Introduction: Cancer disease is one of the causes of death, and the diagnosis of its early stages has greatly affected the survival of patients. Prostate cancer is the second most common cancer in men and the fourth most common cancer. Prostate cancer is an aggressive disease and usually manifests itself in the advanced stage of the disease. The present study aims to analyze and analyze the metabolic amino acid profile as a strategy to identify biomarkers for the diagnosis and early screening of prostate cancer.
Methods: The present study aims to analyze the metabolic amino acid profile as a strategy to identify biomarkers for the diagnosis and early screening of prostate cancer. Biological samples are provided from patients in the early and often asymptomatic stages of cancer. What is important is the observed changes in the amino acid profile of these patients. Among several analytical methods, LC-ESI-MS/MS is a promising tool. In this pilot study, we are dealing with complex biological samples, so a sample prpration method namly magnetic dipersive solid phase extraction is used to purify and concentrate amino acids. The synthesized magnetic adsorbent and the effective factors on the extraction method are investigated and optimized one by one. In the same way, a device analysis method is also launched to determine the amino acid profile. The parameters affecting the device analysis method are optimized one parameter at a time and are used in the next experiments in an optimized way.
Results: The present study was devoted to synthesizing Fe3O4@SiO2 nanocomposites via silica nanoparticles obtained from rice husk waste and then its functionalization using ammonia solution to get Fe3O4@SiO2-NH2 nano-hybrid as a new sorbent by improving selectivity in aminoacids sorption. The structure and morphology of the synthesized sorbent were characterized by X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM), and Fourier transform infrared spectroscopy (FT-IR). As-prepared nano-sorbent was successfully used in magnetic solid phase extraction setup with no hazardous solvents and extraction was performed in 9 min. Some important parameters affecting the extraction of the aminoacids were optimized. Under the optimized experimental conditions, the method provided a linear range of 10-1000 ng mL-1 with the correlation coefficients (R2) of 0.998. The intra-day (n=3) and inter-day precisions (n=3 working days) calculated in the form of percent relative standard deviations (%RSDs) were below 10%. The proposed method was successfully practiced for analyzing aminoacids in several human plasma samples, with recoveries in the range of 84.0–109.5% for the spiked samples.
Conclusion: This study obtains and analyzes the profile of free amino acids in the plasma of prostate cancer patients and healthy men. Considering the contradictory results in changing the characteristics of amino acids in cancers and the high prevalence of prostate cancer in Iran and the lack of epidemiological studies on the level of plasma amino acids in patients with prostate cancer, this study can provide the basis for further studies. Determining amino acid profile using non-invasive HPLC-Tandem MS and new extraction method based on magnetic adsorbent helps in simple separation of amino acids from complex environment and their rapid determination. The developed method is a reliable method for prostate cancer screening, diagnosis and pathogenesis.
Keywords: Biomarker - Prostate cancer - Amino acid profile - Magnetic solid phase extraction