Testing Methodologies

All botanical ingredients are analysed in a laboratory, either in-house or by an independent third party, to check that the active ingredients are at the correct levels, per the published specification.

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There are various methodologies for analytical testing, as far as we are concerned there are three main ones:

1. Spectroscopy (Light & Radiation Interactivity)

These methods measure how matter absorbs, emits, or scatters electromagnetic radiation to decode molecular structures.

• FTIR (Fourier-Transform Infrared): Identifies organic polymers and functional groups by measuring molecular vibrations.
• UV-Vis (Ultraviolet-Visible): Quantifies light absorption to find concentrations of coloured compounds in a solution.
• NMR (Nuclear Magnetic Resonance): Maps the carbon-hydrogen framework of complex organic molecules.
• Raman Spectroscopy: Provides a structural fingerprint through non-destructive laser light scattering.

2. Chromatography (Separation Techniques)

Chromatography forces a liquid or gas sample through a stationary phase to separate individual components based on their physical properties.

• HPLC (High-Performance Liquid Chemistry): Separates non-volatile liquids, widely used in pharmaceuticals.
• GC (Gas Chromatography): Separates volatile gas mixtures, critical for environmental testing.
• TLC (Thin-Layer Chromatography): A fast, low-cost screening method used to track chemical reaction progress.

3. Mass Spectrometry (MS)

MS ionizes chemical species and sorts them based on their mass-to-charge ratio, offering unparalleled sensitivity for unknown identification.

• GC-MS / LC-MS: Couples separation power with mass identification to isolate and name trace contaminants.
• ICP-MS (Inductively Coupled Plasma): Detects ultra-trace metals down to parts-per-trillion (ppt) levels.

Mass Spectrometry coupled with Chromatography (LC-MS or GC-MS) is the most accurate method for identifying and quantifying active ingredients in botanical extracts. Plant extracts are highly complex chemical "matrices" containing hundreds of distinct phytochemicals that must be separated before they can be accurately measured.

Accuracy

We often see test results from UV and HPLC tests which are widely different. This is because stand-alone UV testing (UV-Vis Spectrophotometry) routinely overestimates active ingredients compared to HPLC because UV testing lacks the ability to separate a sample before measuring it. When a raw botanical extract is placed in a UV spectrophotometer, the machine reads the total light absorbed at a specific wavelength. It cannot distinguish between your target active ingredient and dozens of other identical-looking plant molecules. The primary reasons for these drastically different results include:

1. Total Absorbance vs. Separated Absorbance

UV-Vis Testing: The sample is placed directly into a glass cuvette. If you are testing for Saffron at 440 nm, the machine counts everything in that liquid that absorbs light at 440 nm. This includes degraded actives, plant pigments, and irrelevant cellular debris. HPLC Testing: The sample is forced through a chemical column that slows down different molecules at different rates. Your active ingredient is physically separated into its own pure "peak" before it ever hits a detector. HPLC measures only that isolated compound.

2. The Overestimation Effect (False Positives)

Because UV testing tallies up all background chemical "noise," UV test results are frequently 20% to 200% higher than HPLC results for the exact same batch. Dishonest ingredient suppliers often prefer UV-Vis testing because it artificially inflates the apparent potency of an extract on a Certificate of Analysis (CoA).

3. Shared Chromophores

Molecules absorb ultraviolet light using chemical structures called chromophores (such as alternating double bonds). In a complex botanical extract, many entirely different plant sugars, tannins, or flavonoids share almost identical chromophores. A standalone UV meter sees them all as the exact same substance