A rapid and simple high-performance liquid chromatography-ultraviolet method was developed for the separation and quantification of 15 sulfonamides (SAs) in foods of animal origin without the need of clean-up procedure. A mixture of acetonitrile-formic acid-ammonium acetate-water was used as the mobile phase to separate 15 SAs on a C18 column with gradient. The selected SAs were separated completely from the matrix mixture based on different retention behaviors at different concentration of acetonitrile. The effects of the additive of formic acid and ammonium acetate in mobile phases on the separation of SAs were also investigated. The additive can greatly improve the resolution between SAs and impurities, so that the SAs can be quantified directly under the optimized chromatographic condition the sample preparation which does not need extra sample clean-up procedure. Complete baseline separation of 15 SAs was achieved in <40 min, the linear range is 0.01-130 μg/mL with a correlation coefficient R2-value > 0.999. Excellent method reproducibility was found by intra- and inter-day precisions with the relative standard deviation <9.5%. The detection limit was <11.0 ng/mL and it can be used for routine screening of the SA residues in foods of animal origin.
A Tempol compound with an amine group (4-amino-2,2,6,6-tetramethylpiperidine-1-oxyl, NH-Tempol) was cross-linked to hemoglobin in a one-step polymerization reaction to produce a novel hemoglobin-based oxygen carrier (HBOC) designated PolyHb-Tempol. The reaction parameters, including the reaction time, pH, temperature, and ratio of reactants, were optimized, and the physiochemical properties of the resulting product were characterized. PolyHb-Tempol didn't show any toxicity towards endothelial cells. Furthermore, from observations of cell morphology and viability, PolyHb-Tempol showed a significant ability to inhibit or eliminate oxidative stress induced by superoxide free radicals. These results suggest that PolyHb-Tempol may potentially be suitable as an HBOC.
Aims Several studies have reported a potential association between prostate volume (PV) and prostate disease. Here, we classified the risk factors for PV among benign prostatic hyperplasia (BPH) patients. Methods In all, 4293 BPH patients with available clinical information were enrolled. Body mass index (BMI) was obtained as weight divided by height squared. PV was calculated as length × width × height (cm) × π/6. Mann‐Whitney U tests were used to determine the differences between PV subgroups. Univariate and multiple linear regression tests were performed to uncover the connection between clinical features and PV. The differences in the age, BMI, height and fasting blood glucose (FBG) of the subgroups were evaluated by Kruskal‐Wallis tests and adjusted with Bonferroni post hoc correction. A nomogram was created to directly illustrate the mutual interaction of amalgamator parameters. Results PV did not influence the incidence of kidney stones (P = .815), whereas prostate calculi were positively associated with an enlarged prostate (>30 mL) (P < .001). Age (adjusted R = 0.363, P < .001), height (adjusted R = 0.088, P < .001), BMI (adjusted R = 0.039, P = .013) and FBG (adjusted R = −0.034, P = .027) were the independent risk/protective factors related to enlarged PV among BPH patients. The nomogram illustrated the predictive risk of an enlarged prostate (>30 mL) in men. The area under the ROC curve value was 0.659 in the training cohort and 0.677 in an internal validation cohort. Conclusions Age, height and BMI were positive independent risk factors of enlarged PV in BPH patients, and FBG had a protective role.
The factors affecting hemoglobin (Hb) oxidation in its short- and long-term storage were investigated. Porcine Hb with different levels of oxygen saturation was stored at 25°C or 4°C for 40 days. The methemoglobin (metHb) content increased rapidly when Hb was at half saturation with oxygen at 4°C, increased gradually with the increase in Hb oxygenation or deoxygenation, and was almost unchanged when Hb was highly oxygenated or deoxygenated. The Hb oxidation was more intense when stored at 25°C. This indicated that highly oxygenated or deoxygenated Hb can be stored short term at 4°C. The accelerated test showed that metHb content was almost unchanged when Hb was highly deoxygenated and stored at 37°C for 7 days; however, the metHb content rapidly increased when Hb was highly oxygenated and stored at 37°C, which indicated that only highly deoxygenated Hb is suitable for long-term storage. Then, the long-term oxidation stability of highly deoxygenated Hb and polymerized hemoglobin was verified; the metHb content of both did not show significant changes at 4°C for 18 months. Moreover, the results in this work indicated that a temperature increasing from 20 to 40°C clearly increased the partial oxygen pressure (P50), which represents decreased oxygen affinity and is beneficial to deoxygenation. Furthermore, P50 was increased when the pH decreased from 9 to 7. Therefore, we concluded that the appropriate high temperature and neutral condition in vitro are conducive to reducing the oxygen affinity of Hb to achieve efficient deoxygenation, which could promote the development of hemoglobin-based oxygen carriers.
Glutaraldehyde (GA), used medically as a disinfectant and as a crosslinker for haemoglobin (Hb)-based oxygen carriers (HBOCs), was investigated for its ability to inactivate viruses during the preparation of these artificial blood substitutes. Porcine parvovirus (PPV; a non-enveloped DNA virus) and porcine pseudorabies virus (PRV; an enveloped DNA virus) were used as the virus indicators. Upon treatment with 0.1 mM GA, the titer of PRV decreased from 9.62 log to 2.62 log within 0.5 h, whereas that of PPV decreased from 7.00 log to 2.30 log in 5 h. Following treatment with 1.0 mM GA, the titer of PRV decreased from 11.00 log to 1.97 log within 0.5 h, whereas that of PPV decreased from 7.50 log to 3.43 log in 4.5 h. During the polymerization of Hb with GA, the GA concentration decreased to 1.0 and 0.1 mM within 30 and 50 min, respectively, at a GA:Hb molar ratio of 10:1, whereas at a GA:Hb molar ratio of 30:1, GA decreased to those same concentrations in 1.5 and 2.5 h, respectively. This rapid decrease in GA concentration during its polymerization with Hb indicates that GA must be added into the Hb solution in a short time in order to get as high a initial concentration as possible. In this study, the GA can only inactivate PRV effectively, given that a longer time (4.5 h) was required for it to inactivate the PPV titer. This study therefore demonstrates that GA inactivates the enveloped DNA virus only during the preparation of HBOCs.
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