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1.
Sample preparation is the most critical step in proteomics as it directly affects the subset of proteins and peptides that can be reliably identified and quantified. Although a variety of efficient and reproducible sample preparation strategies have been developed, their applicability and efficacy depends much on the biological sample. Here, three approaches were evaluated for the human milk and plasma proteomes. Protein extracts were digested either in an ultrafiltration unit (filter-aided sample preparation, FASP) or in-solution (ISD). ISD samples were desalted by solid-phase extraction prior to nRPC-ESI-MS/MS. Additionally, milk and plasma samples were directly digested by FASP without prior protein precipitation. Each strategy provided inherent advantages and disadvantages for milk and plasma. FASP appeared to be the most time efficient procedure with a low miscleavage rate when used for a biological sample aliquot, but quantitation was less reproducible. A prior protein precipitation step improved the quantitation by FASP due to significantly higher peak areas for plasma and a much better reproducibility for milk. Moreover, the miscleavage rate for milk, the identification rate for plasma, and the carbamidomethylation efficiency were improved. In contrast, ISD of both milk and plasma resulted in higher miscleavage rates and is therefore less suitable for targeted proteomics. 相似文献
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在红外测试中,固体样品通常采用红外压片等方法进行前处理.通过对香豆素分别采用红外压片法、薄膜法和石蜡油糊法进行测试,发现薄膜法比压片法制样更加简便快捷、用量少、可回收,而且薄膜法制样还能避免压片过程中出现的碎片和粘片现象,测试时能够得到高质量的红外谱图. 相似文献
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Determination of Glimepiride in Human Plasma by LC-MS-MS and Comparison of Sample Preparation Methods for Glimepiride 总被引:1,自引:0,他引:1
Hohyun Kim Kyu Young Chang Chang Hun Park Moon Sun Jang Jung-Ae Lee Hee Joo Lee Kyung Ryul Lee 《Chromatographia》2004,60(1-2):93-98
A sensitive and selective method for quantitation of glimepiride in human plasma was established using liquid chromatography-electrospray ionization tandem mass spectrometry. Three different methods for the sample preparation of glimepiride and an internal standard were investigated (liquid-liquid extraction, solid-phase extraction and protein precipitation). Glipizide was used as an internal standard. Compounds were separated on a C18 column with 80% acetonitrile and 20% deionized water (adjusted to pH 3.5 with acetic acid), as mobile phase at a flow rate of 200 L min–1. By use of multiple reaction monitoring mode in MS-MS with liquid-liquid extraction and solid-phase extraction, glimepiride and glipizide were detected without severe interference from the human plasma matrix. Glimepiride produced a protonated precursor ion ([M+H]+) at m/z 491 and a corresponding product ion at m/z 352, and the internal standard produced a protonated precursor ion ([M+H]+) at m/z 446 and a corresponding product ion at m/z 321. The limit of quantitation was 0.1 ng mL–1, 0.5 ng mL–1 and 1.0 ng mL–1 when using liquid-liquid extraction, solid-phase extraction and protein precipitation, respectively. The validation, reproducibility, stability, and recovery of the different sample preparation methods were comparable and all the methods gave reliable results. The method has been successfully applied to pharmacokinetic study of glimepiride in human plasma. 相似文献
4.
