High-performance liquid chromatography of rat and mouse islet polypeptides: potential risk of oxidation of methionine residues during sample preparation

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Standard

High-performance liquid chromatography of rat and mouse islet polypeptides : potential risk of oxidation of methionine residues during sample preparation. / Linde, S; Hansen, B; Welinder, B S; Nielsen, Jens Høiriis.

I: Journal of Chromatography A, Bind 530, Nr. 1, 24.08.1990, s. 29-37.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Harvard

Linde, S, Hansen, B, Welinder, BS & Nielsen, JH 1990, 'High-performance liquid chromatography of rat and mouse islet polypeptides: potential risk of oxidation of methionine residues during sample preparation', Journal of Chromatography A, bind 530, nr. 1, s. 29-37.

APA

Linde, S., Hansen, B., Welinder, B. S., & Nielsen, J. H. (1990). High-performance liquid chromatography of rat and mouse islet polypeptides: potential risk of oxidation of methionine residues during sample preparation. Journal of Chromatography A, 530(1), 29-37.

Vancouver

Linde S, Hansen B, Welinder BS, Nielsen JH. High-performance liquid chromatography of rat and mouse islet polypeptides: potential risk of oxidation of methionine residues during sample preparation. Journal of Chromatography A. 1990 aug 24;530(1):29-37.

Author

Linde, S ; Hansen, B ; Welinder, B S ; Nielsen, Jens Høiriis. / High-performance liquid chromatography of rat and mouse islet polypeptides : potential risk of oxidation of methionine residues during sample preparation. I: Journal of Chromatography A. 1990 ; Bind 530, Nr. 1. s. 29-37.

Bibtex

@article{210a78a194d14c458f54ee0032cf386f,
title = "High-performance liquid chromatography of rat and mouse islet polypeptides: potential risk of oxidation of methionine residues during sample preparation",
abstract = "After preparative high-performance liquid chromatography of mouse islet culture medium, concentrated on disposable C18 cartridges (Sep-Pak), an unexpected insulin immunoreactive peak eluting earlier than mouse insulin I and II was detected. Molecular mass determination by mass spectrometry supported its suspected identity as methionine sulphoxide insulin II. We have examined the formation of Met-O derivatives of insulin II, glucagon and pancreatic polypeptide during sample preparation (Sep-Pak and Speed-Vac concentrating). The oxidation of methionine residues was found to depend very much on the buffer, the organic modifier and the procedure. In particular the use of methanol-trifluoroacetic acid resulted in extensive oxidation. The oxidation could be minimized by adding 2 mM dithiothreitol to the buffer and by degassing and/or nitrogen-bubbling of the buffer. Minimal formation of Met-O derivatives is important for the quantitation of methionine-containing polypeptides.",
keywords = "Animals, Ascorbic Acid, Chromatography, High Pressure Liquid, Dithiothreitol, Glucagon, Insulin, Islets of Langerhans, Methionine, Mice, Molecular Weight, Oxidation-Reduction, Pancreatic Polypeptide, Peptides, Quality Control, Rats, Specimen Handling",
author = "S Linde and B Hansen and Welinder, {B S} and Nielsen, {Jens H{\o}iriis}",
year = "1990",
month = "8",
day = "24",
language = "English",
volume = "530",
pages = "29--37",
journal = "Journal of Chromatography A",
issn = "0021-9673",
publisher = "Elsevier",
number = "1",

}

RIS

TY - JOUR

T1 - High-performance liquid chromatography of rat and mouse islet polypeptides

T2 - potential risk of oxidation of methionine residues during sample preparation

AU - Linde, S

AU - Hansen, B

AU - Welinder, B S

AU - Nielsen, Jens Høiriis

PY - 1990/8/24

Y1 - 1990/8/24

N2 - After preparative high-performance liquid chromatography of mouse islet culture medium, concentrated on disposable C18 cartridges (Sep-Pak), an unexpected insulin immunoreactive peak eluting earlier than mouse insulin I and II was detected. Molecular mass determination by mass spectrometry supported its suspected identity as methionine sulphoxide insulin II. We have examined the formation of Met-O derivatives of insulin II, glucagon and pancreatic polypeptide during sample preparation (Sep-Pak and Speed-Vac concentrating). The oxidation of methionine residues was found to depend very much on the buffer, the organic modifier and the procedure. In particular the use of methanol-trifluoroacetic acid resulted in extensive oxidation. The oxidation could be minimized by adding 2 mM dithiothreitol to the buffer and by degassing and/or nitrogen-bubbling of the buffer. Minimal formation of Met-O derivatives is important for the quantitation of methionine-containing polypeptides.

AB - After preparative high-performance liquid chromatography of mouse islet culture medium, concentrated on disposable C18 cartridges (Sep-Pak), an unexpected insulin immunoreactive peak eluting earlier than mouse insulin I and II was detected. Molecular mass determination by mass spectrometry supported its suspected identity as methionine sulphoxide insulin II. We have examined the formation of Met-O derivatives of insulin II, glucagon and pancreatic polypeptide during sample preparation (Sep-Pak and Speed-Vac concentrating). The oxidation of methionine residues was found to depend very much on the buffer, the organic modifier and the procedure. In particular the use of methanol-trifluoroacetic acid resulted in extensive oxidation. The oxidation could be minimized by adding 2 mM dithiothreitol to the buffer and by degassing and/or nitrogen-bubbling of the buffer. Minimal formation of Met-O derivatives is important for the quantitation of methionine-containing polypeptides.

KW - Animals

KW - Ascorbic Acid

KW - Chromatography, High Pressure Liquid

KW - Dithiothreitol

KW - Glucagon

KW - Insulin

KW - Islets of Langerhans

KW - Methionine

KW - Mice

KW - Molecular Weight

KW - Oxidation-Reduction

KW - Pancreatic Polypeptide

KW - Peptides

KW - Quality Control

KW - Rats

KW - Specimen Handling

M3 - Journal article

C2 - 2277117

VL - 530

SP - 29

EP - 37

JO - Journal of Chromatography A

JF - Journal of Chromatography A

SN - 0021-9673

IS - 1

ER -

ID: 47974007