Determination of Free and Total Myo-Inositol in Infant Formula and Adult/Pediatric Nutritional Formula by High-Performance Anion Exchange Chromatography with Pulsed Amperometric Detection, Including a Novel Total Extraction Using Microwave-Assisted Acid Hydrolysis and Enzymatic Treatment: First Action 2012.12

2013 ◽  
Vol 96 (5) ◽  
pp. 1068-1072 ◽  
Author(s):  
David Ellingson ◽  
Ted Pritchard ◽  
Pamela Foy ◽  
Kathryn King ◽  
Barbara Mitchell ◽  
...  
Keyword(s):  
Acid Hydrolysis ◽  
Anion Exchange ◽  
High Performance ◽  
Amperometric Detection ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Pulsed Amperometric Detection ◽  
Microwave Assisted ◽  
Total Extraction

Analysis of Free and Total Myo-Inositol in Foods, Feeds, and Infant Formula by High-Performance Anion Exchange Chromatography with Pulsed Amperometric Detection, Including a Novel Total Extraction Using Microwave-Assisted Acid Hydrolysis and Enzymatic Treatment

2012 ◽  
Vol 95 (5) ◽  
pp. 1469-1478 ◽  
Author(s):  
David Ellingson ◽  
Ted Pritchard ◽  
Pamela Foy ◽  
Kathryn King ◽  
Barbara Mitchell ◽  
...  
Keyword(s):  
Acid Hydrolysis ◽  
Anion Exchange ◽  
Infant Formula ◽  
High Performance ◽  
Amperometric Detection ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Pulsed Amperometric Detection ◽  
Microwave Assisted ◽  
Total Analysis

Abstract A method for the analysis of free and total myo-inositol in foods, feeds, and infant formulas has been developed and validated using high-performance anion exchange chromatography with pulsed amperometric detection. The option of a free myo-inositol determination or a complete total myo-inositol determination from main bound sources can be achieved. These sources include phytates, lower phosphorylated forms, and phosphatidylinositol. This approach gives the option for subtraction of myo-inositol from nonbioavailable sources when it is quantified using other methods if a total bioavailable myo-inositol result is desired for nutritional labeling of a product. The free analysis was validated in a milk-based infant formula, giving RSDR of 2.29% and RSDr of 2.06%. A mean recovery of 97.9% was achieved from various spike levels of myo-inositol. Certified National Institute of Standards and Technology reference material verified the method's compatibility and specificity. Two different total analyses were validated in a soy-based infant formula and compared. One technique involved using a conventional acid hydrolysis with autoclave incubation for 6 h, while the other used a novel technique of microwave-assisted acid hydrolysis with enzymatic treatment that can minimize extraction to 1 day. The autoclave analysis had RSDR of 2.08% and RSDr of 1.55%, along with a mean spike recovery of 102.1% at various myo-inositol spike levels. The microwave/enzyme total analysis had RSDR of 4.34% and RSDr of 4.70%, along with a mean spike recovery of 104.2% at various spike levels of myo-inositol. Main sources of myo-inositol including phytic acid and phosphatidylinositol were tested with both total analyses. Mean recoveries of phytic acid and phosphatidylinositol through the autoclave total analysis were 90.4 and 98.3%, respectively. Mean spike recoveries for these same sources in soy-based infant formula through the microwave/enzyme total analysis were 97.2 and 96.3%, respectively. Comparison of soy-based infant formula and corn grain samples with high levels of these main sources showed in similar results, indicating both total analyses are acceptable for use. An additional glycerol kinase step was also developed to remove glycerol from the chromatographic elution window of myo-inositol in samples with high levels of glycerol.

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