Water-soluble vitamins are essential dietary components with a multitude of important functions that require quantification from food sources to characterise the nutritional status of food. In this study, we have developed a hydrophilic interaction chromatography (HILIC) based method coupled to single-quadrupole mass spectrometry (MS) for the analysis of selected water-soluble vitamins. Due to their involvement in energy release from macronutrients, the quantification of thiamine (B₁), riboflavin (B₂), nicotinamide (B₃) and pyridoxine (B₆) offers significant value in food analysis.

A commercially available vegetable soup was selected as the food matrix for this study and utilised to develop an efficient extraction procedure for the vitamins of interest. Vitamins were extracted using meta-phosphoric acid coupled with a reducing agent, DL-dithiothreitol (DTT) to produce the parent compound. The extracted vitamins were then analysed using an LC-MS system with electrospray – atmospheric pressure ionization (ES-API) source, operated in positive single ion monitoring (SIM) mode. The MS provided good linearity within the investigated range from 0.005-0.4 µg/mL with coefficient of determination (r²) ranging from 0.98-0.99. Retention times (0.65-9.04 mins) were reproducible and no coelution between vitamins was observed. Limit of detection (LOD) varied from 2.4-9.0 ng/mL and limit of quantification (LOQ) was from 8-30 ng/mL, comparable to previously published studies. The extraction method provided good intra-day (%CV 1.73-5.83) and inter-day precision (%CV 8.07-10.97). Standard injections were used as part of quality control measures and provided excellent reproducibility (%CV 0.9-3.4). The overall runtime of this method was 19 mins, including column reconditioning. Using this method, the quantity of thiamine (67 ± 7 ng/g), riboflavin (475 ± 45 ng/g), nicotinamide (856 ± 77 ng/g) and pyridoxine (133 ± 11 ng/g) was determined from a complex food matrix.

In conclusion, we have developed a rapid and reliable, HILIC-single quad MS method utilising SIM for the low-level quantification of four B vitamins in a vegetable soup matrix in under 20 mins. This method has shown excellent linearity, intra- and inter-day reproducibility and is directly applicable to other plant-based food matrices.

水溶性维生素是必需的饮食成分，具有许多重要功能，需要从食物来源进行定量以表征食物的营养状况。在这项研究中，我们开发了一种基于亲水相互作用色谱（HILIC）的方法，并结合了单四极杆质谱（MS）来分析所选的水溶性维生素。由于它们参与从大量营养素中释放能量，因此硫胺素（B ₁），核黄素（B ₂），烟酰胺（B ₃）和吡ido醇（B ₆）的定量在食品分析中具有重要价值。

选择可商购的蔬菜汤作为本研究的食物基质，并用于开发有效的目标维生素提取工艺。使用间磷酸与还原剂DL-二硫苏糖醇（DTT）结合提取维生素，可生成母体化合物。然后使用带有电喷雾–大气压电离（ES-API）源的LC-MS系统，以正单离子监测（SIM）模式运行，分析提取的维生素。MS在0.005-0.4 µg / mL的测定范围内具有良好的线性，并具有测定系数（r ²），范围从0.98-0.99。保留时间（0.65-9.04分钟）是可重现的，未观察到维生素之间的共洗脱。检出限（LOD）为2.4-9.0 ng / mL，定量限（LOQ）为8-30 ng / mL，与以前发表的研究相当。提取方法提供了良好的日内（％CV 1.73-5.83）和日间精度（％CV 8.07-10.97）。标准注射液用作质量控制措施的一部分，并提供了出色的重现性（％CV 0.9-3.4）。该方法的总运行时间为19分钟，其中包括色谱柱重新处理。使用该方法，可通过以下方法测定硫胺素（67±7 ng / g），核黄素（475±45 ng / g），烟酰胺（856±77 ng / g）和吡ido醇（133±11 ng / g）的量。复杂的食物基质。

总之，我们开发了一种快速，可靠的HILIC单四极杆质谱方法，该方法利用SIM在20分钟内对蔬菜汤基质中的四种B族维生素进行了低含量定量。此方法显示出极好的线性，日内和日间重现性，可直接应用于其他基于植物的食品基质。