Determination of tetramethylpyrazine in animal serum and cerebrospinal fluid by high performance liquid chromatography

A reversed\|phase high performance liquid chromatographic( RP HPLC ) method for the determination of the tetramethylpyrazine(TMP) in Chuanxiong extract, the animal(mouse) serum and cerebrospinal fluid has been developed. The TMP was separated on an ODS column Zorbax SB C 18 (4.6 mm i.d.×250 mm,5 μm ) at room temperature and detected by using UV detector at 270 nm. The mobile phase was methanol water ( 50∶50, V/V ) containing 0.2 mmol/L of NH 4H 2PO 4 flowing at a rate of 0.8 mL/min and 20 μL samples were injected. The detection limit of TMP was 1 mg/L and the calibration curve is linear between 5 and 500 mg/L with a correlation coefficient (r) of 0.999. The recovery of TMP ranged 98% 103%. The extract of Chuanxiong and pretreated serum and cerebrospinal fluid sample are stable for a week at room temperature.