Microflow LC-MS with the turn of a key

Transcription

1 [ ionkey/ms ] Microflow LC-MS with the turn of a key

2 Dramatically enhance your sensitivity for both quantitative and qualitative chromatographic data, with an integrated microflow LC-MS solution that is simple, reliable, and reproducible. The ionkey /MS System integrates micro-scale UPLC separation into the source of a mass spectrometer. Its modern microflow technology delivers exceptional sensitivity, robustness, reproducibility, and ease-of-use. The ionkey/ms System provides: Increased sensitivity A simplified user experience Flexibility for higher throughput/ varying speeds of analysis THE KEY TO SUCCESS ikey SEPARATION TECHNOLOGY The ikey Separation Device fully exploits the power of sub-2-µm UPLC separations with the ionkey/ms System to deliver superior chromatographic resolution. ikey with 15 µm I.D. separation channel for highest sensitivity ikey with 3 µm I.D. separation channel for highest sample throughput on ionkey/ms ikey Post Column Addition (PCA) for addition of modifier solvent post separation The ikey Separation Device simplifies microflow LC-MS and greatly reduces chromatographic variability. 15 µm I.D. ikey up to 4X increase in sensitivity compared to 2.1 mm UPLC columns 3 µm I.D. ikey HT delivers increased sample throughput, reduced cycle times, and increased sensitivity levels 2

3 [ ionkey/ms ] ionkey/ms makes assays which were not possible in a routine lab, possible ionkey/ms SYSTEM The ionkey/ms System is designed to achieve excellent quantitative and qualitative data by integrating the ACQUITY UPLC M-Class System, ionkey Source, ikey Separation Device, and a supported Waters mass spectrometer addressing the need for both high sensitivity and routine applications, on one platform. ionkey/ms technology is available for: ionkey and Xevo TQ-S and TQ-XS Ultimate sensitivity and reliability Market-leading performance delivered by StepWave XS ion guide Enhanced Product Ion Confirmation Scanning from ScanWave collision cell ionkey and Xevo QTof HRMS Maximum robustness with no compromise in performance Flexibility to adapt to changing needs Accessible to experts and non-experts alike ionkey and Xevo TQ-S micro ionkey and SYNAPT G2-Si HRMS Multiple ionization modes Tof coupled to ion mobility analysis Sensitive, reliable, and compact Class-leading performance that can fit into any lab Robust sensitivity driven by proven StepWave ion guide technology 3

4 Sensitivity advantage of microflow LC-MS With microflow LC-MS, the lower flow rate of the mobile phase can dramatically improve the sensitivity of the mass spectrometer: Increased sampling efficiency Increased ionization efficiency Reduced matrix effects 6 75 µm I.D. Selectivity improvement from 2.1 mm scale µm I.D. 3 µm I.D. 1 mm I.D. 2.1 mm I.D Flow rate (µl/min) BALANCING SENSITIVITY AND SPEED 3 µm ikey HT ikey HT (3 µm) is a higher throughput microflow device coupled with the ionkey/ms System that demonstrates improved sensitivity over UPLC methods on a timescale that is practical for routine analysis. Moving a 2.1 mm method to the 3 µm ikey HT increased the signal to noise by up to 6.1X while maintaining a 3 minute injection to injection cycle time in plasma. 4

5 Enabling sensitivity and speed [ ionkey/ms ] 15 µm ikey 1 1 pg/ml Apidra 4.96;3145 MRM of 9 Channels ES >137.1 (Apidra) 6.32e4 Area pg/ml Apidra 4.97;1477 MRM of 9 Channels ES >137.1 (Apidra) 3.14e4 Area pg/ml Apidra MRM of 9 Channels ES >137.1 (Apidra) 2.47e4 Area ~41 amol on column at LOD Blank human plasma MRM of 9 Channels ES >137.1 (Apidra) 1.92e4 Area min High sensitivity analysis of insulin glusine (Apidra) from 1 µl of human plasma sample, 1 µl injection on the ikey 15 µm. Comparison of 3 insulin methods Standard curve range (pg/ml) Analyte 2.1 x 5 mm 3 µm x 5 mm 15 µm x 1 mm Lispro 5 1, 1 1, 25 1, Glargine 5 1, 5 1, 25 1, Detemir 2 1, 1 5, 5 1, Glulisine 5 1, 25 1, 25 1, Aspart 1 1, 5 1, 25 1, Sample volume 25 µl 25 µl 1 µl Injection volume 3 µl 15 µl 1 µl Run time 8 min 7 min 13.5 min A comparison of three methods for therapeutic insulin and four analogs in human plasma demonstrates how the ionkey/ms System enables flexibility to modulate a microflow LC-MS system between ultra-sensitivity analysis and higher throughput by simply switching between the ikey Separations Devices. UPLC 2.1 mm Good sensitivity and run time ikey 3 µm Fastest run time: Same sample volume, 1/2 the injection volume, and 2X more sensitive than the 2.1 mm method ikey 15 µm Greatest sensitivity (2 4X): Using 2.5X less sample, 1/3 injection volume, 2X longer column, and 1.7X longer run time 5

