High-Throughput Serum 25-Hydroxy Vitamin D Testing with Automated Sample Preparation
Elapsed time (min) % Mobile phase B Flow rate (mL/min) 0 30 1.0 0.900 80 1.0 0.901 92 1.0 1.150 92 1.0 1.160 95 1.0 1.170 95 1.7 1.460 95…
Elapsed time (min) % Mobile phase B Flow rate (mL/min) 0 30 1.0 0.900 80 1.0 0.901 92 1.0 1.150 92 1.0 1.160 95 1.0 1.170 95 1.7 1.460 95…
Calibrator Volume of stock standard (μL) Volume of ethanol (μL) Final concentration (μg/mL) Final dilution 6 200 – 6.0 – 5 200 200 3.0 1:2 4 333 667 2.0 1:3…
Calibrator concentration pg/mL Intermediate 50 ng/mL std (mL) Stripped serum (mL) 0 0 250 50 0.25 249.75 100 0.50 249.5 300 1.50 248.5 600 3.0 247.0 1000 5.0 245.0 2000…
To make Use source solution Volume of source solution Volume of 10 mM ammonium phosphate pH 6.5 (mL) Metanephrine concentration (pg/mL) Normetanephrine concentration (pg/mL) Std7 Working solution 100 μL QS…
Pre- Name Synonym Supp. Mol. wt. Acetoacetic Li salt Lithium acetoacetate Sigma 108.00 N– Acetylaspartic Aldrich 175.10 N– Acetyltyrosine Sigma 223.20 Aconitic cis-Aconitic Sigma 174.10 Adipic Sigma 146.14 Benzoic-Na salt …
2.4 Internal Standards 1. 1α,25-Dihydroxyvitamin D3 (26,26,26,27,27,27-d6), Medical Isotopes, Pelham, NH. 2. 1α,25-Dihydroxyvitamin D3 (26,26,26,27,27,27-d6), Medical Isotopes, Pelham, NH. 3. Preparation of individual standard stocks (20 μg/mL):…
Calibrator Vol. of 200 ng/mL stock added (μL) Vol. of MeOH added (μL) Urine added (μL) Total vol. (mL) Approx. final conc. (pg/mL)a 1 0 750 99,250 100 0 2…
Calibrator Tertiary combo standards (μL) Secondary combo standards (μL) Serum volume Final conc. (ng/dL) Blank 10 mL 1 10 μL qs to 10 mL 10 2 20 μL qs to…
Fig. 1 HPLC -ESI-MS/MS ion chromatogram of SAM (m/z 399.1 > 250.1), SAH (m/z 385.1 > 136.1), SAM-D3 (m/z 402.1 > 250.1) and SAH-D4 (m/z 389.1 > 138.1). Concentration of SAM and SAH is 95 nM and 41…
Analyte Stock solution [Expected] pH, Max UV wavelength ε max × 10−3 Adenine 30 mg in 100 mL Buffer A 2.2 mM pH 6–8, 260 nm 13.3 Xanthinea 12 mg in 1…