HPLC法测定复方利血平片中6个成分的含量均匀度

Objective: To establish an HPLC method for the determination of the content uniformity of the six components (hydrochlorothiazide, dihydralazine sulfate, vitamin B₁, vitamin B₆, promethazine hydrochloride and reserpine)in compound reserprine tablets.Methods: The Phenomenex Gemini C₁₈ column (250 mm×4.6 mm, 5 μm)was adopted at the flow rate of 1.0 mL·min^-1 with the column temperature of 40℃.The mobile phase (1)was 0.14% sodium 1-heptanesulfonate solution (adjust pH to 3.5 with 5% phosphoric acid solution)-acetonitrile (82:18)and the detection wavelength was 256 nm for hydrochlorothiazide, dihydralazine sulfate, vitamin B₁ and vitamin B₆.The mobile phase (2)was 0.14% sodium 1-heptanesulfonate solution (adjust pH to 3.5 with 5% phosphoric acid solution)-acetonitrile (60:40), and the detection wavelength was 220 nm for promethazine hydrochloride and reserpine.Results: The averages (A+1.8S)of the content uniformity for the six components (hydrochlorothiazide, dihydralazinesulfate, vitaminB1, vitamin B₆, promethazine hydrochloride and reserpine)were 100.5%-103.4% (2.7-5.7), 101.8%-105.3% (5.6-8.4), 102.8%-104.8% (8.2-9.6), 97.6%-9.8% (2.8-6.7), 102.5%-103.6% (5.4-9.9), 97.5%-103.4% (12.2-14.1).The limits of quantitation of the six components were 1.6, 2.7, 7.8, 26.2, 1.7 and 0.7 ng, respectively.The linear ranges of the six components were 0.036 88-0.086 04 mg·mL^-1 (r=0.999 9), 0.050 95-0.118 9 mg·mL^-1 (r=0.999 6), 0.011 69-0.027 27 mg·mL^-1 (r=0.999 4), 0.013 10-0.030 58 mg·mL^-1 (r=0.999 6), 0.082 65-0.289 3 mg·mL^-1 (r=1.000), 1.221-4.273 μg·mL^-1 (r=1.000).The average recoveries (n=9)from three concentrations of low, medium and high samples for the six components were 99.9% (RSD=0.4%), 100.7% (RSD=1.4%), 100.5% (RSD=0.3%), 101.2% (RSD=0.5%), 100.6% (RSD=0.5%)and 100.5% (RSD=0.7%), respectively.Hydrochlorothiazide, dihydralazine sulfate, vitamin B₁, vitamin B₆, promethazine hydrochloride and reserpine were indentified simultaneously in the method.Conclusion: The method is simple, rapid, reproducible and specific for the quality control of compound reserprine tablets after validation.