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The ratio of the area under the curves of each amino acid to its assigned internal standard was then plotted against a multiple point calibration curve, allowing for the quantitation of the amino acids in the patient samples. Intra-assay coefficient of variation (CV) for all amino acids ranged from 2% to 5% while the inter-assay CV was approximately 6%. Metabolic variables associated with insulin resistance To compare the magnitude of the relationship between insulin resistance and amino acids, we assessed the relationship between SSPG and metabolic variables known to be associated with insulin resistance: fasting glucose, triglyceride, high-density lipoprotein cholesterol (HDL-C), and insulin concentrations (Kim and Reaven high throughput screening 2013). Triglyceride and HDL-C concentrations were measured in the core laboratory at Stanford University Medical Center. Glucose was determined by the oxidase method (Analyzer 2; Beckman, Brea, CA). Fasting plasma insulin concentrations were measured at Washington University (St. Louis, MO) using radioimmunoassay (Millipore, St. Charles, MO); the inter- and intra-assay CV ranged between 4.7% and 9.7%. Insulin concentrations were available on 114 individuals (75 women and 39 men). Statistical analysis All statistical analyses were performed using SPSS (version 22 for Windows, SPSS Inc., Armonk, NY). Data are presented as mean?��?SD or median [interquartile Idelalisib supplier range]. Amino acids were log transformed prior to analysis. Pearson correlations (r-values and 95% confidence intervals) are reported between individual amino acids and SSPG concentration. Confidence intervals are based on bootstrapping with 1000 replications. To adjust for body mass index (BMI) or waist circumference, a linear regression model was used to predict SSPG with individual amino acids as the independent variable. Measurement of waist circumference was only available in 140 of 182 individuals. Amino acid concentrations were also compared between subjects in the lowest (most insulin sensitive) and highest (most insulin resistant) SSPG tertiles. Statistical significance was defined as P?Ficain metabolic characteristics for the study population by sex. Women and men were not different in age, BMI, and degree of insulin resistance (SSPG). However, women had a better metabolic profile with lower fasting glucose and triglyceride concentration and higher HDL-C concentration. As expected, women also had lower creatinine concentration. Table 1 Baseline characteristics of women and men Sex-specific comparisons of amino acid concentrations are listed in Table?Table2.2.