Simultaneous high performance liquid chromatographic separation of purines, pyrimidines, N-acetylated amino acids, and dicarboxylic acids for the chemical diagnosis of inborn errors of metabolism

Tavazzi, Barbara; Lazzarino, Giuseppe; Leone, Paola; Amorini, Angela Maria; Bellia, Francesco; Janson, Christopher G.; Pietro, Valentina Di; Ceccarelli, Lia; Donzelli, Sonia; Francis, Jeremy S.; Giardina, Bruno

doi:10.1016/j.clinbiochem.2005.08.002

Cited by 97 publications

(87 citation statements)

References 40 publications

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“…Patients with Canavan have plasma uric acid concentration as high as ∼1700 μM, which is 3-fold higher than the concentrations in CKD patients. 68 Hippuric acid is a clinically relevant uremic solute that inhibited OAT1 and OAT3 in our study. It can be acquired from consumption of food enriched in phenolic compounds such as wine, grape juice, tea, etc.…”

Section: Clinical Evidence Supporting the Role Of Uremic Solutes In Mmentioning

confidence: 50%

Identification and Quantitative Assessment of Uremic Solutes as Inhibitors of Renal Organic Anion Transporters, OAT1 and OAT3

et al. 2016

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One of the characteristics of chronic kidney disease (CKD) is the accumulation of uremic solutes in the plasma. Less is known about the effects of uremic solutes on transporters that may play critical roles in pharmacokinetics. We evaluated the effect of 72 uremic solutes on organic anion transporter 1 and 3 (OAT1 and OAT3) using a fluorescent probe substrate, 6-carboxyfluorescein. A total of 12 and 13 solutes were identified as inhibitors of OAT1 and OAT3, respectively. Several of them inhibited OAT1 or OAT3 at clinically relevant concentrations and reduced the transport of other OAT1/3 substrates in vitro. Review of clinical studies showed that the active secretion of most drugs that are known substrates of OAT1/3 deteriorated faster than the renal filtration in CKD. Collectively, these data suggest that through inhibition of OAT1 and OAT3, uremic solutes contribute to the decline in renal drug clearance in patients with CKD.

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Section: Clinical Evidence Supporting the Role Of Uremic Solutes In Mmentioning

confidence: 50%

Identification and Quantitative Assessment of Uremic Solutes as Inhibitors of Renal Organic Anion Transporters, OAT1 and OAT3

et al. 2016

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“…Other nucleobases were detected at 4.58 min (5mC), 5.63 min (HX), and 5.89 min (X), respectively. The LOD concentrations for primary nucleobases ranged from 20-32 nmol L -1 (Table 1), and were comparable to or lower than those reported previously for HPLC and UPLC analyses (Berg and Jørgensen 2006;Fan et al 2007;Sotgia et al 2010;Tavazzi et al 2005). The LOQ concentrations ranged from 65 to 110 nmol L -1 .…”

Section: Hplc-dad Methods For Nucleobase Analysismentioning

confidence: 84%

“…A wide variety of separation techniques, including paper chromatography, thin-layer chromatography, bioassays, capillary electrophoresis, gas chromatography, high-performance liquid chromatography (HPLC), and ultra-performance liquid chromatography (UPLC), have been applied for the measurement of purines and pyrimidines (Tavazzi et al 2005) and the detection of oxidative DNA damage (Sotgia et al 2010). Reversed-phase HPLC can be employed for various applications, but most conventional reversed-phase columns (e.g., C 8 and C 18 alkyl columns) are unsuitable for retaining and separating such highly polar, ionic, and hydrophilic compounds under low organic or aqueous mobile phase conditions due to collapse of the bonded-phase (Pesek and Matyska 2005;Sotgia et al 2010).…”

mentioning

confidence: 99%

A simple high performance liquid chromatography method for the measurement of nucleobases and the RNA and DNA content of cellular material

