Determination of the geographic origin of 52 honey samples based on the assessment of anionic content profiling with a new algorithm using monolithic column-based micellar nano-liquid chromatography.

In the present study, a new micellar nano LC-UV was, for the first time, reported for the separation and determination of five anions (chloride, nitrite, bromide, sulfate and nitrate) in 52 honey samples. Based on this approach, a graphene oxide-based monolithic column was prepared and applied for the samples. Various amounts of hexadecyltrimethyl-ammonium bromide (HTAB) in the mobile phase were used in order to optimize the separation conditions. The baseline separation was achieved using mobile phase with 25/75% (v/v) ACN/10 mM phosphate buffer at pH 3.4, while the amount of HTAB was optimized as 0.22 mM in the mobile phase. The whole method was validated and it leads to high sensitivity. The LOD values were found in the range of 0.02-0.22 µg/kg, while LOQ values were found in the range of 0.06-0.18 µg/kg. The method allowed to achieve sensitivity analyses of anionic content in 52 honey samples. All data were evaluated using a new algorithm for geographic origin discrimination. K-nearest neighbor algorithm (K-NN), cubic support vector classifier (K-DVS), and K-Mean cluster analysis were used for geographic origin discrimination of honeys. The accuracy of the whole model was calculated as 94.4% with the K-DVS method. The samples from five provinces were classified 100% correctly, while two of them were classified with one misclassification, with an accuracy of 89.9% and 83.3%, respectively. PRACTICAL APPLICATION: The new platforms and advanced technologies are crucial for advanced food analysis. In this article, a novel methodology was attempted for the determination of geographic origin of 52 honey samples. In this sense, micellar nano LC technique with a homemade monolithic nano-column was, for the first time, applied for the anion analysis using a new algorithm.

Nano-liquid chromatography with a new nano-structured monolithic nanocolumn for proteomics analysis.

Herein, we report the preparation and application of a new nano-structured monolithic nanocolumn based on modified graphene oxide using narrow fused silica capillary column (e.g., 50 µm internal diameter). The nanocolumn was prepared by an in situ polymerization using butyl methacrylate, ethylene dimethacrylate, and methacryloyl graphene oxide nanoparticles. Dimethyl formamide and water were used as the porogenic solvent. After polymerization, the obtained nanocolumn was coated with dimethyloctadecylchlorosilane in order to enhance the hydrophobicity. Both isocratic and gradient nano-liquid chromatographic separations for small molecules (e.g., alkylbenzenes) and macromolecules (e.g., intact proteins) were performed. Theoretical plates number up to 3600 plates/m in isocratic mode for propylbenzene were achieved. It was demonstrated that the feasibility of graphene oxide modified monolithic nanocolumn for high-efficiency and high-throughput nanoscale proteomics analysis. The high resolving power of monolithic nanocolumn yielded sensitive protein separation with narrower peak width while a high-resolution analysis of peptides from trypsin-digested cytochrome C could be obtained. Graphene oxide based monolithic nanocolumns are promising and can allow to powerful tools for trace proteom sample analysis.

Graphene oxide-octadecylsilane incorporated monolithic nano-columns with 50 µm id and 100 µm id for small molecule and protein separation by nano-liquid chromatography.

In this study, graphene oxide-octadecylsilane incorporated monolithic nano-columns were developed for protein analysis by nano liquid chromatography (nano LC). The monolithic column with 100 µm id was first prepared by an in situ polymerization using ethylene dimethacrylate (EDMA), 3-chloro-2-hydroxypropylmethacrylate (HPMA-Cl), and methacryloyl graphene oxide nanoparticles (MGONPs). MGONPs were synthesized by the treatment of 3-(trimethoxysilyl)propylmethacrylate (TMSPM) and GO. Tetrahydrofuran (THF) and dodecanol were used as the porogenic solvent. The resulting column was functionalized by dimethyloctadecylch lorosilane (DODCS) for the enhancement of hydrophobicity. The functionalization greatly improved the baseline separation of hydrophobic compounds such as polyaromatic hydrocarbons (PAHs). The optimized monolith with respect to total polymerization mixture was characterized by using Fourier-transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) X-ray diffraction (XRD) and chromatographic analyses. The blank monoliths without functionalization exhibited poor separation while a good separation performance of MGONPs functionalized monoliths was achieved. The monolith with 100 µm id was evaluated in protein separation in nano LC using RNase A, Cytochrome C, Lysozyme, Trypsin, and Ca isozyme II as the test proteins. It was shown that protein separation mechanism was based on large π-system of GO and hydrophobicity of the monolithic structure. Theoretical plates number up to 57 600 plates were achieved. The nano-column with 50 µm id was also prepared using the same polymerization mixture under the same chemical conditions. These nano-columns were employed for protein separation by nano LC, and the dependence of both nano-column performance on the internal diameter was also discussed.