University of Würzburg, Institute for Pharmacy and Food Chemistry, 97074 Würzburg, Germany.
Thermo Fisher Scientific, 82110 Germering, Germany.
J Chromatogr A. 2018 Nov 16;1576:58-66. doi: 10.1016/j.chroma.2018.09.031. Epub 2018 Sep 18.
The analysis of polysorbate 80 is a challenge because all components lack a chromophore. Here, an ultra-high-performance liquid chromatography system equipped with a charged aerosol detector (UHPLC-CAD) was used to study the effect of systematic variation of the CAD settings, namely evaporation temperature, filter constant and power function value (PFV), on the detector response of fatty acid standards and manufacturing batches of polysorbate. Evaporation temperature and filter constant strongly affect the detection limits described by signal-to-noise (S/N) ratios. Although evaporation temperature can be increased to improve signal to noise ratios, analyte volatility at higher temperatures is an important limiting factor. The PFV was found to be a strong tool for optimizing response linearity, but the optimal PFV differed depending on analyte volatility. Because PFV optimization required some additional measurement time and because double-logarithmic transformation at the default PFV of 1.0 yielded satisfying universal results with less measurement time over a range of two orders of magnitude for every homologue fatty acid from C14 to C18, use of the log-log transformation is the favored linearization strategy. Possible optimization procedures for semi volatile substances are presented. Overall, this new UHPLC method method offers improved detection limits, as well as time savings of over 75% and eluent savings of more than 40% compared to the previously published HPLC-CAD method for polysorbate analysis.
聚山梨酯 80 的分析具有挑战性,因为所有成分都缺乏生色团。在这里,使用配备带电气溶胶检测器(UHPLC-CAD)的超高效液相色谱系统研究了 CAD 设置(即蒸发温度、过滤器常数和幂函数值(PFV))的系统变化对脂肪酸标准和聚山梨酯制造批次的检测器响应的影响。蒸发温度和过滤器常数强烈影响由信噪比(S/N)比描述的检测限。尽管可以提高蒸发温度来提高信噪比,但在较高温度下分析物的挥发性是一个重要的限制因素。发现 PFV 是优化响应线性度的有力工具,但最佳 PFV 因分析物的挥发性而异。由于 PFV 优化需要一些额外的测量时间,并且在默认 PFV 为 1.0 时进行双对数转换可以在从 C14 到 C18 的每个同系脂肪酸的两个数量级范围内以较少的测量时间获得令人满意的通用结果,因此使用对数-对数转换是首选的线性化策略。还提出了针对半挥发性物质的可能优化程序。总体而言,与之前发表的用于聚山梨酯分析的 HPLC-CAD 方法相比,这种新的 UHPLC 方法提高了检测限,同时节省了超过 75%的时间和超过 40%的洗脱剂。