Development of a fast method for direct analysis of intact synthetic insulins in human plasma: the large peptide challenge

Erin E Chambers; Cristina Legido-Quigley; Norman Smith; Kenneth J Fountain

doi:10.4155/bio.12.290

Development of a fast method for direct analysis of intact synthetic insulins in human plasma: the large peptide challenge

Erin E Chambers, Cristina Legido-Quigley, Norman Smith, Kenneth J Fountain

61 Citationer (Scopus)

Abstract

BACKGROUND: Intact insulins are difficult to analyze by LC-MS/MS due to nonspecific binding and poor sensitivity, solubility and fragmentation. This work aims to provide a simpler, faster LC-MS method and focuses on solving the above issues.

RESULTS: A novel charged-surface chromatographic column produced peak widths for insulin that were significantly narrower than traditional C18 columns when using formic acid as mobile phase. Mass spectral fragments m/z >700 provided greater specificity, significantly reducing endogenous background. Detection limits in human plasma were 0.2 ng/ml for insulin glargine, glulisine and detemir, and 0.5 ng/ml for insulin aspart. Average accuracy for standard curve and QC samples was 93.4%.

CONCLUSION: A simple SPE LC-MS analysis was developed for direct, simultaneous quantification of insulin glargine, detemir, aspart and glulisine.

Originalsprog	Engelsk
Tidsskrift	Bioanalysis
Vol/bind	5
Udgave nummer	1
Sider (fra-til)	65-81
Antal sider	17
ISSN	1757-6180
DOI	https://doi.org/10.4155/bio.12.290
Status	Udgivet - jan. 2013
Udgivet eksternt	Ja

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10.4155/bio.12.290

Citationsformater

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title = "Development of a fast method for direct analysis of intact synthetic insulins in human plasma: the large peptide challenge",

abstract = "BACKGROUND: Intact insulins are difficult to analyze by LC-MS/MS due to nonspecific binding and poor sensitivity, solubility and fragmentation. This work aims to provide a simpler, faster LC-MS method and focuses on solving the above issues.RESULTS: A novel charged-surface chromatographic column produced peak widths for insulin that were significantly narrower than traditional C18 columns when using formic acid as mobile phase. Mass spectral fragments m/z >700 provided greater specificity, significantly reducing endogenous background. Detection limits in human plasma were 0.2 ng/ml for insulin glargine, glulisine and detemir, and 0.5 ng/ml for insulin aspart. Average accuracy for standard curve and QC samples was 93.4%.CONCLUSION: A simple SPE LC-MS analysis was developed for direct, simultaneous quantification of insulin glargine, detemir, aspart and glulisine.",

keywords = "Amino Acid Sequence, Analytic Sample Preparation Methods, Blood Chemical Analysis/methods, Calibration, Chromatography, Liquid, Humans, Insulins/blood, Limit of Detection, Mass Screening, Molecular Sequence Data, Molecular Weight, Quality Control, Surface Plasmon Resonance, Time Factors",

author = "Chambers, {Erin E} and Cristina Legido-Quigley and Norman Smith and Fountain, {Kenneth J}",

year = "2013",

month = jan,

doi = "10.4155/bio.12.290",

language = "English",

volume = "5",

pages = "65--81",

journal = "Bioanalysis",

issn = "1757-6180",

publisher = "Future Science Ltd",

number = "1",

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T1 - Development of a fast method for direct analysis of intact synthetic insulins in human plasma

T2 - the large peptide challenge

AU - Chambers, Erin E

AU - Legido-Quigley, Cristina

AU - Smith, Norman

AU - Fountain, Kenneth J

PY - 2013/1

Y1 - 2013/1

N2 - BACKGROUND: Intact insulins are difficult to analyze by LC-MS/MS due to nonspecific binding and poor sensitivity, solubility and fragmentation. This work aims to provide a simpler, faster LC-MS method and focuses on solving the above issues.RESULTS: A novel charged-surface chromatographic column produced peak widths for insulin that were significantly narrower than traditional C18 columns when using formic acid as mobile phase. Mass spectral fragments m/z >700 provided greater specificity, significantly reducing endogenous background. Detection limits in human plasma were 0.2 ng/ml for insulin glargine, glulisine and detemir, and 0.5 ng/ml for insulin aspart. Average accuracy for standard curve and QC samples was 93.4%.CONCLUSION: A simple SPE LC-MS analysis was developed for direct, simultaneous quantification of insulin glargine, detemir, aspart and glulisine.

AB - BACKGROUND: Intact insulins are difficult to analyze by LC-MS/MS due to nonspecific binding and poor sensitivity, solubility and fragmentation. This work aims to provide a simpler, faster LC-MS method and focuses on solving the above issues.RESULTS: A novel charged-surface chromatographic column produced peak widths for insulin that were significantly narrower than traditional C18 columns when using formic acid as mobile phase. Mass spectral fragments m/z >700 provided greater specificity, significantly reducing endogenous background. Detection limits in human plasma were 0.2 ng/ml for insulin glargine, glulisine and detemir, and 0.5 ng/ml for insulin aspart. Average accuracy for standard curve and QC samples was 93.4%.CONCLUSION: A simple SPE LC-MS analysis was developed for direct, simultaneous quantification of insulin glargine, detemir, aspart and glulisine.

KW - Amino Acid Sequence

KW - Analytic Sample Preparation Methods

KW - Blood Chemical Analysis/methods

KW - Calibration

KW - Chromatography, Liquid

KW - Humans

KW - Insulins/blood

KW - Limit of Detection

KW - Mass Screening

KW - Molecular Sequence Data

KW - Molecular Weight

KW - Quality Control

KW - Surface Plasmon Resonance

KW - Time Factors

U2 - 10.4155/bio.12.290

DO - 10.4155/bio.12.290

M3 - Journal article

C2 - 23256473

SN - 1757-6180

VL - 5

SP - 65

EP - 81

JO - Bioanalysis

JF - Bioanalysis

IS - 1

ER -

Development of a fast method for direct analysis of intact synthetic insulins in human plasma: the large peptide challenge

Abstract

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