Abstract A gradient reverse phase high pressure liquid chromatographic method has been proposed for simultaneous determination and separation of curcumin (CUR), bisdemethoxycurcumin (BMC), demethoxycurcumin (DMC), and metanil yellow (MET). The separation was achieved on an Agilent Eclipse XDB-C18 (4.6 mm × 150 mm, 3.5 µm) analytical column using photodiode array (PDA) detection at 425 nm. Gradient elution of solvent A (mixture of 0.1% trifluoro acetic acid and 0.1% formic acid 50:50 v/v) and solvent B (acetonitrile) at a flow rate of 0.8 mL/min was used to accomplish adequate separation of the analytes. The developed method was extensively validated with respect to linearity, detection and quantitation limits, precision, recovery, and robustness testing. Method variables, viz., mobile phase flow rate (0.90 ± 0.05 mL/min), percentage of B at start of gradient (40 ± 2%) and detection wavelength (425 ± 5 nm) were studied by a Box–Behnken Design (BBD) for testing the robustness of the proposed method. The method was found robust, with no significant variations in the method performance, linear in the range of 10–80 µg/mL (r2 = 0.999) and precise (RSD ˂ 2%) satisfying regulatory criteria. The method is selective for rapid determination of turmeric adulteration with a detection limit of 0.37–2.48 µg/mL. Graphical Abstract

中文翻译：

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一种用于测定姜黄掺假的可靠 RP-HPLC 方法

摘要 提出了一种梯度反相高压液相色谱法同时测定和分离姜黄素（CUR）、双去甲氧基姜黄素（BMC）、去甲氧基姜黄素（DMC）和间苯胺黄（MET）的方法。使用 425 nm 光电二极管阵列 (PDA) 检测在 Agilent Eclipse XDB-C18（4.6 mm × 150 mm，3.5 µm）分析柱上实现分离。溶剂 A（0.1% 三氟乙酸和 0.1% 甲酸的混合物 50:50 v/v）和溶剂 B（乙腈）以 0.8 mL/min 的流速梯度洗脱，以实现分析物的充分分离。开发的方法在线性、检测和定量限、精密度、回收率和稳健性测试方面得到了广泛验证。方法变量，即流动相流速 (0.90 ± 0.05 mL/min)，通过 Box-Behnken 设计 (BBD) 研究了梯度开始时 B 的百分比 (40 ± 2%) 和检测波长 (425 ± 5 nm)，以测试所提出方法的稳健性。发现该方法稳健，方法性能没有显着变化，线性范围为 10–80 µg/mL (r2 = 0.999)，精确度 (RSD ˂ 2%) 满足监管标准。该方法具有选择性，可快速检测姜黄掺假，检测限为 0.37–2.48 µg/mL。图形概要 该方法具有选择性，可快速检测姜黄掺假，检测限为 0.37–2.48 µg/mL。图形概要 该方法具有选择性，可快速检测姜黄掺假，检测限为 0.37–2.48 µg/mL。图形概要