Pharmacy College, Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 611137, P.R. China.
Nat Prod Res. 2010 Oct;24(17):1616-29. doi: 10.1080/14786411003757937.
A simple, sensitive and reliable high-performance liquid chromatography (HPLC) method with evaporative light scattering detection (ELSD) has been developed for simultaneous determination of five major alkaloids in Coptidis rhizoma. Simultaneous separation of five alkaloids was achieved on a Kromasil C₁₈ analytical column (250 mm x 4.6 mm, 5 µm) with the isocratic elution of acetonitrile : water (30:70, v/v, the pH was adjusted to 6.0 with 0.2 moL L⁻¹ trichloroacetic acid). The drift tube temperature of the ELSD system was set at 115 °C and the nitrogen flow rate was 2.8 L min⁻¹. All the five calibration curves showed a good linear relationship (r² > 0.9991) within the test ranges. The limit of detections and quantifications for the five alkaloids in ELSD were less than 0.88 and 2.73 ng, respectively. The established method was successfully applied to quantify the five ingredients in different C. rhizoma samples and evaluate the internal quality of C. rhizoma samples from various sources by analysing the chemical fingerprints using similarity analysis (SA), hierarchical clustering analysis (HCA) and principal component analysis (PCA), which provided a useful basis of overall evaluation of the quality of C. rhizoma.
建立了一种同时测定黄连中 5 种主要生物碱的高效液相色谱-蒸发光散射检测（HPLC-ELSD）法。采用 Kromasil C₁₈ 分析柱（250mm×4.6mm，5μm），以乙腈-水（30∶70，v/v，用 0.2mol·L⁻¹ 三氯乙酸调节 pH 值至 6.0）为流动相，在等度洗脱条件下，实现了 5 种生物碱的同时分离。ELSD 系统的漂移管温度设定为 115℃，氮气流量为 2.8L·min⁻¹。5 种生物碱在各自的测定范围内均呈现良好的线性关系（r²＞0.9991）。在 ELSD 中，5 种生物碱的检测限和定量限均小于 0.88 和 2.73ng。该方法成功地应用于不同黄连样品中 5 种成分的定量分析，并通过相似度分析（SA）、层次聚类分析（HCA）和主成分分析（PCA）对不同来源黄连样品的指纹图谱进行分析，评价黄连样品的内在质量，为黄连整体质量评价提供了依据。
