Robust Design Space Development for HPLC Analysis of Five Chemical Components in <i>Panax Notoginseng</i> Saponins
Creators: name: Shengyun Dai
name: Xu, Bing
name: Zhang, Yi
name: Sun, Fei
name: Jianyu Li
name: Yahua Cui
name: Xinyuan Shi
name: Yanjiang Qiao
Publisher: name: Taylor & Francis
Year: 2016
Subjects: subject: Biophysics
subject: Biochemistry
subject: Space Science
subject: Medicine
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: FOS: Chemical sciences
subject: Immunology
subject: FOS: Clinical medicine
subject: FOS: Clinical medicine
subject: 69999 Biological Sciences not elsewhere classified
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: 19999 Mathematical Sciences not elsewhere classified
subject: FOS: Mathematics
subject: FOS: Mathematics
subject: Developmental Biology

description: The present work describes the systematic development of a robust, precise and rapid reversed-phase liquid chromatography method for the simultaneous analysis of five chemical components in <i>Panax Notoginseng</i> Saponins (PNS) using Quality by Design (QbD). The method was developed in two main phases: screening and optimization. During the screening phase, the most suitable stationary phase, column temperature and flow rate were identified, while the secondary influential parameters, such as the gradient slope, the initial concentration of acetonitrile and the initial isocratic hold of the gradient elution system were fine tuned in the later optimization phase. In this phase, a seventeen-run experiment was used to examine multifactorial effects of these parameters on the critical resolution, the analysis time and the peak symmetry. The Monte Carlo simulation method was successfully used to build the chromatographic design space and the process capability index <i>Cp</i> was introduced to evaluate the robustness of the design space. At last, the verification experiment was performed within the working design space by the accuracy profile methodology and model was found to be accurate. A robust combination of separation conditions predicted in the design space was obtained with the gradient slop of 1.6% · min<sup>−1</sup>, the initial concentration of acetonitrile of 20% and the initial isocratic hold of 20 min and the total analysis time was no more than 40 min. The results demonstrated that rapid separation of five components of PNS herbal extract could be achieved on the chromatographic column packed with narrow size particles at backpressures available in ordinary HPLC instruments. <b>Short Abstract</b> A systematic approach applying Quality by Design (QbD) principles was utilized to develop an HPLC method to separate five components of the <i>Panax Notoginseng</i> Saponins extracted from the traditional Chinese medicine <i>Panax Notoginseng</i> for the first time. The novel QbD methodology described here combines Design of Experiments with Monte Carlo simulation to successfully integrate the quantitative robustness metrics <i>Cp</i> into HPLC method development. This method quicken the analytical speed and can be applied in the regular instruments which contributes to the widely utilization.