Anthocyanin detection analysis

Introduction

Anthocyanins are natural pigments belonging to the flavonoid group widely distributed in the plant kingdom, particularly in fruits, vegetables, and flowers. These water-soluble compounds are responsible for the vibrant red, purple, and blue colors observed in many plant tissues. Beyond their visual appeal, anthocyanins play a crucial role in the plant's defense mechanisms and contribute to human health due to their antioxidant properties.

Food sources richest in anthocyanins

Sources of Anthocyanins in Foods

Common dietary sources of anthocyanins include berries such as blueberries, strawberries, and raspberries, as well as grapes, cherries, blackcurrants, and blackberries. Red and purple fruits like pomegranates, plums, and cranberries are also rich in anthocyanins. Vegetables like red cabbage and eggplant, along with certain grains, also contain these compounds. The specific types and concentrations of anthocyanins vary among different plant species.

Importance of Testing Anthocyanins in Food

Testing for anthocyanins in foods is vital for several reasons:

Importance of Testing in Foods	Description
Nutritional Content	Determines and quantifies the anthocyanin content, contributing to the overall nutritional profile of foods.
Quality Control	Ensures consistent color attributes, taste, and overall quality, particularly in products like fruit juices, jams, and confectionery.
Antioxidant Activity	Assesses the antioxidant capacity of foods, providing insights into potential health benefits.
Regulatory Compliance	Ensures adherence to regulatory standards for specific compounds in food products, promoting consumer safety.
Research and Development	Supports exploration of new formulations and processing methods to preserve or enhance anthocyanin content in foods.

Anthocyanin detection

Anthocyanin detection analysis involves various methods to identify, quantify, and characterize the presence of anthocyanins in a given sample. Several analytical techniques are commonly employed for this purpose:

Spectrophotometry: This method measures the absorbance of light by anthocyanins at specific wavelengths. It provides a quick and simple way to estimate total anthocyanin content in a sample.

High-Performance Liquid Chromatography (HPLC): HPLC is a powerful technique that separates and quantifies individual anthocyanin compounds in a mixture. It provides detailed information about the specific types and concentrations of anthocyanins present.

Liquid Chromatography-Mass Spectrometry (LC-MS): LC-MS combines chromatography with mass spectrometry for even higher sensitivity and specificity. It helps identify and quantify anthocyanins with greater precision.

Capillary Electrophoresis (CE): CE separates anthocyanins based on their charge-to-mass ratio. It is a rapid and efficient method for analyzing anthocyanin profiles in complex mixtures.

Nuclear Magnetic Resonance (NMR): NMR spectroscopy provides structural information about anthocyanins, allowing researchers to identify specific compounds and understand their molecular structure.

Colorimetry: This method assesses the color changes associated with anthocyanin presence. The color intensity correlates with the concentration of anthocyanins and can be quantified using a colorimeter or spectrophotometer.

Fluorescence Spectroscopy: Anthocyanins exhibit natural fluorescence, and this method measures the intensity of fluorescence emissions. It can be used for qualitative and quantitative analysis.

Paper Chromatography: This simple and cost-effective method separates anthocyanins based on their affinity for a paper substrate. It provides a quick visual assessment of the different anthocyanin components.

Regulations and Standards of Anthocyanins in Foods: FSSAI, EU, and USFDA

Anthocyanins, with their beautiful colors and potential health benefits, are used as natural food colorants and supplements. However, their inclusion in food products must comply with regulations set by different regions. Here's a breakdown of anthocyanin regulations for FSSAI, EU, and USFDA:

FSSAI (India):

Regulation: Food Safety and Standards (Food Products Standards and Additives) Regulations, 2012 (Regulation No. 38)

Specifications:

Anthocyanins are permitted for use in various categories of food products like beverages, confectionery, baked goods, fruit spreads, etc. (See specific categories and permitted levels in Appendix B of the regulation).
The maximum permitted level (MPL) of anthocyanins varies depending on the food category. For example, in fruit-based beverages, the MPL is 250 mg/kg, while in confectionery it's 100 mg/kg.
Anthocyanins used must be derived from edible sources like fruits, vegetables, or cereals.
Specific analytical methods for anthocyanin quantification are not outlined in the regulations.

