Jeong Won Tae, Lim Heung Bin
Department of Industrial Plant Science & Technology, Chungbuk National University, Cheongju, Chungbuk 28644, Republic of Korea.
Department of Industrial Plant Science & Technology, Chungbuk National University, Cheongju, Chungbuk 28644, Republic of Korea.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Mar 30;1080:27-36. doi: 10.1016/j.jchromb.2018.02.018. Epub 2018 Feb 17.
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 (R > 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超高效液相色谱 - 四极杆飞行时间（UPLC - Q - TOF）质谱方法，该方法能够灵敏、快速且可靠地检测和鉴定六种具有代表性的吲哚生物碱（长春新碱、长春碱、阿吗碱、长春质碱、蛇根碱和文多灵），这些生物碱在长春花中具有生理活性。生物碱在C18柱上以含有0.1%甲酸和10 mM乙酸铵的乙腈和水为流动相进行洗脱，并在13分钟内实现良好分离。采用电喷雾电离 - Q - TOF（ESI - Q - TOF）分析对分子及其碎片离子进行表征，并利用特征离子和碎裂模式来鉴定生物碱。所提出的分析方法依据国际人用药品注册技术协调会（ICH）指南进行了验证，结果显示所有组分均具有出色的线性（R＞0.9988）、检测限（1 ng/mL至10 ng/mL）、定量限（3 ng/mL至30 ng/mL）、日内和日间精密度以及提取回收率（92.8%至104.1%）。经验证的UPLC - Q - TOF方法应用于长春花根、茎和叶提取物的分析，通过与对照品的保留时间、分子离子和碎裂模式进行比较，鉴定出六种生物碱。通过将色谱图与化学数据库及文献报道进行比较，初步鉴定出另外十六种吲哚生物碱。双吲哚生物碱（长春新碱和长春碱）在地上部分含量较高，而单吲哚生物碱（阿吗碱、长春质碱、蛇根碱和文多灵）在根部含量较高。目前的结果表明，所提出的UPLC - Q - TOF方法可用于药用植物植物化学成分的研究。
