Utilization of Vaccinium meridionale S. pomace as an eco-friendly and functional colorant in Greek-style yogurt.

Greek-style yogurt (GSY) has gained reputation as a healthy food because of its high protein content. Vaccinium meridionale S. is a bilberry with a high content of bioactive phytochemicals, whose vaccinium meridionale pomace (VMP) represents about 20% of the fruit weight. However, this byproduct is normally discarded as waste. In this study, VMP was used as a natural colorant in GSY. Coloring before or after the fermentation process resulted in significant increase in anthocyanins, total phenolics content, antioxidant activity, conjugated linoleic acid, and sensory acceptance. These results indicate that VMP is a potential natural, eco-friendly, and functional colorant to improve the nutritional value of GSY. PRACTICAL APPLICATION: Vaccinium meridionale pomace is a waste product with potential to be utilized as a natural, eco-friendly, and functional colorant to obtain value-added Greek-style yogurt. Besides providing bioactive compounds and natural color, this pomace improves the nutritional value, sensory acceptance, and functional properties of Greek yogurt.

Systematic assessment of mechanistic data for FDA-certified food colors and neurodevelopmental processes.

Seven US FDA-batch certified synthetic food colors are approved for use as food additives in the United States. Perceived neurodevelopmental concerns for these colors persist. This study assessed the plausibility of such an association through the evaluation of mechanistic evidence collected from in vitro assays or other alternative models. Mechanisms and molecular targets underlying neurodevelopmental processes associated with attention deficit and hyperactivity disorder (ADHD) and other neurodevelopmental-related symptoms (e.g., cognitive function, learning and memory disorder, etc.) were identified. Publicly available data from the ToxCast/Tox21 high-throughput screening (HTS) program and peer-reviewed literature that measure activity of the colors for such molecular targets were analyzed and reviewed. Erythrosine (Red No. 3) was active in several assays mapped to neurodevelopmental processes - specifically, HTS assays that measure signals in neurotransmitter pathways. The remaining six colors do not appear to alter signaling pathways related to neurodevelopmental processes on the molecular or cellular level. This assessment provides an approach for systematically identifying and mapping mechanistic data to putative neurodevelopmental processes as a means to prioritize substances for possible further investigation. The assessment also provides insights into the lack of activity of synthetic food colors for key events in neurodevelopmental signaling pathways.

Establishing Standards on Colors from Natural Sources.

Color additives are applied to many food, drug, and cosmetic products. With up to 85% of consumer buying decisions potentially influenced by color, appropriate application of color additives and their safety is critical. Color additives are defined by the U.S. Federal Food, Drug, and Cosmetic Act (FD&C Act) as any dye, pigment, or substance that can impart color to a food, drug, or cosmetic or to the human body. Under current U.S. Food and Drug Administration (FDA) regulations, colors fall into 2 categories as those subject to an FDA certification process and those that are exempt from certification often referred to as "natural" colors by consumers because they are sourced from plants, minerals, and animals. Certified colors have been used for decades in food and beverage products, but consumer interest in natural colors is leading market applications. However, the popularity of natural colors has also opened a door for both unintentional and intentional economic adulteration. Whereas FDA certifications for synthetic dyes and lakes involve strict quality control, natural colors are not evaluated by the FDA and often lack clear definitions and industry accepted quality and safety specifications. A significant risk of adulteration of natural colors exists, ranging from simple misbranding or misuse of the term "natural" on a product label to potentially serious cases of physical, chemical, and/or microbial contamination from raw material sources, improper processing methods, or intentional postproduction adulteration. Consistent industry-wide safety standards are needed to address the manufacturing, processing, application, and international trade of colors from natural sources to ensure quality and safety throughout the supply chain.

Food colors: Existing and emerging food safety concerns.

Food colors are added to different types of commodities to increase their visual attractiveness or to compensate for natural color variations. The use of these additives is strictly regulated in the European Union, the United States, and many other countries worldwide. There is a growing concern about the safety of some commonly used legal food colorants and there is a trend to replace the synthetic forms with natural products. Additionally, a number of dyes with known or suspected genotoxic or carcinogenic properties have been shown to be added illegally to foods. Robust monitoring programs based on reliable detection methods are required to assure the food is free from harmful colors. The aim of this review is to present an up to date status of the various concerns arising from use of color additives in food. The most important food safety concerns in the field of food colors are lack of uniform regulation concerning legal food colors worldwide, possible link of artificial colors to hyperactive behavior, replacement of synthetic colors with natural ones, and the presence of harmful illegal dyes-both known but also new, emerging ones in food. The legal status of food color additives in the EU, United States, and worldwide is summarized. The reported negative health effects of both legal and illegal colors are presented. The European Rapid Alert System for Food and Feed notifications and US import alerts concerning food colors are analyzed and trends in fraudulent use of color additives identified. The detection methods for synthetic colors are also reviewed.

