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Quantitation of donepezil and its active metabolite 6-O-desmethyl donepezil in human plasma by a selective and sensitive liquid chromatographytandem mass spectrometric method
2nd International Conference on Analytical & Bioanalytical Techniques
A sensitive and selective LC-MS/MS assay for the simultaneous determination of
donepezil (D) and its pharmacologically active metabolite, 6-
O
-desmethyl donepezil
(ODD) in human plasma is developed using galantamine as internal standard (IS). Th
e
analytes and IS were extracted from 500 μL aliquots of human plasma via solid phase
extraction on Waters Oasis HLB cartridges. Chromatographic separation was achieved
in a run time of 6.0 min on a Waters Novapak C18 (150 mm x 3.9 mm, 4 μm) column
under isocratic conditions. Detection of analytes and IS was done by tandem mass
spectrometry, operating in positive ion and multiple reaction monitoring (MRM)
acquisition mode. Th
e protonated precursor to product ion transitions monitored for
D, ODD and IS were at m/z 380.1 → 91.2, 366.3 → 91.3 and 288.2 → 213.2 respectively.
Th
e method was fully validated for its selectivity, interference check, sensitivity, linearity,
precision and accuracy, recovery, matrix eff
ect, ion suppression/enhancement, cross
specifi
city, stability and dilution integrity. A linear dynamic range of 0.10-50.0 ng mL
-1
for D and 0.02-10.0 ng mL
-1
for ODD was evaluated with mean correlation coeffi
cient
(r) of 0.9975 and 0.9985 respectively. Th
e intra-batch and inter-batch precision (%CV)
across fi
ve validation runs (LLOQ, LQC, MQC, HQC and ULOQ) was less than 7.5 % for
both the analytes. Th
e method was successfully applied to a bioequivalence study of 10 mg
donepezil tablet formulation in 24 healthy Indian male subjects under fasting condition.
Biography
Dr. Bhavin kumar patel has completed his PhD at the age of 31 from School of science, Gujarat
University, is currently working as a Senior Research Scientist-I (project manager) in BA Research India
Ltd. Authored Eight Scienti
fi
c Papers in Reputed International Journals along with Mr. Naveen Sharma,
Dr. (Mrs.) Mallika Sanyal and Dr. Pranav S. Shrivastav.
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