Understanding Botanical and Natural Ingredient Testing: What are phytochemicals?
Phytochemicals are chemicals derived from a plant. Within the plant, phytochemicals support the functions necessary for survival, as well as protection from external stressors, like drought or insects. Any botanical product innately contains phytochemicals. While the term phytochemical broadly covers all plant compounds, it often refers to those which can alter human health and nutrition. In other words, the compounds that are bioactive in the body.
Phytochemicals are responsible for the dietary, medicinal, or nutritional effect of botanical products, and thus are important to understand at all points of product sourcing, production, and sale.
The phytochemical profile of a plant, or its unique set of compounds, can be used for confirmation of the identity, purity, and potency of a botanical product.
Examples of phytochemicals:
- Anthocyanins: Berry-based products are filled with anthocyanins, which support immune health, are strong antioxidants, and have anti-microbial properties. Different berries have different anthocyanin levels, so each product has its own uses and benefits. Click the image to the right to watch a video from our Instagram feed about anthocyanins in botanical prodcuts.
- Flavonoids: Flavonoids are largely responsible for the variety plant and fruit colors. Flavonoids are credited with many health benefits, including
anti-inflammatory, anti-carcinogenic, and cardiovascular benefits.
Subclasses of flavonoids include anthocyanins, flavanones, and chalcones
(Panche, Diwan, and Chandra 2016).
- Carotenoids: Carotenoids are commonly used to add a yellow-orange pigment to food and other products. They are found in carrots, yams, and lots of fruits. This class of compound is important for eye health, antioxidant properties, and other nutritional roles. Examples include lutein and beta-carotene (Johnson 2002). Click the image to the right to learn more about carotenoids in botanical products from out Instagram feed.
Phytochemicals as marker compounds:
Plants of different species, geographic origin, variety, and even plant part (roots vs shoot) have unique sets of phytochemicals. Combined with the relative stability and abundance of most key phytochemicals, their natural variation makes them ideal marker compounds for authentication and identification.
Confirming the presence of the marker compounds, or in some cases the ratio of certain compounds, expected in a given botanical is a standard approach for identification. The presence of unexpected compounds may point to contamination or adulteration. In many cases, the phytochemicals used to identify a product are also responsible for the desired health effect, meaning phytochemical markers can be used to analyze product strength.
How does phytochemical testing help meet cGMP requirements:
Current Good Manufacturing Practices (cGMP) regulations include testing the identity, strength, and purity of dietary supplements. The justification for the use of phytochemicals for cGMP testing for each of these categories are outlined below.
Identity:
There are many ways to confirm the identity of botanicals, including genetic, morphological, and phytochemical screens. However, product processing complicates reliable identification. For example, finished products are often powders or capsules, which hinders morphological identification and makes high quality DNA extractions difficult. Chemical testing, on the other hand, is possible at each stage of production – from raw material to finished products.
As previously mentioned, each botanical product has a unique phytochemical profile. Chemical analysis allows the comparison of the marker compounds expected in a product with what is actually present. If the predetermined phytochemicals are present in the correct ratio unique to the product, the identity can be confirmed. Marker compounds used for product identification are determined through analysis of certified of voucher reference materials, industry guidelines, or fit-for-purpose testing.
Purity:
Phytochemical testing allows detection of a broad range of adulterants and contaminants in botanical products. Contamination includes heavy metal, pesticide, mycotoxin (fungal toxins), solvent, and microbial residues. Ultimately, looking for chemical compounds not found naturally in the material (a pesticide or solvent, for example) allows detection of contamination.
While contamination is typically evaluated by screening for common contaminants, it is important to understand the phytochemical composition of the botanical. This allows detection of unknown or unexpected compounds that may have arisen during the production process. Finding an unwanted compound before product distribution prevents economic and reputation damage in the future.
In many ways, adulteration is harder to detect than contamination. Adulteration includes the intentional replacement, supplementation, or dilution of the target botanical with a different plant. Adulteration can cause the loss of health effects since the specific chemicals in a plant are responsible for bioactivity; replacement of the target plant with an alternative can lead to complete loss of the desired effect.
Phytochemical screening ensures that only the intended plant is present by capitalizing on the specificity of chemical profiles. Often, a screen for marker compounds unique to common adulterants is used to detect non-target plants in a product. Additionally, deviation from the expected phytochemical profile beyond a subset of marker compounds (looking at the entire chemical composition) can indicate the presence of non-target plant material.
Strength:
Since the health effects of botanicals are due to the presence of particular phytochemicals, the potency of products can be evaluated via phytochemical testing. If a product claims a certain level of a chemical, evaluation of the quantity and purity of that compound can be evaluated via chemical testing. Bioactive compounds can be quantified with a variety of chromatography techniques. Most often, liquid chromatography is used to separate the phytochemical of interest from the rest of the compounds and measure the amount present in a sample. Strength testing requires careful method optimization and use of appropriate reference and analytical standards.
Types of testing:
There are many approaches to phytochemical testing.
The most common techniques are chromatography based, including liquid- and gas- chromatography (LC and GC) and thin – layer chromatography (TLC). In some cases, the retention time of compounds in a sample, which are based on the compound’s mass and charge is enough to visualize differences in marker composition. In others, detailed mass spectrometry (MS) data may be required to compare the exact mass of a compound with spectral libraries for accurate identification.
Other phytochemical analysis techniques include absorption tests, like Fourier – transform infrared (FTIR), near – infrared (NIR), or Raman spectroscopy. It is best to use a fit-for-purpose approach to determine the ideal test, or combination of tests, best suits a specific product.
Examples of phytochemical testing at Eurofins:
- Ginger: Eurofins is proud to offer an AOAC accepted method for analyzing the main eight ginger phytochemicals used in dietary supplements. This 12 minute method measures gingerols, shagols, 6-paradol, and zingerone. This method greatly improves ginger phytochemical analysis. Learn more from this paper and poster.
- Cocoa Flavanols: Eurofins Botanical Testing in Brea, CA assisted in validating the AOAC Official Method of AnalysisSM 2020.05, which measures cocoa flavanols and proyanidins with a degree of polymerization DP1-7. This method, offered at Eurofins locations globally, provides appropriate characterization and quantification of the distinct mixture found in cocoa-based products thus offer important scientific and commercial value. Read this paper to learn more.
Our Eurofins team of experts can help you with all of your botanical testing needs.
Eurofins – Your Partner for Laboratory Testing Services
The Eurofins network of laboratories provides its customers with complete analytical solutions supported by a commitment to excellent customer service, high-quality standards, and scientific expertise.
