| Botanical identity | Vaccinium myrtillus L. | Macroscopic and microscopic identity checks, supported by botanical reference material; DNA barcoding may be used as a supplementary test. | Do not accept a specification based only on the generic term “bilberry.” The scientific name should appear on the purchase specification and certificate of analysis. |
| Plant material | Fruit, including fruit skin and pulp; aerial parts and leaves excluded | Visual inspection, microscopy, and supplier batch documentation | Bilberry anthocyanins are concentrated in the dark blue-purple fruit. Require a documented raw-material lot and harvesting season. |
| Anthocyanin content | 25.0% minimum total anthocyanins on the supplier’s declared basis | Validated HPLC or UHPLC method with individual anthocyanin reporting; UV–Vis may be used for screening but should not replace chromatographic confirmation. | The result must state whether it is calculated on an as-is, dry, or anhydrous basis. Do not compare results from different bases without conversion. |
| Anthocyanin expression | Report individual anthocyanins and total content as cyanidin-3-O-glucoside equivalents, where applicable | HPLC-UV/Vis or HPLC-DAD with a documented reference standard and calculation formula | Bilberry contains multiple anthocyanin glycosides, including glucosides, galactosides, and arabinosides. A single UV–Vis number can overestimate or misrepresent the profile if the method is not defined. |
| Anthocyanin fingerprint | Cyanidin, delphinidin, malvidin, peonidin, and petunidin derivatives should be detectable in a profile consistent with bilberry fruit | Chromatographic fingerprint compared with an authenticated bilberry reference profile | Use the fingerprint to identify dilution, substitution, or an extract derived from a different Vaccinium species. |
| Extract ratio | Declare the native plant-to-extract ratio, for example, “equivalent to X kg fresh or dried fruit per 1 kg extract” | Review manufacturing records, mass balance, and batch yield | Do not use extract ratio as a substitute for an anthocyanin assay. Carrier materials and processing losses can materially change the final concentration. |
| Extraction solvent | Food- or pharmaceutical-grade water, ethanol, or a documented water–ethanol system; residual solvent must meet the applicable legal limit | Review process description and test residual solvents when relevant | The supplier should disclose solvent type, concentration range, extraction temperature, concentration method, and use of carriers or processing aids. |
| Appearance and color | Purple to dark purple powder; free-flowing or uniformly agglomerated; no visible foreign matter | Visual inspection under standardized lighting; optional colorimetric measurement for lot-to-lot control | Color alone is not proof of anthocyanin potency because permitted or undeclared colors may alter appearance. |
| Moisture or loss on drying | ≤ 5.0% unless a justified product-specific limit is agreed | Validated loss-on-drying or Karl Fischer method, as appropriate for the matrix | Lower moisture generally improves powder stability, but the selected limit should be confirmed against hygroscopic carriers and packaging performance. |
| Carrier and excipients | Full qualitative and quantitative declaration; no undisclosed colorants or active botanical adulterants | Specification review, label review, and targeted analytical testing when risk-based assessment requires it | State carrier percentage because it affects potency, bulk density, dosage calculations, and the 25% anthocyanin result. |
| Microbiological quality | Set limits according to the intended use and destination-market food or supplement requirements; absence of relevant pathogens required | Validated total aerobic count, yeast and mold, and specified pathogen methods | The final limits are jurisdiction- and product-category-specific. Document the applicable regulation in the purchasing agreement rather than using one universal limit. |
| Contaminants | Comply with the maximum levels applicable in the destination market for heavy metals, pesticides, mycotoxins, and other relevant contaminants | Risk-based multi-residue testing using validated laboratory methods | Testing frequency should reflect origin, harvesting environment, processing controls, and historical supplier performance. |
| Residual solvents | Meet applicable ICH, pharmacopeial, food, or national requirements according to product classification | Headspace GC or another validated residual-solvent method | A “water-extracted” claim does not automatically exclude processing solvents used during concentration, cleaning, or carrier manufacture. |
| Stability and packaging | Protect from light, oxygen, heat, and moisture; sealed, food-contact-compliant packaging with tamper evidence | Accelerated and real-time stability monitoring of anthocyanins, moisture, color, and microbiology | Anthocyanins are sensitive to processing and storage conditions. Require recommended storage temperature, humidity, and maximum time after opening. |
| Certificate of analysis | Batch-specific certificate showing identity, 25% anthocyanins, moisture, microbiology, contaminants, and test methods | Compare certificate results with independent verification testing on initial and periodic lots | The certificate should include batch number, production date, retest or expiry date, analytical basis, laboratory status, and authorized approval. |
| Traceability | One-step-back and one-step-forward traceability from harvested fruit to finished extract | Audit of lot records, supplier declarations, and mass-balance documentation | Record botanical source, country or region of harvest, harvest year, processing site, and any blending between lots. |
| Recommended release decision | Release only when identity is confirmed and total anthocyanins are ≥ 25.0% on the agreed analytical basis | Quality review of the complete dossier plus confirmatory testing according to a documented sampling plan | A passing color or generic “bilberry extract” declaration should not override failure of botanical identity, anthocyanin potency, contaminant, or traceability requirements. |