Design of a molecularly imprinted stir-bar for isolation of patulin in apple and LC-MS/MS detection

Mycotoxins are a very diverse group of natural products produced as secondary metabolites by fungi. Patulin is produced by mold species normally related to vegetable-based products and fruit, mainly apple. Its ingestion may result in agitation, convulsions, edema, intestinal ulceration, inflammation, vomiting, and even immune, neurological or gastrointestinal disorders. For this reason, the European Commission Regulation (EC) 1881/2006 established a maximum content for patulin of 10 ppb in infant fruit juice, 50 ppb for fruit juice for adults and 25 ppb in fruit-derived products. In this work, a rapid and selective method based on magnetic molecularly imprinted stir-bar (MMISB) extraction has been developed for the isolation of patulin, using 2-oxindole as a dummy template. The final extraction protocol consisted of simply pouring in, stirring and pouring out samples and solvents from a beaker with the MMISB acting inside. The magnetic device provided satisfactory recoveries of patulin (60%–70%) in apple samples. The successful MMISB approach has been combined with high performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) to determine patulin.This research was supported by the project EM 2012/153 from Consellería de Cultura, Educación e Ordenacion Universitaria, Xunta de GaliciaS

Development of a novel molecularly imprinted stir-bar for isolation of aflatoxins

The 19th International Electronic Conference on Synthetic Organic Chemistry session Polymer and Supramolecular ChemistryMycotoxins are natural substances produced as secondary metabolites by a wide variety of different species of filamentous fungi. One of the most important groups in terms of occurrence and toxicity, is the group of aflatoxins (AFs). The major members of this group are aflatoxin B1, B2, G1, G2, M1 and M2. Exposure to AFs can cause chronic and acute toxic effects or death, as they can be carcinogenic, mutagenic, teratogenic and immunosuppressive. AFB1 is considered as the strongest carcinogen of natural origin and it is normally predominant in crops as well as in food products. Aflatoxin M1 is a major metabolite of aflatoxin B1 in humans and animals and it may be present in milk from animals fed with contaminated feed. Although the toxicity of M1 is about ten times lower than B1, it is known for its hepatotoxic and carcinogenic effects. Solid phase extraction (SPE) or the clean-up with organic solvents or immunoaffinity columns (IAC) have been frequently applied in the analysis of mycotoxins from cereals and/or milk. IAC in combination with HPLC are increasingly used as reference method due to their high selectivity and good elimination of matrix interferences. However, the cost of analysis is usually high. In recent years, molecularly imprinted polymers (MIP) have been become very popular and promising materials for extracting different analytes. AFs are too toxic to be used in MIP preparation and template bleeding may be an additional problem, especially when dealing with very low levels. For this reason, a dummy template was preferred for MIP synthesis. A rapid and selective extraction method based on magnetic MIP (MMIP) has been developed for the isolation of aflatoxins (B1, B2, G1, G2, M1) using a combination of imprinted polymer and magnetite. The successful MMIP stirring “cake” (used as a stir-bar) has been combined with HPLC-MS/MS for the determination of AFM1 in milk powder (infant formulas) to demonstrate its applicability to real sample

Magnetic molecularly imprinted stirring bar for isolation of patulin using grafting technique

