Department of Analytical Chemistry, Medical University of Lublin, Chodźki 4A, 20-093 Lublin, Poland.
Department of Chemical Technology, Faculty of Chemistry, Maria Curie Skłodowska University, Pl. Maria Curie-Skłodowskiej 3, 20-031 Lublin, Poland.
Molecules. 2024 May 1;29(9):2094. doi: 10.3390/molecules29092094.
The techniques used to detect and quantify cyanocobalamin (vitamin B12) vary considerably in terms of detection sensitivity, from the most sensitive, based on radioisotopes and mass spectrometry (MS) with limits of detection (LOD) in fg mL, to fluorescence (FL) and surface plasmon resonance (SPR) biosensors with LOD values in the range of a few µg mL. For accurate quantification of an analyte present at trace levels in complex biological matrices, a selective separation and enrichment step is required to overcome matrix interferences and ensure sufficient detection sensitivity. In this study, iron oxide magnetic nanoparticles (IONPs) were used for the extraction and initial preconcentration of cyanocobalamin (vitamin B12). In the dependence of the magnetization on the H-field (hysteresis loop), no coercivity and remanence values were found at 300 K, indicating the superparamagnetic properties of the tested IONPs. Perfluorinated acids were used as amphiphilic agents to allow the sorption of cyanocobalamin onto the IONPs. FT-IR/ATR spectroscopy was used to confirm the sorption of cyanocobalamin on the IONPs. The influence of the addition of a homologous series of perfluorinated acids such as trifluoroacetic acid (TFAA), heptafluorobutyric acid (HFBA), and trichloroacetic acid (TCAA) to the extraction mixture was tested considering their type, mass, and time required for effective sorption. The adsorption kinetics and isotherm, described by the Freundlich and Langmuir equations, were analyzed. The maximum adsorption capacity () exceeded 6 mg g and was 8.9 mg g and 7.7 mg g for HFBA and TCAA, respectively, as the most efficient additives. After the desorption process using aqueous KHPO solution, the sample was finally analyzed spectrophotometrically and chromatographically. The IONP-based method was successfully applied for the isolation of cyanocobalamin from human urine samples. The results showed that the developed approach is simple, cheap, accurate, and efficient for the determination of traces of cyanocobalamin in biological matrices.
用于检测和定量氰钴胺素(维生素 B12)的技术在检测灵敏度方面差异很大,从最灵敏的基于放射性同位素和质谱(MS)的方法(检测限(LOD)为 fg/mL)到荧光(FL)和表面等离子体共振(SPR)生物传感器,其检测限(LOD)值在几 µg/mL 范围内。为了准确定量痕量存在于复杂生物基质中的分析物,需要选择性的分离和富集步骤来克服基质干扰并确保足够的检测灵敏度。在这项研究中,使用氧化铁磁性纳米粒子(IONP)来提取和初步预浓缩氰钴胺素(维生素 B12)。在磁化与 H 场(磁滞回线)的关系中,在 300 K 时未发现矫顽力和剩磁值,表明所测试的 IONP 具有超顺磁性。全氟酸被用作两亲试剂,以允许氰钴胺素吸附到 IONP 上。傅里叶变换红外/衰减全反射(FT-IR/ATR)光谱用于证实氰钴胺素在 IONP 上的吸附。测试了添加全氟羧酸(如三氟乙酸(TFAA)、七氟丁酸(HFBA)和三氯乙酸(TCAA))的同系物系列对萃取混合物的影响,考虑了它们的类型、质量和有效吸附所需的时间。采用 Freundlich 和 Langmuir 方程对吸附动力学和吸附等温线进行了分析。吸附容量(qmax)超过 6 mg/g,HFBA 和 TCAA 的最大吸附容量分别为 8.9 和 7.7 mg/g,是最有效的添加剂。在使用 KHPO 水溶液进行解吸过程后,最终采用分光光度法和色谱法对样品进行分析。基于 IONP 的方法成功地用于从人尿样中分离氰钴胺素。结果表明,该方法简单、廉价、准确、高效,可用于测定生物基质中痕量的氰钴胺素。