scholarly journals Evaluation of Saliva Stability for NMR Metabolomics: Collection and Handling Protocols

Metabolites ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 515
Author(s):  
Daniela Duarte ◽  
Beatriz Castro ◽  
Joana Leonor Pereira ◽  
Joana Faria Marques ◽  
Ana Luísa Costa ◽  
...  
Keyword(s):  
Metabolic Profile ◽  
Room Temperature ◽  
Clinical Laboratory ◽  
Sample Collection ◽  
Short Term ◽  
General Stability ◽  
Sample Stability ◽  
Choline Esters ◽  
Different Temperatures ◽  
Term Storage

Maintaining a salivary metabolic profile upon sample collection and preparation is determinant in metabolomics. Nuclear magnetic resonance (NMR) spectroscopy was used to identify metabolite changes during short-term storage, at room temperature (RT)/4 °C/−20 °C, and after sample preparation, at RT/4 °C (mimicking typical clinical/laboratory settings). Interestingly, significant metabolic inter-individual and inter-day variability were noted, probably determining sample stability to some extent. After collection, no changes were noted at −20 °C (at least for 4 weeks). RT storage induced decreases in methylated macromolecules (6 h); lactate (8 h); alanine (12 h); galactose, hypoxanthine, pyruvate (24 h); sarcosine, betaine, choline, N-acetyl-glycoproteins (48 h), while acetate increased (48 h). Less, but different, changes were observed at 4 °C, suggesting different oral and microbial status at different temperatures (with a possible contribution from inter-individual and inter-day variability), and identifying galactose, hypoxanthine, and possibly, choline esters, as potential general stability indicators. After preparation, addition of NaN3 did not impact significantly on saliva stabilization, neither at RT nor at 4 °C, although its absence was accompanied by slight increases in fucose (6.5 h) and proline (8 h) at RT, and in xylose (24 h) at 4 °C. The putative metabolic origins of the above variations are discussed, with basis on the salivary microbiome. In summary, after collection, saliva can be stored at RT/4 °C for up to 6 h and at −20 °C for at least 4 weeks. Upon preparation for NMR analysis, samples are highly stable at 25 °C up to 8 h and at 4 °C up to 48 h, with NaN3 addition preventing possible early changes in fucose, proline (6–8 h), and xylose (24 h) levels.

scholarly journals The effect of short-term storage temperature on the key headspace volatile compounds observed in Canadian faba bean flour

2021 ◽  
pp. 108201322199884
Author(s):  
Rami Akkad ◽  
Ereddad Kharraz ◽  
Jay Han ◽  
James D House ◽  
Jonathan M Curtis
Keyword(s):  
Fatty Acids ◽  
Volatile Compounds ◽  
Room Temperature ◽  
Unsaturated Fatty Acids ◽  
Solid Phase ◽  
Short Term ◽  
Bean Flour ◽  
Odour Activity Value ◽  
Short Term Storage ◽  
Term Storage

The odour emitted from the high-tannin fab bean flour ( Vicia faba var. minor), was characterized by headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC–MS). The relative odour activity value (ROAV) was used to monitor the changes in key volatile compounds in the flour during short-term storage at different temperature conditions. The key flavour compounds of freshly milled flour included hexanal, octanal, nonanal, decanal, 3-methylbutanal, phenyl acetaldehyde, (E)-2-nonenal, 1-hexanol, phenyl ethyl alcohol, 1-octen-3-ol, β-linalool, acetic acid, octanoic acid, and 3-methylbutyric acid; these are oxidative degradation products of unsaturated fatty acids and amino acids. Despite the low lipid content of faba beans, the abundances of aldehydes arising during room temperature storage greatly contributed to the flavour of the flour due to their very low odour thresholds. Two of the key volatiles responsible for beany flavour in flour (hexanal, nonanal) increased greatly after 2 weeks of storage at room temperature or under refrigerated conditions. These volatile oxidation products may arise as a result of enzymatic activity on unsaturated fatty acids, and was seen to be arrested by freezing the flour.

scholarly journals Quality Markers Addressing Preanalytical Variations of Blood and Plasma Processing Identified by Broad and Targeted Metabolite Profiling

2014 ◽  
Vol 60 (2) ◽  
pp. 399-412 ◽  
Author(s):  
Beate Kamlage ◽  
Sandra González Maldonado ◽  
Bianca Bethan ◽  
Erik Peter ◽  
Oliver Schmitz ◽  
...  
Keyword(s):  
Human Plasma ◽  
Metabolite Profiling ◽  
Plasma Processing ◽  
Short Term ◽  
High Quality ◽  
Plasma Metabolome ◽  
Short Term Storage ◽  
Quality Markers ◽  
Different Temperatures ◽  
Term Storage

