 ACE-HILA-5-1003U ACE-HILA-3-1503U ACE-HILA-17-1003U 3.0 x 150 ACE-HILA-5-7503U ACE-HILA-3-1003U ACE-HILA-17-7503U 3.0 x 100 ACE-HILA-5-0503U ACE-HILA-3-7503U ACE-HILA-17-0503U 3.0 x 75 ACE-HILA-5-0303U ACE-HILA-3-0503U ACE-HILA-17-0303U 3.0 x 50 ACE-HILA-5-0203U ACE-HILA-3-0303U ACE-HILA-17-0203U 3.0 x 30 ACE-HILA-5-2502U ACE-HILA-3-0203U - 3.0 x 20 ACE-HILA-5-1502U ACE-HILA-3-2502U - 2.1 x 250 ACE-HILA-5-1002U ACE-HILA-3-1502U ACE-HILA-17-1002U 2.1 x 150 ACE-HILA-5-7502U ACE-HILA-3-1002U ACE-HILA-17-7502U 2.1 x 100 ACE-HILA-5-0502U ACE-HILA-3-7502U ACE-HILA-17-0502U 2.1 x 75 ACE-HILA-5-0302U ACE-HILA-3-0502U ACE-HILA-17-0302U 2.1 x 50 ACE-HILA-5-0202U ACE-HILA-3-0302U ACE-HILA-17-0202U 2.1 x 30 ACE-HILA-3-0202U 2.1 x 20 Semi-Prep and 10 µm also available, please visit lifescience.ca ACE® Microbore HPLC Column Ordering Information - (ACE® HILIC-A) Dimensions ACE® Excel™ 100Å HILIC-A Microbore HPLC Columns ID x Length (mm) 5 µm 3 µm 1.0 x 250 1/16\" Port 1/32\" Port 1/16\" Port 1/32\" Port 1.0 x 150 1.0 x 100 ACE-HILA-5-2501 ACE-HILA-5-2501S - - 1.0 x 75 ACE-HILA-5-1501 ACE-HILA-3-1501S 1.0 x 50 ACE-HILA-5-1001 ACE-HILA-5-1501S ACE-HILA-3-1501 ACE-HILA-3-1001S 0.5 x 250 ACE-HILA-5-7501 ACE-HILA-3-7501S 0.5 x 150 ACE-HILA-5-0501 ACE-HILA-5-1001S ACE-HILA-3-1001 ACE-HILA-3-0501S 0.5 x 100 ACE-HILA-5-25005 0.5 x 75 ACE-HILA-5-15005 ACE-HILA-5-7501S ACE-HILA-3-7501 - 0.5 x 50 ACE-HILA-5-10005 ACE-HILA-3-15005S 0.5 x 30 ACE-HILA-5-75005 ACE-HILA-5-0501S ACE-HILA-3-0501 ACE-HILA-3-10005S ACE-HILA-5-05005 ACE-HILA-3-75005S ACE-HILA-5-03005 ACE-HILA-5-25005S - ACE-HILA-3-05005S ACE-HILA-3-03005S ACE-HILA-5-15005S ACE-HILA-3-15005 ACE-HILA-5-10005S ACE-HILA-3-10005 ACE-HILA-5-75005S ACE-HILA-3-75005 ACE-HILA-5-05005S ACE-HILA-3-05005 ACE-HILA-5-03005S ACE-HILA-3-03005 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-31 LIFE SCIENCE

Colonnes HPLC/UHPLC ACE HILIC-A • An acidic character phase with an ionisable negative surface charge depending upon mobile phase pH. βA-CblEoc®keErsxc(eAClE™AN#H45I30L)IC-B Colonnes HPLC/UHPLC mPmAhUAUysical Properties 2 S60il0ica : Ultra-Inert Base Deactivated P550a000rticle Size 1 : 1.7 µm, 3 µm, 5 µm 1. Propranolol 2. Sotalol SP440uo000rrfeaSceizeA rea : 300 m²/g Methylmalonic Acid and Succinic Acid : 100 Å B330o000nded Phase : Proprietary Aminopropyl using HILIC Mode E220n000d-Capping : No 3 Application UC110a0S00rPbCoondLeo ad : 4.0%4 5 Conditions ACE 5 HILIC-B : N.A. Column: pH00 Range : 2.0-7.0 77..55 mmiinn 3. Acebutalol Dimensions: 4. Salbutamol 150 x 4.6 mm 