Solvent optimization for anthocyanin extraction from Syzygium cumini L. Skeels using response surface methodology.

Anthocyanins are plant pigments that are potential candidates for use as natural food colourant. In this study, Syzygium cumini fruit skin has been used as anthocyanin source. All the six major types of anthocyanins were identified in the sample by ultra performance liquid chromatography studies, and the antioxidant activity was found to be 4.34 ± 0.26 Fe(2+)g(- 1) in the sample with highest anthocyanin content. Optimization of conditions for extracting high amounts of anthocyanin from the fruit peels was investigated by response surface methodology. The results suggested that highest anthocyanin yield (763.80 mg; 100 ml(- 1)), highest chroma and hue angle in the red colour range could be obtained when 20% ethanol was used in combination with 1% acetic acid. Methanol was replaced with ethanol for the extraction of pigments due to its less toxicity and being safe for human consumption. The optimized solvent can be used to extract anthocyanins from the S. cumini fruits and used as natural colourants in the food industries.

Qualitative identification of permitted and non-permitted colour additives in food products.

Colour additives are dyes, pigments or other substances that can impart colour when added or applied to food, drugs, cosmetics, medical devices, or the human body. The substances must be pre-approved by the US Food and Drug Administration (USFDA) and listed in Title 21 of the US Code of Federal Regulations before they may be used in products marketed in the United States. Some also are required to be batch certified by the USFDA prior to their use. Both domestic and imported products sold in interstate commerce fall under USFDA jurisdiction, and the USFDA's district laboratories use a combination of analytical methods for identifying or confirming the presence of potentially violative colour additives. We have developed a qualitative method for identifying 17 certifiable, certification exempt, and non-permitted colour additives in various food products. The method involves extracting the colour additives from a product and isolating them from non-coloured components with a C(18) Sep-Pak cartridge. The colour additives are then separated and identified by liquid chromatography (LC) with photodiode array detection, using an Xterra RP18 column and gradient elution with aqueous ammonium acetate and methanol. Limits of detection (LODs) ranged from 0.02 to 1.49 mg/l. This qualititative LC method supplements the visible spectrophotometric and thin-layer chromatography methods currently used by the USFDA's district laboratories and is less time-consuming and requires less solvent compared to the other methods. The extraction step in the new LC method is a simple and an efficient process that can be used for most food types.

Methods for the determination of European Union-permitted added natural colours in foods: a review.

Coupled to increasing consumer demand, food manufacturers have moved towards increased usage of approved natural colours. There is a legal requirement for governments to monitor the consumption of all food additives in the European Union to ensure the acceptable daily intakes (ADIs) are not exceeded, especially by young children. Validated analytical methods are needed to fulfil this requirement. The aim of this paper is to review the available literature on methods of extraction for approved natural colours in food and drink. Available analytical methods for the determination of European Union-permitted natural food colour additives in foods and beverages have been assessed for their fitness for purpose in terms of their key extraction and analysis procedures, selectivity and sensitivity, especially with regard to maximum permitted levels, and their applicability for use in surveillance and in an enforcement role. The advantages and disadvantages of available analytical methods for each of nine designated chemical classes (groups) of natural colours in different food and beverage matrices are given. Other important factors such as technical requirements, cost, transferability and applicability are given due consideration. Gaps in the knowledge and levels of validation are identified and recommendations made on further research to develop suitable methods. The nine designated natural colour classes covered are: 1. Curcumin (E100), 2. Riboflavins (E101i-ii), 3. Cochineal (E120), 4. Chlorophylls--including chlorophyllins and copper analogues (E140-141), 5. Caramel Classes I-IV (E150a-d), 6. Carotenoids (E160a-f, E161b, E161g), 7. Beetroot red (E162), 8. Anthocyanins (E163), and 9. Other colours--Vegetable carbon (E153), Calcium carbonate (E170), Titanium dioxide (E171) and Iron oxides and hydroxides (E172).

Fungal polyketide azaphilone pigments as future natural food colorants?

