Embed Size (px)
Citation preview
[ ionkey/MS ]
Microflow LC-MS with the turn of a key
2
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
Dramatically enhance your sensitivity for both quantitative and qualitative chromatographic data, with an integrated microflow LC-MS solution that is simple, reliable, and reproducible.
150 µm I.D. iKey up to 40X increase in sensitivity compared to 2.1 mm
UPLC columns
300 µm I.D. iKey HT delivers increased sample throughput,
reduced cycle times, and increased sensitivity levels
THE KEY TO SUCCESS – iKEY SEPARATION TECHNOLOGYThe 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 150 µm I.D. separation channel for highest sensitivity
■■ iKey with 300 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.
[ ionkey/MS ]
3
ionKey/MS makes assays which were not possible in a routine lab, possible
ionKey and SYNAPT® G2-Si HRMS
Multiple ionization modes
Tof coupled to ion mobility analysis
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:
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® TQ-S and TQ-XSionKey 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
Sensitive, reliable, and compact
Class-leading performance that can fit into any lab
Robust sensitivity driven by proven StepWave ion guide technology
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:
Reduced matrix effects
Increased sampling efficiency
Increased ionization efficiency
75 µm I.D.
150 µm I.D.
300 µm I.D. 2.1 mm I.D.
1 mm I.D.
Flow rate (µL/min)
Sel
ectiv
ity im
prov
emen
t fro
m 2
.1 m
m s
cale
60
50
40
30
20
10
00.1 1 10 100 1000
BALANCING SENSITIVITY AND SPEED
300 µm iKey HTiKey HT (300 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 300 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.
[ ionkey/MS ]
5
min2.00 4.00 6.00 8.00 10.00
%
0
100
2.00 4.00 6.00 8.00 10.00
%
0
100
2.00 4.00 6.00 8.00 10.00
%
0
100
2.00 4.00 6.00 8.00 10.00
%
0
100MRM of 9 Channels ES+ 1165.5>1370.1 (Apidra)
6.32e4Area
4.96;3145
MRM of 9 Channels ES+ 1165.5>1370.1 (Apidra)
3.14e4Area
4.97;1477
MRM of 9 Channels ES+ 1165.5>1370.1 (Apidra)
2.47e4Area
Area
4.96764
MRM of 9 Channels ES+ 1165.5>1370.1 (Apidra)
1.92e45.58
6.846.10
50 pg/mL Apidra
100 pg/mL Apidra
Blank human plasma
25 pg/mL Apidra
150 µm iKey
~41 amol on column at LOD
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.
Comparison of 3 insulin methodsStandard curve range (pg/mL)
Analyte 2.1 x 50 mm 300 µm x 50 mm 150 µm x 100 mmLispro 50–10,000 100–10,000 25–10,000
Glargine 50–10,000 50–10,000 25–10,000Detemir 200–10,000 100–5,000 50–10,000Glulisine 50–10,000 25–10,000 25–10,000Aspart 100–10,000 50–10,000 25–10,000
Sample volume 250 µL 250 µL 100 µLInjection volume 30 µL 15 µL 10 µL
Run time 8 min 7 min 13.5 min
UPLC 2.1 mm■■ Good sensitivity
and run time
iKey 300 µm■■ Fastest run time:
Same sample volume, 1/2 the injection volume, and 2X more sensitive than the 2.1 mm method
iKey 150 µm■■ Greatest sensitivity
(2–4X): Using 2.5X less sample, 1/3 injection volume, 2X longer column, and 1.7X longer run time
Enabling sensitivity and speed
High sensitivity analysis of insulin glusine (Apidra) from 100 µL of human plasma sample, 10 µL injection on the iKey 150 µm.
6
Post Column Addition flexibility
The iKey with Post Column Addition (PCA) allows for adding a modifier solvent post chromatographic separation. This affords the decoupling of the enhancement of the micro electrospray ionization process from the chromatography and offers an additional degree of freedom to the scientist which can lead to significant enhancements to the MS signal.
In this example, the number of charge states on the Waters mAb is increased using PCA addition of methanol:water (50:50):0.1% formic acid (top) to further increase sensitivity. Adding Imidazole (lower post column), the charge state distribution shifts by an average of 19 charges and an increase in charge state spacing by 50%. This spacing affords more spectral space to process complex mAbs and ADC’s where charge states could overlap.
