Determination of 12 kinds of components in Tianma Shouwu Pills by HPLC method / 中草药

ABSTRACT

Objective To develop an HPLC combined with gradient elution method for the determination of the contents of paeoniflorin, 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucosid, gastrodin, ferulic acid, acteoside, liquiritin, tanshinone IIA, specnuezhenide, imperatorin, rutinum, wedelolactone, and emod in Tianma Shouwu Pills (TSP) at the same time. Methods The chromatographic separation was achieved on a Hypersil GOLD C18 (250 mm × 4.6 mm, 5 μm) with acetonitrile (A)-0.1% phosphoric acid solution (B) as mobile phase at the flow rate of 1.0 mL/min for gradient elution; sample quantity was 10 μL. Results The linear relation between peak area and concentration of the twelve active components were good, which were 3.12-31.20 μg/mL (r = 0.999 5) paeoniflorin, 4.98-49.80 μg/mL (r = 0.999 4) 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucosid, 1.05-10.50 μg/mL (r = 0.999 5) gastrodin, 0.99-9.90 μg/mL (r = 0.999 2) ferulic acid, 1.11-11.10 μg/mL (r = 0.999 2) acteoside, 3.24-32.40 μg/mL (r = 0.999 2) liquiritin, 3.63-36.30 μg/mL (r = 0.999 2) tanshinone IIA, 1.13-11.30 μg/mL (r = 0.999 3) specnuezhenide, 1.9-19.0 μg/mL (r = 0.999 5) imperatorin, 1.55-15.50 μg/mL (r = 0.999 5) rutinum, 1.48-14.80 μg/mL (r = 0.999 5) wedelolactone, and 102.8-1028.0μg/mL (r = 0.9999) emodin. The precision and repeatability were good, and RSD values were less than 2.0%. Moreover, the average recoveries and the corresponding RSD values were 99.88% (1.55%), 101.25% (0.98%), 99.67% (1.29%), 102.04% (1.17%), 101.17% (1.67%), 98.27% (1.51%), 100.28% (1.20%), 99.11% (0.95%), 98.49% (1.67%), 101.57% (0.94%), 102.37% (0.58%), and 97.89% (0.69%), respectively. The contents of 12 batches of the 12 active components were 1.241-1.261, 2.116-2.133, 0.540-0.558, 0.077-0.099, 0.089-0.110, 1.111-1.134, 0.158-0.183, 1.375-1.399, 0.342-0.372, 0.542-0.571, 0.648-0.672, and 45.05-45.93 mg/g. Conclusion An HPLC combined with gradient elution method has been successfully established for simultaneous determination of 12 components in TSP. The method is simple, quick, accurate, and helpful for the quality control of TSP.