Investigation of application possibilities of novel analytical methods for controlling the addition of natural and artificial dyes in meat products

Kontrola upotrebe boja koje se kao aditivi danas široko koriste u proizvodnji hrane ima veliki značaj za bezbednost i kvalitet proizvoda od mesa. Pored zakonskih regulativa važnu ulogu imaju i analitičke metode za utvrđivanje prisustva i određivanje količine boja. Cilj ove doktorske disertacije je bio da se razviju metode za istovremeno, rutinsko određivanje više boja kod različitih proizvoda od mesa. Analiza podataka iz literature pokazala je da ima malo analitičkih metoda za određivanje boja u proizvodima od mesa, da referentna metoda obuhvata samo veštačke boje, priprema uzoraka je, često, komplikovana i nepodesna za rutinski rad i da su savremene analitičke metode tečne hromatografije primenjene za određivanje boja u bezalkoholnim i alkoholnim pićima, konditirskim proizvodima, želeu, džemovima itd., ali ne i kod proizvoda od mesa. Za ispitivanja su izrađivani eksperimentalni proizvodi od mesa (finousitnjena barena kobasica - „pariska kobasica“ i konzerva od mesa u komadima - „kuvana šunka“), i proizvodi od mesa sa tržišta iz navedenih grupa. Eksperimentalni proizvodi su izrađivani u industrijskoj klanici i radionici Instituta za higijenu i tehnologiju mesa, Beograd. Izrađeni su kontrolni proizvodi bez dodatih boja i proizvodi u koje su u toku postupka izrade posebno dodavane boje Košenila, Ponso 4R i Alura crvena AC. U toku ispitivanja proizvoda od mesa korišćene su hemijske i fizičko-hemijske metode: (određivanje sadržaja azota (proteina): SRPS ISO 937:192, određivanje sadržaja slobodne masti: SRPS ISO 1444:1998, određivanje sadržaja vlage: SRPS ISO 1442:1998, određivanje sadržaja ukupnog pepela: SRPS ISO 936:1999 i određivanje pH vrednosti: SRPS ISO 2917:2004), senzorna ispitivanja boje preseka proizvoda, ujednačenosti boje preseka i postojanost boje 10 minuta nakon narezivanja (kvantitativna deskriptivna analiza, zasnovana na metodi SRPS ISO 6564:2001 i SRPS ISO 6658:2001), originalna metoda visokoefikasne tečne hromatografije sa fotodiodnom detekcijom (HPLC – PDA) sa prethodnim izolovanjem boja iz uzorka ubrzanom ekstrakcijom na visokom pritisku i temperaturi i merenje boje preseka proizvoda CIE L*a*b kolorimetrijom...Proper control of dyes that are widely used as food additives in production of foodstuffs is of considerable significance for safety and quality of meat products. Besides the adoption of legislative regulating this area, analytical methods for qualitative and quantitative determination of dyes also play an important role. The aim of this Doctoral dissertation was to develop routine analytical methods for simultaneous determination of multiple dyes in various meat products. Review of available literature showed that the number of analytical methods for determination of dyes in meat products is scarce, while the current reference method is intended for artificial dyes only. Furthermore, sample preparation techniques are often tedious and time-consuming, while available modern liquid chromatographic methods are mainly focused on dyes determination in alcoholic and non-alcoholic beverages, confectionary products, jellies, marmalades, etc., as opposed to availability of such methods applicable to meat products. For the purposes of investigations, experimental meat products were manufactured (finely grinded cooked sausage – “Parisian sausage” and chopped meat products – “cooked ham”), and a number of products of the same type were obtained from the retail as well. Manufacturing took place in industrial slaughterhouse and within the facilities of the Institute of meat hygiene and technology, Belgrade. Two types of products were made – control batches without addition of dyes and experimental batches – products with addition of the following dyes: Cochineal, Ponceau 4R and Allura red AC. The following chemical and physico-chemical methods were used during the experiments: (determination of nitrogen (protein) content SRPS ISO 937:192, determination of free fats content: SRPS ISO 1444:1998, determination of moisture content: SRPS ISO 1442:1998, determination of total ash content: SRPS ISO 936:1999 and determination of pH: SRPS ISO 2917:2004), sensory investigations of cut-surface color, consistency of cut-surface color and color stability ten minutes after slicing (quantitative descriptive test based on the methods SRPS ISO 6564:2001 and SRPS ISO 6658:2001), original liquid chromatographic method with photodiode array detection (HPLC – PDA) with dyes isolation from the sample using accelerated solvent extraction under high pressure and high temperature and color measuring of products’ cut-surface using CIE L*a*b colorimetry..