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 10 ng (on-column) IgG that coelutes with SILu MAb. Imidazole PCA made it possible to baseline resolve both charge state distributions. The inset shows the baseline comparison, below, showing the overlapping spectra without decharging.
pKa = 14.5 m/z2000 2250 2500 2750 3000 3250 3500 3750 4000 4250 4500 4750 5000 5250 5500 5750
%
0
100
m/z2000 2250 2500 2750 3000 3250 3500 3750 4000 4250 4500 4750 5000 5250 5500 5750
%
0
100322016 mAb Calibration Curve PCA 2 µL min_6 812 (4.405) 1: TOF MS ES+
1.43e52801.1069
2749.1362
2651.0981
2604.5320
2559.7236
2516.3511
2474.54982394.6401
2854.9636
2910.8782
2914.1023
2972.3525
3029.66993032.8464
3092.65263158.4902
322016 mAb Calibration Curve PCA Imidizole_22 672 (3.648) 1: TOF MS ES+ 2.68e44638.55664365.7407
4241.06054123.2295
4011.8774
3906.1653
3806.12923805.9272
3620.7014
3534.4778
4788.0522
4788.5054
4948.14555118.3882
5123.6880
5124.0981
5301.24415307.2930
-19
PCA addition of MeOH:H2O (50:50) 0.1%FA
PCA addition of MeOH:H2O (50:50) 0.1%FA w/ 1 mg/mL Imidazol
m/z4300 4325 4350 4375 4400 4425 4450 4475 4500 4525 4550 4575 4600 4625 4650 4675 4700
%
0
100
Imidazole PCA (10 ng mAb on-column)
No PCA (10 ng mAb on-column)
m/z2720 2730 2740 2750 2760 2770 2780 2790 2800 2810 2820 2830 2840 2850 2860 2870 2880 2890
%
0
100
R1
R2
R3
R4
Reduction in peak overlap
Charge manipulation can enhance value of the transformation algorithm
m/z2000 2200 2400 2600 2800 3000 3200 3400 3600 3800 4000 4200 4400 4600 4800 5000 5200
%
0
100
m/z2000 2200 2400 2600 2800 3000 3200 3400 3600 3800 4000 4200 4400 4600 4800 5000 5200
%
0
100
Imidazole PCA (10 ng mAb on-column)
No PCA (10 ng mAb on-column)
m/z2500 2550 2600 2650 2700 2750
%
0
100
8
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.
QC reproducibility for one iKey with over 500 injections of protein precipitated plasma.
QC reproducibility across multiple users, ionKey/MS systems, and iKeys.
1 user X 1 system X 1 iKey
5 users X 5 systems X 18 iKeys
Peptide from P00924, yeast enolase6.84 min Retention time 6.69 min
0.01 min SD of retention time 0.11 min
3.01 sec Average peak width FWHM 2.72 sec
retention time RSDretention time RSD
Consistent pressure trace before study and after 1000 x 1 µL injections of 2:1 protein precipitated plasma samples.
Peak area reproducibility over 1000 x 1 µL injections of 2:1 protein precipitated plasma samples.
HUNDREDS OF INJECTIONS, IDENTICAL RESULTS
First 5 µL injection
After 1000 5 µL injections of crashed plasma
min
First injection
Peak area RSD: 5.5%
Last injection
0.50 1.50 2.50 3.50 4.50 5.50 6.50
psi
0
1000
2000
3000
4000
5000
6000
7000
Injection Number
120010008006004002000
14000
12000
10000
8000
6000
4000
2000
0
min3.20 3.24 3.28 3.32 3.36 3.40 3.44 3.48
%
0
100
min3.20 3.24 3.28 3.32 3.36 3.40 3.44 3.48
%
0
100
Quality control injections of dextromethorphan and propranolol showing equivalent chromatographic resolution before study and after 1000 x 5 µL injections of 2:1 protein precipitated plasma.
Proven robustness and reproducibility
[ ionkey/MS ]
Waters offers a complete portfolio of standards, certified vials, and certified containers for your ionKey/MS System. For more information, visit www.waters.com/standards.
