Assessment of environmental chemical contamination and histamine levels in the production of Colatura di Alici di Cetara

Summary

D’Anza and colleagues monitored cadmium, lead, total mercury, four PAHs, and histamine across the full 390-day production cycle of Colatura di Alici di Cetara, an Italian PDO fermented anchovy sauce. cadmium (Cd) and lead (Pb) were below the respective limits of quantification (LOQ 0.005 and 0.010 mg/kg) in the raw salt and in all processed matrices. mercury (Hg) was detectable in raw anchovies (0.072 mg/kg) and in anchovy muscle at 30 days but was not detectable in the final colatura (LOQ 0.010 mg/kg). PAHs were not detected at any processing stage. All values were well below EU Regulation 2023/915 maxima for the raw fish matrix. This is a single-batch process-characterization study; it is not a market survey of commercial colatura or fish sauce.

Key numbers

Heavy metals measured by AAS electrothermal atomization (Cd, Pb; Perkin Elmer PinAAcle 900T) and direct mercury analyzer (Hg; Milestone DMA 80 Evo). LOQs: Cd 0.005 mg/kg, Pb 0.010 mg/kg, Hg 0.010 mg/kg. All samples run in duplicate; results corrected for mean recovery. Accredited per UNI EN ISO/IEC 17025:2018.

Table 1 of the paper — heavy metals in production matrices (mean +/- SD; nd = not detected, below LOQ):

PAHs (benzo[a]pyrene, benzo[a]anthracene, benzo[b]fluoranthene, chrysene; HPLC-FLD; LOQ 0.2 µg/kg each): not detected at T0, T1, or T8 (final colatura) in either production protocol.

Histamine (UHPLC-DAD; LOQ 0.20 mg/kg): peak values in final colatura were 51.0 +/- 1.7 mg/kg (N1) and 49.7 +/- 2.1 mg/kg (N2), both well below the EU 400 mg/kg maximum for fermented fish sauces (Regulation 1019/2013).

EU regulatory limits cited (Regulation 2023/915): Pb 0.30 mg/kg (fish muscle), Cd 0.25 mg/kg (Engraulis anchovies), Hg 0.30 mg/kg (Engraulis anchovies). The authors note that Annex I does not set specific maxima for processed fish sauces; Article 3 concentration-change provisions apply to the final product.

Evidence Fitness

Modeled or limited evidence. This is a process-characterization study of one production batch from a single geographic source, not a market survey. The heavy-metal results for the final colatura are all below LOQ, so they establish an upper bound rather than a measured central tendency. The study design is rigorous (accredited laboratory, duplicate measurements, recovery correction) but the single-batch scope limits generalizability across commercial colatura products or other fish sauce types.

Methods (brief)

Cd and Pb: AAS electrothermal atomization, Perkin Elmer PinAAcle 900T, Zeeman effect background correction; acid digestion (0.75 g sample, 70% HNO3 + 30% H2O2, microwave, final volume 50 mL); matrix modifiers (NH4)H2PO4 and Mg(NO3)2; absorption wavelengths 247.6 nm (Pb) and 228.8 nm (Cd); argon inert gas; five-point calibration (R2 > 0.99); duplicates; spiked-sample recovery; z scores not exceeding |2| in proficiency testing.

Hg: Direct mercury analyzer, Milestone DMA 80 Evo; ~0.300 g sample; sublimation, gold amalgam adsorption, thermal desorption, detection at 253.65 nm; duplicates; LOQ 0.010 mg/kg.

PAHs: HPLC-FLD (Waters Alliance); saponification with 2 M ethanolic KOH; liquid extraction with cyclohexane; SPE C18 cleanup; five-point calibration; LOQ 0.2 µg/kg each.

Anchovies (Engraulis encrasicolus L., 9-12 cm) were caught by lampara fishing off the Amalfi-Positano coast in October 2020. Processing began immediately post-catch; average seasoning temperature 12.8 degrees C, relative humidity 76%. Fermentation vessels were traditional wooden terzigni.

Speciation note: only total Hg was measured; methylmercury fraction was not determined.

Implications

Certification: The final colatura data show Cd, Pb, and Hg below LOQ, which is consistent with the EU regulatory picture but does not establish a population distribution for certification limit-setting. The raw-anchovy Hg value (0.072 mg/kg) is well below EU limits (0.30 mg/kg) for Engraulis. This paper is process-characterization evidence, not an occurrence dataset.

Courses: Useful illustration of how heavy metal levels change (or do not change) across a long fermentation process; the disappearance of Hg from raw fish to finished sauce supports discussion of potential volatilization or redistribution during fermentation.

App: No change to contamination_profile blocks; single-batch data with all final-product values below LOQ do not update central-tendency estimates.

Microbiome: No metal-microbiome data in this paper.

Provenance notes

Open-access CC BY-NC publication. PDF obtained via PMC OA (PMC11616580). SHA-256 verified against MANIFEST.csv from HMI-FILL-0034 deposit. Colatura di Alici di Cetara received EU PDO status in 2020. The paper compares two production protocols (eviscerated N1 vs. whole-anchovy N2); no significant difference was found between protocols for heavy metals or final-sauce histamine.

Related evidence

• Fish sauce
• Mercury, Total
• Cadmium
• Lead

Update history

No substantive edit history is available in this build. The full commit record is available in git.