Study on fingerprint of rhizoma chuanxiong by HPLC-DAD-MS / 药学学报

ABSTRACT

<p><b>AIM</b>To establish a high performance liquid chromatographic fingerprint for the quality control of rhizoma Chuanxiong, a traditional Chinese medicine derived from the root of Ligusticum chuanxiong Hort..</p><p><b>METHODS</b>An on-line optimized HPLC-DAD-MS technique was employed. The HPLC analysis was performed on a Waters Symmetry C18 column (150 mm x4. 6 mm ID, 5 microm) with a Waters Spherisorb S5 ODS2 (10 mm x 4.6 mm) guard column. The mobile phase consisted of A (methanol) and B (0.25% acetic acid). Components were separated using the following gradient profile 32% B at 0-3 min, 32%-85% B at 3-33 min, 85%-100% B at 33-52 min; flow rate was 0.7 mL x min(-1). DAD was set from 190 to 400 nm, the fingerprint was monitored at 294 nm. All mass spectra were acquired in the positive ion mode with electrospray ionization; the full scan mass spectrum was recorded over the range of m/z 100-800. Nine samples from three companies were analyzed; the main characteristic peaks were identified based on the comparison of UV and MS spectra of each analyte with that of authentic compounds and literature data.</p><p><b>RESULTS</b>The HPLC fingerprint was established based on the analysis of nine rhizoma Chuanxiong herbal samples supplied by three companies. Twenty-one characteristic peaks were found in all nine samples. These peaks were classified into four groups group I at 0-12 min, three peaks were found, and the marker peak 3 was confirmed as ferulic acid; group II at 12-24 min, four peaks were found, and the marker peaks 4 and 5 were identified as senkyunolide I and senkyunolide H; group III at 24-32 min, there were seven peaks, and the marker peaks 9, 11, 13 and 14 were elucidated as senkyunolide A, coniferylferulate, ligustilide and 3-butylidenephthalide, respectively; group IV at 32-50 min, seven peaks were observed, and the marker peaks 15 and 17 were identified as riligustilide and levistolide A. The peak areas of 13 main peaks with normalized peak area (1% were determined. Using the most abundant peak 13 as the reference peak, the calculated relative retention times (tR of the characteristic peak/tR of the reference peak) among nine samples were consistent (RSD < or = 1%), while the calculated relative peak areas (peak area of the characteristic peak/peak area of the reference peak) among nine samples were significantly different (P < 0.001), indicating that all nine samples tested contain similar 13 main components with different quantities.</p><p><b>CONCLUSION</b>The established HPLC fingerprint is very specific, and can be used to evaluate the quality consistency of different rhizoma Chuanxiong herbs.</p>