🍎 Detection of cocoa butter equivalents (CBEs) in cocoa butter and plain chocolate — fat from 10–40 g of grated chocolate is extracted with a non-chlorinated solvent, a 0.5 mg/ml solution is separated by high-resolution capillary gas chromatography with cold on-column injection and FID, the triacylglycerols POP, POS, POO, SOS and SOO are identified against the certified cocoa butter IRMM-801, and from the proportions of POP, POS and SOS (normalized to 100 %) it is checked whether POP < 44.025 − 0.733 × SOS; qualitative result: CBEs present or absent, PN-EN ISO 23275-1

Food chemistry PN-EN ISO 23275-1

Overview

WHAT THE STANDARD COVERS. We read the text of the standard in the iTeh sample of ISO 23275-1:2006 in the French version (“Corps gras d’origines animale et végétale — Équivalents au beurre de cacao dans le beurre de cacao et dans le chocolat de ménage — Partie 1: Détermination de la présence d’équivalents au beurre de cacao”): title page, contents, foreword, introduction, Clauses 1–9 and Clause 10 up to 10.2 b) (detector response factors) — the sample ends on page 5. The end of 7.2 and the denominator of formula (2) are missing in the text. We do not read the rest of the requirements, precision, test report or Annex A with the results of the interlaboratory test; we know the titles from the contents.

ACCORDING TO THE TEXT OF ISO 23275-1:2006. The standard was prepared by ISO/TC 34 “Food products”, Subcommittee SC 11 “Animal and vegetable fats and oils”; the ISO 23275 series has two parts: 1 — detection of the presence of cocoa butter equivalents, 2 — their quantification. Introduction: cocoa butter equivalents (CBEs) is the general name for fats replacing cocoa butter in chocolate; their composition and properties are very similar to cocoa butter, which makes them extremely difficult to quantify and sometimes even to detect; by definition they are fats low in lauric acid, rich in symmetrical monounsaturated triacylglycerols of the POP, POS and SOS type, miscible with cocoa butter and obtained only by refining and fractionation; in the European Union under Directive 2000/36/EC fats from illipe (Borneo tallow, tengkawang, Shorea spp.), oil palm (Elaeis guineensis, E. olifera), sal (Shorea robusta), shea (Butyrospermum parkii), kokum gurgi (Garcinia indica) and mango kernel (Mangifera indica) may be used, alone or in blends. Part 1 describes the detection of these fats (limitations apply only to samples of pure illipe fat), and Part 2 — reliable quantification at the 5 % level, consistent with the limit of Directive 2000/36/EC. A “CoCal-1” toolkit was prepared for both parts: validated methods, the certified reference material of cocoa butter IRMM-801 for calibration and a spreadsheet for calculating the result (information on the IRMM website). Scope (1): detection of CBEs in cocoa butter and plain chocolate by triacylglycerol analysis using high-resolution capillary gas chromatography (HR-GC) and regression analysis; the method detects 2 % CBE in cocoa butter, which corresponds to about 0.4 % CBE in chocolate assuming 30 % fat. Definition (2.1): CBEs — fats detected by the procedure of this part; qualitative result: present/absent (YES/NO). Principle (3): cocoa butter or fat extracted from chocolate is separated by HR-GC into triacylglycerol fractions according to molecular mass and degree of unsaturation; the presence of CBEs is detected by linear regression on the proportions of individual triacylglycerols. Reagents (4): certified reference material of cocoa butter IRMM-801 for calibration and system suitability check; extraction solvent — non-chlorinated, e.g. diethyl ether, n-heptane, isooctane. Apparatus (5): balance reading to 0.1 mg; oven at 55 °C (or heating block); food processor with the motor above the bowl so as not to melt the sample; rotary evaporator; 1 ml pipettes; 20 ml volumetric flasks; microsyringe up to 10 µl or autosampler; chromatograph with cold on-column injection and FID — other injectors (split, PTV, moving needle) are acceptable if they give the results of 10.2; proven conditions: 25–30 m × 0.25 mm fused silica column with 50 % polyphenylmethylsiloxane, film 0.1–0.15 µm, programme 100 °C → 340 °C, 30 °C/min, carrier gas helium or hydrogen (≥ 99.999 %); data system. Sampling (6): not part of the method, recommended procedure in ISO 5555. Preparation (7): the IRMM-801 ampoule is heated in the oven until melted, mixed for at least 20 s by inversion and transferred to a tightly closed vial kept in the cold; about 200 g of chocolate is cooled until hard and grated into fine shavings, mixed and kept in the cold. Procedure (8): fat from 10–40 g of grated chocolate is extracted with two to three portions of 100 ml of solvent, centrifuged, decanted, evaporated and dried in a stream of nitrogen (other methods — Soxhlet, supercritical CO2, microwaves — acceptable if they give the same results); samples (cocoa butter, chocolate fat, IRMM-801) are melted completely in the oven, filtered in the oven if a sediment is present, and the pipettes are warmed to about 55 °C to avoid partial fractionation of the fat; about 0.2 g in 20 ml of solvent, 1 ml of this solution to 20 ml, injection of 0.5–1.0 µl (0.5 mg/ml) on-column; identification of the five main triacylglycerols — POP (1,3-dipalmitoyl-2-oleoylglycerol), POS, POO, SOS and SOO — by comparing retention times with IRMM-801 (elution in order of increasing carbon number and, at the same number, increasing unsaturation). Calculations (9): response factors Fi from the analysis of IRMM-801 — the area proportions of the five triacylglycerols in the reference material are compared with their mass proportions from the certificate; the proportions of POP, POS and SOS in the sample wtest,i = Fi × Atest,i / Σ(Fi × Atest,i) × 100 %, to two decimal places. Decision (9.3): the variability of cocoa butter is described by the line POP = 43.734 − 0.733 × SOS (residual standard deviation 0.125) with POP + POS + SOS = 100 %; the POS content of cocoa butter is almost constant with large changes of POP and SOS, and mixtures with CBEs deviate from this “cocoa butter line” the more, the more their POS differs from that of cocoa butter; the line was established from the profiles of 74 samples of genuine cocoa butter; in 99 % of analyses pure cocoa butter satisfies POP < 44.025 − 0.733 × SOS, and a higher POP value means that the sample is not pure cocoa butter; calibration with IRMM-801 links the laboratory’s results to this database without establishing its own line. Requirements (10): chromatographic details depend on the equipment, column, injection technique and sample size — they may be changed if the system suitability requirements are met; these are checked mandatorily on IRMM-801: resolution of the pairs POS/POO and SOS/SOO at least 1.0 (otherwise optimization of conditions), and the response factors of the five main triacylglycerols in a properly working system lie within 0.80–1.20.