Abstract BACKGROUND Metabolomics is a valuable tool with applications in almost all life science areas. There is an increasing awareness of the essential need for high-quality biospecimens in studies applying omics technologies and biomarker research. Tools to detect effects of both blood and plasma processing are a key for assuring reproducible and credible results. We report on the response of the human plasma metabolome to common preanalytical variations in a comprehensive metabolomics analysis to reveal such high-quality markers. METHODS Human EDTA blood was subjected to preanalytical variations while being processed to plasma: microclotting, prolonged processing times at different temperatures, hemolysis, and contamination with buffy layer. In a second experiment, EDTA plasma was incubated at different temperatures for up to 16 h. Samples were subjected to GC-MS and liquid chromatography–tandem mass spectrometry–based metabolite profiling (MxP™ Broad Profiling) complemented by targeted methods, i.e., sphingoids (as part of MxP™ Lipids), MxP™ Catecholamines, and MxP™ Eicosanoids. RESULTS Short-term storage of blood, hemolysis, and short-term storage of noncooled plasma resulted in statistically significant increases of 4% to 19% and decreases of 8% to 12% of the metabolites. Microclotting, contamination of plasma with buffy layer, and short-term storage of cooled plasma were of less impact on the metabolome (0% to 11% of metabolites increased, 0% to 8% decreased). CONCLUSIONS The response of the human plasma metabolome to preanalytical variation demands implementation of thorough quality assurance and QC measures to obtain reproducible and credible results from metabolomics studies. Metabolites identified as sensitive to preanalytics can be used to control for sample quality.

Use of a Maggot Motility Index to Evaluate Survival of Therapeutic Larvae

2004 ◽  
Vol 94 (4) ◽  
pp. 353-355 ◽  
Author(s):  
Anthony Rosales ◽  
Jefferey R. Vazquez ◽  
Brian Short ◽  
Heather R. Kimbriel ◽  
Matthew J. Claxton ◽  
...  
Keyword(s):  
Room Temperature ◽  
Wound Care ◽  
In Vivo Studies ◽  
Clinical Use ◽  
Short Term ◽  
Motility Index ◽  
Term Storage ◽  
Over Time ◽  
Surrogate Method

Maggot debridement therapy is rapidly increasing in popularity at major diabetic foot and wound care centers worldwide. However, we are unaware of specific guidelines on the short-term storage of larvae. We sought to evaluate differences in maggot motility over time in larvae refrigerated versus those stored at room temperature. We also introduce a simple surrogate method for evaluating maggot vitality that may be useful for in vivo studies if validated in future works. We randomly selected ten larvae from the same shipment at ten different times in 9 days. Larvae were placed on a translucent acetate grid, and their total excursion in 30 sec was measured. This was converted into a Maggot Motility Index. In the refrigerated group, the index remained at or above 40 mm/min for approximately 60 hours from baseline, when there was a significant decrease. This same phenomenon occurred during the first 12 hours in the nonrefrigerated group. There were significant differences in motility between refrigerated and nonrefrigerated larvae immediately after baseline until day 8. Larvae are more practical for repeated clinical use if kept refrigerated between applications. (J Am Podiatr Med Assoc 94(4): 353–355, 2004)

scholarly journals Stability of Hemocytometry Parameters Under Different Sample Storage Conditions

Medicina ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 54-74
Author(s):  
A. S. Shulga ◽  
◽  
N. N. Kraynova ◽  
D. V. Burtsev ◽  
◽  
...  
Keyword(s):  
Room Temperature ◽  
Clinical Laboratory ◽  
Hematological Parameters ◽  
Storage Conditions ◽  
The Other ◽  
Sample Collection ◽  
Hematological Indices ◽  
Temperature Regimes ◽  
Time Changes ◽  
The One

Sample stability is essential for reliable results in clinical laboratory practice. The aim of the work was to investigate the change in values of hematological indices in samples stored for up to 72 hours under different temperature regimes. A total of 60 whole blood samples stored under different conditions were analyzed: at room temperature (25°C), heated to 35°C and cooled to 4°C. Analysis was performed at different time points: immediately after blood sampling and then consecutively after 3, 6, 12, 24, 48 and 72 hours. K2EDTA anticoagulant tubes were used, and results were obtained using a UniCel DxH 800 hematology analyzer. The median shift of the parameters relative to baseline for each combination of time and temperature was assessed using the Wilcoxon matched pairs test. The shift in hemogram values obtained using Bland-Altman plots was compared with the maximum permissible error specified in the quality specification for the desirable error. Hemoglobin, erythrocyte count, mean erythrocyte hemoglobin content and platelet content were stable for at least 72 hours at all temperatures used in the experiment. For the other tested parameters, the first unacceptable changes in hemogram values were observed after 3 hours when the samples were stored at 25°C and 35°C. In samples cooled to 4°C, the first statistically significant differences were recorded after 6 hours. As a result, storage of samples for 72 hours at room temperature led to reliable unacceptable changes in 6 hemogram parameters of the 11 studied, at 4°C 5 parameters changed unacceptably, and at 35°C – 7 parameters. The obtained results, on the one hand, indicate that when analyzing the results of hematological tests is performed with a delay after sample collection, changes in hematological parameters should be considered; on the other hand, they provide information about the list of parameters subject to temperature-time changes, as well as about the intensity of these changes.