Huang

Kaiser

Benner

2012

Limnology & Ocean Methods

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Current methods are not generally suitable for the quantitative analysis of nucleobases in cellular material and more complex matrices that are often encountered with environmental samples. Simple and reliable quantitative methods for nucleobase analysis in purified ribonucleic acid (RNA) and deoxyribonucleic acid (DNA), microorganisms, and environmental samples are presented. Novel microwave-assisted vapor phase and liquid phase hydrolyses with HCl (6 mol L -1 ) were developed for the analysis of nucleobases in soluble and particulate samples, respectively. Optimal conditions for the release of purines (130°C, 10 min) and pyrimidines (160°C, 40 min) from DNA and RNA were applicable to both the vapor phase and liquid phase hydrolysis. Residual hydrochloric acid in hydrolysate can be removed easily by drying under a stream of nitrogen gas. Nucleobases were separated and quantified using high performance liquid chromatography with diode array detection (HPLC-DAD). The limit of detection for primary nucleobases ranged from 20 to 32 nmol L -1 . Total hydrolysable nucleobases were analyzed in salmon testes DNA, yeast RNA, cultures of marine microalgae and cyanobacteria, and suspended particles from an estuary. The RNA and DNA contents and RNA/DNA ratios in cellular material were estimated from analyses of hydrolysable nucleobases. AcknowledgmentsWe thank Si Chen for the collection and preparation of estuarine suspended particles. We thank the editor and anonymous reviewers for helpful comments on the manuscript. This research was supported by US National Science Foundation (Grant 0850653) and National Natural Science Foundation of China (Grant 41071301).DOI 10.4319/lom.2012.10.608 Limnol. Oceanogr.: Methods 10, 2012, 608-616 © 2012, by the American Society of Limnology and Oceanography, Inc. LIMNOLOGY and OCEANOGRAPHY: METHODSand some of their derivatives, but the quantitative release of nucleobases from nucleic acids remains challenging. Various methods have been used for the hydrolysis of nucleic acids, but it is still difficult to recover high yields of nucleobases using published methods. The commonly used hydrolysis protocols for the determination of purines and pyrimidines in DNA are 98% formic acid at 175°C for 30 min (Vischer and Chargaff 1948;Wyatt 1951) and 72% perchloric acid at 100°C for 1 h (Marshak and Vogel 1951;Wyatt 1951). Several attempts were made to use these hydrolyses for the analysis of RNA (Narurkar and Sahasrabudhe 1957;Vischer and Chargaff 1948;Wyatt 1951), but they were largely unsuccessful as were early attempts to develop hydrolyses with hydrochloric acid (Smith and Markham 1950;Vischer and Chargaff 1948). There have been surprisingly few improvements in methods for the hydrolysis of RNA in the years since these earlier studies (Fan et al. 2007).In the present study, novel methods were developed for the hydrolysis and analysis of nucleobases in organisms and environmental samples. Two separate hydrolyses with HCl were developed for the efficient release of purines and pyrimidine...

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“…We have recently demonstrated that this nonlinear time-course of post-traumatic NAA recovery may be due to the cerebral energy imbalance, assessed by high-energy phosphate quantification (ATP, ADP, AMP, etc. ), caused mainly by mitochondrial malfunctioning, as indicated by altered mitochondrial phosphorylating capacity (measured by the ATP/ADP ratio) (Signoretti et al, 2010;Tavazzi et al, 2005). Under these conditions, the remarkable decrease in cerebral NAA, which mirrors the changes in brain ATP, may possibly be attributed to the general energy depression consequent to impaired mitochondrial functions (Lifshitz et al, 2003;Robertson et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

The Importance of Restriction from Physical Activity in the Metabolic Recovery of Concussed Brain

Lazzarino¹,

Vagnozzi²,

Signoretti³

et al. 2012

Brain Injury - Pathogenesis, Monitoring, Recovery and Management

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Simultaneous high performance liquid chromatographic separation of purines, pyrimidines, N-acetylated amino acids, and dicarboxylic acids for the chemical diagnosis of inborn errors of metabolism

Cited by 97 publications

References 40 publications

Identification and Quantitative Assessment of Uremic Solutes as Inhibitors of Renal Organic Anion Transporters, OAT1 and OAT3

Identification and Quantitative Assessment of Uremic Solutes as Inhibitors of Renal Organic Anion Transporters, OAT1 and OAT3

A simple high performance liquid chromatography method for the measurement of nucleobases and the RNA and DNA content of cellular material

The Importance of Restriction from Physical Activity in the Metabolic Recovery of Concussed Brain

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