scholarly journals Stability of Folate and Vitamin B12in Human Serum after Long-Term Storage: A Follow-Up after 13 Years

2018 ◽  
Vol 2018 ◽  
pp. 1-4 ◽  
Author(s):  
Eugène H. J. M. Jansen ◽  
Piet K. Beekhof

In epidemiological and nutrition research, it is very important to evaluate the stability of biomarkers as function of both storage time and temperature. In this study, the stability of folate and vitamin B12in human serum samples has been tested after long-term storage at −80°C up to 13 years. Serum samples of 16 individuals were used in this study. The concentration of folate and vitamin B12has been determined att=0and at 1, 8, and 13 years after storage at −80°C. The folate concentrations in serum samples remained stable at −80°C. The concentration of vitamin B12was decreasing during the time of the study to about 50%. The correlation of the folate and also of the vitamin B12concentrations in the stored samples compared with the starting values was still good. Therefore, although the concentration of vitamin B12decreased upon storage, reliable comparative analyses can still be performed.

Metabolites ◽  
2018 ◽  
Vol 8 (4) ◽  
pp. 93 ◽  
Author(s):  
Cora McHugh ◽  
Thomas Flott ◽  
Casey Schooff ◽  
Zyad Smiley ◽  
Michael Puskarich ◽  
...  

Background: Though blood is an excellent biofluid for metabolomics, proteins and lipids present in blood can interfere with 1d-1H NMR spectra and disrupt quantification of metabolites. Here, we present effective macromolecule removal strategies for serum and whole blood (WB) samples. Methods: A variety of macromolecule removal strategies were compared in both WB and serum, along with tests of ultrafiltration alone and in combination with precipitation methods. Results: In healthy human serum, methanol:chloroform:water extraction with ultrafiltration was compared to methanol precipitation with and without ultrafiltration. Methods were tested in healthy pooled human serum, and in serum from patients with sepsis. Effects of long-term storage at −80 °C were tested to explore the impact of macromolecule removal strategy on serum from different conditions. In WB a variety of extraction strategies were tested in two types of WB (from pigs and baboons) to examine the impact of macromolecule removal strategies on different samples. Conclusions: In healthy human serum methanol precipitation of serum with ultrafiltration was superior, but was similar in recovery and variance to methanol:chloroform:water extraction with ultrafiltration in pooled serum from patients with sepsis. In WB, high quality, quantifiable spectra were obtained with the use of a methanol: chloroform precipitation.


2020 ◽  
Vol 100 (10) ◽  
pp. 1345-1355 ◽  
Author(s):  
Stefaniya Boneva ◽  
Anja Schlecht ◽  
Daniel Böhringer ◽  
Hans Mittelviefhaus ◽  
Thomas Reinhard ◽  
...  

Abstract This study aims to compare the potential of standard RNA-sequencing (RNA-Seq) and 3′ massive analysis of c-DNA ends (MACE) RNA-sequencing for the analysis of fresh tissue and describes transcriptome profiling of formalin-fixed paraffin-embedded (FFPE) archival human samples by MACE. To compare MACE to standard RNA-Seq on fresh tissue, four healthy conjunctiva from four subjects were collected during vitreoretinal surgery, halved and immediately transferred to RNA lysis buffer without prior fixation and then processed for either standard RNA-Seq or MACE RNA-Seq analysis. To assess the impact of FFPE preparation on MACE, a third part was fixed in formalin and processed for paraffin embedding, and its transcriptional profile was compared with the unfixed specimens analyzed by MACE. To investigate the impact of FFPE storage time on MACE results, 24 FFPE-treated conjunctival samples from 24 patients were analyzed as well. Nineteen thousand six hundred fifty-nine transcribed genes were detected by both MACE and standard RNA-Seq on fresh tissue, while 3251 and 2213 transcripts were identified explicitly by MACE or RNA-Seq, respectively. Standard RNA-Seq tended to yield longer detected transcripts more often than MACE technology despite normalization, indicating that the MACE technology is less susceptible to a length bias. FFPE processing revealed negligible effects on MACE sequencing results. Several quality-control measurements showed that long-term storage in paraffin did not decrease the diversity of MACE libraries. We noted a nonlinear relation between storage time and the number of raw reads with an accelerated decrease within the first 1000 days in paraffin, while the numbers remained relatively stable in older samples. Interestingly, the number of transcribed genes detected was independent on FFPE storage time. RNA of sufficient quality and quantity can be extracted from FFPE samples to obtain comprehensive transcriptome profiling using MACE technology. We thus present MACE as a novel opportunity for utilizing FFPE samples stored in histological archives.


