A Simple Gas Chromatographic Method for the Simultaneous Determination and Pharmacokinetic Study of Tetramethylpyrazine Phosphate and Borneol in Mouse Plasma and Brain Tissue After Oral Administration of the Fufang Tetramethylpyrazine Phosphate Tablets
Journal of Chromatographic Science
A Simple Gas Chromatographic Method for the Simultaneous Determination and Pharmacokinetic Study of Tetramethylpyrazine Phosphate and Borneol in Mouse Plasma and Brain Tissue After Oral Administration of the Fufang Tetramethylpyrazine Phosphate Tablets
Xiao Yan-yu 0
Ping Qi-neng 0
Chen Zhi-peng 0
0 Department of Pharmacy, China Pharmaceutical University , 210009 Nanjing , China
A rapid, sensitive, and simple gas chromatographic method with flame ionization detection is developed for the simultaneous determination of tetramethylpyrazine phosphate (TMPP) and borneol in mouse plasma and brain tissue. Sample preparations are carried out by deproteinization with an internal standard solution in methanol. The analytes and internal standard (dimethyl sulfoxide) are well-separated on an HP-5 MS capillary column. The analytical curves are linear over a wide concentration range of 0.02-40 μg/mL for both TMPP and borneol in plasma and brain tissue, with the intra- and inter-day precision (the relative standard deviation values) at less than 15%. TMPP and borneol are both stable under different conditions. The method described is successfully applied to the pharmacokinetic study of mouse plasma and brain tissue after oral administration of the Fufang TMPP and TMPP tablets to mice.
Introduction
Tetramethylpyrazine phosphate (TMPP) (Figure 1A) has
been widely used in China for the treatment of
cardiovascular and cerebrovascular disorders (
1–3
). It has been
found to block calcium channels, reduce the bioactivity of
platelets and platelet aggregation, inhibit free radicals,
increase cerebral blood flow (
4–6
), and improve blood
viscosity (
7
). However, TMPP possesses a low oral
bioavailability (10–30%) after oral administration, due to hepatic
first pass metabolism (
8
).
Borneol (Figure 1B and 1C), a monoterpenoid alcohol, is
isolated from Fructus amoni. Recently, it has been reported
that borneol can increase the oral bioavailability of TMPP and
enhance its distribution in brain tissue. To increase oral
bioavailability of TMPP and to enhance its distribution in brain
tissue, the Fufang TMPP tablets containing TMPP and borneol
were developed for the treatment of cerebral thrombosis.
Therefore, it was necessary to establish a sensitive and simple
method to simultaneously determinate TMPP and borneol in
biological fluids.
To date, several analytical methods have been developed
for the determination of tetramethylpyrazine (TMP) and
borneol in biological fluids. Peng et al. (
9
) determined the
concentrations of TMP and its metabolite in dog plasma using
a liquid chromatographic–mass spectrometric (LC–MS)
method. Tung-Hu et al. (
10
) researched the pharmacokinetics
of TMP in rat blood and brain using microdialysis. Liang et al.
(
11–12
) measured the pharmacokinetics of TMP in rat blood
and regional brain tissue by high-performance LC with UV
detection, and also studied the pharmacokinetics of borneol in
rat blood and brain tissue by gas chromatography (GC). Guo
et al. (13) studied the simultaneous determination of TMP
and bornoel in human plasma, administrated with Suxiao
Jiuxin Wan. But most of those methods require complicated
sample preparation procedures involving basification of the
plasma sample, extraction, acidification, preconcentration,
and reconstitution of the analytes. These procedures are rather
time-consuming and more complex than the procedure
described in this paper. With this method, samples using
dimethyl sulfoxide as an internal standard could be applied to
GC with flame ionization detection (FID) with simple
deproteinization and without multiple pretreatment steps.
The present method was also successfully applied to the
pharmacokinetic study of mouse plasma and brain tissue after
oral administration of the Fufang TMPP tablets and TMPP
tablets to mice.
Experimental
Materials
Tetramethylpyrazine phosphate (TMPP, 99.8% purity) was
purchased from Beijing Yanjing Pharmaceutical Factory
(Beijing, China). Borneol (99.4% purity) and dimethyl sulfoxide
(internal standard, I.S.) was obtained from Shanghai Chemical
Reagent Co. (Shanghai, China). The other chemical reagents
were of analytical grade or better.
The reference formulation consisted of TMPP tablets
purchased from Beijing Yanjing Pharmaceutical Factory
(Beijing, China). Each tablet contains 50 mg of TMPP. The
Fufang TMPP tablets were prepared in our laboratory. Each
tablet contains 50 mg of TMPP and 40 mg of borneol.
Instrument and chromatographic conditions
The GC–FID system consisted of an HP 4890 gas
chromatographic system, an FID (Agilent 4890, Palo Alto, CA), and
Agilent Chemstation software for data analysis. Separation of
TMPP, borneol, and I.S. from endogenous substances was
performed on an HP-5 MS capillary column (30.0 m × 250 μm,
A
B
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