STATUS (as of 03.10.2026). In the PKN search (query “23275-1”, 03.10.2026) PN-EN ISO 23275-1:2009 (Polish version, “Wprowadza: EN ISO 23275-1:2008 [IDT], ISO 23275-1:2006 [IDT]”) carries no withdrawal note; PN-ISO 23275-1:2007 (Polish version) — “Wycofana i zastąpiona przez PN-EN ISO 23275-1:2009 – wersja angielska”, and that one — withdrawn and replaced by PN-EN ISO 23275-1:2009 in the Polish version. Card of the Polish version: publication date 03.12.2009, 19 pages, KT 92 Nasion Roślin Oleistych, Tłuszczów Roślinnych i Zwierzęcych oraz ich Produktów Ubocznych, no withdrawal note. In the iTeh catalogue (read 03.10.2026) ISO 23275-1:2006 has the status “Published”, stage 90.92 (International Standard to be revised), dated 24.09.2026.

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 23275-1 appears in 4 records at 3 laboratories: AB 285, AB 370 (2), AB 705. In the current PCA documents (read 03.10.2026; AB 285 issue no. 28 of 13.01.2026, AB 705 issue no. 21 of 06.05.2026, AB 370 issue no. 25 of 04.08.2026) the number appears in 3 lines at the same 3 laboratories (the two AB 370 records — the result in fat and the calculated result in the product — share one line with the reference in the document); in each document the number of page markers equals the number of PDF pages, without repetitions; a search of all PCA documents downloaded from the BIP on 03.10.2026 found no laboratories outside the database copy. The form is “PN-EN ISO 23275-1:2009” — alone (AB 705) or together with PN-EN ISO 23275-2:2010 and the report “Report EUR 22666 EN:2007” (AB 285, AB 370). None of these items is suspended. The “Laboratories” tab (query “PN-EN ISO 23275-1”) shows all 3 laboratories — checked by a tab query on the labcoda.pl production server on 03.10.2026.

