Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume 45,
Number 2, February 2007, pp.91-96
Rapid and Sensitive Method for the Determination
of Sibutramine Active Metabolites in Human Plasma by Reversed-Phase
Liquid Chromatography–Tandem Mass Spectroscopy
Jignesh Bhatt[1,2], Bhavin Shah[1], Sandeep Kambli[1],
Gunta Subbaiah[1], Sadhana Singh[2], and Suresh Ameta[3]
[1]Torrent Research Centre, Gandhinagar-382428, Gujarat State,
India;
[2]Department of Chemistry, B.N.P.G. College, Mohanlal Sukhadia
University, Udaipur, Rajasthan State, India; and
[3]Department of Chemistry, College of science, Mohanlal Sukhadia
University, Udaipur, Rajasthan State, India
A new, rapid, and sensitive liquid chromatography–tandem
mass spectrometry method is developed and validated to quantitate
the sibutramine active metabolites mono desmethyl sibutramine
(M1) and di-desmethyl sibutramine (M2) using imipramine as the
internal standard in human plasma samples for routine bioequivalence
studies. The method involves rapid solid-phase extraction from
plasma, eliminating the drying and reconstitution steps. The analytes
are chromatographed on a C8 reversed-phase chromatographic column
and analyzed by mass spectrometry in the multiple reaction monitoring
mode, which enables a quantitation limit at the sub-nanogram level.
The method has a chromatographic run time of 2.8 min. The proposed
method is validated with a linear range of 0.1–8.0 and 0.2–16.0
ng/mL for M1 and M2, respectively, with a correlation coefficient
of regression ≥ 0.9990. The method is sensitive and reproducible,
having intra- and inter-assay precision at the lower limit of
quantitation (0.1 ng/mL for M1 and 0.2 ng/mL for M2) < 10.0%.
The overall recovery for M1 and M2 is 93.5% and 77.9%, respectively.
The method has been applied to a bioequivalence clinical study
with great success.
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