摘要
目的 研究川芎药材的高效液相色谱指纹谱 ,为科学评价和有效控制其内在质量提供可靠方法。方法应用HPLC DAD MS联用技术测定四川GAP示范基地 3个公司的 9个川芎样品 ,鉴定共有峰和特征峰 ,指定参照峰 ,计算共有峰的相对保留时间和相对峰面积。结果 9个样品的色谱指纹图谱有 2 1个共有峰。色谱图分为 4个区 ,第1区 (0 12min)主要分布酚酸类和生物碱类化合物 ,有 3个共有峰 ,峰 3被确证为ferulicacid ;第 2区 (12 2 4min)主要分布羟基化苯酞类化合物 ,有 4个共有峰 ,特征峰 4和 5被确证为senkyunolideI和senkyunolideH ;第 3区 (2 4 32min)主要分布烷基化苯酞类化合物 ,有 7个共有峰 ,特征峰 9,11,13和 14分别被确证为senkyunolideA ,coniferylferulate ,Z ligustilide和 3 butylidenephthalide。峰 13Z ligustilide被指定为参照峰 ;第 4区 (32 5 0min)分布苯酞二聚体化合物 ,有 7个共有峰 ,峰 15和 17分别被确证为riligustilide和levistolideA。所有样品中 2 1个共有峰的相对保留时间稳定 (RSD≤1% ) ,同一公司同一年采收的样品 13个主要共有峰 (归一化法峰面积≤ 1% )的相对峰面积稳定 ,而不同公司同一年采收的样品 13个主要共有峰的相对峰面积差异非常显著 (P <0 0 0 1)。
Aim 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.. Methods An on-line optimized HPLC-DAD-MS technique was employed. The HPLC analysis was performed on a Waters Symmetry C 18 column (150 mm×4.6 mm ID, 5 μm) with a Waters Spherisorb S5 ODS2 (10 mm×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·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. Results 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 (t R of the characteristic peak/ t R of the reference peak) among nine samples were consistent (RSD≤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. Conclusion The established HPLC fingerprint is very specific, and can be used to evaluate the quality consistency of different rhizoma Chuanxiong herbs.
出处
《药学学报》
CAS
CSCD
北大核心
2004年第8期621-626,共6页
Acta Pharmaceutica Sinica
基金
香港创新科技署创新及科技基金项目 (UIM 0 3 4)