The recent approval of fungal carotenoids as food colorants by the European Union has strengthened the prospects for fungal cell factories for the production of polyketide pigments. Fungal production of colorants has the main advantage of making the manufacturer independent of the seasonal supply of raw materials, thus minimizing batch-to-batch variations. Here, we review the potential of polyketide pigments produced from chemotaxonomically selected non-toxigenic fungal strains (e.g. Penicillium and Epicoccum spp.) to serve as food colorants. We argue that the production of polyketide azaphilone pigments from such potentially safe hosts is advantageous over traditional processes that involve Monascus spp., which risks co-production of the mycotoxin citrinin. Thus, there is tremendous potential for the development of robust fungal production systems for polyketide pigments, both to tailor functionality and to expand the color palette of contemporary natural food colorants.

Revised procedures for the certification of carmine (C.I. 75470, Natural red 4) as a biological stain.

Carmine is one of the original dyes certified by the Biological Stain Commission (BSC). Until now it has lacked both an assay procedure for dye content and a means to positively identify the dye. The methods for testing carmine in the laboratory of the BSC have been revised to include spectrophotometric examination at pH 12.5-12.6 to determine that the dye is carmine (lambda(max)=530-335 nm). The maximum absorbance of a solution containing 100 mg of dye per liter of water, adjusted to pH 12.5-12.6, which provides a relative measure of dye content, should lie in the range 1.2 to 1.8. If the dye is not carmine, spectrophotometry at pH 1.9-2.1 shows whether it is carminic acid (lambda(max)=490-500 nm) or 4-aminocarminic acid (lambda(max)=525-530 nm). The latter two dyes, which are also called carmine when sold as food colorants, have physical properties different from those of true carmine. The functional tests for carmine as a biological stain are Orth's lithium-carmine method for nuclei, Southgate's mucicarmine method for mucus, and Best's carmine method for glycogen.

[Identification and determination of the isomer and subsidiary color in food blue no. 2 (indigo carmine) by LC/MS and HPLC].

CFR and JECFA specify that the total color in FD&C Blue No. 2 (B2; Indigo Carmine, Indigotine, Food Blue No. 2) is not less than 85%, its isomer (B2iso) in B2 is not more than 18%, and its subsidiary color (B2sub) in B2 is not more than 2% (CFR) or 1% (JECFA). Japan's Specifications and Standards for Food Additives, 7th Edition, specifies that the total color in B2 is not less than 85.0% and other color materials in B2 are not detected by paper chromatography. LC/MS and HPLC were employed to identify and determine the main component (B2m) of B2, B2iso, and B2sub. The pseudo molecular ions (B2m and B2iso: [M-2Na+H]-, m/z=421; B2sub: [M-Na]-, m/z=341) of each color material were obtained and identified by LC/MS based on their absorptions and mass spectra. The contents of B2iso and B2sub in B2 samples (certified samples from fiscal year 1998 to fiscal year 2002) were determined by HPLC using calibration curves for the standards of B2m and B2iso. The contents of B2iso in most samples were less than 10%, and the contents of B2sub in all samples were not more than 1%. All of them were within the regulatory limits set by the CFR and JECFA.

[Regulatory aspects in the use of natural colorants]. / Regulamentação de uso de corantes naturais.

A number of preparations of natural colours from vegetable, insect and algae sources are presently used in various foods, although many of them have not been evaluated in relation to their safety of use. In evaluating the toxicity of food additives the concept of the Acceptable Daily Intake (ADI) has been used to provide an indication of safety for use and to enable regulatory authorities to take adequate legislative measures for their control. This paper will focus on the principles for the safety assessment of food additives, with emphasis on the guidelines that have been established by the Joint Expert Committee on Food Additives and Contaminants (JECFA) for evaluating natural food colours. Recent data on the potential intake of annato extracts in Brazil and current aspects of regulation of food colours at the level of MERCOSUR will also be presented.

Growth performance and intestinal microbial populations of growing pigs fed diets containing sucrose thermal oligosaccharide caramel.

