Sample preparation for inositol measurement: Sep-Pak C18 use in detergent removal

Susan E. Laursen; Harvey R. Knull; John K. Belknap

doi:10.1016/0003-2697(86)90107-7

Sample preparation for inositol measurement: Sep-Pak C₁₈ use in detergent removal

Susan E. Laursen, Harvey R. Knull, John K. Belknap

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

6 Scopus citations

Abstract

The enzymatic-fluorometric method of myo-inositol quantitation by L. C. MacGregor and F. M. Matschinsky (1984, Anal. Biochem. 141, 382-389) has proved to be very useful in accurately measuring small amounts of myo-inositol. Although this procedure is quite satisfactory, relatively high concentrations of detergents, salts, NADH, and malate interfere. To extend the usefulness of the MacGregor and Matschinsky method we report here an extraction procedure which removed the interfering substances, yet allowed the recovery of close to 100% of the inositol. The procedure involved first passing the sample through a Sep-Pak C₁₈ cartridge to remove detergent and then through a mixed-bed resin to remove the ionic constituents. The procedure with the Sep-Pak C₁₈ cartridge is applicable to a wide variety of biological samples requiring detergent removal.

Original language	English (US)
Pages (from-to)	387-390
Number of pages	4
Journal	Analytical Biochemistry
Volume	153
Issue number	2
DOIs	https://doi.org/10.1016/0003-2697(86)90107-7
State	Published - Mar 1986
Externally published	Yes

Keywords

NADH
Sep-Pak C
detergent extraction
fluorometry
malate extraction
myo-inositol

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

Access to Document

10.1016/0003-2697(86)90107-7

Cite this

@article{ec703f988b8542c198ad8c35eb56cad5,

title = "Sample preparation for inositol measurement: Sep-Pak C18 use in detergent removal",

abstract = "The enzymatic-fluorometric method of myo-inositol quantitation by L. C. MacGregor and F. M. Matschinsky (1984, Anal. Biochem. 141, 382-389) has proved to be very useful in accurately measuring small amounts of myo-inositol. Although this procedure is quite satisfactory, relatively high concentrations of detergents, salts, NADH, and malate interfere. To extend the usefulness of the MacGregor and Matschinsky method we report here an extraction procedure which removed the interfering substances, yet allowed the recovery of close to 100% of the inositol. The procedure involved first passing the sample through a Sep-Pak C18 cartridge to remove detergent and then through a mixed-bed resin to remove the ionic constituents. The procedure with the Sep-Pak C18 cartridge is applicable to a wide variety of biological samples requiring detergent removal.",

keywords = "NADH, Sep-Pak C, detergent extraction, fluorometry, malate extraction, myo-inositol",

author = "Laursen, {Susan E.} and Knull, {Harvey R.} and Belknap, {John K.}",

note = "Funding Information: {\textquoteright} This research was supported by NIH Grants NS I77 I 1, AA06342, and DA02723.",

year = "1986",

month = mar,

doi = "10.1016/0003-2697(86)90107-7",

language = "English (US)",

volume = "153",

pages = "387--390",

journal = "Analytical Biochemistry",

issn = "0003-2697",

publisher = "Academic Press Inc.",

number = "2",

}

TY - JOUR

T1 - Sample preparation for inositol measurement

T2 - Sep-Pak C18 use in detergent removal

AU - Laursen, Susan E.

AU - Knull, Harvey R.

AU - Belknap, John K.

N1 - Funding Information: ’ This research was supported by NIH Grants NS I77 I 1, AA06342, and DA02723.

PY - 1986/3

Y1 - 1986/3

N2 - The enzymatic-fluorometric method of myo-inositol quantitation by L. C. MacGregor and F. M. Matschinsky (1984, Anal. Biochem. 141, 382-389) has proved to be very useful in accurately measuring small amounts of myo-inositol. Although this procedure is quite satisfactory, relatively high concentrations of detergents, salts, NADH, and malate interfere. To extend the usefulness of the MacGregor and Matschinsky method we report here an extraction procedure which removed the interfering substances, yet allowed the recovery of close to 100% of the inositol. The procedure involved first passing the sample through a Sep-Pak C18 cartridge to remove detergent and then through a mixed-bed resin to remove the ionic constituents. The procedure with the Sep-Pak C18 cartridge is applicable to a wide variety of biological samples requiring detergent removal.

AB - The enzymatic-fluorometric method of myo-inositol quantitation by L. C. MacGregor and F. M. Matschinsky (1984, Anal. Biochem. 141, 382-389) has proved to be very useful in accurately measuring small amounts of myo-inositol. Although this procedure is quite satisfactory, relatively high concentrations of detergents, salts, NADH, and malate interfere. To extend the usefulness of the MacGregor and Matschinsky method we report here an extraction procedure which removed the interfering substances, yet allowed the recovery of close to 100% of the inositol. The procedure involved first passing the sample through a Sep-Pak C18 cartridge to remove detergent and then through a mixed-bed resin to remove the ionic constituents. The procedure with the Sep-Pak C18 cartridge is applicable to a wide variety of biological samples requiring detergent removal.

KW - NADH

KW - Sep-Pak C

KW - detergent extraction

KW - fluorometry

KW - malate extraction

KW - myo-inositol

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JO - Analytical Biochemistry

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