Comparison of sugars, lipids and phenolics content in the grains of organically and conventionally grown soybean in Serbia

The aim of the current study was to determine the content of several primary metabolites: total soluble sugars, starch and lipids, soluble sugars, fatty acids and triacylglycerols profile, and secondary metabolites: total phenolics and flavonoids, in the grains of soybean (Glycine max (L.) Merr.) cultivar ‘Kaća’. Additionally, grain antioxidant properties were assessed using ABTS•+ scavenging capacity and ferric reducing power (FRP) assays. Soybean was developed and grown in Serbia under two cultivation systems (conventional and organic) during two growing seasons (2016 and 2017). In both growing seasons and cultivation systems, soybean grains were characterised by reduced lipid content (8.16–14.34%) and as an excellent source of polyunsaturated fatty acids. Triacylglycerols (TAGs) with 44 equivalent carbon numbers (ECN44) represented the main fraction (30.95–32.79%) followed by ECN46 TAGs (23.27–26.36%). Low total soluble sugars (2.36–11.51%) content was determined. High-performance liquid chromatography (HPLC) analysis revealed a significant prevalence of non-reducing disaccharides (1.41–6.57%) among the individual sugars. Soybean grains were proved as a good source of phenolic (2493.9–4419.5 mg kg-1) and flavonoid (292.7–500.9 mg kg-1) compounds with the dominance of free (extractable) fractions. Strong positive correlations were observed between both cultivation systems and growing seasons indicating no clear differences for the majority of analysed parameters. All examined extracts possessed a significant ability (27.6–38.2%) to neutralize ABTS•+ radicals, while in the case of FRP assay a significant ability for iron ions (Fe3+) reduction was recorded for the samples from the second growing season

Investigation of application possibilities of novel analytical methods for controlling the addition of natural and artificial dyes in meat products

Kontrola upotrebe boja koje se kao aditivi danas široko koriste u proizvodnji hrane ima veliki značaj za bezbednost i kvalitet proizvoda od mesa. Pored zakonskih regulativa važnu ulogu imaju i analitičke metode za utvrđivanje prisustva i određivanje količine boja. Cilj ove doktorske disertacije je bio da se razviju metode za istovremeno, rutinsko određivanje više boja kod različitih proizvoda od mesa. Analiza podataka iz literature pokazala je da ima malo analitičkih metoda za određivanje boja u proizvodima od mesa, da referentna metoda obuhvata samo veštačke boje, priprema uzoraka je, često, komplikovana i nepodesna za rutinski rad i da su savremene analitičke metode tečne hromatografije primenjene za određivanje boja u bezalkoholnim i alkoholnim pićima, konditirskim proizvodima, želeu, džemovima itd., ali ne i kod proizvoda od mesa. Za ispitivanja su izrađivani eksperimentalni proizvodi od mesa (finousitnjena barena kobasica - „pariska kobasica“ i konzerva od mesa u komadima - „kuvana šunka“), i proizvodi od mesa sa tržišta iz navedenih grupa. Eksperimentalni proizvodi su izrađivani u industrijskoj klanici i radionici Instituta za higijenu i tehnologiju mesa, Beograd. Izrađeni su kontrolni proizvodi bez dodatih boja i proizvodi u koje su u toku postupka izrade posebno dodavane boje Košenila, Ponso 4R i Alura crvena AC. U toku ispitivanja proizvoda od mesa korišćene su hemijske i fizičko-hemijske metode: (određivanje sadržaja azota (proteina): SRPS ISO 937:192, određivanje sadržaja slobodne masti: SRPS ISO 1444:1998, određivanje sadržaja vlage: SRPS ISO 1442:1998, određivanje sadržaja ukupnog pepela: SRPS ISO 936:1999 i određivanje pH vrednosti: SRPS ISO 2917:2004), senzorna ispitivanja boje preseka proizvoda, ujednačenosti boje preseka i postojanost boje 10 minuta nakon narezivanja (kvantitativna deskriptivna analiza, zasnovana na metodi SRPS ISO 6564:2001 i SRPS ISO 6658:2001), originalna metoda visokoefikasne tečne hromatografije sa fotodiodnom detekcijom (HPLC – PDA) sa prethodnim izolovanjem boja iz uzorka ubrzanom ekstrakcijom na visokom pritisku i temperaturi i merenje boje preseka proizvoda CIE L*a*b kolorimetrijom...Proper control of dyes that are widely used as food additives in production of foodstuffs is of considerable significance for safety and quality of meat products. Besides the adoption of legislative regulating this area, analytical methods for qualitative and quantitative determination of dyes also play an important role. The aim of this Doctoral dissertation was to develop routine analytical methods for simultaneous determination of multiple dyes in various meat products. Review of available literature showed that the number of analytical methods for determination of dyes in meat products is scarce, while the current reference method is intended for artificial dyes only. Furthermore, sample preparation techniques are often tedious and time-consuming, while available modern liquid chromatographic methods are mainly focused on dyes determination in alcoholic and non-alcoholic beverages, confectionary products, jellies, marmalades, etc., as opposed to availability of such methods applicable to meat products. For the purposes of investigations, experimental meat products were manufactured (finely grinded cooked sausage – “Parisian sausage” and chopped meat products – “cooked ham”), and a number of products of the same type were obtained from the retail as well. Manufacturing took place in industrial slaughterhouse and within the facilities of the Institute of meat hygiene and technology, Belgrade. Two types of products were made – control batches without addition of dyes and experimental batches – products with addition of the following dyes: Cochineal, Ponceau 4R and Allura red AC. The following chemical and physico-chemical methods were used during the experiments: (determination of nitrogen (protein) content SRPS ISO 937:192, determination of free fats content: SRPS ISO 1444:1998, determination of moisture content: SRPS ISO 1442:1998, determination of total ash content: SRPS ISO 936:1999 and determination of pH: SRPS ISO 2917:2004), sensory investigations of cut-surface color, consistency of cut-surface color and color stability ten minutes after slicing (quantitative descriptive test based on the methods SRPS ISO 6564:2001 and SRPS ISO 6658:2001), original liquid chromatographic method with photodiode array detection (HPLC – PDA) with dyes isolation from the sample using accelerated solvent extraction under high pressure and high temperature and color measuring of products’ cut-surface using CIE L*a*b colorimetry..

