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21.
22.
Kirtikumar D. Bharwad Priyanka A. Shah Pranav S. Shrivastav Vinay S. Sharma Puran Singhal 《Biomedical chromatography : BMC》2020,34(1):e4708
A rapid, simple and sensitive ultra-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) method has been developed to quantify fenoprofen, a nonsteroidal anti-inflammatory drug in human plasma for a pharmacokinetic study in healthy subjects. Owing to high levels of protein binding, protein precipitation followed by solid-phase extraction was employed for the extraction of fenoprofen and fenoprofen-d3 (used as internal standard) from 200 μL human plasma. Separation was performed on a BEH C18 (50 × 2.1 mm, 1.7 μm) column using methanol−0.2% acetic acid in water (75:25, v/v) under isocratic elution. Electrospray ionization was operated in the negative mode for sample ionization. Ion transitions used for quantification in the selected reaction monitoring mode were m/z 241/197 and m/z 244/200 for fenoprofen and fenoprofen-d3, respectively. Under the optimized conditions, fenoprofen showed excellent linearity in the concentration range 0.02–20 μg/mL (r2 ≥ 0.9996), adequate sensitivity, favorable accuracy (96.4–103.7%) and precision (percentage coefficient of variation ≤4.3) with negligible matrix effect. The validated method was successfully applied to a pharmacokinetic study of fenoprofen in healthy subjects. The significant features of the method include higher sensitivity, small plasma volume for processing and a short analysis time. 相似文献
23.
Sujan Dilly Penchala Jasmini Alagaratnam Elizabeth Challenger Alieu Amara Laura Else Alan Winston Saye Khoo 《Biomedical chromatography : BMC》2020,34(1):e4711
A high-performance liquid chromatography tandem mass spectrometric method was developed and validated for cenicriviroc (CVC) quantification in human plasma and cerebrospinal fluid (CSF). The method involved precipitation with acetonitrile and injecting supernatants onto the column. Separation was achieved on an XBridge C18 column with a gradient elution of 0.1% formic acid in water and acetonitrile. Analyte detection was conducted in positive ion mode using selected reaction monitoring. The m/z transitions were: CVC (697.3 → 574.3) and CVC-d7 (704.4 → 574.3). Calibration curve ranged from 5 to 1000 ng/mL for plasma and from 0.241 to 15.0 ng/mL for CSF. The intra- and inter-day precision and accuracy were <15% for both plasma and CSF across four different concentrations. CVC recovery from plasma and artificial CSF was >90%. The method was utilized for the measurement of patients’ plasma and CSF samples taking a dose of 50, 150 and 300 mg q.d. 相似文献
24.
Nile red bound to human serum albumin (HSA) shows an order of magnitude increase in the probe's fluorescence intensity. Here, we report on the fluorescence characteristics of the probe-protein complex in Trizma buffer (pH 7.1), urea, guanidine hydrochloride, and AOT/isooctane/buffer reverse micelles using both steady—state and time-resolved fluorescence techniques. With a view to illustrating the use of extrinsic probe fluorescence spectroscopy in protein research, we demonstrate that protein unfolding can be observed through measurements of the probe's time-resolved anisotropy and steady-state fluorescence spectrum. Moreover, this shows that thermal unfolding is fundamentally different from using denaturant, with respect to changes in both the nanosecond diffusional rotation of the probe at intermediate stages and in the denatured protein's structure. Also, the large Stokes shift of Nile red allows the changes in the environment of the probe-protein complex in reverse micelles of varying waterpool size to be easily identified in the steady-state fluorescence. This was not seen in earlier work exploiting the intrinsic tryptophan fluorescence of HSA and further demonstrates the complementary information that extrinsic fluorescence probe studies can offer protein science. We discuss the complex acrylamide quenching characteristics of Nile red bound to HSA in terms of the possibility of at least two binding sites for the probe and the effect of acrylamide on the probe-protein structure at very high quencher concentrations. 相似文献
25.
