Liquid Extraction Surface Analysis
(LESA)
Analyzing whole body animal sections
Daniel Eikel Advion BioSciences
Lecture 5, Page 60
Outline LESA is a new surface sampling technology based on liquid extraction (Liquid Extraction Surface Analysis) and nano-electrospray mass spectrometry (nESI-MS)
Example 1: LESA drug distribution profiling from whole body mouse sections: Terfenadine and its metabolite Fexofenadine
Lecture 5, Page 61
Introduction to LESA Liquid Junction Surface Sampling
Mass
spectrometer
Sample
Solvent
Sampling tip
Aspirate solvent Dispense solvent on sample Aspirate sample solution Transfer sample Spray sample
Nozzle
96- or 384-well plate
Side view Lecture 5, Page 62
Introduction to LESA – Schematic Workflow –
Step 1 – Solvent Delivery
Disposable Tip picks up extraction solvent at reservoir
Step 2 – Analyte Extraction
Robot places extraction solvent on target and initiates aspirate/dispense
cycles for analyte extraction
Step 3 – Analyte Ionization
Robot aspirates extracted analytes from target and initiates electrospray at a
400 nozzle nESI chip
Chip-based nano ESI
Kertesz V. and Van Berkel GJ:
Fully automated liquid extraction-based surface sampling and ionization using a chip-based
robotic nano electrospray platform. Journal of Mass Spectrometry 2010 45(3) p252-260 Lecture 5, Page 63
Introduction to LESA – Schematic Workflow –
LESA enabled Advion TriVersa NanoMate
Nozzle dimensions A chip:
5.5 m ID x 28 m OD x 55 m height
Lecture 5, Page 64
Example 1: LESA drug distribution profiling from whole body mouse sections: Terfenadine and its metabolite, Fexofenadine
Eikel D, Vavrek M, Korfmacher WA and Henion JD: Liquid Extraction Surface Analysis Mass Spectrometry (LESA-MS) as a Novel Profiling Tool for Drug Distribution and Metabolism Analysis: The Terfenadine Example
Lecture 5, Page 65
Example 1 - Background Information -
Terfenadine and its active metabolite, Fexofenadine, are both antihistamines
Animal experiment:
50 mg/kg PO dosed mice put down in CO2, frozen and sectioned 1 and 4 h post dose
MS experiment:
2 min Infusion
Q3 survey MS/MS and SRM analysis on AB/Sciex 5500 Qtrap
or
Exact mass experiment on Thermo Exactive Benchtop Orbitrap
Lecture 5, Page 66
Whole Body Distribution Analysis
Liver Kidney
Cecum of Intestine
Stomach
Spleen
Brain Skeletal muscle
Small Intestine
Lecture 5, Page 67
Lecture 5, Page 68
LESA spatial resolution
0.8 mm
0.8 mm
1.1 mm
0.9 mm
Lecture 5, Page 69
Example of Single Organ Profiling
Brain
Lecture 5, Page 70
Terfenadine MALDI-MS
1 h post dose
LESA comparison to other ‘imaging’ technologies (MALDI-MS comparison data, Chen et al. 2008)
Fexofenadine MADLI-MS
Optical microscope pictures
from mouse tissue
Lecture 5, Page 71
LESA Comparison to other ‘imaging’ technologies (Whole Body Autoradiography comparison data, Leeson et al. 1982)
Lecture 5, Page 72
Comparing different Analysis Approaches regarding Sensitivity
14C-terfenadine* 10 mg/kg PO rat
[Ref. 22]
MALDI-IMS 50 mg/kg PO mouse
[Ref. 23]
LESA-MS-Profiling 50 mg/kg PO mouse
(Orbitrap-Exact Mass)
LESA-MS-Profiling 50 mg/kg PO mouse
(QQQ-SRM)
Terfenadine Fexofenadine Terfenadine Fexofenadine Terfenadine Fexofenadine
Blood
Brain
Liver
Lung n.a. n.a. n.a. n.a.
Kidney
Spleen
Skeletal Muscle n.a.
Stomach
Cecum of Intestine
Small Intestine
Large Intestine
Lecture 5, Page 73
Summary Example 1
LESA …
… a profiling technique with a maximum spatial resolution of 1 mm on target
… can analyze drug distribution within a single organ or within a whole body
… is at least 20-fold more sensitive than MALDI-imaging MS and only ca. 5 fold less sensitive than QWBA in the given Terfenadine/ Fexofenadine example
Lecture 5, Page 74
References
• Sheerin Khatib-Shahidi, Malin Andersson, Jennifer L. Herman, Todd A. Gillespie, and Richard M. Caprioli, Anal. Chem., 2006, 78 (18), pp 6448–6456
• Hsieh et al. RCMS, 20, 965-72 (2006).
• Angus C. Grey, Pierre Chaurand, Richard M. Caprioli and Kevin L. Schey, J. Proteome Res., 2009, 8 (7), pp 3278–3283
• Nemes and Vertes, Anal. Chem., 2007, 79, 8098.
• D. Eikel and J. D. Henion, Liquid Extraction Surface Analysis (LESA) of Food Surfaces Employing Chip-Based Nano Electrospray Mass Spectrometry, RCMS 2011. Rapid Commun. Mass Spectrom. 2011, 25, 1–10.
• Girod, M.; Shi, Y.; Cheng, J.-X.; Cooks, R. G., Desorption Electrospray Ionization Imaging Mass Spectrometry of Lipids in Rat Spinal Cord. Journal of the American Society for Mass Spectrometry 2010, 21 (7), 1177-1189.
• Li, H.; Hummon, A. B., Imaging Mass Spectrometry of Three-Dimensional Cell Culture Systems. Analytical Chemistry 2011, 83 (22), 8794-8801.
• Luo, G.; Marginean, I.; Vertes, A., Internal Energy of Ions Generated by Matrix-Assisted Laser Desorption/Ionization. Analytical Chemistry 2002, 74 (24), 6185-6190.
. Lecture 5, Page 75