Masnokiselinski profil organskog i konvencionalnog semena spelte

Usled izuzetno povoljnih prehrambenih, medicinskih i agronomskih karakteristika, kao i povoljnog hemijskog sastava semena koje je u skladu sa najsavremenijim nutritionističkim zahtevima, proizvodnja spelte dobija sve veći značaj, naročito u organskoj proizvodnji hrane. Veoma važan nutritivni faktor namirnica predstavlja sadržaj masnih kiselina u njima, neophodnih za pravilno funkcionisanje ljudskog organizma. Cilj ovog rada bio je da GC-FID metodom ispita sadržaj masnih kiselina u organskom i konvencionalnom semenu spelte, poreklom iz 2015 i 2016. godine. Sadržaj masnih kiselina izražen je kao % sadržaj u odnosu na ukupne masne kiseline. Detektovano je deset masnih kiselina. Kod obe vrste semena, u najvećoj količini se nalaze linolna (C18:2n-6) i oleinska (C18:1n-9) kiselina. Najveći udeo polinezasićenih masnih kiselina sadrži seme konvencionalne spelte (59.46 %), dok se u organskom semenu iz 2016. godine nalazio najveći udeo zasićenih (18.33 %) i mononezasićenih masnih kiselina (26.71 %).Due to the extremely favorable nutritional, medical and agronomic characteristics, as well as the favorable chemical composition of seed which is in accordance with the current nutritional requirements, spelt production is gaining increasing importance, especially in organic food production. The fatty acids (FAs) content is a very important nutritional factor. Adequate FAs daily intake is necessary for the proper functioning of the human body. The aim of this paper was to examine fatty acids content in organic and conventional spelt seeds produced during 2015 and 2016 year. GC-FID method was used for FAs determination. FAs content is expressed as % of content in relation to the total fatty acids. Ten fatty acids were detected. Both types of seeds contained linoleic (C18:2n-6) and oleic acid (C18:1n-9) as predominant. Conventional spelt seed possessed higher content of polyunsaturated fatty acids (59.46 %) while organic seed from 2016 contained higher amount of saturated (18.33 %) and monounsaturated fatty acids (26.71 %)

Effect of modified atmosphere and vacuum packaging on selected chemical parameters of rainbow trout (Oncorhynchus mykiss) and carp (Cyprinus carpio) cuts freshness

