An efficient reverse-phase high-performance liquid chromatographic method, based on the design of the experiment approach, was developed for the simultaneous determination of capsiate isomers. Critical method parameters, i.e., flow rate and mobile phase composition, demarcated during preliminary screening were optimized using central composite design. Chromatographic separation was achieved on a Nucleodur C18 column (250 × 4.6 mm, 5 μm), with the mobile phase consisting of water–acetonitrile (40:60), both acidified with 0.1% v/v formic acid, passed at a flow rate of 1 mL/min. E- and Z-capsiates were eluted from the column at the retention times of 18.56 ± 0.09 and 17.30 ± 0.08 min, respectively, with a resolution factor of 1.693 ± 0.046. The method was found to be linear within the concentration range of 1.054–5.270 and 8.623–43.115 μg/mL for Z- and E-isomers, respectively, with an R2 of >0.99. Recovery of the individual values was in range of 96.15–101.92%, with a relative standard deviation of <2%. The developed method was used to quantify capsiate isomers extracted from sweet pepper fruits. Therefore, the proposed method is presented for optimum isomeric resolution of these closely related E- and Z-isomers with convenient sample preparation, acceptable run-time, cost effectiveness and use of conventional instruments. Highlights

中文翻译：

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质量源于设计 – 通过反相 HPLC 和 LC-MS 对衣壳的几何异构体进行受控色谱分离和鉴定


在实验方法设计的基础上，建立了一种高效的反相高效液相色谱方法，用于同时测定衣壳酸异构体。使用中心复合材料设计优化了初步筛选期间确定的关键方法参数，即流速和流动相组成。在 Nucleodur C18 色谱柱（250 × 4.6 mm，5 μm）上实现色谱分离，流动相由水-乙腈 （40：60） 组成，两者均用 0.1% v/v 甲酸酸化，流速为 1 mL/min。从色谱柱中洗脱 E 和 Z 果壳酸盐，保留时间为 18.56 ± 0.09 和 17.30 ± 0.08 min，分离因子为 1.693 ± 0.046。发现该方法在 Z- 和 E-异构体的浓度范围分别为 1.054–5.270 和 8.623–43.115 μg/mL，R2 为 >0.99。单个值的回收率在 96.15–101.92% 的范围内，相对标准偏差为 <2%。采用所开发的方法定量从甜椒果实中提取的辣椒酸异构体。因此，提出了所提出的方法，以实现这些密切相关的 E 和 Z 异构体的最佳异构体分离度，同时具有方便的样品制备、可接受的运行时间、成本效益和常规仪器的使用。突出