Effect of pH on 1H-NMR Spectroscopy of Mouse Urine

Miyataka, Hideki; Ozaki, Toshihiro; Himeno, Seiichiro

doi:10.1248/bpb.30.667

Cited by 20 publications

(13 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Rat urine typically covered a pH range from 5.5 to 8.5 (mean ± SD was 6.8 ± 1.1) depending on the time of day and on the metabolic status of the animal, and can reach extreme values after treatment of rats with different pharmaceuticals between pH 4.0 and 9.1 (Supplement Figure 2). Similar ranges were also described for mice, covering an even slightly broader range from 3 to 9 [3], which is supported by own unpublished data showing a pH range in mice from 3.9 to 7.8, with a mean ± SD of 7.2 ± 0.7 ( n = 32) for NMRI mice and of 5.4 ± 0.7 ( n = 77) for a different breed, showing in addition breed-specific normal ranges. In men, including healthy volunteers and patients with different treatments and for different illnesses, we found a pH range from 3.0 to 8.7 with a mean ± SD of 5.1 ± 1.3 ( n = 265 samples) in treated patients and of 5.0 ± 1.2 in healthy volunteers ( n = 104 samples).…”

Section: Discussionsupporting

confidence: 58%

“…If changes in these parameters are not taken into account during sample preparation, they may affect the nuclear magnetic resonance (NMR) analysis of a single metabolite. Only relatively few recommendations for sample collection, preparation, and storage have been published, especially on stabilization of the pH, which is the most important parameter in 1 H NMR spectroscopy [1–3]. Other studies give recommendations for buffer compositions to stabilize plasma [4], bile, and urine samples [1] or urine samples only [11], or for the maximum time for storing samples at 4°C [2, 5] or room temperature (RT) [6].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Reproducibility of NMR Analysis of Urine Samples: Impact of Sample Preparation, Storage Conditions, and Animal Health Status

Schreier¹,

Kremer

Huber³

et al. 2013

BioMed Research International

View full text Add to dashboard Cite

Introduction. Spectroscopic analysis of urine samples from laboratory animals can be used to predict the efficacy and side effects of drugs. This employs methods combining 1H NMR spectroscopy with quantification of biomarkers or with multivariate data analysis. The most critical steps in data evaluation are analytical reproducibility of NMR data (collection, storage, and processing) and the health status of the animals, which may influence urine pH and osmolarity. Methods. We treated rats with a solvent, a diuretic, or a nephrotoxicant and collected urine samples. Samples were titrated to pH 3 to 9, or salt concentrations increased up to 20-fold. The effects of storage conditions and freeze-thaw cycles were monitored. Selected metabolites and multivariate data analysis were evaluated after 1H NMR spectroscopy. Results. We showed that variation of pH from 3 to 9 and increases in osmolarity up to 6-fold had no effect on the quantification of the metabolites or on multivariate data analysis. Storage led to changes after 14 days at 4°C or after 12 months at −20°C, independent of sample composition. Multiple freeze-thaw cycles did not affect data analysis. Conclusion. Reproducibility of NMR measurements is not dependent on sample composition under physiological or pathological conditions.

show abstract

Section: Discussionsupporting

confidence: 58%

Section: Introductionmentioning

confidence: 99%

Reproducibility of NMR Analysis of Urine Samples: Impact of Sample Preparation, Storage Conditions, and Animal Health Status

Schreier¹,

Kremer

Huber³

et al. 2013

BioMed Research International

View full text Add to dashboard Cite

show abstract

“…[66] Miyataka et al, however, saw similar increases in TMA while studying effects of pH on mouse urine and ascribed it to the aging of the mice rather than to the effects of administering a drug. [67] Therefore, it is likely that the overall increase in TMA in antibiotic-treated mice is not directly associated with reduction of microbial flora, but rather due to aging over the course of the study.…”

Section: Discussionmentioning

confidence: 99%

NMR‐based metabonomics analysis of mouse urine and fecal extracts following oral treatment with the broad‐spectrum antibiotic enrofloxacin (Baytril)

