The development and assessment of high-throughput mass spectrometry-based methods for the quantification of a nanoparticle drug delivery agent in cellular lysate.
The safe use of lipid-based drug delivery agents requires fast and sensitive qualitative and quantitative assessment of their cellular interactions. Many mass spectrometry (MS) based analytical platforms can achieve such task with varying capabilities. Therefore, four novel high-throughput MS-based quantitative methods were evaluated for the analysis of a small organic gene delivery agent: N,N-bis(dimethylhexadecyl)-1,3-propane-diammonium dibromide (G16-3). Analysis utilized MS instruments that detect analytes using low-resolution tandem MS (MS/MS) analysis (i.e. QTRAP or linear ion trap in this work) or high-resolution MS analysis (i.e. time of flight (ToF) or Orbitrap). Our results indicate that the validated fast chromatography (FC)-QTRAP-MS/MS, FC- LTQ-Orbitrap-MS, desorption electrospray ionization-collision-induced dissociation (CID)-MS/MS and matrix assisted laser desorption ionization-ToF/ToF-MS MS methods were superior in the area of method development and sample analysis time to a previously developed liquid chromatography (LC)-CID-MS/MS. To our knowledge, this is the first evaluation of the abilities of five MS-based quantitative methods that target a single pharmaceutical analyte. Our findings indicate that, in comparison to conventional LC-CID-MS/MS, the new MS-based methods resulted in a (1) substantial reduction in the analysis time, (2) reduction in the time required for method development and (3) production of either superior or comparable quantitative data. The four new high-throughput MS methods, therefore, were faster, more efficient and less expensive than a conventional LC-CID-MS/MS for the quantification of the G16-3 analyte within tissue culture. When applied to cellular lysate, no significant change in the concentration of G16-3 gemini surfactant within PAM212 cells was observed between 5 and 53 h, suggesting the absence of any metabolism/excretion from PAM212 cells.
College of Pharmacy and Nutrition, University of Saskatchewan, 110 Science Place, Saskatoon, SK, S7N 5C9, Canada.
Joshua
J
Buse
Randy W
RW
Purves
Ronald E
RE
Verrall
Ildiko
I
Badea
Haixia
H
Zhang
Christopher C
CC
Mulligan
Kerry M
KM
Peru
Jonathan
J
Bailey
John V
JV
Headley
Anas
A
El-Aneed
D000818:Animals; D002457:Cell Extracts / Q000737:chemistry; D002460:Cell Line; D002853:Chromatography, Liquid / Q000379:methods; D016503:Drug Delivery Systems / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D016014:Linear Models; D051379:Mice; D053758:Nanoparticles / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D004364:Pharmaceutical Preparations / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014162:Transfection
D002457:Cell Extracts; D004364:Pharmaceutical Preparations
desorption electrospray ionization; drug delivery; fast chromatography; gemini surfactants; liquid chromatography; mass spectrometry; matrix assisted laser desorption ionization; quantification