Matthew Arnold (Presenter)
Gerstel Inc.
Bio: Matthew Arnold received a degree in Chemistry/Biology minor from the University of San Diego. He worked for 5 years developing analytical HPLC and GCMS methods to support the pharmaceutical development teams of Metabasis Therapeutics and Isis Pharmaceuticals. After earning an MBA at the University of Southern California he joined GERSTEL as the West Coast Regional Representative.
Authorship: Fred Foster, John Stuff, Edward Pfannkoch and Matthew Arnold
GERSTEL, Inc., 701 Digital Dr. Suite J, Linthicum, MD 21090, USA
| Short Abstract In this report, the complete automation of dried blood spot (DBS) analysis is demonstrated. DBS cards are inserted automatically into a flow through cell in which individual blood spots are rapidly and effectively desorbed. The DBS elution is integrated into a complete cleanup and analysis system using online SPE with replaceable cartridges combined with automated injection to an LC/MS/MS system. Automated DBS extraction methods were optimized for a variety of analytes from rat and bovine blood. The resulting precision and accuracy data are provided. |
Long Abstract
Introduction
The extraction of dried blood spots (DBS) typically involves manual intervention. First, a small disc is punched out of the center of a dried blood spot placed on a DBS card. Following solvent extraction of the sample, it is also common to include further cleanup steps, using solid phase extraction (SPE) to improve detection limits or exchanging solvents for compatibility with subsequent chromatographic separations. Modern analytical labs are looking to automate the process to help reduce solvent usage and to increase sample throughput while ensuring the high quality of the resulting data.
In this report, the complete automation of dried blood spot analysis is demonstrated and the results evaluated. A novel autosampler automatically inserts DBS cards into a flow through cell in which individual blood spots are rapidly and effectively desorbed. The DBS elution is integrated into a complete cleanup and analysis system using online SPE with replaceable cartridges combined with automated injection to an LC/MS/MS system. Automated DBS extraction methods were optimized for a variety of analytes from rat and bovine blood.
Methods
Automated DBS extraction and cleanup was performed with a GERSTEL MPS 2 autosampler with DBSA and SPExos options. Separation of the drug compounds was performed using an Agilent 1290 HPLC with a Poroshell 120, EC-C18, (3.0 x 50mm, 2.7um) column. Analysis of target compounds was performed on an Agilent 6460 Triple Quadrupole Mass Spectrometer with Jet stream electrospray source.
Results
Drug compounds of veterinary interest (ketamine, amitriptyline and ketoprophen) were successfully extracted from dried blood spot samples using an automated DBS-SPExos procedure coupled to LC/MS/MS analysis using the Agilent 6460 Triple Quadrupole Mass Spectrometer.
Linear calibration curves were established using both dried rat blood and dried bovine blood resulting in R2 values 0.99 or greater.
The DBS-SPExos-LC/MS/MS method proved to be accurate and precise. For dried rat blood spots, accuracy data averaged 91.7% (range: 82.6% - 104%) and precision data averaged 5.75%CV (range: 2.47% -11.5%) for all compounds analyzed. For dried bovine blood spots, accuracy data averaged 108% (range: 98.6% - 119%) and precision data averaged 5.31%CV (range: 0.288% -10.2%) for all compounds analyzed.
References & Acknowledgements:
| Description | Y/N | Source |
| Grants | no | |
| Salary | yes | Gerstel Inc. |
| Board Member | no | |
| Stock | no | |
| Expenses | no |
IP Royalty: no
| Planning to mention or discuss specific products or technology of the company(ies) listed above: | yes |