Patent ID: 8334110

Claim:
A method for detecting amygdalin in the blood plasma of an animal after administering a botanical composition comprising Strengthening Body Resistance and Dissipating Blood Stasis Botanical to the animal, comprising the steps of: (a) collecting the blood plasma from the animal after the administration of Strengthening Body Resistance and Dissipating Blood Stasis Botanical, wherein said botanical composition comprises Salviae miltiorrhizae , peach kernel, and Schisandrae chinensis; (b) mixing the blood plasma with phosphoric acid with a concentration ranging from 2-5 mol/L at a blood plasma to phosphoric acid volume ratio of 1:2-3 (v/v); (c) applying the mixture of step (b) to a chromatography column, wherein the column is pre-activated by methanol and water prior to application of the mixture; (d) washing the column with water and 80-100% methanol; (e) eluting the column with 0.2-1% ammonia in methanol to obtain an eluent, (f) drying the eluent at 25-30° C.; (g) dissolving the dried eluent of step (f) in a mobile phase A solution to form a dissolved eluent, wherein the mobile phase A solution comprises water, acetonitrile and formic acid at a water/acetonitrile/formic acid ratio (v/v/v) of 95:5:0.1; and (h) separating and detecting amygdalin in the dissolved eluent by ultra performance liquid chromatography/mass spectrometry (UPLC-MS), wherein the UPLC separation conditions comprise separating amygdalin on a 2.1×100 mm, C 18 UPLC column with an elution gradient comprising the mobile phase A, and the an increasing amount of a mobile phase B, wherein the mobile phase B comprises acetonitrile and formic acid at an acetonitrile/formic acid ratio (v/v) of 100:0.1; and wherein the MS detection conditions comprise electrospray ionization (ESI) ion source detecting in a positive ion mode, and a mass scanning range of m/z 150-800.