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

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

59 Citations (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.

Original language	English
Pages (from-to)	65-81
Number of pages	17
Journal	Bioanalysis
Volume	5
Issue number	1
DOIs	https://doi.org/10.4155/bio.12.290
Publication status	Published - 1 Jan 2013

Access to Document

10.4155/bio.12.290

Cite this

@article{84bb6dbb7fa34ba8bc25a58a2b393c5a,

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. ",

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

note = "{\textcopyright} 2013 Future Science Ltd.",

year = "2013",

month = jan,

day = "1",

doi = "10.4155/bio.12.290",

language = "English",

volume = "5",

pages = "65--81",

journal = "Bioanalysis",

issn = "1757-6180",

publisher = "Future Science",

number = "1",

}

TY - JOUR

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 - 2013/1/1

Y1 - 2013/1/1

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.

UR - http://www.scopus.com/inward/record.url?scp=84871539893&partnerID=8YFLogxK

U2 - 10.4155/bio.12.290

DO - 10.4155/bio.12.290

M3 - Article

C2 - 23256473

AN - SCOPUS:84871539893

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

Access to Document

Other files and links

Fingerprint

Cite this