Simultaneous Determination of Five Constituents in "Tzyy-Yun-Gau" Medicine by High Performance Liquid Chromatography
HORNG-LIANG LAY2*, I-JEN SHIH1, CHIH-HO YEH1, CHWAN-FWU LIN1 AND J-WEN LIANG1
1. Pharmaceutical Industry Technology and Development Center, 5F, No.101, Lane 169 Kang Ning St., Hsi Chih Chen, Taipei County, Taiwan 221, R.O.C.
2. National Ping-Tung University of Science & Technology, Department of Plant Industry, No.1, Hseuh Fu Rd., Nei-Pu Hsiang, Pingtung County, Taiwan 912, R.O.C.
(Received: May 11, 2000; Accepted: August 7, 2000)
ABSTRACT
A 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, b,b-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 I.D. ´ 250 mm) at 30℃ 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 min, 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"
Key words: Tzyy-Yun-Gau, Arnebiae Radix, Angelicae Radix, shikonin, deoxyshikonin, b,b-dimethylacrylshikonin, acetylshikonin, ferulic acid