A novel sequential electrostatic-HILIC SPE within a quantitative method for bivalirudin in human plasma by LC-SRM.
OBJECTIVE
This work set out to realize an idea for a novel means of extracting the peptide therapeutic bivalirudin from human plasma in what would be a uniquely selective means of SPE, a mixed-mode protocol involving electrostatic interactions followed by HILIC.


RESULTS
Inter and intra-assay relative error ranged from 3.52 to 8.23%, and 2.37 to 6.90%, respectively. Inter and intra-assay precision ranged from 2.64 to 7.12%, and 0.855 to 2.90%, respectively. Recoveries of 80% were attained, and there was no hint of discernible manifestation of matrix effects.


CONCLUSIONS
The method was shown to perform excellently in the assessment tantamount to method validation. The essence of the extraction method presents a new option for highly selective extraction of peptides from biological matrices.
Department of Bioanalytical Services, Envigo, Mettlers Road, East Millstone, NJ 08875-2360, USA.
Robert
R
MacNeill
Department of Bioanalytical Services, Envigo, Mettlers Road, East Millstone, NJ 08875-2360, USA.
Vishva
V
Acharya
Department of Bioanalytical Services, Envigo, Mettlers Road, East Millstone, NJ 08875-2360, USA.
Neha
N
Patel
Department of Bioanalytical Services, Envigo, Mettlers Road, East Millstone, NJ 08875-2360, USA.
Gregory
G
Parisi
Department of Bioanalytical Services, Envigo, Mettlers Road, East Millstone, NJ 08875-2360, USA.
Kim
K
Katto
Department of Bioanalytical Services, Envigo, Mettlers Road, East Millstone, NJ 08875-2360, USA.
Robert
R
Lovich
D001774:Blood Chemical Analysis / Q000295:instrumentation / Q000379:methods*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D006629:Hirudins / Q000097:blood* / Q000302:isolation & purification / Q000592:standards; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D010446:Peptide Fragments / Q000097:blood* / Q000302:isolation & purification / Q000592:standards; D011994:Recombinant Proteins / Q000097:blood / Q000302:isolation & purification / Q000592:standards; D052616:Solid Phase Extraction; D055672:Static Electricity; D053719:Tandem Mass Spectrometry / Q000592:standards*
D006629:Hirudins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C074619:bivalirudin
bivalirudin; electrostatic; hydrophilic