ligustilide and tetramethylpyrazine

An automated on-line SPE and innovative fast polarity switch bioanalysis method employing dual-gradient liquid chromatography (DGLC) coupled with mass spectrometry (DGLC-MS/MS) was established and validated for the simultaneous determination of ferulic acid, ligustrazine and ligustilide in rat plasma after administration of Rhizoma Chuanxiong, Angelica sinensis extract or monomer. The proteins in plasma samples were precipitated using acetonitrile: methanol (1:1, v/v). Sulfamethoxazole was used as an internal standard. The DGLC system contains two high-pressure pumps. The first pump was used for on-line solid phase extraction with a Cyclone™ SPE column. Chromatographic separations were performed with the other pump on a Syncronis C18 rapid analytical column. The analytical column was eluted by a gradient program that featured an acetonitrile/methanol/water gradient (flow-rate, 0.4ml/min). DGLC afforded greater convenience for bioanalysis. All analytes were simultaneously monitored in positive- and negative-ion mode by SRM (selective reaction monitoring) using the fast polarity switch speed of TSQ Vantage™. Method validation of the assay was implemented. No significant matrix effect was observed. The LLOQ of all analytes were <1.0ng/ml. The precision, recovery and linearity of the analysis met the pre-established requirements. The method was applied to the pharmacokinetics of ferulic acid, ligustrazine and ligustilide in Rhizoma Chuanxiong or Angelica sinensis extracts or monomers.

Topics: 4-Butyrolactone; Absorption; Administration, Oral; Animals; Automation; Calibration; Chromatography, High Pressure Liquid; Chromatography, Liquid; Coumaric Acids; Male; Medicine, Chinese Traditional; Plant Extracts; Pressure; Pyrazines; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Sulfamethoxazole; Tandem Mass Spectrometry

To develop a RP-HPLC method for the simultaneous determination of five components including ligustrazine, ferulic acid, butylphthalide, ligustilide and butene-NBP in Rhizoma Chuanxiong, a Chinese herbal medicine.. The chromatographic analysis was carried out by using a grace smart RP C18 column with the mobile phase consisted of acetonitrile and 0.1% phosphoric acid (volume fraction), which was in gradient elution. A DAD detector was used to detect the components. The peak area was chosen under their maximum absorption wavelength for different components, and quantitated by using the external standard method.. The calibration curves were linear within the range of 0.008 35-0.668, 0.020 6-1.648, 0.012 2-0.976 , 0.050 75-4.06, 0.015 7-1.256 microg for ligustrazine, ferulic acid, butylphthalide, ligustilide and butene-NBP, respectively. The average recoveries of the five components were 101.98%, 99.91%, 96.94%, 100.85% and 99.04%, respectively.. This method is simple, quick, reproducible, with high recovery, and has been successfully applied to the simultaneous determination of the five components in Rhizoma Chuanxiong. This method can be used to control the quality of Rhizoma Chuanxiong and its preparations.

Topics: 4-Butyrolactone; Benzofurans; Calibration; Chromatography, High Pressure Liquid; Coumaric Acids; Drugs, Chinese Herbal; Pyrazines