Table 2.
Batch‐to‐batch analysis of the NF
| Parameter | Batch number | Method of analysis | ||||
|---|---|---|---|---|---|---|
| #1 | #2 | #3 | #4 | #5 | ||
| Composition | ||||||
| 3‐FL (% w/w DM) | 96.0 | 96.0 | 98.8 | 94.1 | 97.4 | HPAEC‐PAD (validated internal method) 1 |
| d‐Lactose (% w/w DM) | 0.7 | 0.5 | 0.6 | 1.1 | 0.5 | |
| d‐Glucose (% w/w DM) | < LOQ | < LOQ | < LOQ | < LOQ | < LOQ | |
| d‐Galactose (% w/w DM) | < LOQ | < LOQ | < LOQ | < LOQ | < LOQ | |
| l‐Fucose (% w/w DM) | 0.7 | 0.5 | 0.6 | < LOQ | 0.9 | |
| Sum of other carbohydrates (% w/w DM) | 2.0 | 2.4 | 0.0 | 4.2 | 0.8 | Calculation 2 |
| Protein (%) | < 0.001 | < 0.001 | < 0.001 | < 0.001 | < 0.001 | Nanoquant (modified Bradford) |
| Ash (%) | 0.36 | 0.33 | 0.25 | 0.02 | 0.12 | ASU L 06.00‐4 |
| Water (%) | 7.6 | 8.0 | 8.5 | 7.7 | 7.5 | Karl‐Fischer titration |
| Contaminants | ||||||
| Arsenic 3 (mg/kg) | < LOQ | < LOQ | < LOQ | < LOQ | < LOQ | ASU L 00.00‐135: 2011‐01 – ICP‐MS |
| Cadmium 3 (mg/kg) | < LOQ | < LOQ | < LOQ | < LOQ | < LOQ | |
| Lead 3 (mg/kg) | < LOQ | < LOQ | < LOQ | < LOQ | < LOQ | |
| Mercury 3 (mg/kg) | < LOQ | < LOQ | < LOQ | 0.006 | < LOQ | |
| Aflatoxin M1 (µg/kg) | < 0.025 | < 0.025 | < 0.025 | < 0.025 | < 0.025 | DIN EN ISO 14501: 2008‐01 – IAC‐HPLC‐FD |
| Aflatoxin B1 (µg/kg) | < 0.10 | < 0.10 | < 0.10 | < 0.10 | < 0.10 | DIN EN ISO 16050:2011‐09 – IAC‐HPLC – post column derivatisation |
| Aflatoxin B2 (µg/kg) | < 0.10 | < 0.10 | < 0.10 | < 0.10 | < 0.10 | |
| Aflatoxin G1 (µg/kg) | < 0.10 | < 0.10 | < 0.10 | < 0.10 | < 0.10 | |
| Aflatoxin G2 (µg/kg) | < 0.10 | < 0.10 | < 0.10 | < 0.10 | < 0.10 | |
| Sum of aflatoxins B1, B2, G1 and G2 (µg/kg) | < 0.40 | < 0.40 | < 0.40 | < 0.40 | < 0.40 | |
| Microbial parameters | ||||||
| Standard Plate Count (CFU/g) | < 10 | < 10 | < 10 | < 10 | < 10 | ISO 4833‐2 |
| Yeast and mould (CFU/g) | < 10 | < 10 | < 10 | < 10 | < 10 | ISO 21527‐2: 2008‐07 |
| Enterobacteriaceae (CFU/g) | < 10 | < 10 | < 10 | < 10 | < 10 | ISO 21528‐2: 2019‐05 |
| Enterobacteriaceae (in 10 g) | ND | ND | ND | ND | ND | ISO 21528‐1: 2017‐09 |
| Salmonella (in 25 g) | ND | ND | ND | ND | ND | DIN EN ISO 6579‐1: 2017‐07 |
| Cronobacter spp. (in 10 g) | ND | ND | ND | ND | ND | ISO/TS 22964: 2017‐04 |
| Listeria monocytogenes (in 25 g) | ND | ND | ND | ND | ND | DIN EN ISO 11290‐1: 2017‐09 |
| Bacillus cereus (CFU/g) | < 50 | < 50 | < 50 | < 50 | < 50 | DIN EN ISO 7932: 2020‐11 |
| Endotoxins (EU/mg) | 0.007 | 0.009 | 0.023 | < 0.005 | < 0.005 | Ph. Eur. 2.6.14 |
3‐FL: 3‐Fucosyllactose; ASU: Official collection of analysis methods according to § 64 of the German Food and Feed Code (LFGB); CFU: Colony forming unit; DIN: German Institute for Standardisation e. V.; DM: Dry matter; EN: European norm; EU: Endotoxin unit; HPAEC‐PAD: High‐performance anion‐exchange chromatography – pulsed amperometric detection; IAC‐HPLC‐FD: Immunoaffinity chromatography – high‐performance liquid chromatography – fluorescence detection; ICP‐MS: Inductively coupled plasma – mass spectrometry; ISO: International Organisation for Standardisation; LOQ: Limit of quantification; ND: Not detected; Ph. Eur.: European Pharmacopeia; TS: Technical specification.
LOQs: d‐Lactose = 0.14% w/w DM; d‐Galactose = 0.14% w/w DM; d‐Glucose = 0.14% w/w DM; l‐Fucose = 0.28% w/w DM.
Sum of other carbohydrates = 100 (% w/w DM) – 3‐FL (% w/w DM) – Quantified carbohydrates (% w/w DM) – Ash (% w/w DM). For those batches of the NF where the levels of any carbohydrate by‐product were below the respective limit of quantification (LOQ), the concentration of the corresponding compound has been considered to be equal to the respective LOQ value for the purpose of calculating the sum of other carbohydrates in the corresponding batch.
LOQs: Arsenic = 0.05 mg/kg; Cadmium = 0.010 mg/kg; Lead = 0.010 mg/kg; Mercury = 0.005 mg/kg.