Separation of Major Safflowers from Carthamus Yellow using High‐Speed Countercurrent Chromatography

Abstract

High‐speed countercurrent chromatography (HSCCC) has been successfully applied to the separation of safflower A (SF‐A) and safflower B (SF‐B) from Carthamus yellow, Carthamus tinctorius L. A 25 mg quantity of Carthamus yellow was separated using a two‐phase solvent composed of tert‐butyl methyl ether/n‐butanol/acetonitrile/0.5% aqueous trifluoroacetic acid solution (2/2/1/5, V/V). The total separation time was 6 hr with the total elution volume of 720 mL. HSCCC fractions were analyzed using LC/MS/MS with scan and daughter scan modes, and DAD monitored by wide absorbance from 200 to 500 nm. The separation yielded 1.5 mg of SF‐A (95%>purity) and 1.1 mg of SF‐B (95%>purity). These results present a successful application of HSCCC to the preparative purification of major safflowers.

Keywords:

• Carthamus yellow
• Safflower
• High‐speed countercurrent chromatography
• Liquid chromatography‐tandem mass spectrometry