An HPLC method was applied for the simultaneous determination of five marker substances in the traditional Chinese medicine preparation "Tzyy-Yun-Gau". These substances included shikonin, deoxyshikonin, β,β-dimethylacrylshikonin, and acetylshikonin in Arnebiae Radix; and ferulic acid in Angelicae Radix. Tzyy-Yun-Gau was partitioned in a mixture of n-hexane and methanol, and a fraction taken into the methanol layer was analyzed. The samples were run through an HPLC column (Inertsil ODS-2, 4.6 mm ID. x 250 mm) at 30°C with a mobile phase consisting of methanol, acetonitrile and 2% acetic acid with linear gradient elution at a flow-rate of 1.0 mL/min. A UV detector was used for detection. The detection wavelength varied with time, which was 325 nm during 0∼25 min, 525 nm during 25∼58.5 rein, 440 nm during 58.5∼62 min, and 525 nm during 62-80 min. This could be a successful separation method for the simultaneous determination of five marker substances in "Tzyy-Yun-Gau".
Lay, H.-L.; Shih, I.-J.; Yeh, C.-H.; Lin, C.-F.; and Liang, J.-W.
"Simultaneous determination of five constituents in "Tzyy-Yun-Gau" medicine by high performance liquid chromatography,"
Journal of Food and Drug Analysis: Vol. 8
, Article 3.
Available at: https://doi.org/10.38212/2224-6614.2816