The 19th International Electronic Conference on Synthetic Organic Chemistry session Polymer and Supramolecular ChemistryMycotoxins are low-molecular-weight natural products with great structural diversity produced as secondary metabolites by fungi. One of the principal toxic fungal metabolites is patulin (PAT), produced by over 30 genera of mold including species as Penicillium expansum or Penicillium griseofulvum, and normally related to vegetable-based products and fruit, mainly apple. These mold grow easily in damaged fruit or in derived-product as juices if storage conditions are deficient. Some of the most serious effects of PAT ingestion are agitation, convulsions, edema, ulceration intestinal, inflammation and vomiting. Thus, European Regulation 1881/2006 established a maximum content of 10 ppb in infant fruit juices, 50 ppb for fruit juices in adults and 25 ppb in fruit-derived products. Nowadays, the official analytical method for food adopted by AOAC International is HPLC with UV detection, using clean-up with ethyl acetate and sodium carbonate. However, the diverse drawbacks of this method (poor stability of PAT under alkaline extraction, poor resolution between PAT and co-extracted hydroxymethylfurfural) have originated interest in alternative options, such as LC methods coupled to mass spectrometry. In the last years, purification with molecularly imprinted polymers (MIP) started to be used, and are becoming promising materials for extracting different analytes present in food. Mycotoxins are too toxic or too expensive to be used as template molecules in MIP preparation. Template “bleeding” may be an additional problem, especially when dealing with very low detection levels. In the present work, a rapid and selective method based on magnetic molecularly imprinted stir-bar (MMIB) extraction has been developed for the isolation of PAT. A structural analogue, 2-oxindole, was used as dummy template. The polymer was grafted to the silanized glass surface of the stir ba

Association between Breast Milk Mineral Content and Maternal Adherence to Healthy Dietary Patterns in Spain: A Transversal Study

The composition of breast milk is influenced by many factors, some of which dependent on the mother and others on the child. Changes in lactation and other factors depending on the mother’s physiology and anthropometric characteristics, as well as her nutritional status and diet, are of key importance. Breast milk minerals have been extensively studied with highly uneven results. In this work, a comparison will made with data across the world. To understand the factors that might explain the disparity, several minerals (Na, K, Ca, P, Mg, Fe, Se and I) have been analyzed using ICP-MS in a set of human milk samples (n = 75). The samples had an identical geographical origin (Galicia, in northwestern Spain) but different lactation circumstances, including maternal anthropometric data, lactating time, newborn sex and maternal adherence to healthy dietary patterns (Mediterranean Diet, MD, or Atlantic Diet, AD). The required concentrations of essential elements reported in the literature are similar to those found in these Spanish women. A univariate approach revealed that factors such as lactating time, body mass index (BMI) and newborn sex have a significant influence in breastmilk mineral content. According to multivariate linear regression analysis, minerals in milk are particularly associated with lactating time, but also with newborn sex, maternal BMI, age and diet pattern in some cases. More precisely, these results suggest that the iron and selenium concentrations in the milk of Galician donors may be positively influenced by maternal adherence to AD and MD, respectivelyS

Fatty acids contribution of bovine and maternal milk to humans: assessment and influential factors

The fatty acids occurring in mammalian milk are one of the most studied components for their effects in human health. Several endogenous and exogenous factors can vary their levels in milk, making this food more beneficial or harmful to the human organism. In this Doctoral Thesis project, the fatty acid profiles of the three types of milk most consumed by humans, i.e. bovine milk, breast milk and/or infant formula, were analysed in detail. Diverse influential factors have been investigated, and their composition has been compared and discussed to assess the adaptation of each type of milk to the specific dietary requirements of humans.2024-03-0

Fattyacidomics and minerals: profiling the effects of recombinant bovine somatotropin on milk composition

SELECTED ABSTRACTS OF THE III INTERNATIONAL CAPARICA SYMPOSIUM ON PROFILING (ISPROF 2017), Costa de Caparica, Portugal, 4th – 7th September 2017The aim of this study was profiling the measurable effects or recombinant bovine somatotropin (rbST) on the nutritional properties of milk. A group of cows was treated with this peptide hormone and milk samples were collected for their analysis. The implementation of univariate and multivariate statistics showed a tendency towards a less saturated fatty-acidome in the milk collected from animals treated with rbST, with higher concentrations of omega-6 and monounsaturated fatty acids. In addition, less calcium and protein content and more potassium was observed in milk from treated animals, in comparison to control population. Thanks to this multi-component profiling of milk, a clear impact of somatotropin treatment on milk qualities was observedMinisterio de Economía y Competitividad. Programa Estatal de Investigación, Desarrollo e Innovación Orientada a los Retos de la Sociedad (proyecto AGL2014-58881-R)S