The purpose of food packing in modified atmosphere is to extend its sustainability by preventing both biochemical processes and growth of spoilage bacteria. Gases or their mixtures which are mostly used in the modified atmosphere food packing technology are carbon-dioxide (CO2), oxygen (O2) and nitrogen (N2). The aim of our research was to examine the influence of packaging in modified atmosphere and vacuum on the total volatile basic nitrogen (TVB-N) content and pH in muscle of rainbow trout (Oncorhynchus mykiss) and common carp (Cyprinus carpio), as well as to determine the most suitable gas mixtures for packing of these freshwater species. Three sample groups of trout and carp cuts were investigated. The first two groups were packaged in modified atmosphere with different gas ratios: 60%CO2+40%N2 (I group) and 40%CO2+60%N2 (II group), whereas the samples from third, control group, (III group) were vacuum packaged. During trials samples were stored in refrigerator at +3°C. Determination of TVB-N and pH was performed on 1st, 7th and 14th day of storage. The obtained results indicate that the investigated mixtures of gases and vacuum as well had a significant influence on the values of TVB-N in trout and carp cuts samples. The lowest increase in TVB-N was established in trout and carp cuts samples from the group I, whereas the highest increase was established in samples from group III. Statistical significant difference (p < 0,001) between the average values of TVB-N for trout (I group: 18,17 ± 0,93; II group: 20,90 ± 0,81 and III group: 36,18 ± 2,65 mg N/100 g ) and carp cuts (I group: 26,74 ± 1,48; II group: 30,02 ± 0,31 and III group: 35,10 ± 1,75 mg N/100 g) was established on 14th day. The lowest pH value was established in samples packaged in modified atmosphere with 60% CO2 +40% N2 (I group). On 14th day of testing the obtained value was 6,15 ± 0,09 for trout and 5,94 ± 1,11 for carp samples. Increase in pH value in trout samples packed in vacuum was established during the whole period of investigation (p < 0,001), while in carp cuts samples packaged in vacuum the increase in pH value (p < 0,05) was established up to 7th day of testing. Based on the obtained results it can be concluded that gas mixture consisting of 60% CO2 and 40% N2 was the most suitable for packaging of fresh trout and carp cuts in terms of selected chemical parameters, such as TVB-N and pH. [Projekat Ministarstva nauke Republike Srbije, br. TR 31011 i br. TR 31075

Hemometrijski pristup razvoju kolorimetrijske metode za procenu količine prehrambenih boja u proizvodima od mesa

The aim of this research was to develop a novel colorimetric method based on mathematical models, by multiple linear regression (MLR), from the CIE L*a*b* measurements and data of the HPLC determination of food colorants. Calibration set of 10 production batches of finely grinded cooked sausage with food colorants added was manufactured in industrial conditions as follows: one control batch and 9 products with various quantities of added food colorants: E120 (3.4, 7.5 and 12.5 mg/kg), E 124 (5.0, 15.0, 25.0 mg/kg) and E 129 (5.0, 15.0, 25.0 mg/kg). The estimation of the added food colorants was assessed by measuring L*, a*, b* parameters of cross-section. The quantification of food colorants was achieved by HPLC-PDA. Food colorants were extracted from meat products using Accelerated Solvent Extraction (ASE). Quantification of food colorants was achieved in the range from 1 to 100 mg / kg, and recovery values were from 76.15% to 107.04%, for E 120, from 97.61% to 101.03%, for E 124 and from 99.91% to 101.67%, for E 129. Correlation of the results obtained using HPLC and colorimetric measuring data was assessed by Multiple Linear Regression (MLR). The results from colorimetric and chromatographic determinations in four experimental batches (three batches with different quantities of food colorants and one control batch) were used for calibration. Coefficients of determination (R2) for linear models in experimental batches were 0.954, for E 124, 0.987, for E 120 and 0.993, for E 129. Correlation functions of food colorant quantities and corresponding L*a*b* values were established. The obtained mathematical models were tested for the estimation of the content of dyes in 21 samples of finely grinded cooked sausages purchased in retail stores. Food colorants were confirmed in 20 samples (95.24 %), and one sample (4.76 %) did not contain any of these compounds. Out of the positive samples, sixteen samples (80.00 %) contained E 120, while four samples (20.00 %) contained E 129. Food colorant E124 was not established in any of the analyzed samples. Colorimetric CIE L*a*b* method might be used during sensory evaluation of meat products for the assessment of the added food colorants.Cilj ovog rada je bio da se, merenjem vrednosti parametara boje preseka proizvoda od mesa u CIE (Commission Internationale de l'Eclairage, Međunarodna komisija za osvetljenje) L*a*b* prostoru (L* – svetloća, a* – udeo crvene boje i b* – udeo žute boje) ispita mogućnost procene sadržaja dodate prehrambene boje u proizvodima od mesa. Količina prehrambene boje u uzorcima od mesa određena je metodom visokoefikasne tečne hromatografije sa detekcijom na fotodiodnom sloju (HPLC-PDA), a međusobna zavisnost dobijenih vrednosti i kolorimetrijskih parametara ispitana je višestrukom linearnom regresijom (MLR). Kalibracioni set od 10 proizvodnih partija fino usitnjene barene kobasice, sa tri dodate boje, izrađen je u industrijskim uslovima, i to: kontrolni proizvod bez boje i 9 proizvoda sa različitim količinama dodatih boja E 120 (3,4; 7,5 i 12,5 mg/kg), E 124 (5, 15 i 25 mg/kg) i E 129 (5, 15 i 25 mg/kg). Postavljene su jednačine funkcije zavisnosti količine boje od L*, a* i b* vrednosti. Dobijeni matematički modeli su provereni i primenjeni za procenjivanje sadržaja boje u 21 uzorku fino usitnjenih barenih kobasica sa tržišta. Utvrđeno je da se kolorimetrijskom CIE L*a*b* metodom mogu, u toku senzorskog ocenjivanja boje proizvoda, proceniti količine dodatih boja u proizvodima od mesa i da se ova metoda može primeniti kao komplementarna HPLC-PDA

