Development of a high-performance liquid chromatography – Tandem mass spectrometry urinary pterinomics workflow

Abstract

Pteridines have evoked considerable interest from the scientific community owing to their prominent roles in human health and disease. The availability of analytical methodologies suitable for comprehensive pteridine profiling, termed here as "pterinomics", has been limited by inconsistent sample preparation and the exclusion of lesser studied pteridine derivatives. In response, the present study describes a new pterinomics workflow using a high-performance liquid chromatography - tandem mass spectrometry (HPLC-MS/MS) methodology for the simultaneous analysis of 15 pteridine derivatives including four structural isomers, marking the largest quantitative pteridine panel that has been studied to-date. The validated method possessed excellent sensitivity with method detection limits (0.025 μg L(-1) to 0.5 μg L(-1)) that were comparable or superior to existing techniques. Spiked recovery studies demonstrated the technique was both accurate (88-112%) and precise (RSD: 0-6%). A comparative study of commonly used oxidative pretreatments, including triiodide, permanganate, and manganese dioxide, revealed that the oxidative mechanisms were inefficient, complex, and concentration dependent. Finally, 50 clinical urine specimens were examined with the new technique wherein 10 pteridine derivatives were quantified and population ranges have been given. This technique can be used to examine pteridine molecular epidemiology and biochemistry to support related research applications, and may further be readily extended to include additional pteridine derivatives and biological matrices for specific applications.

Development of an HPLC-MS method for the determination of natural pteridines in tomato samples

“…These data are contradictory with the results reported by Van Daele and co-workers. 27 However, results similar to ours have been recently described by Burton et al, 28 who found negligible in-source oxidation of 7,8-dihydroxanthopterin to xanthopterin. a Fragmentor voltage: 100 V, capillary voltage: 4500 V, nebulizer pressure: 30 psi, temperature: 300 C.…”

“…27 In the above mentioned paper the authors report that dihydropterins are subjected to interconversion, on column, in source and auto oxidation, and they are degraded into non-pterin products during boiling. Later, Burton et al 28 established the pterinomic workow for 15 pteridin derivatives in urine using HPLC-MS/MS revealing that previous oxidative steps were inefficient. The elimination of the preoxidation step in the analysis of pteridinic derivatives was previously recommended by Cañada et al 29 It is remarkable that Burton et al showed that 7,8-dihydroxanpthopterin exhibited negligible in-source oxidation to xanthopterin.…”

Urinary neopterin: A novel biomarker of disease progression in amyotrophic lateral sclerosis

“…More recently, HPLC‐mass spectrometry provides a means to detect neopterin more accurately. This technique has been shown to be robust in measuring urinary neopterin [53] and will be pursued in the future.…”

Analysis of Catecholamines and Pterins in Inborn Errors of Monoamine Neurotransmitter Metabolism—From Past to Future

“…The concentration of BH 4 present in the sample is calculated by determining the difference in biopterin content between the two different oxidation procedures. Most protocols are based on HPLC coupled to fluorescence detection, but there are also newer protocols available combining the separation via HPLC with MS/MS [126,128]. Analysis of urinary sepiapterin content has been shown to be a useful non-invasive tool to identify patients suffering from SRD [103].…”