6 Post Column Addition flexibility The ikey with Post Column Addition mab Calibration Curve PCA 2 µl min_6 812 (4.45) 1 (PCA) allows for adding a modifier 1: TOF MS ES+ 1.43e solvent post chromatographic electrospray ionization process from mab Calibration Curve PCA Imidizole_ (3.648) additional degree of freedom to the 1: TOF MS ES+ 2.68e4 scientist which can lead to significant enhancements to the MS signal the chromatography and offers an of the enhancement of the micro PCA addition of MeOH:H2O (5:5).1FA separation. This affords the decoupling In this example, the number of charge 55 states on the Waters mab is increased pka = using PCA addition of methanol:water PCA addition of MeOH:H2O (5:5).1FA w/ 1 mg/ml Imidazol (5:5):.1 formic acid (top) to further increase sensitivity. Adding Imidazole R2 (lower post column), the charge state Imidazole PCA (1 ng mab on-column) 1 distribution shifts by an average of 19 R1 charges and an increase in charge state Reduction in peak overlap spacing by 5. This spacing affords more spectral space to process complex mabs and ADC s where charge states could overlap. R4 No PCA (1 ng mab on-column) Charge manipulation can enhance value of the transformation algorithm 1 R3 The increase in the charge state spacing could be used to further accommodate more complex spectra with multiple charge state distributions. The top spectrum illustrates the charge state distribution that is generated with a 1 ng (on-column) IgG that coelutes with SILu MAb. Imidazole PCA made Imidazole PCA (1 ng mab on-column) it possible to baseline resolve both 1 charge state distributions. The inset shows the baseline comparison, below, showing the overlapping spectra without decharging No PCA (1 ng mab on-column)

7 Proven robustness and reproducibility Achieve reproducible results from user to user and system to system. Each ikey Separation Device is LC-MS tested to ensure performance with intra- and inter-ikey reproducibility. 1 user X 1 system X 1 ikey Peptide from P924, yeast enolase 6.84 min Retention time 6.69 min.1 min 3.1 sec SD of retention time Average peak width FWHM.11 min 2.72 sec 5 users X 5 systems X 18 ikeys retention time RSD QC reproducibility for one ikey with over 5 injections of protein precipitated plasma. retention time RSD QC reproducibility across multiple users, ionkey/ms systems, and ikeys. HUNDREDS OF INJECTIONS, IDENTICAL RESULTS 1 First 5 µl injection 1 After 1 5 µl injections of crashed plasma min min Quality control injections of dextromethorphan and propranolol showing equivalent chromatographic resolution before study and after 1 x 5 µl injections of 2:1 protein precipitated plasma. 7 First injection Peak area RSD: psi 3 2 Last injection min Injection Number 12 Consistent pressure trace before study and after 1 x 1 µl injections of 2:1 protein precipitated plasma samples. Peak area reproducibility over 1 x 1 µl injections of 2:1 protein precipitated plasma samples. 8