5. Atenolol Part Number: HILB-5-1546U 22..55 55 Mobile Phase: 10 mM ammonium formate pH 3.0 in MeCN/H2O (90:10 v/v depCFeloonmlwVudmiRnnag:teA:uC1pE.5oHmnILLICm/m-Aoi,nb15ilI0nejxepc4ht.i6oanms:em2p,µ5HL µT.mempPearrattNuurem: b2e5rº:CACDE-eHteILcAti-o5n-1: 524360UnmMobile Phase: 12 mM ammFolonwiumRaftoer:mate pH 4.7 in MeCN/H21O.5(8m8:L1/2mvi/nv) charge Injection: 5 µL Temperature: 25 °C 120 Detection: ELSD Evaporator temperature: 30 °C ACE HILIC-B Nebuliser temperature: 30 °C • A100 basic character phase with an ionisable positive surface charge depending upon mobile phase pH. Gas speed: 1 SLM 1. PropranSMoul8o0eclcthinyilcmaacliodnaincdAmcide2t.ahSnoytdlamlSoaul lcocninicicacAidcid using(AHCIELAINC#4M52o0)de mV Solvent front C6o600nditions: Column: ACE 5 HILIC-B D4i4m00 ensions: 150 x 4.6 mm Part Number: ACE-HILB-5-1546U formate pH 3.0 12 120 Mobile Phase: 10 mM ammonium in MeCN/H2O (90:10 v/v) Fl2o200w Rate: 1.5 mL/min 1. Succinic acid 2. Methylmalonic acid bil3e .PAhcaeseb:u1tCF2NDGEITalnoeovmleaelojma0wu0etsbMlpmec0upsR0octpnalaietirom:tiesraoeAneaetm:nCtr:od1uo:E5r.t:5rneEH1teμimLme2u2I:LSLSmmL2IpDLC/5eMpm-fBore°iar,nCrmt1au5taT4ru04etee4rmx:e.pp43:SH.e03a6r40la°mb.tC7°uum66Cirtne,a:5Mm2µe5omClºNC8/8PHaIn2rOtjeN(c8tu8imo:1n11b002:e5vr:/µvAL)CED1-1eH225tIeL. BcAt-it5oe-n1n:15o14E44lL6oSUlD 100 MeCN/H2O (90:10 v/v) 1. Suc 1166 1188 mmin in speed: 1 SLM) Mobile Phase: 10 mM ammo8n0ium formate pH 3.0 in (Evaporator temp: 30 ºC, Nebuliser temp: 30 ºC, Gas rge depenAdA1i.nnCSagulEycutcepiHnso:icnIaLmcIidCob-iNle phase pH. 60 •2.AMpeothlayrlmnaeluotnricalacchidaracter phase with the capability of H-bonding amongst other 4m0echanisms of interaction. 12 2. Methyl 10 12 14 16 18 Nucleobases and nucleosides (ACE AN#4550) mAU 12 20 1.0E+1.01E+1 3 00 2 4 6 8 8.0E+8.00E+0 6.0E+6.00E+0Int ensity (mAU) 1. Succinic acid 2. Methylmalonic acid 4.0E+4.00E+0 66 1188 2.0E+2.0m0mEi+n0 in 1. Deoxyadenosine 2. Adenine 3. Adenosine bile Phase: 100.m0EM+0.00E+0ammonium formate pH 3.0 in sMpee33 eCdN:/1H2SOLM(494)0:10 v/v) min porator temp: 30 ºC, N00 ebuliser 1t1 emp: 30 º22C, Gas 55 Column: ACE HILIC-N, 150 x 4.6 mm,Time (min) 5 µm Part Number: ACE-HILN-5-1546U Mobile Phase: 10 mM ammonium formate pH 4.7 in MeCN/H2O (90:10 v/v) Flow Rate: 1.5 mL/min Temperature: 25 ºC Injection: 5 µL Detection: 254 nm ing amongst other mechanisms of interaction. A-32 1. Deoxyadenosine CAN2. AAdeDninIAe N 3. Adenosine 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE® HILIC-B) Dimensions 5 µm ACE® Excel™ 100Å HILIC-B Analytical