Produced in Madeira Island from regional sugarcane cultivars through a traditional manufacturing and storage process, sugarcane honey (SCH) is a black syrup recognized by its excellent quality. Its economic value has led to the emergence of adulterated SCH, whereby the identification of molecular markers became an essential task in order to overcome the fraudulent activities, protect its authenticity, and guarantee the consumer safety. In the present study, an analytical strategy based on ultrasound-assisted liquid-liquid extraction (USA-LLE) followed by reversed-phase liquid chromatography with a refractive index detector (LC-RI) was developed for the determination of sugars (glucose, fructose, sucrose, xylose, and mannose) in SCH samples from certified producers, supported on analytical quality-by-design (AQbD) approach, as a useful tool to establish its typicality. The application of AQbD was based on analytical risk assessment, multivariate statistics and quality control procedures for definition of the Method Operable Design Region (MODR). The optimal conditions into MODR were accomplished using BEH Amide column operating at a temperature of 80 °C and a flow rate of 300 μL min⁻¹, with a mobile phase composed by acetone and water (85:15, v v⁻¹) at a flow rate of 1.0 mL min⁻¹. The robustness was determined by Monte Carlo simulation and capability analysis. The concentration-response function for all sugars was described by polynomial models. Accuracy was presented by recovery values between 98.2 and 119.5%. The analytical figures of merit validated the utility of AQbD in the systematic design of a LC-RI method with fine sensitivity for sugar analysis in SCH.

甘蔗蜂蜜（SCH）是一种黑糖浆，以其卓越的品质而闻名，是由马德拉岛地区的甘蔗品种通过传统的制造和存储过程生产而成的。它的经济价值导致了掺假SCH的出现，由此，分子标记的鉴定成为克服欺诈活动，保护其真实性并确保消费者安全的一项基本任务。在本研究中，开发了一种基于超声辅助液-液萃取（USA-LLE），然后使用反相液相色谱仪和折射率检测器（LC-RI）进行分析的测定糖类（葡萄糖，果糖）的分析策略。认证生产者的SCH样品中的蔗糖，木糖和甘露糖），并获得了设计质量分析（AQbD）方法的支持，作为确立其典型性的有用工具。AQbD的应用是基于分析风险评估，多元统计和质量控制程序来定义方法可操作设计区域（MODR）。使用BEH Amide色谱柱在80°C的温度和300μLmin的流速下运行可实现进入MODR的最佳条件^-1，流动相由丙酮和水（85：15，v v ^-1）组成，流速为1.0 mL min ^-1。鲁棒性是通过蒙特卡洛模拟和能力分析确定的。通过多项式模型描述了所有糖的浓度响应函数。回收率值介于98.2和119.5％之间，显示了准确性。品质因数的分析证实了AQbD在LC-RI方法系统设计中的实用性，该方法对SCH中的糖分析具有良好的灵敏度。