Determination of Vitamins A and E in Infant Formula and Adult/Pediatric Nutritional Formula by HPLC with UV and Fluorescence Detection: First Action 2012.09

2013 ◽  
Vol 96 (6) ◽  
pp. 1407-1413 ◽  
Author(s):  
Linda Butler Thompson ◽  
Karen Schimpf ◽  
Steve Baugh
Keyword(s):  
Vitamin E ◽  
Vitamin A ◽  
Infant Formula ◽  
Methylene Chloride ◽  
Lipid Fraction ◽  
Intermediate Precision ◽  
Silica Column ◽  
Vitamin E Acetate ◽  
Uv And Fluorescence Detection

Abstract The method presented is for quantification of α-tocopherol (vitamin E), vitamin E acetate, vitamin A acetate, and vitamin A palmitate in infant formula and adult/pediatric nutritionals. The entire lipid fraction, including vitamins A and E, is extracted from product with iso-octane after products are mixed with methanol, which precipitates proteins and disrupts micelles freeing lipids for extraction. Vitamin A palmitate, vitamin A acetate, and vitamin E acetate are separated from α-tocopherol on a 3 cm silica column with a 1% methylene chloride, 0.06% isopropanol in iso-octane mobile phase; eluted onto a 20 cm silica column; and, after a column switch, further separated on the 20 cm column before UV detection at 325 nm (vitamin A palmitate and vitamin A acetate) and 285 nm (vitamin E acetate). α-Tocopherol is further separated from other extraneous compounds on the 3 cm silica column and detected by fluorescence at excitation and emission wavelengths of 295 and 330 nm, respectively. Quantification limits in ready-to-feed products were estimated to be 80 IU/L for vitamin A palmitate, 207 International Units (IU)/L for vitamin A acetate, 2.4 mg/L for vitamin E acetate, and <0.15 mg/L for α-tocopherol. Over-spike recoveries and intermediate precision averaged 100.4 and 2.09% RSD for vitamin A palmitate, 100.4 and 1.52% RSD for vitamin E acetate, and 99.6 and 3.02% RSD for α-tocopherol. Vitamin A acetate spike recovery data averaged 96.6%, and the intermediate precision for the only product fortified with vitamin A acetate was 2.75% RSD.

Simultaneous Determination of 13-cis and all-trans Vitamin A Palmitate (Retinyl Palmitate), Vitamin A Acetate (Retinyl Acetate), and Total Vitamin E (α-Tocopherol and DL-α-Tocopherol Acetate) in Infant Formula and Adult Nutritionals by Normal Phase HPLC: First Action 2012.10

2013 ◽  
Vol 96 (5) ◽  
pp. 1073-1081 ◽  
Author(s):  
Adrienne McMahon ◽  
Scott Christiansen ◽  
Lynsey Shine ◽  
Calvin Loi ◽  
Dawn Dowell
Keyword(s):  
Vitamin E ◽  
Vitamin A ◽  
Simultaneous Determination ◽  
Infant Formula ◽  
Normal Phase ◽  
Total Vitamin ◽  
Validation Data ◽  
Tocopherol Acetate ◽  
Selection Of

Abstract This HPLC method, with both variable UV and fluorescence detection, allows for the simultaneous determination of vitamin A palmitate, vitamin A acetate, and total vitamin E in infant, pediatric, and adult nutritional formulas. The concentration of each vitamin form is calculated by comparison with standards of known concentration. Following hydrolysis, the vitamins are extracted into iso-octane and analyzed by normal phase (NP) HPLC. The method was evaluated for linearity, precision, and accuracy using a selection of the Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN) matrixes, including milk-based, soy-based, and hydrolyzed protein, as well as high- and low-fat products. A single-laboratory validation has been completed for all analytes using a selection of SPIFAN matrixes. Performance parameters included a working range of 2–450 μg/100 g ready-to-feed for vitamin A and 0.03–8.0 mg/100 g reconstituted final product for vitamin E. LOD was <1.0 μg and <0.1 mg/100 g reconstituted final product for vitamins A and E, respectively; RSD was 1.08–8.70% over a range of concentration; and average recoveries of 97.4–101.3%. Repeatability of <4% for vitamin A and <8% for vitamin E was calculated from five laboratories using this method. Results indicate that this method is suitable for the analysis of vitamins A and E in all forms of infant, adult, and pediatric formulas (powders, ready-to-feed liquids, and liquid concentrates). The Expert Review Panel (ERP) of Infant Formula reviewed this method separately for vitamins A and E, including all available method validation data at the AOAC INTERNATIONAL Annual Meeting on September 29, 2012. Following evaluation of the data for both methods, the ERP agreed that both methods met the standard method performance requirements articulated by SPIFAN. The ERP granted First Action status to both methods, and recommended that a single method be published for the simultaneous determination of vitamin A palmitate, vitamin A acetate, and total vitamin E (DL-α-tocopherol and DL-α-tocopherol acetate) in infant formula and adult nutritionals by NP HPLC.