9
Ordering InformationPart No. Description Particle Size I.D. Length UOM
iKey Separations Devices
186006764 iKey Peptide BEH C18 130Å 1.7 µm 150 µm 50 mm 1/pkg
186007256 iKey BEH C18 130Å 1.7 µm 150 µm 50 mm 1/pkg
186007557 iKey PCA Peptide BEH C18 130Å 1.7 µm 150 µm 50 mm 1/pkg
186007580 iKey PCA BEH C18 130Å 1.7 µm 150 µm 50 mm 1/pkg
186006766 iKey Peptide BEH C18 130Å 1.7 µm 150 µm 100 mm 1/pkg
186007258 iKey BEH C18 130Å 1.7 µm 150 µm 100 mm 1/pkg
186006969 iKey Peptide BEH C18 300Å 1.7 µm 150 µm 50 mm 1/pkg
186006970 iKey Peptide BEH C18 300Å 1.7 µm 150 µm 100 mm 1/pkg
186006765 iKey Protein BEH C4 300Å 1.7 µm 150 µm 50 mm 1/pkg
186006968 iKey Protein BEH C4 300Å 1.7 µm 150 µm 100 mm 1/pkg
186007257 iKey Peptide CSH C18 130Å 1.7 µm 150 µm 50 mm 1/pkg
186007244 iKey CSH C18 130Å 1.7 µm 150 µm 50 mm 1/pkg
186007259 iKey Peptide CSH C18 130Å 1.7 µm 150 µm 100 mm 1/pkg
186007245 iKey CSH C18 130Å 1.7 µm 150 µm 100 mm 1/pkg
186007260 iKey HSS T3 100Å 1.8 µm 150 µm 50 mm 1/pkg
186007261 iKey HSS T3 100Å 1.8 µm 150 µm 100 mm 1/pkg
186008725 iKey Peptide BEH C18 130Å 1.7 µm 300 µm 50 mm 1/pkg
186008726 iKey Peptide CSH C18 130Å 1.7 µm 300 µm 50 mm 1/pkg
186008727 iKey HSS T3 100Å 1.8 µm 300 µm 50 mm 1/pkg
Trap Columns
186007498 ACQUITY UPLC M-Class Symmetry C18 Trap Column, 100Å 5 µm 300 µm 50 mm 1/pkg
186007471 ACQUITY UPLC M-Class Peptide BEH C18 Trap Column, 130Å 5 µm 300 µm 50 mm 1/pkg
186008029 ACQUITY UPLC M-Class HSS T3 Trap Column, 100Å 5 µm 300 µm 50 mm 1/pkg
186008470 ACQUITY UPLC M-Class BEH C4 Trap Column, 300Å 5 µm 300 µm 50 mm 1/pkg
Utility iKey
186007049 iKey Infusion Device n/a 85 µm 50 mm 1/pkg
186007051 iKey Flow Injection Analysis Device n/a 85 µm 50 mm 1/pkg
186008450 iKey Diagnostic Device V3 n/a n/a n/a 1/pkg
[ ionkey/MS ]
7
Choose your chemistries
WATERS GLOBAL SERVICES
Delivering world renowned services and supportWaters 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.
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 2001 for providing best-in-class technical knowledge, issue resolution, and process support.
Hybrid particles Silica-based particles
130Å 300Å 300Å 300Å1.7 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 (80 °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 silica-based selectivities are desired.
■■ High retentivity for polar organic compounds and metabolites
■■ Balanced retention of polar and hydrophobic analytes
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: www.waters.com/ionKey
For your local sales office, please visit www.waters.com/contact
Waters Corporation 34 Maple Street Milford, MA 01757 U.S.A. T: 1 508 478 2000 F: 1 508 872 1990 www.waters.com
www.waters.com/ionKey
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.
©2017 Waters Corporation. Printed in the U.S.A.December 2017 720006092EN LM-SIG
Sales Offices:
Australia 61 2 9933 1777
Austria 43 1 877 18 07
Belgium and Luxembourg 32 2 726 1000
Brazil 55 11 4134 3788
Canada 1 800 252 4752
China 86 21 6156 2666
Czech Republic 420 2 617 11384
Denmark 45 46 59 8080
Finland 358 9 5659 6288
France 33 1 30 48 72 00
Germany 49 6196 400 600
Hong Kong 852 2964 1800
Hungary 36 1 350 5086
India 91 080 49292200 03
Ireland 353 1 448 1500
Israel 9723 3731391
Italy 39 02 265 0983
Japan 81 3 3471 7191
Korea 82 2 6300 9200
Malaysia 603 7841 1688
Mexico 52 55 52 00 1860
The Netherlands 31 76 508 7200
Norway 47 6 384 6050
Poland 48 22 101 5900
Portugal 351 21 893 61 77
Puerto Rico 1 787 747 8445
Singapore 65 6593 7100
Spain 34 93 600 9300
Sweden 46 8 555 115 00
Switzerland 41 56 676 7000
Taiwan 886 2 2508 5500
UAE 971 4 214 62 38
UK 44 208 238 6100
US 1 800 252 4752