Ji-Eun Lee Hyonchol Jang Eun-Jung Cho Hong-Duk Youn 《Experimental & molecular medicine》2009,41(7):471-477
Down syndrome critical region 1 (DSCR1), an oxidative stress-response gene, interacts with calcineurin and represses its phosphatase activity. Recently it was shown that hydrogen peroxide inactivates calcineurin by proteolytic cleavage. Based on these facts, we investigated whether oxidative stress affects DSCR1-mediated inactivation of calcineurin. We determined that overexpression of DSCR1 leads to increased proteolytic cleavage of calcineurin. Convertsely, knockdown of DSCR1 abolished calcineurin cleavage upon treatment with hydrogen peroxide. The PXIIXT motif in the COOH-terminus of DSCR1 is responsible for both binding and cleavage of calcineurin. The knockdown of overexpressed DSCR1 in DS fibroblast cells also abrogated calcineurin proteolysis by hydrogen peroxide. These results suggest that DSCR1 has the ability to inactivate calcineurin by inducing proteolytic cleavage of calcineurin upon oxidative stress. 相似文献
26.
High-level expression of soluble human β-defensin-2 fused with green fluorescent protein in Escherichia coli cell-free system 总被引:2,自引:0,他引:2
Human β-defensin-2 (hBD2), a small cationic peptide, exhibits a broad range of antimicrobial activity and does not acquire
any microbial resistance. To produce this uneasily detectable, degradable, and toxic polypeptide efficiently, an alternative
approach based on the Escherichia coli cell-free biosynthesis system was proposed. The approach implies that a polypeptide of interest is synthesized as a fusion
protein linked to a green fluorescent protein (GFP) through a cleavable spacer. With batch-mode operation, a significant amount
of hBD2 fused with GFP (0.25 mg/mL) can be expressed in this cell-free system. The productivity of the fusion protein can
be improved up to 1.2 mg/mL by employing a continuous-exchange cell-free system. Furthermore, the GFP moiety provides directly
visible and quantitative monitoring of the polypeptide synthesis, and the product is soluble and stable. This work will be
helpful in allowing the rapid and visible expression of other similar defensins using an in vitro cell-free system. 相似文献
27.
The objective of this article was to design the selective molecularly imprinted sorbent dedicated to the solid‐phase extraction of S‐pramipexole from the complex matrix such as human urine. For that purpose, S‐2,6‐diamino‐4,5,6,7‐tetrahydrobenzothiazole was used as the template acting as the structural analog of S‐pramipexole and five various monomers were employed in the presence of ethylene glycol dimethacrylate to produce molecularly imprinted polymers. The binding capabilities of resulted polymers revealed that the highest imprinting effect was noted for polymer prepared from the itaconic acid. The comprehensive analysis of morphology and the characterization of binding sites showed not only negligible differences in the extension of surfaces of imprinted and nonimprinted polymers but also higher heterogeneity of binding sites in the imprinted material. Comprehensive optimization of the molecularly imprinted solid‐phase extraction allowed to select the most appropriate solvents for loading, washing, and elution steps. Subsequent optimization of mass of sorbent and volumes of solvents allowed to achieve satisfactory total recoveries of S‐pramipexole from the model multicomponent real sample of human urine that equals to 91.8 ± 3.2% for imprinted sorbent with comparison to only 37.1 ± 1.1% for Oasis MCX. 相似文献
28.
Subaqueous deltaic deposits with approximately 30 radiocarbon ages show that the Changjiang River delta was strongly affected by tides and that the delta progradation rate after 2 kaBP was almost double the rate before 2 kaBP. This change in the progradation rate correlates well with the active extension of the subaerial delta plain shown by previous work. Widespread human activities, such as farming, deforestation, and dike construction, probably resulted in an increase in sediment discharge to the river-mouth area. 相似文献
29.
《Analytical letters》2012,45(10):1827-1841
Abstract The determination of bromine in human serum by inductively coupled plasma-mass spectrometry (ICP-MS) is discussed. Sample preparation was kept as limited as possible: serum samples were only diluted with nitric acid (five- or ten-fold). Indium was added as internal standard. In order to avoid overlap of 40Ar 40Ar with 相似文献
30.
糖蛋白及与蛋白质结合的糖链在生物体内的丰度一般极低,为实现其准确定性及定量,去除高丰度非糖蛋白、非糖肽的干扰,通常需要进行富集前处理操作。该文设计并制备了一种基于点击化学反应和烯丙基氯多功能化位点聚合物的葡萄糖改性核-壳-壳型磁性纳米颗粒,并建立了相应的用于糖肽亲水富集的固相微萃取方法。以人免疫球蛋白G酶解液为样品,共测得32个糖肽信号。该方法检出限可达10 fmol,每克磁性分离介质的富集容量达100 mg。 相似文献