scholarly journals Effect of Extender, Storage Time and Temperature on Kinetic Parameters (CASA) on Bull Semen Samples

Biology ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 806
Author(s):  
Aitor Fernandez-Novo ◽  
Sergio Santos-Lopez ◽  
Clara Barrajon-Masa ◽  
Patricia Mozas ◽  
Eduardo de Mercado ◽  
...  
Keyword(s):  
Kinetic Parameters ◽  
Room Temperature ◽  
Storage Time ◽  
Storage Conditions ◽  
Short Term ◽  
Bull Semen ◽  
The Impact ◽  
Term Storage ◽  
Diagnostic Centre ◽  
Holding Temperature

CASA kinetic parameters are often evaluated in a diagnostic centre. How storage conditions affect ejaculates up to evaluation is unclear. We assessed, in 25 commercial bulls electroejaculated in the field, the impact of time until evaluation (0–2 h, 4–6 h, and 24 h post-ejaculation), holding temperature (5 °C vs. room temperature), and extender (AndroMed®, BIOXcell® or INRA96®) on CASA kinetic parameters. Total and progressive motility, VCL, VAP, VCL, ALH, BCF, STR, LIN, and WOB were assessed. CASA kinetic parameters were preserved for up to 4–6 h post-ejaculation, except for AndroMed®. Regardless of extender or temperature, motility decreased from 4–6 h up to 24 h, with the best values obtained with BIOXcell® at 5 °C. Our results suggest that BIOXcell® can preserve sperm motility for up to 6 h, either at 5 °C or room temperature, and also INRA96® at room temperature, with motility assessments and the percentage of the most rapid sperms being the lowest with INRA96® at 5 °C. The kinetic parameters decreased when analyses were performed at 24 h. Therefore, we suggest evaluating seminal quality as soon as possible, before 6 h after collection. These results help to fix adequate protocols for the short-term storage and shipment of bovine semen collected under field conditions.

Evaluation of sample stability for a quantitative faecal immunochemical test and comparison of two sample collection approaches

2018 ◽  
Vol 55 (6) ◽  
pp. 657-664 ◽  
Author(s):  
Samantha Mellen ◽  
Maria de Ferrars ◽  
Claire Chapman ◽  
Sarah Bevan ◽  
James Turvill ◽  
...  
Keyword(s):  
Room Temperature ◽  
False Negative ◽  
Haemoglobin Concentration ◽  
Sample Collection ◽  
Negative Results ◽  
Collection Device ◽  
Sample Stability ◽  
Haemoglobin Measurement ◽  
Standard Collection ◽  
The Stability

Background Faecal immunochemical testing is increasingly being used to triage symptomatic patients for suspected colorectal cancer. However, there are limited data on the effect of preanalytical factors on faecal haemoglobin when measured by faecal immunochemical testing. The aim of this work was to evaluate the stability of faecal haemoglobin in faeces and to compare two methods of faecal haemoglobin sampling for faecal immunochemical testing. Methods Six patients provided faeces for faecal haemoglobin measurement which were transferred into specialized collection devices at baseline and at 1, 2, 3 and 7 days after storage at either room temperature or 4°C. A total of 137 patients returned both faeces transferred into the specialized collection device and faeces in a standard collection pot. A quantitative immunoturbidometric method was used to measure faecal haemoglobin and results were compared categorically. Discrepant results were assessed against diagnosis. Results Faecal haemoglobin concentration declined rapidly within a day of storage at room temperature but results remained ≥10 μg Hb/g faeces in 5/6 patients after two days. A faecal haemoglobin result ≥10 μg Hb/g faeces was obtained in 4/6 patients after storage for seven days at 4°C. Results obtained when patients used specialized collection devices were significantly different from results obtained when faeces was transferred into the specialized collection device in the laboratory. Conclusion There is considerable heterogeneity in the sample stability of faecal haemoglobin; therefore, samples should be transferred rapidly into specialized collection devices to prevent false-negative results. Use of collection devices by patients can lead to false-positive results compared with their use in a laboratory.

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