Detection of urinary metabolites of metabolic pathway disorders by using VTGE and LC-HRMS techniques

Abstract

Background:In recent, various human health disorders including cancer, diabetes, neurodegenerative and metabolic diseases are noticed among human populations. Currently, genetic and proteomic approaches are highly reported to detect metabolic disorders that also include inborn error of metabolisms. These existing detection methods are faced with cost issue and time consuming factors. Therefore, metabolites as biomarkers are one of potential avenues to detect metabolic disorders. Further, exploitation of urine as potential source of metabolite biomarkers, there are limitation in this area of research due to abundance of non-metabolite components such as proteins and nucleic acids. Hence, methods and processes are required to precisely fractionate metabolites from urine of inborn error of metabolism patients and then identified by analytical tools such as LC-HRMS and GC-MS.Methods: Sterile filtered urine samples (750 µl) mixed with (250 µl) loading buffer were electrophoresed on VTGE that uses acrylamide gel (acrylamide:bisacrylamide, 30:1) as matrix of 15%. Further, vertical tube gel electrophoresis (VTGE) technique combined with LC-HR-MS to identify metabolites that are known as the biomarkers of metabolic disorders was carried out. Results and Discussion:The authors provide evidence on the use of novel VTGE coupled with LC-HRMS to detect metabolites among metabolic disorders. Data suggest the applicability of VTGE coupled with LC-HRMS technique to detect metabolites such as 2-methyluridine, 2-Methylglutaric acid, 2-Methyl citric acid, 2-Hydroxyglutaric acid in case of metabolic disorders. Conclusion:This preliminary work is suggested to be extended to large clinical samples to validate application of this method to detect metabolic disorders including inborn error of metabolisms.

Detection of Nail Oncometabolite SAICAR in Oral Cancer Patients and Its Molecular Interactions with PKM2 Enzyme

“…Additionally, the importance of SAICAR has relevance as a biomarker and a source of future anticancer drugs as metabolic mimetics that can block the pro-cancer enzymes. In line with existing views, the authors report on a novel approach to the detection of SAICAR in the nails of oral cancer patients with the assistance of a novel and in-house vertical tube gel electrophoresis (VTGE) tool [ 26 , 29 ] (27).…”

“…Altered metabolism and survival under stressed conditions are some of the hallmarks of cancer. During metabolic reprogramming, cancer cells and cancer supporting stroma including microbiome establish a source and link partnership for the exchange of key oncometabolites including SAICAR, for growth and proliferation [ 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 ]. SAICAR is known as an important intermediate metabolite that is catalyzed by SAICAR synthetase in both cancer cells and the microbiome [ 30 , 31 , 32 , 33 ].…”

Detection of Nail Oncometabolite SAICAR in Oral Cancer and Its Molecular Interactions with PKM2 Enzyme

“…Recent studies on oral cancer, metabolite biomarker is a focus for the early diagnosis of oral cancer. By taking these findings in consideration, we explored nail metabolome by the purification of metabolites from nails of oral cancer subjects with the help of novel VTGE tools (27) (28).…”