Patent ID: 7993928
Filing Date: 2011-08-09
Classification: G01N

Abstract:
1. A method for detecting the presence of Danshensu and salvianolic acid B in the blood plasma of an animal after administering the botanical extract composition Strengthening Body Resistance and Dissipating Blood Stasis Botanical to said animal, comprising the steps of: a. collecting the blood plasma from said animal 0.5 hours after administration of Strengthening Body Resistance and Dissipating Blood Stasis Botanical to said animal; b. adding 2-5 mol/L of phosphoric acid to said blood plasma to form a blood plasma solution at a ratio (v/v) of blood plasma:phosphoric acid equals to 1: 2-3; c. apply said blood plasma solution to a chromatography column, wherein said column is activated by methanol and water prior to applying said blood plasma solution; d. washing said column with water and 80-100% methanol; e. eluting said column with 0.2-1% methanol in ammonia to obtain an eluent; f. drying said eluent at 25-30° to obtain a dried eluent; g. redissolving said dried eluent in a mobile phase solution comprising a mobile phase A and a mobile phase B at a ratio of 80 (mobile phase A)/20 (mobile phase B) to form a redissolved eluent, wherein said mobile phase A comprises water, acetonitrile, and formic acid at a ratio (v/v/v) of water/acetonitrile/formic acid equals to 95/5/0.1 and said mobile phase B comprises acetonitrile and formic acid at a ratio (v/v/v) of acetonitrile/formic acid equals to 100/0.1; h. preparing conditions for detection by ultra performance liquid chromatography electrospray ionization tandem mass spectrometry (HPLC/MS/ESI), wherein the ultra performance liquid chromatography (HPLC) conditions comprise a 2.1×100 mm chromatography column, said mobile phase A, and said mobile phase B; and wherein the mass spectrometry (MS) conditions comprise electrospray ionization (ESI) ion source in a negative ion mode; and i. detecting the presence of Danshensu and salvianolic acid B in said redissolved eluent under the conditions in step h at a mass scan range of m/z 150-800.