HPLC Columns 1.7 µm HPLC/UHPLC Columns 3 µm ID x Length (mm) ACE-HILB-5-2546U - ACE-HILB-5-1546U ACE-HILB-3-2546U - 4.6 x 250 ACE-HILB-5-1046U ACE-HILB-3-1546U - 4.6 x 150 ACE-HILB-5-7546U ACE-HILB-3-1046U - 4.6 x 100 ACE-HILB-5-0546U ACE-HILB-3-7546U - 4.6 x 75 ACE-HILB-5-0346U ACE-HILB-3-0546U - 4.6 x 50 ACE-HILB-5-0246U ACE-HILB-3-0346U - 4.6 x 30 ACE-HILB-5-2503U ACE-HILB-3-0246U - 4.6 x 20 ACE-HILB-5-1503U ACE-HILB-3-2503U - 3.0 x 250 ACE-HILB-5-1003U ACE-HILB-3-1503U ACE-HILB-17-1003U 3.0 x 150 ACE-HILB-5-7503U ACE-HILB-3-1003U ACE-HILB-17-7503U 3.0 x 100 ACE-HILB-5-0503U ACE-HILB-3-7503U ACE-HILB-17-0503U 3.0 x 75 ACE-HILB-5-0303U ACE-HILB-3-0503U ACE-HILB-17-0303U 3.0 x 50 ACE-HILB-5-0203U ACE-HILB-3-0303U ACE-HILB-17-0203U 3.0 x 30 ACE-HILB-5-2502U ACE-HILB-3-0203U - 3.0 x 20 ACE-HILB-5-1502U ACE-HILB-3-2502U - 2.1 x 250 ACE-HILB-5-1002U ACE-HILB-3-1502U ACE-HILB-17-1002U 2.1 x 150 ACE-HILB-5-7502U ACE-HILB-3-1002U ACE-HILB-17-7502U 2.1 x 100 ACE-HILB-5-0502U ACE-HILB-3-7502U ACE-HILB-17-0502U 2.1 x 75 ACE-HILB-5-0302U ACE-HILB-3-0502U ACE-HILB-17-0302U 2.1 x 50 ACE-HILB-5-0202U ACE-HILB-3-0302U ACE-HILB-17-0202U 2.1 x 30 ACE-HILB-3-0202U 2.1 x 20 Semi-Prep and 10 µm also available, please visit lifescience.ca ACE® Microbore HPLC Column Ordering Information - (ACE® HILIC-B) Dimensions ACE® Excel™ 100Å HILIC-B Microbore HPLC Columns ID x Length (mm) 5 µm 3 µm 1.0 x 250 1/16\" Port 1/32\" Port 1/16\" Port 1/32\" Port 1.0 x 150 1.0 x 100 ACE-HILB-5-2501 ACE-HILB-5-2501S - - 1.0 x 75 ACE-HILB-5-1501 ACE-HILB-3-1501S 1.0 x 50 ACE-HILB-5-1001 ACE-HILB-5-1501S ACE-HILB-3-1501 ACE-HILB-3-1001S 0.5 x 250 ACE-HILB-5-7501 ACE-HILB-3-7501S 0.5 x 150 ACE-HILB-5-0501 ACE-HILB-5-1001S ACE-HILB-3-1001 ACE-HILB-3-0501S 0.5 x 100 ACE-HILB-5-25005 0.5 x 75 ACE-HILB-5-15005 ACE-HILB-5-7501S ACE-HILB-3-7501 - 0.5 x 50 ACE-HILB-5-10005 ACE-HILB-3-15005S 0.5 x 30 ACE-HILB-5-75005 ACE-HILB-5-0501S ACE-HILB-3-0501 ACE-HILB-3-10005S ACE-HILB-5-05005 ACE-HILB-3-75005S ACE-HILB-5-03005 ACE-HILB-5-25005S - ACE-HILB-3-05005S ACE-HILB-3-03005S ACE-HILB-5-15005S ACE-HILB-3-15005 ACE-HILB-5-10005S ACE-HILB-3-10005 ACE-HILB-5-75005S ACE-HILB-3-75005 ACE-HILB-5-05005S ACE-HILB-3-05005 ACE-HILB-5-03005S ACE-HILB-3-03005 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-33 LIFE SCIENCE

