Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume
46, Number 5, May/June 2008, pp. 445-449
Sensitive HPLC–ESI-MS Method for the Determination
of Tiotropium in Human Plasma
Li Ding1, Wenming Tan1,
Yindi Zhang2, Jianping
Shen2, and Zhengxing Zhang1
1Department of Pharmaceutical Analysis, China Pharmaceutical
University, 24 Tongjiaxiang, Nanjing 210009, China and
2Organization
for State Drug Clinical Trial, The First Hospital Affiliated
to Nanjing Medical University, Nanjing 210029, China
A sensitive high-performance liquid chromatography–electrospray
ionization-mass spectrometry (HPLC–ESI-MS) assay is established
for the determination of tiotropium in human plasma using benzyltriethylammonium
chloride as the internal standard (IS). After being treated with
C18 cartridges, plasma samples are separated by HPLC
on a reversed-phase C18 column with a mobile phase
of 40mM ammonium acetate buffer–methanol
(56:44, v/v). Tiotropium is determined in a single-quadrupole
MS. HPLC–ESI-MS is performed in the selected ion monitoring
mode using target ions at m/z 392.0 for tiotropium and
m/z 192.3 for the IS. The calibration curve is linear
over the range 1.5–30
pg/mL. The intra- and inter-assay variability values are less
than 10.1% and 13.6%, respectively. The mean plasma extraction
recovery of tiotropium is 92.3 ± 5.0%. The method has
been successfully applied to studying the pharmacokinetics of
tiotropium in healthy Chinese volunteers.
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