Separation optimization of long porous‐layer open‐tubular columns for nano‐<scp>LC</scp>–<scp>MS</scp> of limited proteomic samples

Røgeberg, Magnus; Vehus, Tore; Grutle, Lene; Greibrokk, Tyge; Wilson, Steven Ray; Lundanes, Elsa

doi:10.1002/jssc.201300499

Cited by 41 publications

(45 citation statements)

References 49 publications

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“…Rogeberg et al . compared the performance of silica-based and polymer-based monolithic columns and discovered a peak capacity of 950 from a 10 μm × 8 m PS-DVB PLOT column, which meets or surpasses peak capacities of previously reported using packed nanoLC or long silica monolithic columns [45]. With the PLOT column, about 500 and 1200 proteins were identified from an extract corresponding to 1000 and 10,000 cells, respectively [45].…”

Section: Lc or Ce For Nanoproteomicsmentioning

confidence: 92%

Advances in microscale separations towards nanoproteomics applications

Lee

Piehowski

Shi

et al. 2017

Journal of Chromatography A

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Microscale separation (e.g., liquid chromatography or capillary electrophoresis) coupled with mass spectrometry (MS) has become the primary tool for advanced proteomics, an indispensable technology for gaining understanding of complex biological processes. In recent decades significant advances have been achieved in MS-based proteomics. However, the current proteomics platforms still face an analytical challenge in overall sensitivity towards nanoproteomics applications for starting materials of less than 1 µg total proteins (e.g., cellular heterogeneity in tissue pathologies). Herein, we review recent advances in microscale separation techniques and integrated sample processing strategies that improve the overall sensitivity and proteome coverage of the proteomics workflow, and their contributions towards nanoproteomics applications.

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Section: Lc or Ce For Nanoproteomicsmentioning

confidence: 92%

Advances in microscale separations towards nanoproteomics applications

Lee

Piehowski

Shi

et al. 2017

Journal of Chromatography A

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“…However, substantial effort is still required in column manufacture to significantly improve chromatographic separation and miniaturization [8][9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Separation of intact proteins on γ‐ray‐induced polymethacrylate monolithic columns: A highly permeable stationary phase with high peak capacity for capillary high‐performance liquid chromatography with high‐resolution mass spectrometry

Simone

Pierri

Foglia

et al. 2015

J of Separation Science

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Polymethacrylate-based monolithic capillary columns, prepared by γ-radiation-induced polymerization, were used to optimize the experimental conditions (nature of the organic modifiers, the content of trifluoroacetic acid and the column temperature) in the separation of nine standard proteins with different hydrophobicities and a wide range of molecular weights. Because of the excellent permeability of the monolithic columns, an ion-pair reversed-phase capillary liquid chromatography with high-resolution mass spectrometry method has been developed by coupling the column directly to the mass spectrometer without a flow-split and using a standard electrospray interface. Additionally, the high working flow and concomitant high efficiency of these columns allowed us to employ a longer column (up to 50 cm) and achieve a peak capacity value superior to 1000. This work is motivated by the need to develop new materials for high-resolution chromatographic separation that combine chemical stability at elevated temperatures (up to 75°C) and a broad pH range, with a high peak capacity value. The advantage of the γ-ray-induced monolithic column lies in the batch-to-batch reproducibility and long-term high-temperature stability. Their proven high loading capacity, recovery, good selectivity and high permeability, moreover, compared well with that of a commercially available poly(styrene-divinylbenzene) monolithic column, which confirms that such monolithic supports might facilitate analysis in proteomics.

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“…Nos últimos anos, as colunas OT do tipo PLOT emergiram como ferramentas promissoras em cromatografia líquida (33,92). OT têm um consumo extremamente baixo de solventes, e permite maior sensibilidade ao usar espectrometria de massas por ionização por electrospray (ESI-MS) (93).…”

