Citation:

Billa, N., D. Hubin-Barrows, T. Lahren, AND L. Burkhard. Evaluation of micro-colorimetric lipid determination method with samples prepared using sonication and accelerated solvent extraction methods. TALANTA. Elsevier Science Ltd, New York, NY, 119:620-622, (2014).

Impact/Purpose:

The suitability of a commonly used colorimetric method for measuring lipid content in small tissue samples (10-20 mg) was evaluated for with extracts prepared by sonication and accelerated solvent extraction techniques. The extracts from the accelerated solvent extraction process had significant bias and without correction, inaccurate measurements of lipid content would occur. The extracts from the sonication process had no biases. Investigators using this technique need to be aware of the biases associated with the accelerated solvent extraction technique.

Description:

Two common laboratory extraction techniques were evaluated for routine use with the micro-colorimetric lipid determination method developed by Van Handel (1985) [E. Van Handel, J. Am. Mosq. Control Assoc. 1(1985) 302] and recently validated for small samples by Inouye and Lotufo (2006) [L.S. Inouye, G.R. Lotufo. Talanta 70(2006) 584]. With accelerated solvent extraction method using chloroform:methanol solvent and the colorimetric lipid determination method, 16 of 19 samples had significant proportional bias (á=1%, determined using standard additions) and 0 of 19 samples had significant constant bias (á=1%, determined using Youden Blank measurements). With sonic extraction, 0 of 9 samples had significant proportional bias (á=1%) and 2 of 9 samples had significant constant bias (á=1%). These demonstrate that accelerate solvent extraction method with chloroform:methanol solvent system creates an interference with the colorimetric assay method, and without accounting for the bias in the analysis, inaccurate measurements would be obtained.

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