22..55 55 77..55 mmiinn 3. Acebutalol 4. Salbutamol 5. Atenolol ComlVumn: ACE HILIC-A, 150 x 4.6 mm, 5 µm Part Number: ACE-HILA-5-1546U Mobile Phase: 12 mM ammonium formate pH 4.7 in MeCN/H2O (88:12 v/v) Colonnes HPLC/UHPLCFlow Rate: 1.5 mL/min Injection: 2 µL Temperature: 25 ºC Detection: 230 nm 120 ACE HILIC-B ACE®• A100 basicEchxacraectle™r pHhaIsLe IwCit-hNan ionisable positive surface charge depending upon mobile phase pH. Succinic acid and methylmalonic acid (ACE AN#4520) Colonnes HPLC/UHPLC 80 PmhVysical PSroolvepnet frrtoinet s Si6l600ica : Ultra-Inert Base Deactivated Particle Size : 1.7 µm, 3µm, 5 µm Su4400rface Area : 300 m²/g Pore Size : 100 Å 12 B2o200nded Phase : Prioprietary Polyhydroxyl 1. Succinic acid 2. Methylmalonic acid End-Capping : No CUaS00rPb00 CoondLeo a22 d : 7N..0A%.66 44 : 88 1100 1122 1144 1166 1188 mmin in pCFlooHlwumRRnaa:tneA:gC1Ee.5H mILLIC/m-Bi,n15T0e:mx2p4..e60ram-t7um.re0, :52µ5mºC Part Number: ACE-HILB-5-1546U Mobile Phase: 10 mM ammonium formate pH 3.0 in MeCN/H2O (90:10 v/v) Injection: 5 µL Detection: ELSD (Evaporator temp: 30 ºC, Nebuliser temp: 30 ºC, Gas speed: 1 SLM) ACE HILIC-N • A polar neutral character phase with the capability of H-bonding amongAstdotehenr imneechaaninsmds oNf inutecralcetioons. ides using HILIC-MS Nucleobases and nucleosides Mode (ACE AN#4550) mAU 1 2 1.0E+1.01E+1 8.0E+8.00E+0 3 Applicat 6.0E+6.00E+0Int ensity (mAU) Conditions ACE 5 HILIC-N Column: 4.0E+4.00E+0 Dimensions: 150 x 4.6 mm Part Number: HILN-5-1546U 2.0E+2.00E+0 Mobile Phase: 10 mM ammonium formate pH 4.7 in MeCN/H2O (90:10 0.0E+0.00E+0 1. DeoxFInylaojedwcetniRooanst:ien:e 2. Adenine 1.5 mL3/m. Aindenosine 00 11 22 33 44 55 min 5 µL Temperature: 25 °C Column: ACE HILIC-N, 150 x 4.6 mm,Time (min) 5 µm Part Number: ACE-HILN-5-1546U Mobile Phase: 10 mM Dametmeocntiiounm: formate pH 4.7 in MeCN/H((ba2O)) U(9V0:,1205v4/vn) m MSD Flow Rate: 1.5 mL/min Temperature: 25 ºC Injection: 5 µL Detection: 254 nm Chromaster SIM Positive ion mode [M + H]+ Instrument: Chromaster Adenine and Nucleosides using HILIC-MS Mode 12 Conditions: 1.0E+1 (a) 3 Column: ACE 5 HILIC-N 8.0E+0 Dimensions: 150 x 4.6 mm Intensi ty(mAU) 1. D Part Number: ACE-HILN-5-1546U 3 6.0E+0 Mobile Phase: 10 mM ammonium formate pH 4.7 in MeCN/H2O (90:10 v/v) Flow Rate: 1.5 mL/min 4.0E+0 Injection: 5 μL Temperature: 25 °C 2.0E+0 Detection: (a) UV, 254 nm (b) Chromaster MSD 0.0E+0 SIM Positive ion mode [M + H]+ 0 1 2 345 Instrument: Chromaster Time (min) Analytes: Intensi ty 1.0E+4 (b) 1. Deoxyadenosine (m/z 252.4) 2. Adenine (m/z 136.1) 8.0E+3 1 3. Adenosine (m/z 268.3) 6.0E+3 4.0E+3 2.0E+3 2345 0.0E+0 Time (min) 0 A-34 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns ACE® Analytical HPLC Column Ordering Information - (ACE® HILIC-N) Dimensions 