Four experiments were conducted to determine growth performance and changes in intestinal microbial populations of growing pigs fed diets containing sucrose thermal oligosaccharide caramel (STOC). Ninety-six barrows and 96 gilts were group-fed experimental nursery diets for 32 d after weaning in both Exp. 1 and 2. For each experiment, pigs were divided into four groups of 48 pigs and were fed either control, antibiotic (Apramycin sulfate, 34 mg/kg), 1% STOC, or 2% STOC diets for 32 d after weaning. Each diet was replicated six times with eight pigs per replication. Pigs were either orally gavaged (Exp 1) with water of STOC (2 g per pig) or pigs were creep-fed (Exp 2) either a control diet or a 2% STOC diet for 5 d before weaning (33 d). At the end of Exp 1 and 2, cecal material was collected for enumeration of total aerobes, total anaerobes, coliforms, lactobacilli, and bifidobacteria. Gilts (96 per experiment) used in Exp. 3 and 4 were weaned at 26 d and fed experimental nursery diets for 32 d. They were fed either a control or 1% STOC diet and were otherwise treated as previously described. There were no significant effects of STOC or antibiotic on ADG, ADFI, feed efficiency, or cecal microbial populations in pigs in this study. Feeding diets containing either antibiotic of STOC did not improve animal performance or change intestinal bacterial populations in the present study.

[Determination and survey of starting materials, intermediates, and subsidiary colors in food color of azo dye by high performance liquid chromatography].

A method for determination of starting materials, intermediates and subsidiary colors in food color of azo dye was developed by use of HPLC. The following conditions were used for analysis: column, L-column ODS (4.6 mm phi x 250 mmL); mobile phase, 0.02 M ammonium acetate (A), acetonitrile (B); concentration gradient, perform the linear concentration gradient from A:B (100:0) to (60:40) for 40 min; detection, starting materials and intermediates at 239 nm, and subsidiary colors at 510 nm. Standard material, domestic product and imported product were analyzed by the present HPLC method and impurities were measured. Recoveries of each impurity from azo dye averaged 99.1-103.5%. The detection limit was 0.05 microgram/g for each impurity.

Health & nutritional implications of food colours.

Colour is a vital constituent of food which imparts distinct appearance to the food product. Artificial colouring becomes a technological necessity as foods tend to lose their natural shade during processing and storage. Most of the food colours tested in the conventional toxicity experiments showed toxic effects at a very high level of intake i.e., 1-5 per cent in the diet. However, such levels of intake are not normally encountered. Human studies indicated that food colours, (natural or synthetic) can induce wide range of allergic reactions only in sensitive or atopic individuals. Most of the foodborne diseases reported are due to the consumption of non-permitted textile colours or abuse of colours. The Government is pressurised periodically to place a total ban on the use of food colours due to their possible ill effects. It should be realised that surveillance should go hand in hand with legal actions.

In vivo cytogenetic studies on male mice exposed to Ponceau 4R and beta-carotene.

Chromosomal aberrations induced by Ponceau 4R (an azo food dye) and beta-carotene (a natural food colour) were studied on bone marrow cells of mice in vivo. The results indicated that Ponceau 4R was more clastogenic than beta-carotene. Ponceau 4R was found to have a minimum effective dose of 4 mg which induced a significant number of chromosome aberrations. This dose is also the recommended 'admissible daily intake'. Chromosome aberrations induced by beta-carotene were not significantly higher than those of the control (olive oil) in the dose range 0.27 to 27 mg/kg body weight. The genotoxicity of these dyes can be attributed to their chemical composition. In so far as genotoxicity is concerned the carotenoid beta-carotene can be safely used as a food colorant whereas Ponceau 4R should be delisted as a food dye.

[Studies on "Sunset Yellow FCF Standard" for the Dye Standard of the National Institute of Health Sciences].

The "Sunset Yellow FCF Standard (C.I. 15985)" for the Dye Standard of the National Institute of Health Sciences was prepared. The content of this Dye Standard determined by the titanium trichloride titration method was 96.9% on the average. The ultraviolet-visible absorption and infrared spectra of the Dye Standard were also measured.

Development of specifications for caramel colours.

Specifications have been developed to define each of the four classes of caramel colour. The specifications were based on analysis of a large database generated during the course of characterization studies of each of the classes. A series of simple and practical tests was developed for the analysis of caramel colour samples to ensure conformity to the specifications.

[Studies on "new coccine standard" for the dye standard of the National Institute of Hygienic Sciences].

The "New Coccine Standard (C.I. 16255)" for the Dye Standard of the National Institute of Hygienic Sciences was prepared. The content of this Dye Standard determined by the titanium trichloride method was 97.5% on the average. The ultraviolet-visible absorption and infrared spectra of the Dye Standard were also measured.

[Studies on "allura red AC standard", dye standards of National Institute of Hygienic Sciences].

"Allura Red AC Standard (C.I. 16035)", Dye Standards of National Institute of Hygienic Sciences was prepared. The content of this Dye Standards was determined by the titanium trichloride method. This content averaged 95.5%. Ultra violet-visible absorption and infrared spectra of the Dye Standard were also determined.