WHAT THE LABORATORIES TEST (items in PCA). Object: plain chocolate (AB 705), chocolate without additives (AB 285), chocolates, couvertures, cocoa (AB 370). Characteristic: presence of cocoa butter equivalents (AB 705 — with Part 1 only); presence and content of cocoa butter equivalents (CBE) and milk fat (MF) based on the triacylglycerol composition — (4–50) g/100 g of fat for CBE and for MF, also in g/100 g of product by calculation (AB 370); presence of CBE with content (1.30–5.00) g CBE/100 g of chocolate and milk fat (3.80–28.27) g/100 g of fat (AB 285). Technique: GC-FID.

WHERE A SEEMINGLY CORRECT RESULT IS EASY TO GET. Nature of the result: Part 1 gives only YES/NO (2.1); a number is given by Part 2 — at AB 705 presence is listed with Part 1 and content with Part 2, i.e. the standards divide the tasks, they are not substitutes. Limit: 2 % CBE in cocoa butter is detected, i.e. about 0.4 % in chocolate only assuming 30 % fat (1) — in chocolate with a different fat content this figure changes; in the note to Part 2 the same limit is given as 0.6 %. Matrix: the standard covers cocoa butter and plain chocolate (1); AB 370 also tests couvertures and cocoa, and AB 285 and AB 370 also milk fat — this characteristic is not described by the standard (at both, the report EUR 22666 EN:2007, which we have not read, is also cited for it). Illipe fat: the introduction indicates limitations for samples of pure illipe fat. Calibration: IRMM-801 is mandatory for checking the system (10.2) — resolution POS/POO and SOS/SOO ≥ 1.0, response factors 0.80–1.20; without this the decision from the POP–SOS line loses its link with the database of 74 samples. Preparation: the fat is melted completely and the pipettes warmed to about 55 °C, because partial fractionation changes the triacylglycerol profile (8.2).

WHAT WE DO NOT GIVE. Precision data, the test report and the results of the interlaboratory test of Annex A are not quoted — they lie outside the sample. We do not give the content of Directive 2000/36/EC or of the report EUR 22666 EN:2007.

Method principle

Cocoa butter or fat extracted from chocolate (0.5 mg/ml solution) is separated by high-resolution capillary gas chromatography with FID into the triacylglycerols POP, POS, POO, SOS and SOO, identified and calibrated against the certified cocoa butter IRMM-801. The proportions of POP, POS and SOS are normalized to 100 %; pure cocoa butter satisfies in 99 % of analyses the condition POP < 44.025 − 0.733 × SOS, and a higher POP value means that cocoa butter equivalents are present (YES/NO result).

Applications

• Detection of cocoa butter equivalents in cocoa butter and plain chocolate — from 2 % CBE in cocoa butter (about 0.4 % in chocolate at 30 % fat) (1)
• Checking the purity of cocoa butter (9.3)
• Plain chocolate, chocolate without additives, chocolates, couvertures and cocoa — items in the accreditation scopes of 3 laboratories in PCA

Key parameters

Standard

Standard number

PN-EN ISO 23275-1:2009

Title (PL)

Oleje i tłuszcze roślinne oraz zwierzęce — Odpowiedniki tłuszczu kakaowego w tłuszczu kakaowym i w czekoladzie naturalnej — Część 1: Oznaczanie obecności odpowiedników tłuszczu kakaowego

Title (EN)

Animal and vegetable fats and oils — Cocoa butter equivalents in cocoa butter and plain chocolate — Part 1: Determination of the presence of cocoa butter equivalents

Step-by-step procedure

1. Preparation

   IRMM-801 melted and mixed; about 200 g of chocolate cooled and grated (7). PN-EN ISO 23275-1:2009 with no withdrawal note in PKN (03.10.2026).

2. Fat extraction

   10–40 g of chocolate, 2–3 × 100 ml of solvent, evaporation, drying under nitrogen (8.1).

3. Chromatography

   About 0.2 g / 20 ml, 1 ml / 20 ml (0.5 mg/ml), injection 0.5–1.0 µl; identification of POP, POS, POO, SOS, SOO against IRMM-801 (8.2, 8.3).

4. Calculations

   Response factors from IRMM-801; proportions of POP, POS, SOS to two decimal places (9.1, 9.2).

5. Decision

   POP < 44.025 − 0.733 × SOS — pure cocoa butter; higher — CBEs present (9.3).

Required equipment and apparatus

Reagents, media and consumables

Health and safety (OHS)

• Diethyl ether and n-heptane — flammable; work in a fume hood, away from ignition sources (warning in 4)
• Hydrogen as carrier gas — flammable; leak-tight installation

Frequently asked questions