Section: Colunas Plotunclassified

“…OT têm um consumo extremamente baixo de solventes, e permite maior sensibilidade ao usar espectrometria de massas por ionização por electrospray (ESI-MS) (93). OT colunas têm recebido atenção devido à sua bem sucedida cromatografia de compostos de elevada massa molecular, tais como proteínas e peptídeos com alta capacidade de pico em sistemas de alta sensibilidade nano-LC tandem MS (MS / MS) (32,92,94,95,96,97). No entanto, as colunas do tipo OT estão associadas a uma fraca retenção e é praticamente não favorável para a análise de moléculas pequenas, o que está muito mais associado a colunas maiores e de alta capacidade.…”

Section: Colunas Plotunclassified

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Novas estratégias de preparo de amostras complexas para a determinação de sulfonamidas

Silva¹

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AGRADECIMENTOPrimeiramente a Deus.Aos meus pais, que diante das dificuldades da vida me proporcionaram a oportunidade de estudar sem medir esforços, pelo amor, puxão de orelha, pelo colo, ombro e por ser minha torcida organizada, obrigada é muito pouco! Ao meu irmão por ser o meu porto seguro sempre dizendo qualquer coisa eu estou aqui, obrigada pelo ombro, força e apoio incondicional! Ao Professor Dr. Fernando M. Lanças por me receber no laboratório de cromatografia, por me capacitar tecnicamente, pelo exemplo de profissional dedicado e eficiente, pela atenção, conselhos e incentivos ao longo desses anos, muito obrigada! Ao Alcimar (in memoria) pelo apoio e ensinamentos.Ao Professor Dr. Álvaro por me motivar através de uma conversa informal a trabalhar com líquidos iônicos e além disso, pelo exemplo de profissional, dedicação e humildade, obrigada.Ao Grupo de Cromatografia que tornaram a minha vida científica muito mais prazerosa não só pelos cafés, bolos e churrascos, mas pela companhia e inúmeras risadas, muito obrigada. Á Odete que agiu como meio mãe, amiga nestes anos e pelas inúmeras risadas, obrigada! Á Elaine pelo exemplo de dedicação, organização e pelas conversas, obrigada.Ao Guilherme que sempre foi solícito e disponível para discutir e dividir seu conhecimento, e lógico pelo cafezinho e pela companhia, obrigada.Ao grupo de Bioanalítica da UiO pelo conhecimento adquirido e por tornarem os dias frios um pouco mais amenos para uma brasileira, obrigada! E por fim, agradecer aqueles que cruzaram meu caminho e foram de extrema importância para chegar até aqui, muito obrigada Carlinhos, Maurinha, Maraíssa, Rodrigo, Raquel, Scarlet, Andreinha, Tanare, Pipe, Fumes, Arley, Camilinha, Van, Dani, Paty, Carol, Gabi, Jiraia, Lari, Naty, Stella, Josi e Kenia.O sucesso nasce do querer, da determinação e persistência em se chegar a um objetivo. Mesmo não atingindo o alvo, quem busca e vence obstáculos, no mínimo fará coisas admiráveis. José de Alencar RESUMO ABSTRACTNowadays health regulatory agencies are focusing on the presence of residues of antimicrobials, pesticides among other that are harmful to the environment, human and animal health that are found on food, river water among others, as example, the residues of sulfonamides (SAs) in bovine milk. The quantification of these compounds in complex matrices requires the sample preparation, extraction and preconcentration of the analytes, besides the elimination of the interferents for the development of chromatographic methods high sensitivity and analytical selectivity. Therefore, it is advantageous to use simple, fast methods that exhibit high selectivity, low reagent consumption, high sensitivity, higher analytical precision and automation of analyzes. In this context, this study explores one of the most employed sample preparation techniques, solid phase extraction (SPE) on automatization approach, the on-line SPE.Recently Compared to the use of monolithic / particulate SPE columns, SPE-OTLC mPLOT significantly improved the preconcentration and peak forms of ...

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Separation optimization of long porous‐layer open‐tubular columns for nano‐LC–MS of limited proteomic samples

Cited by 41 publications

References 49 publications

Advances in microscale separations towards nanoproteomics applications

Advances in microscale separations towards nanoproteomics applications

Separation of intact proteins on γ‐ray‐induced polymethacrylate monolithic columns: A highly permeable stationary phase with high peak capacity for capillary high‐performance liquid chromatography with high‐resolution mass spectrometry

Novas estratégias de preparo de amostras complexas para a determinação de sulfonamidas

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