5 µm ACE® Excel™ 100Å HILIC-N Analytical HPLC Columns 1.7 µm HPLC/UHPLC Columns 3 µm ID x Length (mm) ACE-HILN-5-2546U - ACE-HILN-5-1546U ACE-HILN-3-2546U - 4.6 x 250 ACE-HILN-5-1046U ACE-HILN-3-1546U - 4.6 x 150 ACE-HILN-5-7546U ACE-HILN-3-1046U - 4.6 x 100 ACE-HILN-5-0546U ACE-HILN-3-7546U - 4.6 x 75 ACE-HILN-5-0346U ACE-HILN-3-0546U - 4.6 x 50 ACE-HILN-5-0246U ACE-HILN-3-0346U - 4.6 x 30 ACE-HILN-5-2503U ACE-HILN-3-0246U - 4.6 x 20 ACE-HILN-5-1503U ACE-HILN-3-2503U - 3.0 x 250 ACE-HILN-5-1003U ACE-HILN-3-1503U ACE-HILN-17-1003U 3.0 x 150 ACE-HILN-5-7503U ACE-HILN-3-1003U ACE-HILN-17-7503U 3.0 x 100 ACE-HILN-5-0503U ACE-HILN-3-7503U ACE-HILN-17-0503U 3.0 x 75 ACE-HILN-5-0303U ACE-HILN-3-0503U ACE-HILN-17-0303U 3.0 x 50 ACE-HILN-5-0203U ACE-HILN-3-0303U ACE-HILN-17-0203U 3.0 x 30 ACE-HILN-5-2502U ACE-HILN-3-0203U - 3.0 x 20 ACE-HILN-5-1502U ACE-HILN-3-2502U - 2.1 x 250 ACE-HILN-5-1002U ACE-HILN-3-1502U ACE-HILN-17-1002U 2.1 x 150 ACE-HILN-5-7502U ACE-HILN-3-1002U ACE-HILN-17-7502U 2.1 x 100 ACE-HILN-5-0502U ACE-HILN-3-7502U ACE-HILN-17-0502U 2.1 x 75 ACE-HILN-5-0302U ACE-HILN-3-0502U ACE-HILN-17-0302U 2.1 x 50 ACE-HILN-5-0202U ACE-HILN-3-0302U ACE-HILN-17-0202U 2.1 x 30 ACE-HILN-3-0202U 2.1 x 20 Semi-Prep and 10 µm also available, please visit lifescience.ca ACE® Microbore HPLC Column Ordering Information - (ACE® HILIC-N) Dimensions ACE® Excel™ 100Å HILIC-N Microbore HPLC Columns ID x Length (mm) 5 µm 3 µm 1.0 x 250 1/16\" Port 1/32\" Port 1/16\" Port 1/32\" Port 1.0 x 150 1.0 x 100 ACE-HILN-5-2501 ACE-HILN-5-2501S - - 1.0 x 75 ACE-HILN-5-1501 ACE-HILN-3-1501S 1.0 x 50 ACE-HILN-5-1001 ACE-HILN-5-1501S ACE-HILN-3-1501 ACE-HILN-3-1001S 0.5 x 250 ACE-HILN-5-7501 ACE-HILN-3-7501S 0.5 x 150 ACE-HILN-5-0501 ACE-HILN-5-1001S ACE-HILN-3-1001 ACE-HILN-3-0501S 0.5 x 100 ACE-HILN-5-25005 0.5 x 75 ACE-HILN-5-15005 ACE-HILN-5-7501S ACE-HILN-3-7501 - 0.5 x 50 ACE-HILN-5-10005 ACE-HILN-3-15005S 0.5 x 30 ACE-HILN-5-75005 ACE-HILN-5-0501S ACE-HILN-3-0501 ACE-HILN-3-10005S ACE-HILN-5-05005 ACE-HILN-3-75005S ACE-HILN-5-03005 ACE-HILN-5-25005S - ACE-HILN-3-05005S ACE-HILN-3-03005S ACE-HILN-5-15005S ACE-HILN-3-15005 ACE-HILN-5-10005S ACE-HILN-3-10005 ACE-HILN-5-75005S ACE-HILN-3-75005 ACE-HILN-5-05005S ACE-HILN-3-05005 ACE-HILN-5-03005S ACE-HILN-3-03005 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-35 LIFE SCIENCE

