Regular ArticleMeasurement of Nitrite and Nitrate in Biological Fluids by Gas Chromatography–Mass Spectrometry and by the Griess Assay: Problems with the Griess Assay—Solutions by Gas Chromatography–Mass Spectrometry
Abstract
Assay methods based on the Griess reaction are frequently used to measure nitrite and nitrate in urine, plasma, and other biological fluids. With minor exceptions, careful attention has not been paid in extending the Griess assay from aqueous solutions to biological fluids. In the present study, parallel measurements of nitrite and nitrate were performed in urine, plasma, and aqueous solutions with a published batch assay based on the Griess reaction and with gas chromatography–mass spectrometry (GC–MS). We report here further interferences by free reduced thiols, proteins, and other plasma constituents in the Griess assay but not in GC–MS. The best correlation (r2= 0.985) between the Griess assay and GC–MS was observed for aqueous solutions in the absence of thiols. Unlike GC–MS, the Griess assay was not applicable to whole human plasma and urine samples. For the measurement of nitrate in diluted human urine samples, reduction by cadmium was performed both under acidic (pH 2 or 5) and alkaline (pH 8.8) conditions. The mean recovery rate of nitrate from urine samples was quantitative in the GC–MS but amounted to only 30–80% in the Griess assay. Measurement of nitrate in human urine samples (n= 33) resulted in an excellent correlation between two GC–MS techniques (r2= 0.979) but only in a poor correlation (r2< 0.64) between the Griess assay and GC–MS. Unlike GC–MS, the batch Griess assay is associated with many problems in measuring nitrate in biological fluids.
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Rapid-Response Nitrite Probes: Intramolecular Griess Reaction for Nitrite Detection at Picogram Level
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Mate tea reduces high fat diet-induced liver and metabolic disorders in mice
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Atorvastatin dose-dependently promotes mouse lung repair after emphysema induced by elastase
2018, Biomedicine and PharmacotherapyEmphysema results in a proteinase – antiproteinase imbalance, inflammation and oxidative stress. Our objective was to investigate whether atorvastatin could repair mouse lungs after elastase-induced emphysema. Vehicle (50 μL) or porcine pancreatic elastase (PPE) was administered on day 1, 3, 5 and 7 at 0.6 U intranasally. Male mice were divided into a control group (sham), PPE 32d (sacrificed 24 h after 32 days), PPE 64d (sacrificed 24 h after 64 days), and atorvastatin 1, 5 and 20 mg treated from day 33 until day 64 and sacrificed 24 h later (A1 mg, A5 mg and A20 mg, respectively). Treatment with atorvastatin was performed via inhalation for 10 min once a day. We observed that emphysema at day 32 was similar to emphysema at day 64. The mean airspace chord length (Lm) indicated a recovery of pulmonary morphology in groups A5 mg and A20 mg, as well as recovery of collagen and elastic fibers in comparison to the PPE group. Bronchoalveolar lavage fluid (BALF) leukocytes were reduced in all atorvastatin-treated groups. However, tissue macrophages were reduced only in the A20 mg group compared with the PPE group, while tissue neutrophils were reduced in the A5 mg and A20 mg groups. The redox balance was restored mainly in the A20 mg group compared with the PPE group. Finally, atorvastatin at doses of 5 and 20 mg reduced nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and matrix metalloproteinase-12 (MMP-12) compared with the PPE group. In conclusion, atorvastatin was able to induce lung tissue repair in emphysematous mice.
Analysis of Nitrite and Nitrate in Foods: Overview of Chemical, Regulatory and Analytical Aspects
2017, Advances in Food and Nutrition ResearchIn this chapter, several factors that should be considered for selecting and developing suitable analytical methods for determining nitrite/nitrate are presented. Nitrite and nitrate occurrence and suitability are a controversial issue. Nitrite is an approved additive considered a foremost curing ingredient for the preservation of meat products. Nitrate is a natural constituent of the human diet that, however, raises fears for its suggested potential harmfulness related to carcinogenesis and environmental contamination. Chemical, regulatory, and analytical aspects are discussed in the light of the need to obtain reliable data of nitrite and nitrate for law enforcement purposes, exposure estimates, and investigation of their physiological role in the human body. In addition, current metrological aspects to ensure the “fitness for purpose” of the selected method are suggested and discussed.
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