Romick-Rosendale

Goodpaster

Hanwright

et al. 2009

Magnetic Reson in Chemistry

View full text Add to dashboard Cite

The human gastrointestinal tract is home to hundreds of species of bacteria and the balance between beneficial and pathogenic bacteria plays a critical role in human health and disease. The human infant, however, is born with a sterile gut and the complex gastrointestinal host/bacterial ecosystem is only established after birth by rapid bacterial colonization. Composition of newborn gut flora depends on several factors including type of birth (Ceasarian or natural), manner of early feeding (breast milk or formula), and exposure to local, physical environment. Imbalance in normal, healthy gut flora contributes to several adult human diseases including inflammatory bowel (ulcerative colitis and Crohn's disease) and Clostridium difficile associated disease, and early childhood diseases such as necrotizing enterocolitis. As a first step towards characterization of the role of gut bacteria in human health and disease, we conducted an 850 MHz (1)H nuclear magnetic resonance spectroscopy study to monitor changes in metabolic profiles of urine and fecal extracts of 15 mice following gut sterilization by the broad-spectrum antibiotic enrofloxacin (also known as Baytril). Ten metabolites changed in urine following enrofloxacin treatment including decreased acetate due to loss of microbial catabolism of sugars and polysaccharides, decreased trimethylamine-N-oxide due to loss of microbial catabolism of choline, and increased creatine and creatinine due to loss of microbial enzyme degradation. Eight metabolites changed in fecal extracts of mice treated with enrofloxacin including depletion of amino acids produced by microbial proteases, reduction in metabolites generated by lactate-utilizing bacteria, and increased urea caused by loss of microbial ureases.

show abstract

“…All other measurement and processing details were performed as described previously (3). As the addition of acidic substances may cause changes in pH and, consequently, chemical shifts, sample pH was assessed using the citrate signal at 2.62 ppm in the 1D spectra (8). Mean and standard deviation of the calculated pH were 7.13 0.01, indicating minimal changes in chemical shift.…”

Section: Methodsmentioning

confidence: 99%

MetaboQuant: A tool Combining individual Peak Calibration and Outlier Detection for Accurate Metabolite Quantification in 1D ¹ H and ¹ H- ¹³ C HSQC NMR Spectra

Klein¹,

Oefner²,

Gronwald³

2013

BioTechniques

View full text Add to dashboard Cite

Solution nuclear magnetic resonance (NMR) spectroscopy is widely used to analyze complex mixtures of organic compounds such as biological fluids and tissue extracts. Targeted profiling approaches with reliable compound quantitifcation are hampered, however, by signal overlap and other interferences. Here, we present a tool named MetaboQuant for automated compound quantification from pre-processed 1D and 2D heteronuclear single quantum coherence (HSQC) NMR spectral data and concomitant validation of results. Performance of MetaboQuant was tested on a urinary spike-in data set and compared with other quantification strategies. The use of individual calibration factors in combination with the validation algorithms of MetaboQuant raises the reliability of the quantification results. MetaboQuant can be downloaded at http://genomics.uni-regensburg.de/site/institute/software/metaboquant/ as stand-alone software for Windows or run on other operating systems from within Matlab. Separate software for peak fitting and integration is necessary in order to use MetaboQuant.

show abstract

Effect of pH on 1H-NMR Spectroscopy of Mouse Urine

Cited by 20 publications

References 16 publications

Reproducibility of NMR Analysis of Urine Samples: Impact of Sample Preparation, Storage Conditions, and Animal Health Status

Reproducibility of NMR Analysis of Urine Samples: Impact of Sample Preparation, Storage Conditions, and Animal Health Status

NMR‐based metabonomics analysis of mouse urine and fecal extracts following oral treatment with the broad‐spectrum antibiotic enrofloxacin (Baytril)

MetaboQuant: A tool Combining individual Peak Calibration and Outlier Detection for Accurate Metabolite Quantification in 1D ¹ H and ¹ H- ¹³ C HSQC NMR Spectra

Contact Info

Product

Resources

About

Effect of pH on 1H-NMR Spectroscopy of Mouse Urine

Cited by 20 publications

References 16 publications

Reproducibility of NMR Analysis of Urine Samples: Impact of Sample Preparation, Storage Conditions, and Animal Health Status

Reproducibility of NMR Analysis of Urine Samples: Impact of Sample Preparation, Storage Conditions, and Animal Health Status

NMR‐based metabonomics analysis of mouse urine and fecal extracts following oral treatment with the broad‐spectrum antibiotic enrofloxacin (Baytril)

MetaboQuant: A tool Combining individual Peak Calibration and Outlier Detection for Accurate Metabolite Quantification in 1D 1 H and 1 H- 13 C HSQC NMR Spectra

Contact Info

Product

Resources

About

MetaboQuant: A tool Combining individual Peak Calibration and Outlier Detection for Accurate Metabolite Quantification in 1D ¹ H and ¹ H- ¹³ C HSQC NMR Spectra