🍎 Determination of sucralose in foodstuffs by HPLC — soluble samples are dissolved in water, incompletely soluble ones (pastries, chewing gum, yoghurt, ketchup, mayonnaise) are mixed with water at 40–60 °C and, if necessary, clarified with modified Carrez solutions; clean-up on C18 SPE cartridges 500 mg, elution with methanol/water 1 + 3, separation on an RP-18 column 250 × 4 mm at 35 °C, refractive index (RI) detector at 30 °C or ELSD; at least 5 standard solutions (e.g. 20–100 mg/l), result in mg/kg or mg/l; validation 83–737 mg/kg; PN-EN 16155:2012

Food chemistry PN-EN 16155

Overview

WHAT THE STANDARD COVERS. We read the text of the standard in the iTeh sample of SIST EN 16155:2012 (Slovenian adoption of EN 16155:2012 of April 2012, English text) — PN-EN 16155:2012 adopts EN 16155:2012 as identical (PKN search): contents, foreword, Clauses 1–7 and Clause 8 up to 8.2 — the sample ends on page 7 of the standard. We do not read the rest of the precision clause (8.2–8.3: repeatability and reproducibility values), the test report (9) or Annex A (statistical data of the interlaboratory trial); we know the titles from the contents.

ACCORDING TO THE TEXT OF EN 16155:2012. The standard was prepared by CEN/TC 275 “Food analysis — Horizontal methods” (secretariat DIN); CEN approved it on 25.02.2012, and national adoption was due by October 2012 at the latest. Scope (1): determination of sucralose in foodstuffs by HPLC with elution from a reversed-phase column with aqueous methanol and RI detection; the method was validated in an interlaboratory study for sucralose from 83 to 737 mg/kg in spiked samples: ketchup, mayonnaise, biscuits, yoghurt, instant beverage powder, sweets. Reference (2): EN ISO 3696:1995 (water). Principle (3): depending on consistency, samples are dissolved or diluted with water and, if appropriate, filtered or clarified with modified Carrez solutions; then extracted on an SPE column and eluted with methanol/water; sucralose is determined by HPLC on an RP column with aqueous methanol and RI detector, alternatively ELSD; quantification by external standard; the sucralose content is the content of 1,6-dichloro-1,6-dideoxy-β-D-fructofuranosyl-4-chloro-4-deoxy-α-D-galactopyranoside determined by this method. Reagents (4): sucralose C12H19Cl3O8, molecular weight 397.63; potassium hexacyanoferrate(II) trihydrate; zinc nitrate hexahydrate; methanol for HPLC; stock solution approx. 1000 mg/l (approx. 100 mg in 100 ml of water, fresh every day, taking account of water content and purity of the standard); at least five standard solutions, e.g. 20, 40, 60, 80 and 100 mg/l, fresh every day, without exceeding the linear range of the detector; Carrez solution A — 53.45 g of hexacyanoferrate to 500 ml; solution B — 148.75 g of zinc nitrate to 500 ml; eluent — methanol and water 1 + 3 (by volume). Apparatus (5): membrane filter up to 0.45 µm; C18 SPE cartridges 500 mg; HPLC with pump, injector, temperature-controlled RI detector or ELSD, column oven; C-18 RP column 250 mm × 4 mm (e.g. 5 µm) with guard column. Procedure (6): homogenization (liquids — stirring, hard candies — crushing, chewing gum — deep-freezing and crushing, yoghurt and ketchup — stirring and clarification if needed); soluble samples (e.g. hard candies) — approx. 5 g to the nearest 1 mg into a 50 ml flask with water, filtration if turbid; incompletely soluble samples (pastries, chewing gum, yoghurt, ketchup, mayonnaise) — approx. 5 g with approx. 25 ml of water, 30 min stirring at about 40–60 °C or ultrasonic bath, protein-containing samples — 1 ml each of Carrez solutions A and B, made up after cooling, filtered through a pleated filter and a membrane filter; SPE — conditioning with methanol and water, 5 ml of sample solution, washing three times with 5 ml of water without letting the cartridge run dry, elution with 5 ml of eluent into a 10 ml flask, made up, filtered into vials; HPLC (for the column of 5.5): injection up to 100 µl, eluent 4.8, oven 35 °C, RI detector 30 °C, flow approx. 1.2 ml/min; identification by retention time and by adding standard to the sample (peak overlap); quantification from the peak area through the calibration function y = a + bx, whose suitability must be checked. Calculation (7): ρ or w = (R − a) · V / (b · m), in mg/l or mg/kg, e.g. V = 50 ml, m = 5 g; the result is rounded to a whole number. Precision (8.1–8.2): data from two interlaboratory trials in 2007/2008 — ketchup, mayonnaise with high and low content, biscuits, yoghurt, 2 beverage powders, 2 compressed tablets (sweets); the values may not apply to other concentration ranges and matrices; the repeatability limit r is exceeded in not more than 5 % of cases and depends on the concentration level.

