New techniques of on-line biological sample processing and their application in the field of biopharmaceutical analysis

Peng, Jie; Tang, Fang; Zhou, Rui; Xie, Xin; Li, Sanwang; Xie, Feifan; Yu, Ping; Mu, Lingli

doi:10.1016/j.apsb.2016.05.016

Cited by 50 publications

(21 citation statements)

References 64 publications

(66 reference statements)

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“…The introduction of a peptide purification step prior to LC-MS/MS analysis (Peng et al, 2016) in routine in proteomics laboratories opened an opportunity to accurately determine peptide concentrations. Such a peptide purification step, typically based on Solid Phase Extraction (SPE), removes buffer components (like chaotropes, certain detergents, reducing/alkylating reagents and trypsin) that interfere with LC-MS/MS analysis.…”

Section: Resultsmentioning

confidence: 99%

“…Furthermore, automated sample preparation workflows (Fu et al, 2018;Guryča et al, 2014) or robotized systems start to become available, which allow accurate liquid handling, temperature control and precise timing of processing steps (Kulak, Geyer, & Mann, 2017;Zhu et al, 2018). Some of these methods can be performed in miniaturized settings, which are advantageous when working with clinical or other low abundance samples where analyte amounts are scarce (Peng et al, 2016;Zhu et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

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A simple approach for accurate peptide quantification in MS-based proteomics

Staes

Plasman

et al. 2019

Preprint

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Despite its growing popularity and use, bottom-up proteomics remains a complex analytical methodology. Its general workflow consists of three main steps: sample preparation, liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) and computational data analysis. Quality assessment of the different steps and components of this workflow is instrumental to identify technical flaws and to avoid loss of precious measurement time and sample material. However, assessment of the extent of sample losses along the sample preparation protocol, in particular after proteolytic digestion, is not yet routinely implemented because of the lack of an accurate and straightforward method to quantify peptides. Here, we report on the use of a microfluidic UV/visible spectrophotometer to quantify MS-ready peptides directly in MS loading solvent, consuming only 2 µl of sample. We determined the optimal peptide amount for LC-MS/MS analysis on a Q Exactive HF mass spectrometer using a dilution series of a commercial K562 cell digest. Careful evaluation of selected LC and MS parameters allowed us to define 3 µg as an optimal peptide amount to be injected on this particular LC-MS/MS system. Finally, using tryptic digests from human HEK293T cells, we showed that injecting equal peptide amounts, rather than approximated ones, results into less variable LC-MS/MS and protein quantification data. The obtained quality improvement together with easy implementation of the approach makes it possible to routinely quantify MS-ready peptides as a next step in daily proteomics quality control.This material is available free of charge via the Internet at

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A simple approach for accurate peptide quantification in MS-based proteomics

Staes

Plasman

et al. 2019

Preprint

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“…The pretreatment of pharmaceutical and biological samples in flow is an important step in closed systems. Due to the complexity of these samples, the determination of chemical species presents significant challenges [40]. Thus, different types of procedures can be developed for the preparation of on-line samples for each matrix, according to its characteristics, such as solid phase extraction, solid phase microextraction, liquid-liquid microextraction, and chemical derivatization.…”

Section: On-line Sample Processing Methods For In Flow Analysismentioning

confidence: 99%

Application of Flow-Injection Spectrophotometry to Pharmaceutical and Biomedical Analyses

Costa¹,

Rezende²,

Tavares³

et al. 2017

Spectroscopic Analyses - Developments and Applications

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The discovery of new drugs, especially when many samples have to be analyzed in the minimum of time, demand the improvement or development of new analytical methods. Various techniques may be employed for this purpose. In this context, this chapter gathers the collection of paper and represents the review of past work on spectrophotometric technique coupled to a continuous flow system to determine low concentrations of several chemical species in different kinds of pharmaceutical and biological samples. A short historical background of the flow-injection analysis technique and a brief discussion of the basic principles and potential are presented. Part of this chapter is devoted to describing the sample preparation techniques, principles, and figures of merit of analytical methods. Representative applications of flow-injection spectrophotometry to pharmaceutical and biomedical analysis are also described.

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“…During the past decades, some methods and tools have been developed for identification and characterization of saffron and its adulterants, which exhibit their own advantages and merits, as well as a lot of shortcomings. For example, laboratory chromatography and spectroscopy techniques are the most commonly used, but may suffer from heavy equipments, tedious sample preparation steps, along with high costs and long time of analysis . DNA barcoding technique needs to be combined with other tools and requires trained experts for the authentication of saffron .…”

Section: Introductionmentioning

confidence: 99%

Development of a high-performance liquid chromatography with tandem mass spectrometry method for identifying common adulterant content in saffron (Crocus sativus L.)

Kong

Zhang

et al. 2019

Journal of Pharmacy and Pharmacology

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Objectives This study aimed to develop an efficient and reliable method for estimating common adulterants in saffron by detecting their characteristic components to warrant its efficacy and regular use as a highly valuable medicinal herb. Methods A selective and sensitive high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS) method was developed to estimate the common adulterants in saffron from corn stigma, chrysanthemum and safflower through the simultaneous determination of specific constituents including allantoin, chlorogenic acid (ChA) and hydroxysafflor yellow A (HSYA). Peak identification of each target compound was confirmed from product ions obtained using multiple reaction monitoring triggered enhanced product ions mass chromatogram. Method validation in terms of linearity, sensitivity, reproducibility, accuracy and stability was systematically performed according to official guidelines. Key findings Satisfactory separation of the three components was achieved on a C 18 column (4.6 9 250 mm, 5 lm) with methanol-acetonitrile-ammonium acetate (3.0 mM) as the mobile phase at gradient elution. The identification of these specific constituents was accomplished using the multiple reaction monitoring mode in combination with enhanced product ion supplementary confirmation. The established method was validated in terms of linearity, sensitivity, reproducibility, accuracy and recovery, which were found satisfactory for sensitive detection of the three target compounds. Conclusions By detecting the specific constituents allantoin, ChA and HSYA in one run, the adulterants of corn stigma, chrysanthemum and safflower can be effectively identified and estimated in saffron. This is the first report on developing a simple, sensitive and operational method for the identification and estimation of common adulterants of saffron, that was forwarded for broaden application.

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New techniques of on-line biological sample processing and their application in the field of biopharmaceutical analysis

Cited by 50 publications

References 64 publications

A simple approach for accurate peptide quantification in MS-based proteomics

A simple approach for accurate peptide quantification in MS-based proteomics

Application of Flow-Injection Spectrophotometry to Pharmaceutical and Biomedical Analyses

Development of a high-performance liquid chromatography with tandem mass spectrometry method for identifying common adulterant content in saffron (Crocus sativus L.)

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