Method for simultaneous determination of chlorogenic acid, ferulic acid, dracohodin, cinnamaldehyde, ligustilide, and osthole in xiangdu huoxue plaster

A method for simultaneous determination of chlorogenic acid, ferulic acid, dracohodin, cinnamaldehyde, ligustilide, and osthole in Xiangdu Huoxue Plaster, the method comprising:

• dissolving a first cinnamaldehyde reference standard in methanol to obtain a cinnamaldehyde reference standard solution;
• subjecting the cinnamaldehyde reference standard solution to first high-performance liquid chromatography (HPLC) detection, thereby obtaining a first cinnamaldehyde reference standard peak area;
• removing a protective film from a test sample of the Xiangdu Huoxue Plaster, crushing the test sample, and subjecting an obtained crushed test sample to extraction with a first phosphoric acid-methanol solution to obtain a test sample solution;
• wherein the first phosphoric acid-methanol solution comprises phosphoric acid at a mass percentage of 2% to 4%;
• subjecting the test sample solution to second HPLC detection, thereby obtaining a test sample peak area, and calculating a concentration of the cinnamaldehyde in the test sample solution by an external standard method based on a concentration of the cinnamaldehyde reference standard solution, the first cinnamaldehyde reference standard peak area, and the test sample peak area;
• and
• calculating concentrations of the chlorogenic acid, the ferulic acid, the dracohodin, the ligustilide, and the osthole in the test sample solution based on the first cinnamaldehyde reference standard peak area, the test sample peak area, the concentration of the cinnamaldehyde in the test sample solution, and relative correction factors;
• wherein the relative correction factors comprise a chlorogenic acid correction factor, a ferulic acid correction factor, a dracohodin correction factor, an osthole correction factor, and a ligustilide correction factor;
• the first HPLC detection and the second HPLC detection independently comprise following conditions:
• a chromatographic column is a C18 chromatographic column;
• a mobile phase system comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is an aqueous phosphoric acid solution, the mobile phase B is acetonitrile, and the aqueous phosphoric acid solution comprises phosphoric acid at a mass percentage of 0.1% to 0.4%;
• an elution mode is gradient elution, wherein a program of the gradient elution comprises:
• 0 min, a volume fraction of the mobile phase A is 95%;
• 0 min to 5 min, the volume fraction of the mobile phase A is decreased from 95% to 90%;
• 5 min to 15 min, the volume fraction of the mobile phase A is 90%;
• 15 min to 17 min, the volume fraction of the mobile phase A is decreased from 90% to 82%;
• 17 min to 30 min, the volume fraction of the mobile phase A is 82%;
• 30 min to 31 min, the volume fraction of the mobile phase A is decreased from 82% to 75%;
• 31 min to 41 min, the volume fraction of the mobile phase A is decreased from 75% to 70%;
• 41 min to 45 min, the volume fraction of the mobile phase A is 70%;
• 45 min to 50 min, the volume fraction of the mobile phase A is decreased from 70% to 65%;
• 50 min to 55 min, the volume fraction of the mobile phase A is decreased from 65% to 52%;
• 55 min to 85 min, the volume fraction of the mobile phase A is 52%;
• 85 min to 86 min, the volume fraction of the mobile phase A is decreased from 52% to 0%;
• 86 min to 90 min, the volume fraction of the mobile phase A is 0%;
• 90 min to 91 min, the volume fraction of the mobile phase A is increased from 0% to 95%;
• 91 min to 95 min, the volume fraction of the mobile phase A is 95%;
• and
• the Xiangdu Huoxue Plaster comprises a backing material, a protective film, a Chinese herbal composition, and a matrix;
• wherein the matrix comprises following components in parts by mass:
• 10 parts to 45 parts of a framework material, 1 part to 3 parts of a cross-linking agent, 30 parts to 70 parts of a humectant, 1 part to 3 parts of a pH adjuster, 15 parts to 35 parts of a tackifier, and 50 parts to 120 parts of purified water;
• the Chinese herbal composition comprises following components in parts by mass:
• 5 parts to 24 parts of Radix Angelicae Pubescentis, 5 parts to 24 parts of Rhizoma Chuanxiong, 5 parts to 24 parts of Radix Vladimiriae, 4 parts to 14 parts of Fructus Foeniculi, 4 parts to 14 parts of Cortex Cinnamomi, 4 parts to 14 parts of Frankincense, 4 parts to 14 parts of Myrrh, 1 part to 12 parts of Radix Aconiti Preparata, 1 part to 12 parts of Radix Aconiti Kusnezoffii Preparata, 1 part to 12 parts of Resina Draconis, and 1 part to 8 parts of Borneol.