STATUS (as of 03.10.2026). The PKN search engine (query “PN-EN 16155”, 03.10.2026) lists PN-EN 16155:2012 — English version, “Introduces: EN 16155:2012 [IDT]”, without a withdrawal note; there is no Polish version in the results.

HOW MANY LABORATORIES AND IN WHAT FORM (copy of the accreditation scope database, data up to 17.09.2026, read 03.10.2026). The number 16155 appears in 3 records at 2 laboratories: AB 370 (2) and AB 592 (1). In the current PCA documents (read 03.10.2026; AB 592 issue no. 24 of 06.02.2026, AB 370 issue no. 25 of 04.08.2026) the number appears at the same 2 laboratories; in every document the number of page markers equals the number of PDF pages, without repetitions; a search of all PCA documents downloaded from BIP on 03.10.2026 found no laboratories outside the database copy. The form “PN-EN 16155:2012” at both. None of these items is suspended. The “Laboratories” tab (query “PN-EN 16155”) shows both laboratories — checked by a tab query on the labcoda.pl production server on 03.10.2026.

WHAT THE LABORATORIES TEST (items in PCA). AB 592: fish and other aquatic animals and their products, fruit and vegetable products, soft drinks, confectionery, food additives — sucralose (5–1800) mg/kg (mg/l), HPLC-RID. AB 370: soft drinks and juices — presence and content of sucralose (10–1000) mg/l; vegetable products (ketchup) — (50–1000) mg/l, HPLC-RID.

WHERE A SEEMINGLY CORRECT RESULT IS EASY TO GET. Validation range: the method was validated from 83 to 737 mg/kg in the listed matrices, and the precision data may not apply to other concentrations and matrices (1, 8.1) — the PCA items reach from 5 to 1800 mg/kg and include e.g. fish and food additives, i.e. beyond these limits. RI detector: refractive index detection is not selective — the standard bases identification on retention time and on adding standard to the sample (6.4); the detector needs a stable temperature (30 °C) (6.3). Linearity: the linear range of the detector must not be exceeded, and the suitability of the calibration function must be checked (4.6, 6.5). SPE: the cartridge must not run dry during washing (6.2.3). Standard: the purity and water content of the standard are taken into account in the stock solution, which is prepared daily (4.5). Units: a result for ketchup in mg/l (AB 370) is a different basis than mg/kg, in which the standard gives the validation range (1, 7).

WHAT WE DO NOT GIVE. We do not quote the repeatability and reproducibility limit values or the data of Annex A — they lie outside the sample. We do not summarize EN ISO 3696 in this entry. We do not give permitted contents of sucralose in foodstuffs.

Method principle

The sample is dissolved or extracted with water (protein-containing samples are clarified with Carrez solutions), and sucralose is retained on a C18 cartridge and eluted with methanol/water. The extract is separated on a reversed-phase column with aqueous methanol and measured with a refractive index detector (or ELSD). Sucralose is identified by retention time and standard addition, and the content is calculated from the peak area through a linear calibration function from at least five standards.

Applications

• Sucralose in foodstuffs: ketchup, mayonnaise, biscuits, yoghurt, instant beverage powders, sweets — validation range 83–737 mg/kg (1)
• Soluble samples (hard candies) and incompletely soluble samples (pastries, chewing gum, yoghurt, ketchup, mayonnaise) (6.2)
• Beverages, juices, fruit and vegetable products, confectionery, fish products, food additives — 3 accreditation scope records of 2 laboratories in PCA

Key parameters

Standard

Standard number

PN-EN 16155:2012

Title (PL)

Artykuły żywnościowe — Oznaczanie sukralozy — Metoda wysokosprawnej chromatografii cieczowej

Title (EN)

Foodstuffs — Determination of sucralose — High performance liquid chromatographic method

Step-by-step procedure

1. Homogenization

   Stirring, crushing, deep-freezing of chewing gum (6.1). PN-EN 16155:2012 — no withdrawal note (PKN, 03.10.2026).

2. Sample solution

   Approx. 5 g to 50 ml; incompletely soluble samples — 30 min at 40–60 °C, Carrez for protein-containing samples (6.2.1, 6.2.2).

3. SPE

   C18: conditioning, 5 ml of solution, washing with water, elution to 10 ml, filtration (6.2.3).

4. HPLC

   RP-18, 35 °C, RI 30 °C, approx. 1.2 ml/min; identification by retention time and standard addition (6.3, 6.4).

5. Result

   (R − a) · V / (b · m), mg/kg or mg/l, rounded to a whole number (7).

Required equipment and apparatus

Reagents, media and consumables

Health and safety (OHS)

• The part of the standard read contains no separate safety requirements; methanol and Carrez solutions (potassium hexacyanoferrate, zinc nitrate) are used (4)

Frequently asked questions