8 [ ionkey/ms ] Ordering Information Part No. Description Particle Size I.D. Length UOM ikey Separations Devices ikey Peptide BEH C 18 13Å 1.7 µm 15 µm 5 mm 1/pkg ikey BEH C 18 13Å 1.7 µm 15 µm 5 mm 1/pkg ikey PCA Peptide BEH C 18 13Å 1.7 µm 15 µm 5 mm 1/pkg ikey PCA BEH C 18 13Å 1.7 µm 15 µm 5 mm 1/pkg ikey Peptide BEH C 18 13Å 1.7 µm 15 µm 1 mm 1/pkg ikey BEH C 18 13Å 1.7 µm 15 µm 1 mm 1/pkg ikey Peptide BEH C 18 3Å 1.7 µm 15 µm 5 mm 1/pkg ikey Peptide BEH C 18 3Å 1.7 µm 15 µm 1 mm 1/pkg ikey Protein BEH C 4 3Å 1.7 µm 15 µm 5 mm 1/pkg ikey Protein BEH C 4 3Å 1.7 µm 15 µm 1 mm 1/pkg ikey Peptide CSH C 18 13Å 1.7 µm 15 µm 5 mm 1/pkg ikey CSH C 18 13Å 1.7 µm 15 µm 5 mm 1/pkg ikey Peptide CSH C 18 13Å 1.7 µm 15 µm 1 mm 1/pkg ikey CSH C 18 13Å 1.7 µm 15 µm 1 mm 1/pkg ikey HSS T3 1Å 1.8 µm 15 µm 5 mm 1/pkg ikey HSS T3 1Å 1.8 µm 15 µm 1 mm 1/pkg ikey Peptide BEH C 18 13Å 1.7 µm 3 µm 5 mm 1/pkg ikey Peptide CSH C 18 13Å 1.7 µm 3 µm 5 mm 1/pkg ikey HSS T3 1Å 1.8 µm 3 µm 5 mm 1/pkg Trap Columns ACQUITY UPLC M-Class Symmetry C 18 Trap Column, 1Å 5 µm 3 µm 5 mm 1/pkg ACQUITY UPLC M-Class Peptide BEH C 18 Trap Column, 13Å 5 µm 3 µm 5 mm 1/pkg ACQUITY UPLC M-Class HSS T3 Trap Column, 1Å 5 µm 3 µm 5 mm 1/pkg ACQUITY UPLC M-Class BEH C 4 Trap Column, 3Å 5 µm 3 µm 5 mm 1/pkg Utility ikey ikey Infusion Device n/a 85 µm 5 mm 1/pkg ikey Flow Injection Analysis Device n/a 85 µm 5 mm 1/pkg ikey Diagnostic Device V3 n/a n/a n/a 1/pkg Waters offers a complete portfolio of standards, certified vials, and certified containers for your ionkey/ms System. For more information, visit 9

9 [ ionkey/ms ] Choose your chemistries Hybrid particles Silica-based particles 13Å 3Å 3Å 3Å 1.7 µm 1.7 µm 1.7 µm 1.8 µm C₁₈ C₁₈ C₄ C₁₈ T3 The BEH stationary phase is a universal media choice, suitable for a diverse range of analytes. High retentivity for basic compounds under elevated ph Suitable across a wide ph range For separations at high temperatures (8 C) CSH Technology utilizes a controlled, low-level surface charge to provide enhanced selectivity and exceptional peak shape, particularly for basic compounds under low ph conditions. Increased mass loads for peptides Excellent MS performance with formic acid as a modifier solvent Fast ph switching and column equilibration The High Strength Silica [HSS] particle technology is designed for UPLC separations where silicabased selectivities are desired. High retentivity for polar organic compounds and metabolites Balanced retention of polar and hydrophobic analytes WATERS GLOBAL SERVICES Delivering world renowned services and support Waters Service and Support offerings are tailored to optimize your laboratory productivity while addressing your budget realities. Our offerings help maintain system peak performance, minimize down time, address scientific application challenges, and support stringent compliance requirements. As your services and support provider, we are committed to the success of your laboratory and business. Download the ionkey/ms Application Compendium to review work performed by scientists that demonstrate the benefits of using the ionkey/ms System. The application notes are from the fields of: Bioanalysis, Lipidomics, Metabolomics, and Proteomics Clinical Research Forensic Toxicology Food Research To download, please visit: Waters quality support and consultative services ensure your success wherever your laboratory is located in the world. Waters has consecutively earned the ACE Award since 21 for providing best-in-class technical knowledge, issue resolution, and process support. 7

10 Sales Offices: Australia Austria Belgium and Luxembourg Brazil Canada China Czech Republic Denmark Finland France Germany Hong Kong Hungary India Ireland Israel Italy Japan Korea Malaysia Mexico The Netherlands Norway Poland Portugal Puerto Rico Singapore Spain Sweden Switzerland Taiwan UAE UK US For your local sales office, please visit Waters Corporation 34 Maple Street Milford, MA 1757 U.S.A. T: F: Waters, The Science of What s Possible, ACQUITY, ionkey/ms, SYNAPT, UPLC, and Xevo are registered trademarks of Waters Corporation. ikey, ionkey, ScanWave, and StepWave are trademarks of Waters Corporation. All other trademarks are the property of their respective owners. 217 Waters Corporation. Printed in the U.S.A. December EN LM-SIG