Lasse Neset Gracious Takayidza Frode S. Berven Maria Hernandez-Valladares 《Molecules (Basel, Switzerland)》2022,27(11)
The use of a proper sample processing methodology for maximum proteome coverage and high-quality quantitative data is an important choice to make before initiating a liquid chromatography–mass spectrometry (LC–MS)-based proteomics study. Popular sample processing workflows for proteomics involve in-solution proteome digestion and single-pot, solid-phase-enhanced sample preparation (SP3). We tested them on both HeLa cells and human plasma samples, using lysis buffers containing SDS, or guanidinium hydrochloride. We also studied the effect of using commercially available depletion mini spin columns before SP3, to increase proteome coverage in human plasma samples. Our results show that the SP3 protocol, using either buffer, achieves the highest number of quantified proteins in both the HeLa cells and plasma samples. Moreover, the use of depletion mini spin columns before SP3 results in a two-fold increase of quantified plasma proteins. With additional fractionation, we quantified nearly 1400 proteins, and examined lower-abundance proteins involved in neurodegenerative pathways and mitochondrial metabolism. Therefore, we recommend the use of the SP3 methodology for biological sample processing, including those after depletion of high-abundance plasma proteins. 相似文献
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用XPS分析固体粉末样品时通常将粉末直接撒在双面胶带上进行测试.这种方法粉末容易脱落,并导致仪器污染或损坏.本文介绍一种将粉末样品铺在胶带上压成薄层进行测试的制样方法.后一种方法不需模具,简单快捷,实验表明它不仅有利于保护仪器,而且有利于提高XPS分析的灵敏度和分辨率. 相似文献
6.
Optimization of a liquid-liquid extraction method for HPLC-DAD determination of penicillin-V in human plasma 总被引:1,自引:0,他引:1
Andrei MedvedoviciMihaela Ionescu Constantin MircioiuVictor David 《Microchemical Journal》2002,72(1):85-92
Optimization of a liquid-liquid extraction procedure used for isolation of penicillin-V in human plasma samples and the subsequent HPLC-DAD analysis are reported. Some aspects related to the stability of penicillin-V in plasma and according to pH are also given. From seven tested solvents, the highest extraction yield was obtained with methyl-t-buthyl ether. Influence of the extraction parameters, such as pH of aqueous phase, sample/solvent volumetric ratio, and extraction duration, is discussed. HPLC separation was achieved on an Eclipse XDB-C8 column, using a mobile phase composed by aqueous phosphate buffer (pH=7.3), methanol and acetonitrile in the ratio 8:1:1. Quantitation limits of 50 ng/ml were obtained. The optimal sample preparation method and HPLC-DAD separations were used for a bioequivalence study, made on 36 volunteers. 相似文献
7.
扫描电子显微镜对样品的要求及样品的制备 总被引:7,自引:0,他引:7
李剑平 《分析测试技术与仪器》2007,13(1):74-77
扫描电子显微镜对样品的要求很严,要求样品必须是固体,且做到五无:无毒、无放射性、无污染、无磁、无水分,成分稳定,块状样品大小要适中,粉末样品要进行特殊处理,对不导电和导电性能差的样品要进行镀膜,且要选择适当的镀膜仪,方能达到理想的分析效果. 相似文献
8.
Ivan Liakh Tomasz Sledzinski Lukasz Kaska Paulina Mozolewska Adriana Mika 《Molecules (Basel, Switzerland)》2020,25(22)
Obesity is associated with alterations in the composition and amounts of lipids. Lipids have over 1.7 million representatives. Most lipid groups differ in composition, properties and chemical structure. These small molecules control various metabolic pathways, determine the metabolism of other compounds and are substrates for the syntheses of different derivatives. Recently, lipidomics has become an important branch of medical/clinical sciences similar to proteomics and genomics. Due to the much higher lipid accumulation in obese patients and many alterations in the compositions of various groups of lipids, the methods used for sample preparations for lipidomic studies of samples from obese subjects sometimes have to be modified. Appropriate sample preparation methods allow for the identification of a wide range of analytes by advanced analytical methods, including mass spectrometry. This is especially the case in studies with obese subjects, as the amounts of some lipids are much higher, others are present in trace amounts, and obese subjects have some specific alterations of the lipid profile. As a result, it is best to use a method previously tested on samples from obese subjects. However, most of these methods can be also used in healthy, nonobese subjects or patients with other dyslipidemias. This review is an overview of sample preparation methods for analysis as one of the major critical steps in the overall analytical procedure. 相似文献
9.
