Simultaneous Determination of Vitamin A and E in Infant Formula by HPLC with Photodiode Array Detection

Abstract

The objective of this study was to develop a method to simultaneously quantify vitamins A and E in infant formula. To determine the vitamin A and E content, vitamin A and four different vitamin E isomers (α-, β-, γ-, and δ-tocopherol) were separated by high performance liquid chromatography with a photodiode array detector using a Develosil RPAQUEOUS RP-C 3 0 column (4.6×250 mm, 5 µm). The vitamin A and E contents in the certified reference material determined using this method were within the certified range of standard values. The limits of detection (LODs) and limits of quantitation (LOQs) for vitamin A were 0.02 and 0.06 µg/L, respectively. LODs and LOQs for the vitamin E isomers ranged from 0.20 to 0.55 and from 0.67 to 1.81 µg/L, respectively. Linear analyses indicated that the square of the correlation coefficient for the vitamin A and E isomers was 0.9997-0.9999. The recovery of vitamins ranged from 96.69 to 97.79%. The results demonstrate that this novel method could be used to reliably analyze vitamin A and E content in infant formula.

Simultaneous and accurate determination of water- and fat-soluble vitamins in multivitamin tablets by using an RP-HPLC method

“…30 Many of the studies developed for the quantification of vitamins in various matrices also include extraction procedures, and the solvents used differ according to the type of vitamin (fat-soluble or water-soluble). For the separate extraction of fat-and water-soluble vitamins in various matrices, solvents such as methanol:water, 31 methanol, 32 ethanol:orthophosphoric acid:hexane, 33 and ethanol:water 34,35 have been used. In addition, for the extraction of all of the fat-and water-soluble vitamins, a mixture of TFA:methanol 18 and a column technique with SPE extraction 24 have been used.…”

Liquid Chromatography Analysis of Common Nutritional Components, in Feed and Food

“…Since these compounds are involved in metabolic pathways, and are paramount in health promotion in animals and humans, it is crucial to determine their content in food and feed to comply with daily requirements and quality control. That is why several studies have been conducted regarding the extraction and quantitative analysis of these vitamins, either individually or simultaneously [359,363,364,365,366,367,368,369,370,371,372,373,374,375,376,377,378,379,380,381,382].…”

“…Qian and Sheng developed the assay procedure without saponification, but nevertheless it is a widespread procedure used in the analysis of vitamins since it is an efficient way of removing interferences of lipid origin that can be found in the matrix [363,365,368,374,378,379,382]. Since it is based on an alkaline digestion (i.e., heated KOH or NaOH aqueous or alcoholic solutions) there is a disadvantage, the saponification could generate oxidation of the vitamins, which translate into a loss for vitamin degradation and low recovery percentage [366,368]. Some researchers have made use of the antioxidant ability of some compounds such as BHT, BHA, TBHQ, ascorbic acid or pyrogallol to reduce oxidation losses [363,365,367,368,371,374,379,380].…”

“…Since it is based on an alkaline digestion (i.e., heated KOH or NaOH aqueous or alcoholic solutions) there is a disadvantage, the saponification could generate oxidation of the vitamins, which translate into a loss for vitamin degradation and low recovery percentage [366,368]. Some researchers have made use of the antioxidant ability of some compounds such as BHT, BHA, TBHQ, ascorbic acid or pyrogallol to reduce oxidation losses [363,365,367,368,371,374,379,380]. Nevertheless, saponification procedures take time, and the extractions procedures are not always straightforward, because emulsions are generated, as Lim and co-worker mentioned in their comparison of extraction methods for determining tocopherols in soybeans [369], these inconveniences introducing considerable variation, low recovery, and reproducibility, situations that we have also seen in the development of this type of methodologies.…”