EU:

Regulation: Commission Regulation (EC) No 1333/2008 on food additives (with subsequent amendments)

Specifications:

Anthocyanins are listed as food additive E 163 (Anthocyanins) in the regulation.
E 163 is approved for use in a wide range of food products, including beverages, bakery products, confectionery, dairy products, etc. (See Annex II of the regulation for specific categories and MPLs).
The MPLs for E 163 vary depending on the food category. For example, in fruit-based beverages, the MPL is 250 mg/kg, while in jams and jellies it's 200 mg/kg.
Anthocyanins used must comply with specific purity criteria related to heavy metals, arsenic, and other contaminants.
Analytical methods for E 163 quantification are outlined in the Commision Regulation (EC) No 1060/2013.

USFDA:

Regulation: 21 CFR Part 73 - Listing of Food Additives permitted for Direct Addition to Food for human consumption

Specifications:

Anthocyanins are not directly listed as a food additive in the CFR.
Grape skin extract (GSE) derived from Concord grapes is considered a color additive exempt from certification (21 CFR 73.169).
GSE can be used in non-beverage foods except those with established standards of identity. Specific MPLs are not defined.
Grape skin extract (enocianina) is also exempt from certification for use in still and carbonated beverages, beverage bases, and alcoholic beverages (21 CFR 73.170). Specific MPLs are not defined.
Anthocyanins from other sources may be used as food additives but require pre-market approval via a food additive petition with detailed safety and analytical data.

Eurofins role in anthocyanin detection analysis

Eurofins, a global leader in analytical testing and scientific services, plays a significant role in anthocyanin detection analysis across various industries, including:

Food and Beverage:

Analysis for regulatory compliance: Eurofins laboratories can analyze food and beverage products for anthocyanin content to ensure compliance with regulations like FSSAI, EU, and USFDA. They offer validated analytical methods based on HPLC, UV-Vis spectroscopy, and other techniques to quantify anthocyanins accurately.

Authentication and quality control: Eurofins helps identify the source and authenticity of food and beverage products by analyzing their anthocyanin profiles. This helps detect adulteration or mislabeling and ensures product quality.

Development and optimization of products: Eurofins can assist in developing new food and beverage products with desired anthocyanin levels for nutritional purposes or color enhancement. They can also help optimize existing formulations for optimal stability and shelf life of anthocyanins.

Cosmetics and Pharmaceuticals:

Quantification of anthocyanins in extracts and formulations: Eurofins offers analysis of anthocyanin content in cosmetic and pharmaceutical ingredients and finished products. This information is crucial for product claims, label accuracy, and quality control.

Stability and safety testing: Eurofins can assess the stability of anthocyanins in cosmetic and pharmaceutical formulations under various storage conditions. They also offer safety testing to comply with relevant regulations.

Identification and characterization of anthocyanins: Eurofins can perform advanced analysis techniques like mass spectrometry and chromatography to identify and characterize specific anthocyanin types present in samples. This is valuable for research and development purposes.

Other Industries:

Analysis of anthocyanins in agricultural products: Eurofins can analyze fruits, vegetables, and other agricultural products for anthocyanin content to assess their nutritional value and quality.

Environmental monitoring: Eurofins can measure anthocyanin levels in environmental samples like water and soil to assess the health of ecosystems and potential contamination.

Eurofins' Advantages:

Global network of laboratories: Eurofins has a vast network of laboratories equipped with advanced analytical instruments and expert scientists to cater to diverse needs.

Accredited and validated methods: Eurofins uses accredited and validated analytical methods for accurate and reliable results.

Expertise in regulatory compliance: Eurofins has experience navigating complex regulations and can help ensure compliance with relevant standards.

Comprehensive solutions: Eurofins offers a one-stop solution for all your anthocyanin detection analysis needs, from sample preparation to data interpretation and reporting.

References

Albuquerque, T. G., Silva, M. A., Oliveira, M. B. P., & Costa, H. S. (2017). Analysis, Identification, and Quantification of Anthocyanins in Fruit Juices. Fruit Juices, 693-737. https://doi.org/10.1016/B978-0-12-802230-6.00034-5
https://www.mdpi.com/2673-4583/5/1/43
https://www.fssai.gov.in/upload/uploadfiles/files/Compendium_Food_Additives_Regulations_08_09_2020-compressed.pdf
https://eurlex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2008:354:0016:0033:en:PDF
https://www.ecfr.gov/current/title-21/chapter-I/subchapter-A/part-73