2008 ◽  
Vol 1 (1) ◽  
pp. 67-78 ◽  
Author(s):  
M. Hafner ◽  
M. Sulyok ◽  
R. Schuhmacher ◽  
C. Crews ◽  
R. Krska

In this paper the stability and degree of epimerisation of six major ergot alkaloids at three different temperature levels (-20 °C, +4 °C and +20 °C) over periods of 18 hours and six weeks is reported for the first time. The behaviour of ergometrine, ergocornine, ergocristine, α-ergocryptine, ergosine and ergotamine was thoroughly studied in seven solvents which are employed for the preparation of calibrants and extraction mixtures, respectively. Moreover, the stability of the ergot alkaloids was tested in different cereal extracts (rye, wheat, barley, oats) for 1, 2 and 6 days. Of the toxins tested, the ergopeptide-type toxins ergosine, ergotamine, ergocornine, α-ergocryptine and ergocristine showed similar behaviour patterns. The simple lysergic acid derivative ergometrine was more stable and showed hardly any epimerisation to ergometrinine, with the sum of both epimers remaining constant in all seven solvents. The ergopeptides tested show variable epimerisation tendencies, and were also less stable during six weeks at 20 °C. Ergosine showed the highest degree of epimerisation (43% after 6 weeks at 20 °C). In general, the order of epimerisation promotion was methanol/dichloromethane > acetonitrile/buffer > extraction mix > stabilising solution > acetonitrile >> chloroform. Long-term storage at room temperature can only be carried out in chloroform, which showed no epimerisation for all toxins even at 20 °C and also kept the sum of R and S forms constant, which indicates no formation of aci-epimers or other degradation products. Long-term storage of ergot alkaloids in acetonitrile, the most convenient solvent with respect to HPLC analysis, should be carried out at temperatures of -20 °C or below. The constant epimer ratio of all ergot alkaloids in the extraction mixture acetonitrile/ammonium carbonate buffer (200 mg/l; 92:8, v/v) during an HPLC run (18 hours) demonstrates the stability of the toxins in this extraction mixture.


2019 ◽  
Vol 974 ◽  
pp. 187-194 ◽  
Author(s):  
Nikolay V. Lyubomirskiy ◽  
Tamara A. Bakhtina ◽  
Alexander S. Bakhtin ◽  
Sergey I. Fedorkin

This paper presents the lime binding forced carbonate-hardening materials properties formation study and determins the stability of these properties during long-term storage and use under normal conditions. The tests showed these materials stability properties over time, confirming the strength and density growth of the test samples after long storage due to the calcium hydroxide recrystallization completion into calcium carbonate processes. Also, the results of the samples carbonate hardening study under natural conditions during 18 months are presented. An efficiency assessment of forced carbonate hardening as one of the methods of recycling technogenic CO2 in order to reduce its emissions in the atmosphere, and, in the result, to obtain high-quality construction materials has been made.


EBioMedicine ◽  
2016 ◽  
Vol 12 ◽  
pp. 309-314 ◽  
Author(s):  
Stefan Enroth ◽  
Göran Hallmans ◽  
Kjell Grankvist ◽  
Ulf Gyllensten

2020 ◽  
Vol 20 (11) ◽  
pp. 6855-6861
Author(s):  
Wooram Kim ◽  
Mijeong Park ◽  
Jong-Ki Jeon ◽  
Youngmin Jo

Dinitramide anion [−N(NO2)2] salt composed of resonance structure is a plausible oxidizing agents, as efficient propellant. Among them, guanidinium dinitramide (GDN) is an organic compound improving the stability against moisture, as well long term storage. An additional advantage composed guanidinium ion is the reaction efficient via the decomposed by-product during pyrognostics, maximum yield of 99%. The types of GDN (GDN-I, II, III, IV, V) were synthesized using several starting material such as guanidine acetate, chloride, carbonate, nitrate and sulfate under hydrodeprivation. In this work, the intermediates formed in these processes were closely identified and their thermal properties, and chemical structure were examined. The absorption peaks by Fourier transform infrared (FT-IR) were found guanidinium infrared frequencies (3452, 3402, 3354, 3278, 1642 cm−1) and dinitramide infrared frequencies (3208, 1570, 1492, 1416, 1337, 1179, 1000 cm−1). The activation energy of GDN samples were obtained Ea = 53.26 Kcal/mole (GDN-I), 50.94 Kcal/mole (GDN-II), 52.34 Kcal/mole (GDN-III), 62.19 Kcal/mole (GDN-IV), 55.32 Kcal/mole (GDN-V) from exothermic at over 153°C.


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