کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5136716 1494022 2016 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Quantitative UPLC-MS/MS assay of urinary 2,8-dihydroxyadenine for diagnosis and management of adenine phosphoribosyltransferase deficiency
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Quantitative UPLC-MS/MS assay of urinary 2,8-dihydroxyadenine for diagnosis and management of adenine phosphoribosyltransferase deficiency
چکیده انگلیسی


- Design of experimental strategy for an efficient method development of an UPLC-MS/MS-based clinical assay.
- Absolute quantification of DHA in urine using an isotopically labeled internal standard is reported for the first time.
- Improved detection of urinary DHA excretion utilizing UPLC-MS/MS.
- Robust method for clinical diagnosis and monitoring of pharmacotherapy in patients with APRT deficiency.

Adenine phosphoribosyltransferase (APRT) deficiency is a hereditary disorder that leads to excessive urinary excretion of 2,8-dihydroxyadenine (DHA), causing nephrolithiasis and chronic kidney disease. Treatment with allopurinol or febuxostat reduces DHA production and attenuates the renal manifestations. Assessment of DHA crystalluria by urine microscopy is used for therapeutic monitoring, but lacks sensitivity. We report a high-throughput assay based on ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) for quantification of urinary DHA.The UPLC-MS/MS assay was optimized by a chemometric approach for absolute quantification of DHA, utilizing isotopically labeled DHA as an internal standard. Experimental screening was conducted with D-optimal design and optimization of the DHA response was performed with central composite face design and related to the peak area of DHA using partial least square regression. Acceptable precision and accuracy of the DHA concentration were obtained over a calibration range of 100 to 5000 ng/mL on three different days. The intra- and inter-day accuracy and precision coefficients of variation were well within ±15% for quality control samples analyzed in replicates of six at three concentration levels. Absolute quantification of DHA in urine samples from patients with APRT deficiency was achieved wihtin 6.5 min. Measurement of DHA in 24 h urine samples from three patients with APRT deficiency, diluted 1:15 (v/v) with 10 mM ammonium hydroxide (NH4OH), yielded a concentration of 3021, 5860 and 10563 ng/mL and 24 h excretion of 816, 1327 and 1649 mg, respectively. A rapid and robust UPLC-MS/MS assay for absolute quantification of DHA in urine was successfully developed. We believe this method will greatly facilitate diagnosis and management of patients with APRT deficiency.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography B - Volumes 1036–1037, 15 November 2016, Pages 170-177
نویسندگان
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