Abstract:

A rapid and accurate method for the simultaneous separation and determination of protocatechuic acid, protocatechuic aldehyde, caffeic acid and tanshinone ⅡA in Fufang Danshen Tablets by high performance liquid chromatography has been established. The former three constituents were detected by electrochemical array detector (ECD) and the last one by ultraviolet diode array detector (DAD). The separation was performed on a Zorbax SB-C18 column (150 mm×4.6 mm i.d., 5.0 μm) by gradient elution with methanol and 0.4% aqueous phosphoric acid as mobile phase at a flow rate of 1.0 mL/min. The detections were done at 0.7 V, 270 nm and 30 ℃. The results showed that the linear ranges were 0.3-9.0 mg/L for protocatechuic acid, 1.1-54.0 mg/L for protocatechuic aldehyde, 1.1-11.1 mg/L for caffeic acid and 5.2-52.0 mg/L for tanshinone ⅡA. The average recoveries (n= 6) were 97.8% with relative standard deviation (RSD) of 2.15% for protocatechuic acid, 98.2% with RSD of 2.07% for protocatechuic aldehyde, 97.6% with RSD of 2.18% for caffeic acid and 97.2% with RSD of 2.07% for tanshinone ⅡA. These four components were separated successfully from each other. The sensitivities of detection of protocatechuic acid, protocatechuic aldehyde and caffeic acid with ECD were 11.0-25.3 times greater than those with DAD. The study provides a scientific basis for the quality control of Fufang Danshen Tablets.

Key words: tanshinone ⅡA, caffeic acid, Fufang Danshen Tablets , protocatechuic acid, protocatechuic aldehyde, high performance liquid chromatography-electrochemical and ultraviolet detections