ABSTRACT
Objectives: AICAR (5-amino-4-imidazolecarboxyamide ribonucleoside) was reported as the first pharmacological AMPK (adenosine 5'-monophosphate (AMP)-activated protein kinase) activator, and it has been confirmed to exhibit a significant endurance enhancement effect and prohibited for doping by the World Anti-Doping Agency. Due to the fact that the human body can produce such substances, in order to ensure fairness in sports competition, methods for rapid detection and multi-type identification of AICAR drugs taken orally should be established. Methods: to assess AICAR levels, a new rapid, sensitive, efficient, and selective method was reported for the quantitative detection of AICAR in urine using LC-MS/MS. The method was validated for quantitative purposes based on the elemental selectivity, intra- (1.0-15.6%) and inter-day precision (1.3-16.3%), accuracy (99.9-112.8%), matrix effects (88.9-103.6%), recovery (87.4-106.5%), and stability at four different concentrations. The calibration curve was linear over a wide concentration range of 10-10,000 ng mL-1 with a high coefficient of determination (R 2 > 0.998). The limit of detection (LOD) and limit of quantification (LOQ) for the experiment were determined to be 1 and 10 ng mL-1, respectively. Simultaneously, metabolomics analysis was used to obtain the metabolic fingerprint of different populations and biomarkers to distinguish administration cases through partial least squares discriminant analysis (PLS-DA) and a receiver operating characteristic (ROC) curve. Results: the method enables easy quantitation for LC-MS/MS analysis with the best recovery yield maintained, and the method was applied to 122 Asian biological samples with an average concentration of 1310.5 ± 1031.4 ng mL-1. Through drug metabolism research, 734 and 294 variables were extracted for data analysis respectively in the positive and negative ion modes, and more than 100 metabolites with significant up- and down-regulation were found after the test. Conclusions: this research developed a fast, precise, effective, and specific approach for the qualitative and quantitative identification of AICAR in urine. Meanwhile, administration metabolism studies found that there were significant changes in AICAR levels and other compounds, such as PC types PC(18:1/16:0), PC(16:0/18:0), and PC(16:0/16:0), PE types PE(18:0/20:4), and LPE-type 18:1, which could better distinguish samples before and after AICAR administration. The analysis provides a multi-perspective reference for WADA to determine a positive criterion.
ABSTRACT
PURPOSE: To evaluate measurement precision and to compare the Pentacam AXL (Oculus Optikgeräte, Wetzlar, German), a new optical biometer based on Scheimpflug imaging and partial coherence interferometry (PCI) with that of the OA-2000 biometer (Tomey, Nagoya, Japan), which combines swept-source optical coherence tomography (SS-OCT) and Placido-disk topography. METHODS: Axial length (AL), central corneal thickness (CCT), anterior chamber depth (ACD), aqueous depth (AQD), mean keratometry (Km), astigmatism vectors J0, J45, and corneal diameter (CD) were measured in triplicate by two technical operators. Within-subject standard deviation (Sw), repeatability and reproducibility (2.77 Sw), coefficient of variation (CoV), and intraclass correlation coefficient (ICC) were used to assess the Pentacam AXL intra-observer repeatability and inter-observer reproducibility. Paired t-test and Bland-Altman plots were used to determine the agreement between the two biometers. RESULTS: The new optical biometer had high intra-observer repeatability [all parameters evaluated had low CoV (<0.71%) and high ICC (>0.88)]. Inter-observer reproducibility was also excellent, with high ICC (>0.95) and low CoV (<0.52%). The 95% LoA between the new biometer and OA-2000 were insignificant for most of the parameters evaluated, especially for AL. However, the measurement agreement was moderate for CCT. CONCLUSIONS: Intra-observer repeatability and inter-observer reproducibility were excellent for all parameters evaluated using the new optical biometer based on Scheimpflug imaging and PCI. There was a high agreement between the two devices and hence could be clinically interchangeable for the measurement of most ocular parameters.
