Optimization of lutein extraction method from real samples

Lutein je pigment svijetložute boje koji pripada skupini ksantofila. Karakteriziraju ga hepatoprotektivna, neuroprotektivna aktivnost te protuupalno i antioksidacijsko djelovanje. U radu je optimirana ekstrakcija luteina ispitivanjem utjecaja različitih ekstrakcijskih smjesa, utjecaj ultrazvuka, omjera suhe tvari (uzorka) i ekstrakcijske smjese, veličine čestica uzorka te vremena ekstrakcije na uspješnost ekstrakcije luteina iz uzorka. Veći sadržaj luteina određen je u usitnjenim uzorcima, boljom se pokazala direktna ultrazvučna ekstrakcija a od ispitivanih ekstrakcijskih smjesa, smjesa metanol:aceton=1:1. Najbolji omjer suhe tvari i uzorka bio je 1:20 kod metode 1 u kojoj je kao ekstrakcijska smjesa upotrebljena metanol: aceton = 1:1, dok je kod metode 4 u kojoj je heksan dodan kao krajnje otapalo bio 1:40 uz 1 sat stajanja uzorka na tamnom. Za analizu realnih uzoraka korištena je metoda 4 koja se pokazala kao najuspješnija. Ovom metodom određivan je sadržaj luteina u osam vrsta hrane za koke nesilice. Najveći sadržaj luteina određen je u hrani obogaćenoj luteinom, selenom, omega-3 masnim kiselinama i vitaminom E (75,314 mg/kg hrane). Dok je najniži sadržaj luteina određen u hrani dostupnoj na tržištu koja nije pomiješana sa žitaricama, 8,482 mg/ kg hrane.Lutein is a light yellow pigment belonging to the xanthophyll group. It is characterized by hepatoprotective, neuroprotective, anti-inflammatory and antioxidant activity. This paper present the optimisation of lutein extraction by examining the influence of different extraction mixtures, the influence of ultrasound, ratio of dry matter (sample) and extraction mixture, sample particle size and extraction time on the success of lutein extraction from the complex sample. The highest lutein content was determined in grinded samples, the direct ultrasonic extraction proved to be better, and the extraction mixture of methanol:acetone= 1:1 showed best results. The best dry matter to sample ratio was 1:20 in Method 1 in which methanol: acetone= 1:1 was used as the extraction mixture, while in Method 4 it was 1:40 in which hexane was added as the final solvent with 1 hour of sample resting in the dark place. Method 4, which proved to be the most successful, was used to analyze the real samples. This method was used to determine the content of lutein in eight types of food for laying hens. The highest lutein content was determined in food samples enriched with lutein, selenium, omega-3 fatty acids and vitamin E (75.314 mg / kg of sample), while the lowest lutein content was determined in food available on the market that was not mixed with cereals, 8.482 mg / kg of sample