Colonnes HPLC/UHPLCColonnes HPLC/UHPLC ACE Step-by-Step HILIC Method Development Protocol Start Are analyte NO STEP 1 properties known? YES Select two pH values Use pH 3.0, 4.7 and 6.0 from pH 3.0, 4.7 and 6.0 based on logD/pKa data Run either the gradient or isocratic screening protocol using the appropriate pH values on the three ACE HILIC phases (Table 1) For Gradient: For Isocratic: Assess if the separation YES Is the retention Is the retention YES Assess if the separation STEP 2 can be achieved using window narrow? window wide? can be achieved using isocratic conditions gradient conditions NO NO Did isocratic YES Select the most promising YES Did gradient analysis improve the column/mobile phase analysis improve the separation? separation? combination from screening data NO Optimise separation by NO Return to investigating other parameters Return to gradient mode i.e. temperature, ionic strength isocratic mode STEP 3 Assess method robustness Figure 1: Flow diagram for step-by-step NO HILIC Method Development protocol. Is method robust? YES Method Developed A-36 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns ACE Step-by-Step HILIC Method Development Protocol HPLC/UHPLC Columns HILIC stationary phase and mobile phase pH are two of the most powerful parameters for altering HILIC selectivity. Assessing these two critical parameters is therefore the optimum starting point for method development. The recommended approach uses stationary and mobile phase screening data to identify a column/mobile phase combination that is most promising for the sample. Once selected, the method can then be fine-tuned using other parameters such as buffer strength and temperature. Figure 1 on page A-36 shows a flow diagram summarising the following step-by-step HILIC method development protocol. Step-by-Step Rational Method Development Step 3: The effects of other parameters such as temperature Step 1: If analyte properties are known, select two appropriate and buffer concentration can be used to fine-tune the method. mobile phase pH’s for screening. If unknown, use pH 3.0, 4.7 Once development is complete, the method robustness can be and 6.0 (these pH values are designed to maximize selectivity assessed as required. differences). Step 2: The sample is screened on the three ACE HILIC phases at the specified pH values using either isocratic or gradient conditions as specified in Table 1. If retention times are too short or too long in isocratic mode, the percentage of strong solvent (water) may require adjustment. If the retention window is wide leading to excessive resolution (i.e. some analytes show much stronger retention than others) in isocratic mode, a gradient screen should