Determination of Vitamin E and Vitamin A in Infant Formula and Adult Nutritionals by Normal-Phase High-Performance Liquid Chromatography: Collaborative Study, Final Action 2012.10

2016 ◽  
Vol 99 (1) ◽  
pp. 223-241 ◽  
Author(s):  
Adrienne McMahon ◽  
S Christiansen ◽  
F-F Chee ◽  
A Chua ◽  
H Braddock ◽  
...  
Keyword(s):  
Vitamin E ◽  
Vitamin A ◽  
Infant Formula ◽  
Collaborative Study ◽  
Retinyl Palmitate ◽  
Normal Phase ◽  
Retinyl Acetate ◽  
Total Vitamin ◽  
Normal Phase Hplc

Abstract The main objective of the AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN) project is to establish international consensus methods for infant formula and adult nutritionals, which will benefit intermarket supply and dispute resolution. A collaborative study was conducted on AOAC First Action Method 2012.10 Simultaneous Determination of 13-cis and All-trans Vitamin A Palmitate (Retinyl Palmitate), Vitamin A Acetate (Retinyl Acetate), and Total Vitamin E (α-Tocopherol and D-α-tocopherol acetate) in Infant Formula and Adult Nutritionals by Normal-Phase HPLC. Fifteen laboratories from 11 countries participated in an interlaboratory study to determine 13-cis and all-trans vitamin A palmitate (retinyl palmitate), vitamin A acetate (retinyl acetate), and total vitamin E (α-tocopherol and D-α-tocopherol acetate) in infant formula and adult nutritionals by normal-phase HPLC and all laboratories returned valid data. Eighteen test portions of nine blind duplicates of a variety of infant formula and adult nutritional products were used in the study. The matrixes included milk-based and soy-based hydrolyzed protein as well as a low fat product. Each of the samples was prepared fresh and analyzed in singlicate. As the number of samples exceeded the recommended number to be prepared in a single day, analysis took place over 2 days running 12 samples on day one and 10 samples on day two. The reference standard stock was prepared once and the six-point curve diluted freshly on each day. Results obtained from all 15 laboratories are reported. The RSDR for total vitamin A (palmitate or acetate) ranged from 6.51 to 22.61% and HorRat values ranged from 0.33 to 1.25. The RSDR for total vitamin E (as tocopherol equivalents) ranged from 3.84 to 10.78% and HorRat values ranged from 0.27 to 1.04. Except for an adult low fat matrix which generated reproducibility RSD >40% for some isomers, most SPIFAN matrixes gave results within the acceptance criteria of <16% RSD as stated in the respective Standard Method Performance Requirements.

Gas-Liquid Chromatographic Determination of Vitamin D2 in Multiple Vitamin Tablets Containing Minerals and Vitamin E Acetate

1974 ◽  
Vol 57 (5) ◽  
pp. 1089-1091
Author(s):  
David O Edlund ◽  
Florido A Filippini ◽  
James K Datson
Keyword(s):  
Vitamin E ◽  
Chromatographic Determination ◽  
Vitamin D2 ◽  
Vitamin E Acetate ◽  
Chromatographic Procedure ◽  
Ether Solution ◽  
Liquid Chromatographic Determination ◽  
Liquid Chromatographic ◽  
Column Chromatographic

Abstract A gas-liquid chromatographic procedure used to determine vitamin D2 in multiple vitamin tablets has been modified to make it applicable for analysis of multiple vitamin tablets containing minerals and vitamin E acetate. The procedure modifications involve pre-extraction with ether, solution in an alcoholic sulfuric acid-pyridine mixture, and column chromatographic separation on phosphate-treated alumina. The modified procedure has been statistically evaluated. A 2.2% coefficient of variation and 100.3% average recovery were obtained for the samples evaluated.

scholarly journals Direct Determination of Free Methionine in Soy-Based Infant Formula

2004 ◽  
Vol 87 (1) ◽  
pp. 123-128
Author(s):  
Paul Johns ◽  
Rosalyn Phillips ◽  
Lobat Dowlat
Keyword(s):  
Infant Formula ◽  
Mobile Phase ◽  
Method Comparison ◽  
Water System ◽  
Direct Determination ◽  
Reversed Phase ◽  
Intermediate Precision ◽  
Relative Standard ◽  
Limit Of Quantitation

Abstract A method was developed for the direct determination of free methionine in soy-based infant formula, with analyte separation and quantitation by reversed-phase liquid chromatography (LC), and UV absorbance at 214 nm, respectively. Sample preparation required only dilution with mobile phase and syringe filtration. Using a 0.02M KH 2 PO 4 mobile phase (pH adjusted to 2.9 with 85% o-phosphoric acid) and 0.7 mL/min flow rate, methionine eluted at approximately 8 min, and total run time was 14 min after column regeneration with acetonitrile–water. System linearity was demonstrated as peak area versus analyte concentration, ranging from 80 to 120% of the formula specification for free methionine (r > 0.999, and all residuals <0.45%). Intermediate precision relative standard deviation values were <1.5% for ready-to-feed and reconstituted powder samples, and recoveries ranged from 98.0 to 103.5% for inter-method comparison with an amino acid analyzer method. The limit of quantitation was 3 mg methionine/L in the “as fed” infant formula. Despite the relatively weak UV absorptivity of methionine, the 214 nm signal was sufficiently intense in the 30–65 mg/L (201–436 μM) range to afford quantitation by peak area proportionation versus a 2-point external standard calibration. This direct UV detection after reversed-phase LC separation provides a simple and accurate method for determining free methionine without derivatization.

Determination of vitamin A and vitamin E esters in infant formulae and fortified milk powders by HPLC: Use of internal standardisation

2016 ◽  
Vol 197 ◽  
pp. 457-465 ◽  
Author(s):  
David C. Woollard ◽  
Anja Bensch ◽  
Harvey Indyk ◽  
Adrienne McMahon
Keyword(s):  
Vitamin E ◽  
Vitamin A ◽  
Infant Formulae ◽  
Fortified Milk
Sign in / Sign up

Export Citation Format

Share Document