A rapid and comprehensive sample preparation method used to extract adriamycin from plasma has been investigated. The samples were passed through an Adsorbex extraction column with an elution solvent. The eluate was directly analyzed by reversed-phase high performance liguid chromatography (HPLC) with fluorescence detector. This solid/liquid extraction method is simpler than the conventional liquid/liquid extraction method. Different elution solvents were used to extract the adriamycin from samples with and without serum. The mobile phase was found to be the optimal elution solvent: 5 mM orthophosphoric acid 62.5% in methanol; acetonitrile; isopropanol = 15:15:7.5, pH 3.2. This method has the merits of better recovery (98%), higher reproducibility, and reguires shorter time and consumes less solvent. 相似文献
10.
Sample preparation is an essential step for nearly every type of biochemical analysis in use today. Among the most important of these analyses is the diagnosis of diseases, since their treatment may rely greatly on time and, in the case of infectious diseases, containing their spread within a population to prevent outbreaks. To address this, many different methods have been developed for use in the wide variety of settings for which they are needed. In this work, we have reviewed the literature and report on a broad range of methods that have been developed in recent years and their applications to point-of-care (POC), high-throughput screening, and low-resource and traditional clinical settings for diagnosis, including some of those that were developed in response to the coronavirus disease 2019 (COVID-19) pandemic. In addition to covering alternative approaches and improvements to traditional sample preparation techniques such as extractions and separations, techniques that have been developed with focuses on integration with smart devices, laboratory automation, and biosensors are also discussed. 相似文献
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《Analytical letters》2012,45(11):767-771
Abstract We have compared the results obtained for plasma cortisol when two methods were used to measure this steroid. The first method, a Competitive protein binding (CPB) assay, used transcortin as binding protein for cortisol. The second method is a radioimmunoassay (RIA) utilizing as binding reagent, a specific antibody against cortisol. Nineteen samples were tested covering a wide range of values. The CPB assay consistently overestimated the levels of plasma cortisol compared to the RIA method. 相似文献
15.
Fiber-in-Tube Solid Phase Extraction (FIT-SPE) for Miniaturized Sample Preparation Process 总被引:2,自引:0,他引:2
A typical analytical separation procedure has several important steps: sample preparation, isolation, identification, quantitation, statistical evaluation and final decision. Each step is alwayscritical to obtain correct results to fulfill the analytical purpose. In these various steps sample 相似文献
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A new sample insertion device for the stabilized capacitive plasma (SCP) has been developed, which enables it to analyze
dry residues of micro amounts of liquid samples. Insertion was applied into an SCP as plasma source because of its good stability
and excitation properties as well as its low instrument and operation costs. The plasma is sustained at a frequency of 27.12 MHz
and an RF power of 150 W. For analysis the liquid samples are positioned at the tip of a quartz rod with the aid of a μL syringe.
Then the sample is dried and the sampling rod inserted into the plasma. After optimization of the carrier gas flow (5 L/h)
and the sample volume (20 μL) the detection limit for Pb with Ar as plasma gas is 200 pg.
By further improving the guidance of the insertion detection limits for Pb, Cu, Cd and Mg in the 1 to 30 ng/mL range or 20
to 600 pg range absolute were obtained. It was found that the detection limits in the case of He are better than those obtained
with Ar. The matrix interferences caused by changes in the concentration of the easily ionizable element Na were found to
be below 10% for Na concentrations of up to 0.45 μg/mL. Ethanol concentrations of up to 14% in the analyte solutions did not
cause any interferences.
Received December 17, 1998. Revision June 4, 1999. 相似文献
18.
《Analytical letters》2012,45(15):1359-1371
Abstract A sensitive method for the determination of metoprolol in plasma has been developed. The procedure is based on gas chromatographic measurements of derivatized metoprolol, using 9-bromophenanthrene as internal standard. Metoprolol is derivatized with pentafluoropropionic anhydride. The resulting derivative gives a four-fold increase in sensitivity compared to the published methods where trifluoroacetic anhydride was used for derivatization. 相似文献
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临床样品微量元素分析过程中样品制备技术的进展 总被引:1,自引:0,他引:1
对临床样品微量元素分析过程中的样品制备技术的最近进展进行了评述,主要从直接稀释、湿法消解、干式灰化及微量元素形态分析的样品预处理技术等几方面进行了分别介绍。 相似文献