Chemometric approach in the development of the colorimetric method for the estimation of food colorants in meat products

The aim of this research was to develop a novel colorimetric method based on mathematical models, by multiple linear regression (MLR), from the CIE L*a*b* measurements and data of the HPLC determination of food colorants. Calibration set of 10 production batches of finely grinded cooked sausage with food colorants added was manufactured in industrial conditions as follows: one control batch and 9 products with various quantities of added food colorants: E120 (3.4, 7.5 and 12.5 mg/kg), E 124 (5.0, 15.0, 25.0 mg/kg) and E 129 (5.0, 15.0, 25.0 mg/kg). The estimation of the added food colorants was assessed by measuring L*, a*, b* parameters of cross-section. The quantification of food colorants was achieved by HPLC-PDA. Food colorants were extracted from meat products using Accelerated Solvent Extraction (ASE). Quantification of food colorants was achieved in the range from 1 to 100 mg / kg, and recovery values were from 76.15% to 107.04%, for E 120, from 97.61% to 101.03%, for E 124 and from 99.91% to 101.67%, for E 129. Correlation of the results obtained using HPLC and colorimetric measuring data was assessed by Multiple Linear Regression (MLR). The results from colorimetric and chromatographic determinations in four experimental batches (three batches with different quantities of food colorants and one control batch) were used for calibration. Coefficients of determination (R2) for linear models in experimental batches were 0.954, for E 124, 0.987, for E 120 and 0.993, for E 129. Correlation functions of food colorant quantities and corresponding L*a*b* values were established. The obtained mathematical models were tested for the estimation of the content of dyes in 21 samples of finely grinded cooked sausages purchased in retail stores. Food colorants were confirmed in 20 samples (95.24 %), and one sample (4.76 %) did not contain any of these compounds. Out of the positive samples, sixteen samples (80.00 %) contained E 120, while four samples (20.00 %) contained E 129. Food colorant E124 was not established in any of the analyzed samples. Colorimetric CIE L*a*b* method might be used during sensory evaluation of meat products for the assessment of the added food colorants

The fatty acid and triacylglycerol profiles of conventionally and organically produced grains of maize, spelt and buckwheat

Triacylglycerols (TAGs) and free fatty acids (FAs) present an important part of the grain lipid fraction. Knowing about its composition can be important information for both chemotaxonomic and nutritional parameters. The aim of the present study was to determine the TAG and FA profiles (and their similarities/differences) of several important crops (maize, spelt, buckwheat) grown under dissimilar agroecological systems - conventional and organic. Fatty acid composition was determined by applying the GC-FID method while HPLC method was used for the determination of the triacylglycerol profile of selected grains. To the best of our knowledge, the TAG profile of spelt grains was determined for the first time. Ten different FAs were identified in examined samples with a predominant presence of linoleic acid, except in the case of buckwheat (oleic acid) grains. Uniformly, buckwheat grains stood out due to the predominance of triglycerides with an equivalent carbon number of fifty (ECN 50) or forty-eight (ECN 48). In contrast, other samples contained ECN 44 and ECN 46 TAGs as the most represented triglycerides. Based on the principal component analysis (PCA) and cluster statistical analysis results, there is no uniformity in the differences between organic and conventional production growing systems