![Page 1: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/1.jpg)
Unlocking the Benefits of MicroLC with AB SCIEX and Eksigent Technology
Anthony Romanelli, Sr. Applications Manager
![Page 2: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/2.jpg)
2 © 2014 AB SCIEX
What is Micro Flow LC?
0.5 mm
area = 18 area = 1area = 85
0.5 mm
area = 18 area = 1area = 85
HPLC using columns with inner diameter (ID) ≤ 1 mm
Typical Flow Rates: 5-200µL/min
Column diameter (mm) 0.3 0.5 1.0 2.1
Typical flow rate (mL/min) 10 35 140 600
![Page 3: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/3.jpg)
3 © 2014 AB SCIEX
• Improve throughput
• Improve sensitivity (LOQ)
• Consume less sample
• Reduce LC and MS downtime
• Reduced costs
Why Use Micro LC?
![Page 4: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/4.jpg)
4 © 2014 AB SCIEX
microLC 200plus System
• Microflow UHPLC (10,000 psi)
• System delay (void) volume: 1 – 3 µL
• Total system volume: ~ 4.5 µL
• Supports 1.0, 0.5, and 0.3 mm ID columns
• Flow rates 5 – 200 µL/min
• Programmable injection - from 15 nL
(metered) to 10 µL (without changing loop)
• Compact, Close coupling with MS source
- Small volume hybrid electrodes for Turbo VTM
and IonDrive™ Turbo V sources
Compact, high-throughput UHPLC system for LC/MS
![Page 5: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/5.jpg)
5 © 2014 AB SCIEX
Electronically
Controlled
Pressure Source
Flow Meter
Control
Processor
Microfluidic Flow Control™ (MFC)
Microfluidic Flow Control Precise, stable flow for reproducible high-speed chromatography
• Proprietary Microfluidic Flow Control (MFC)
technology for rapid flow adjustment
• Continuous flow monitoring
• Operates up to 10,000 psi
• MFC and high pressure capability combine to
ensure precise and stable flow for reproducible,
high-speed chromatography
![Page 6: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/6.jpg)
6 © 2014 AB SCIEX
MicroLC for Exenatide Quantitation
Assay Development Goals:
• Achieve sensitivity levels that enable
quantitation in plasma when slow release
exenatide is administered.
• Develop a selective, robust, and
reproducible method
• Demonstrate the ability to quantitate over a
wide dynamic range
QTRAP® 6500 System Nano LC400 System
IonDrive™ ion source microflow electrodes
![Page 7: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/7.jpg)
7 © 2014 AB SCIEX
MicroLC for Exenatide Quantitation
Samples prepared by spiking standards into a crashed rat plasma matrix
![Page 8: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/8.jpg)
8 © 2014 AB SCIEX
Stable Isotope Labeled Universal Monoclonal Antibody Standard
XIC chromatogram of signature peptide transitions in plasma matrix; SILUMab Standard in the insight
Heavy Chain DTLMISR
Heavy Chain FNWYVDGVEVHNAK
Heavy Chain VVSVLTVLHQDWLNGK
Light Chain YAASSYLSLTPEQWK
Light Chain AGVETTTPSK
![Page 9: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/9.jpg)
9 © 2014 AB SCIEX
Quantitation of Signature Peptide AGEVETTPSK (light chain: +2y6 light)
• Excellent accuracy and precision over 4 orders of linear dynamic range
• Improved sensitivity compared to previous high flow data
• Over a 2 fold increase in throughput
LOQn=1
LOQn=2
LOQn=3
![Page 10: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/10.jpg)
10 © 2014 AB SCIEX
J Proteome Res. 2014 Apr 4;13(4):1905-10
Peptide Quantitation with Micro Flow Publication
![Page 11: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/11.jpg)
11 © 2014 AB SCIEX
MicroLC for Lipid Profiling
![Page 12: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/12.jpg)
12 © 2014 AB SCIEX
MicroLC for Lipid Profiling - Chromatography
Improvement in chromatographic resolution with the microLC 200
![Page 13: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/13.jpg)
13 © 2014 AB SCIEX
MicroLC for Lipid Profiling - Sensitivity
Significant gains
in sensitivity
achieved with
only 1/10th the
amount of
analyte loaded
on column
![Page 14: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/14.jpg)
14 © 2014 AB SCIEX
MicroLC for Lipid Profiling - Reproducibility
Excellent reproducibility was achieved in regards to retention time,
chromatographic resolution, and signal intensity.
![Page 15: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/15.jpg)
15 © 2014 AB SCIEX
Clin/Tox Robustness - Long Term Retention Time Stability
Retention time plot of quality control sample over 1000 injections in
pooled human urine
Guard column change (every ~250 injections)
![Page 16: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/16.jpg)
16 © 2014 AB SCIEX
Pooled 8 Human Urine
10 ng/mL
Injection #20
1st Guard
Pooled 8 Human Urine
10 ng/mL
Injection #1000
4th Guard
Pooled 8 Human Urine
10 ng/mL
Injection #1001
No Guard
Clin/Tox Long Term Robustness – Column Performance and Peak Shape
Note: Micro flow method increased throughput ~2x
![Page 17: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/17.jpg)
17 © 2014 AB SCIEX
• Improve throughput
• Improve sensitivity (LOQ)
• Consume less sample
• Reduce LC and MS downtime
• Reduced costs
Benefits of Micro LC with AB SCIEX and Eksigent Technology
![Page 18: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/18.jpg)
18 © 2014 AB SCIEX
Trademarks/Licensing
• For Research Use Only. Not for use in diagnostic procedures.
• SCIEX and Eksigent are part of AB SCIEX. The trademarks
mentioned herein are the property of AB Sciex Pte. Ltd. or their
respective owners. AB SCIEX™ is being used under license.
• © 2014 AB SCIEX. All rights reserved. Information subject to change
without notice.
![Page 19: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/19.jpg)
Questions and Answers
![Page 20: AB SCIEX Presentation · AB SCIEX Presentation Author: Microsoft account Created Date: 7/1/2014 11:31:26 AM](https://reader035.vdocuments.mx/reader035/viewer/2022071019/5fd2bb86bdf1b719d7175764/html5/thumbnails/20.jpg)
20 © 2013 AB SCIEX
AB SCIEX and Eksigent
What Can be Achieved When Technology Challenges Science