Improved Extraction of Saturated Fatty Acids but not Omega‐3 Fatty Acids from Sheep Red Blood Cells Using a One‐Step Extraction Procedure

Clayton, Edward; Gulliver, Catherine; Piltz, John; Taylor, Robert D.; Blake, Robert R.; Meyer, Richard G.

doi:10.1007/s11745-012-3674-1

Cited by 32 publications

(16 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Adjusting BHT concentrations in the hexane solvent present during derivatization in the direct method had no effect, suggesting that extraction differences rather than lipid peroxidation might be responsible for differences in PUFA yields (data not shown). In a similar comparison for red blood cells, more saturated fatty acids were extracted with a direct method than with Folch but not MUFAs or PUFAs (Clayton et al 2012). Conversely, two comparative studies using marine animal material reported improved recovery of PUFAs in direct methods relative to conventional two-step methods, however (Abdulkadir and Tsuchiya 2008; Indarti et al 2005).…”

Section: Discussionmentioning

confidence: 99%

Comparison of screening methods for high-throughput determination of oil yields in micro-algal biofuel strains

et al. 2012

View full text Add to dashboard Cite

The phenotypic and phylogenetic diversity of micro-algae capable of accumulating triacylglycerols provides a challenge for the accurate determination of biotechnological potential. High-yielding strains are needed to improve economic viability and their compositional information is required for optimizing biodiesel properties. To facilitate a high-throughput screening programme, a very rapid direct-derivatization procedure capable of extracting lyophilized material for GC analysis was compared with a scaled-down Folch-based method. This was carried out on ten micro-algal strains from 6 phyla where the more rapid direct-derivatization approach was found to provide a more reliable measure of yield. The modified Folch-based procedure was found to substantially underestimate oil yield in one Chlorella species (P < 0.01). In terms of fatty acid composition however, the Folch procedure proved to be slightly better in recovering polyunsaturated fatty acids, in six out of the ten strains. Therefore, direct-derivatization is recommended for rapid determination of yields in screening approaches but can provide slightly less compositional accuracy than solvent-based extraction methods.

show abstract

Section: Discussionmentioning

confidence: 99%

Comparison of screening methods for high-throughput determination of oil yields in micro-algal biofuel strains

et al. 2012

View full text Add to dashboard Cite

show abstract

“…Previous studies have shown that fatty acid analysis of human red blood cell can be determined by a two-step extraction and methylation or a combined one-step extraction and methylation procedure [17,20,25,26].…”

Section: Methods Of Determining Fatty Acid Compositionmentioning

confidence: 99%

“…A vivid picture of the long-term dietary intake, with particular emphasis on long-chain omega-3 polyunsaturated fatty acids, can be obtained by deducing the concentrations of fatty acids in red blood cells (RBCs) [17,18]. The fatty acids present in erythrocyte membrane phospholipids can be incorporated during de novo synthesis or via exchange from the pool of fatty acids in the plasma [12].…”

Section: Fatty Acid Composition In Biological Systemsmentioning

confidence: 99%

“…Of notable interest is the use of 14% boron trifluoride/methanol which is widely used in the transmethylation of fatty acids in red blood cells [1,17,27,28]. Other methylating reagents used include acetyl chloride [29], 2% sulfuric acid in methanol [30] and sodium methoxide [16,31].…”

Section: Methylation Proceduresmentioning

confidence: 99%

“…Boron trifluoride in methanol (12-14%) is the most often used for transesterification of RBC lipids [26]. Several other reagents used for trans-esterification of human and plant samples include acetyl chloride [17,26,36,37] and aluminum chloride [26,38].…”

Section: Methylation Proceduresmentioning

confidence: 99%

See 2 more Smart Citations

A Rapid and Simple Method for Fatty Acid Profiling and Determination of ω-3 Index in Red Blood Cells

Akinyemi¹,

Bruckner²,

Johnson³

et al. 2017

TONUTRJ

View full text Add to dashboard Cite

Fatty acid profiling has become a very useful and effective tool in the diagnosis, prevention and treatment of several diseases with cardiovascular disease being particularly important. In order to arrive at accurate conclusions that would help promote the health of individuals plagued by such diseases, not only excellent laboratory methods are required, but also very important monitoring responses to treatment. Improvements in methods of fatty acid profiling in biological systems regarding safety of extraction, precision and time for analysis are valuable. The ω-3 index (a measure of the amount of eicosapentaenoic acid, EPA, and docosahexaenoic acid, DHA, in Red Blood Cell membranes expressed as the percent of total fatty acids) is of growing interest because it has been reported to provide prognostic information regarding the risk for heart diseases. Sodium methoxide has been widely used for the determination of ω -3 fatty acids in food samples. This study demonstrates that sodium methoxide can be used effectively in RBC fatty acid profiling and determination of the ω-3 index. Briefly, the fatty acid profiles and ω-3 index of red blood cell samples were analyzed and results compared using three different methods: a two-step extraction and methylation method described by Hara and Radin, a single step extraction and methylation method described by Harris et al. and the sodium methoxide method.Our results revealed that there were no statistically significant differences (p<0.05) between the three methods for the representative fatty acids, [16:0 (p = 0.10), 18:0 (p=0.40), 18:1(ω9) (p = 0.29), 18:2(ω6) (p = 0.95), 18:3(ω3) (p = 0.50), 20:5(ω3) (p=0.56), 22:6(ω3) (p = 0.06)] and ω-3 index (p = 0.11) except for 20:4(ω6), (P = 0.02). In conclusion, we show that sodium methoxide can be used effectively in a one-step extraction and methylation procedure for high throughput analysis of fatty acids in red blood cell membranes. It is rapid (10 minute extraction and methylation), simple, safer than and as accurate as other commonly reported methods.

show abstract

Simple Methodology for the Quantitative Analysis of Fatty Acids in Human Red Blood Cells

et al. 2015

View full text Add to dashboard Cite

methods, in which lipid extraction and derivatization were performed simultaneously. For the one-step methods, different methylation periods and the inclusion of a saponification reaction were evaluated. Differences in absolute FA concentrations were observed among the tested methods, in particular for some metabolically relevant FAs such as trans elaidic acid and eicosapentaenoic acid. The one-step method with saponification and 60 min of methylation time was selected since it allowed the identification of a higher number of FAs, and was further submitted to in-house validation. The proposed methodology provides a simple, fast and accurate tool to quantitatively analyse FAs in human RBCs, useful for clinical and nutritional studies.

show abstract

Improved Extraction of Saturated Fatty Acids but not Omega‐3 Fatty Acids from Sheep Red Blood Cells Using a One‐Step Extraction Procedure

Cited by 32 publications

References 26 publications

Comparison of screening methods for high-throughput determination of oil yields in micro-algal biofuel strains

Comparison of screening methods for high-throughput determination of oil yields in micro-algal biofuel strains

A Rapid and Simple Method for Fatty Acid Profiling and Determination of ω-3 Index in Red Blood Cells

Simple Methodology for the Quantitative Analysis of Fatty Acids in Human Red Blood Cells

Contact Info

Product

Resources

About