be attempted to assess whether this provides a better option. Likewise, if analyte peaks are clustered too closely in gradient mode, an isocratic separation may be required. From this data, the stationary phase/mobile phase combination that gives the most promising result is selected for further development. Column Table 1: Suggested conditions for HILIC screening Isocratic Screening ACE HILIC-A, ACE HILIC-B and ACE HILIC-N, 150 x 4.6 mm, 5 μm Gradient Screening 10 mM ammonium formate in MeCN/H2O (90:10 v/v) Ammonium formate at pH 3.0, 4.7 or 6.0. Flow Rate Temperature Line A: 10 mM ammonium formate in MeCN/H2O (94:6 v/v) Detection Line B: 10 mM ammonium formate in MeCN/H2O (50:50 v/v) Ammonium formate at pH 3.0, 4.7 or 6.0. Gradient: Time %B 00 15 100 20 100 21 0 41 0 1.5 mL/min 25 °C Dependent on sample 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-37 LIFE SCIENCE

Colonnes HPLC/UHPLC ACE HILIC Method Development Colonnes HPLC/UHPLC • ACE HILIC columns provide alternative selectivity to each other. • The power of systematic screening of different phase chemistries for HILIC method development is seen below. • Maximize your HILIC method development success by following the ACE HILIC method development protocol using three optimised ACE HILIC column chemistries Advantages of using ACE HILIC Method Development Kits mAmUAU 2 8080 7070 1,3 6060 5,6 5050 ACE HILIC-A 4040 9 3030 4 2020 1010 7 8 00 00 22..55 55 77..55 1100 1122..55 1155 1177..55 mmiinn mmiinn ACE HILIC-B mAmUAU 5 92 mmiinn Different Elution 5050 77..55 1100 Orders and Retention 4 4040 31 Times Observed 3030 2020 78 1122..55 6 1177..55 1010 22..55 55 1155 00 00 mAmUAU 1,4 6060 5050 5 2 4040 ACE HILIC-N 3030 3 9 2020 1010 7 6,8 00 22..55 55 77..55 1100 1122..55 1155 1177..55 00 1. p-Aminobenzoic acid 2. 4-Hydroxybenzoic acid 3. Nicotinamide 4. Acebutolol 5. Adenine 6. Mandelic acid 7. Tyramine 8. Atenolol 9. 2-Deoxyguanosine Columns: All 150 x 4.6 mm, 5 µm (Part numbers: ACE HILIC-A: HILA-5-1546U, ACE HILIC-B: HILB-5-1546U, ACE HILIC-N: HILN-5-1546U) Mobile Phase: 10 mM ammonium formate pH 4.7 in MeCN/H2O (90:10 v/v) Flow Rate: 1.5 mL/min Temperature: 25 °C Detection: 254 nm Sample: 1) p-Aminobenzoic acid 2) 4-Hydroxybenzoic acid 3) Nicotinamide 4) Acebutolol 5) Adenine 6) Mandelic acid 7) Tyramine 8) Atenolol 9) 2-Deoxyguanosine AACCEE HHIILLIICC ccoolluummnnss pprroovviiddee aalltteerrnnaattiivvee sseelleeccttiivviittyy ttoo eeaacchh ootthheerr –- iiddeeaallffoorrHHIILLIICCmmeetthhooddddeevveellooppmmeenntt.. A-38 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE

