We developed a novel ultra performance liquid chromatography-quadrupole time-of-flight (UPLC-Q-TOF) mass spectrometry method that allows sensitive, rapid, and reliable detection and identification of six representative indole alkaloids (vincristine, vinblastine, ajmalicine, catharanthine, serpentine, and vindoline) that exhibit physiological activity in Catharanthus roseus (C. roseus). The alkaloids were eluted on a C18 column with acetonitrile and water containing 0.1% formic acid and 10 mM ammonium acetate, and separated with good resolution within 13 min. Electrospray ionization-Q-TOF (ESI-Q-TOF) analysis was performed to characterize the molecules and their fragment ions, and the characteristic ions and fragmentation patterns were used as to identify the alkaloids. The proposed analytical method was verified in reference to the ICH guidelines and the results showed excellent linearity (R² > 0.9988), limit of detection (1 ng/mL to 10 ng/mL), limit of quantification (3 ng/mL to 30 ng/mL), intra-day and inter-day precisions, and extraction recovery rates (92.8% to 104.1%) for all components. The validated UPLC-Q-TOF method was applied to the analysis of extracts from the root, stem, and leaves of C. roseus, allowing the identification of six alkaloids by comparison of retention times, molecular ions, and fragmentation patterns with those of reference compounds. Sixteen additional indole alkaloids were tentatively identified by comparison of chromatograms to chemical databases and literature reports. The contents of bis-indole alkaloids (vincristine and vinblastine) were high in the aerial parts, while the contents of mono-indole alkaloids (ajmalicine, catharanthine, serpentine, and vindoline) were high in the roots. The present results demonstrate that the proposed UPLC-Q-TOF method can be useful for the investigation of phytochemical constituents of medicinal plants.

我们开发了一种新型的超高效液相色谱-四极杆飞行时间（UPLC-Q-TOF）质谱方法，该方法可灵敏，快速，可靠地检测和鉴定六种代表性的吲哚生物碱（长春新碱，长春碱，阿马利辛，蛇纹石和长春花碱）在长春花（C. roseus）中表现出生理活性）。用乙腈和含有0.1％甲酸和10 mM乙酸铵的水在C18柱上洗脱生物碱，并在13分钟内以良好的分离度分离。进行电喷雾电离-Q-TOF（ESI-Q-TOF）分析以表征分子及其片段离子，并使用特征离子和片段化模式鉴定生物碱。参考ICH指南验证了所提出的分析方法，结果显示出优异的线性度（R ² > 0.9988），检出限（1 ng / mL至10 ng / mL），定量限（3 ng / mL至30 ng / mL），日内和日间精度以及提取回收率（92.8％）到104.1％）。经验证的UPLC-Q-TOF方法用于分析玫瑰假花的根，茎和叶的提取物，通过与参考化合物的保留时间，分子离子和片段化模式进行比较，可以鉴定出六个生物碱。通过色谱图与化学数据库和文献报告的比较，初步确定了另外16种吲哚生物碱。地上部分的双吲哚生物碱（长春新碱和长春碱）含量高，而根部的单吲哚生物碱（长春花碱，卡特兰汀，蛇纹石和长春花碱）含量高。目前的结果表明，所提出的UPLC-Q-TOF方法可用于研究药用植物的植物化学成分。