HPLC/UHPLC Columns ACE HILIC Method Development Kits Ordering Information Dimensions 5 µm ACE® HILIC Method Development kits 1.7 µm HPLC/UHPLC Columns 3 µm ID x Length (mm) ACE-MDKH-5-2546U - ACE-MDKH-5-1546U ACE-MDKH-3-2546U - 4.6 x 250 ACE-MDKH-5-1046U ACE-MDKH-3-1546U - 4.6 x 150 ACE-MDKH-5-7546U ACE-MDKH-3-1046U - 4.6 x 100 ACE-MDKH-5-0546U ACE-MDKH-3-7546U - 4.6 x 75 ACE-MDKH-5-0346U ACE-MDKH-3-0546U - 4.6 x 50 ACE-MDKH-5-0246U ACE-MDKH-3-0346U - 4.6 x 30 ACE-MDKH-5-2503U ACE-MDKH-3-0246U - 4.6 x 20 ACE-MDKH-5-1503U ACE-MDKH-3-2503U - 3.0 x 250 ACE-MDKH-5-1003U ACE-MDKH-3-1503U ACE-MDKH-17-1003U 3.0 x 150 ACE-MDKH-5-7503U ACE-MDKH-3-1003U ACE-MDKH-17-7503U 3.0 x 100 ACE-MDKH-5-0503U ACE-MDKH-3-7503U ACE-MDKH-17-0503U 3.0 x 75 ACE-MDKH-5-0303U ACE-MDKH-3-0503U ACE-MDKH-17-0303U 3.0 x 50 ACE-MDKH-5-0203U ACE-MDKH-3-0303U ACE-MDKH-17-0203U 3.0 x 30 ACE-MDKH-5-2502U ACE-MDKH-3-0203U - 3.0 x 20 ACE-MDKH-5-1502U ACE-MDKH-3-2502U - 2.1 x 250 ACE-MDKH-5-1002U ACE-MDKH-3-1502U ACE-MDKH-17-1002U 2.1 x 150 ACE-MDKH-5-7502U ACE-MDKH-3-1002U ACE-MDKH-17-7502U 2.1 x 100 ACE-MDKH-5-0502U ACE-MDKH-3-7502U ACE-MDKH-17-0502U 2.1 x 75 ACE-MDKH-5-0302U ACE-MDKH-3-0502U ACE-MDKH-17-0302U 2.1 x 50 ACE-MDKH-5-0202U ACE-MDKH-3-0302U ACE-MDKH-17-0202U 2.1 x 30 ACE-MDKH-3-0202U 2.1 x 20 Dimensions ACE® HILIC Microbore Method Development kits ID x Length (mm) 5 µm 3 µm 1.0 x 250 1/16\" Port 1/32\" Port 1/16\" Port 1/32\" Port 1.0 x 150 1.0 x 100 ACE-MDKH-5-2501 ACE-MDKH-5-2501S - - 1.0 x 75 ACE-MDKH-5-1501 ACE-MDKH-3-1501S 1.0 x 50 ACE-MDKH-5-1001 ACE-MDKH-5-1501S ACE-MDKH-3-1501 ACE-MDKH-3-1001S 0.5 x 250 ACE-MDKH-5-7501 ACE-MDKH-3-7501S 0.5 x 150 ACE-MDKH-5-0501 ACE-MDKH-5-1001S ACE-MDKH-3-1001 ACE-MDKH-3-0501S 0.5 x 100 ACE-MDKH-5-25005 0.5 x 75 ACE-MDKH-5-15005 ACE-MDKH-5-7501S ACE-MDKH-3-7501 - 0.5 x 50 ACE-MDKH-5-10005 ACE-MDKH-3-15005S 0.5 x 30 ACE-MDKH-5-75005 ACE-MDKH-5-0501S ACE-MDKH-3-0501 ACE-MDKH-3-10005S ACE-MDKH-5-05005 ACE-MDKH-3-75005S ACE-MDKH-5-03005 ACE-MDKH-5-25005S - ACE-MDKH-3-05005S ACE-MDKH-3-03005S ACE-MDKH-5-15005S ACE-MDKH-3-15005 ACE-MDKH-5-10005S ACE-MDKH-3-10005 ACE-MDKH-5-75005S ACE-MDKH-3-75005 ACE-MDKH-5-05005S ACE-MDKH-3-05005 ACE-MDKH-5-03005S ACE-MDKH-3-03005 1 888-226-2775 :: [email protected] :: www.lifescience.ca CANADIAN A-39 LIFE SCIENCE

Colonnes HPLC/UHPLC Contact your sales representative for any of these ACE brochures. Colonnes HPLC/UHPLC A-40 CANADIAN 1 888-226-2775 :: [email protected] :: www.lifescience.ca LIFE SCIENCE