Sweetener Analysis — EN 12856 / HPLC / (EC) 1333/2008

What Is Sweeteners?

Sweetener analysis is the determination of the amount of intense sweeteners used instead of sugar in food. The sample is extracted accordingly and the sweeteners separated by HPLC are measured individually. A separate chromatographic method is applied for steviol glycosides.

Purpose of Sweeteners

The aim is to verify that the sweeteners used comply with the permitted substances and maximum levels in Annex II (EC) 1333/2008 and to check the label declaration.

Which Products Is Sweeteners Applied To?

Sugar-free and light drinks
Chewing gum and candies
tabletop sweeteners
Diet products and desserts
Dairy products and sweetened yoghurts

In Which Sectors Is Sweeteners Used?

FoodAgricultureRetail

Standards Related to Sweeteners

TS EN 12856 (food — determination of acesulfame K, aspartame and saccharin, HPLC)
TS EN 12857 (food — determination of cyclamate)
JECFA monographs (steviol glycosides purity and determination)
AOAC related HPLC methods

Sweeteners Conditions

Extraction: Aqueous or aqueous-organic; carbonated drinks require degassing
Measurement: HPLC-DAD or LC-MS/MS
Unit: mg/kg veya mg/L

Equipment for Sweeteners

HPLC-DAD
LC-MS/MS
Ultrasonic bath
Analytical balance and filtration devices

Sample Requirements for Sweeteners

Homogenized representative sample
Carbonated products must be degassed
Product category and label declaration must be declared
The list of sweeteners to be analyzed should be specified.

Duration of Sweeteners

Extraction and chromatographic measurement together take several working days.

Sweeteners Procedure

1The sample is homogenized and weighed.
2In carbonated products, gas is removed.
3It is extracted with the appropriate solvent.
4The extract is cleaned and filtered.
5Measurement is made by HPLC or LC-MS/MS.
6Results are reported on a sweetener basis.

Sweeteners Acceptance Criteria

Annex II of (EC) 1333/2008 defines permitted sweeteners and maximum levels for each food category. In addition, labeling obligations such as warning of phenylalanine source in products containing aspartame are required within the scope of (EU) 1169/2011.

Evaluation of Results

When more than one sweetener is used together, the maximum level of each is evaluated separately, as well as the combination rules stipulated in the regulation. The consistency of the analysis result with the label declaration is also checked.

Sweeteners Report

Sample description and food category
List of sweeteners analyzed
Method applied
Result and detection limit for each sweetener
Compared level of legislation

Accreditation

EUROLAB holds internationally recognised accreditation obtained from accreditation bodies party to the IAF and ILAC multilateral recognition arrangements, under ISO/IEC 17025 for laboratory testing, ISO/IEC 17020 for inspection and surveillance, and ISO/IEC 17021-1 and ISO/IEC 17065 for system and product certification. This chain ensures the reports and certificates issued are accepted worldwide.

Whether this test can be offered within accredited scope depends on your product and the standard applied; since accreditation is method-based, the scope is confirmed in writing at the quotation stage.

Scope and authorisations

Related Regulations

Food Additives Regulation (EC) 1333/2008
Food Information Regulation (EU) 1169/2011

Food Testing — What Does This Testing Field Cover?

The section below describes the field that the Sweeteners test belongs to; the conditions and acceptance criteria of this particular test are given in the sections above. The scope specific to your product is confirmed in writing at the quotation stage.

Food testing shows that a product is safe for consumption and contains what its label states. The family works along three axes: microbiological safety, chemical safety and nutritional composition. All three require different laboratory infrastructure, a different body of methods and a different turnaround time.

The most distinctive feature of the field is TIME PRESSURE. A steel specimen can wait on a shelf; a raw milk or fresh meat sample cannot. The time the sample was taken, the transport temperature and how long it takes to reach the laboratory directly determine the validity of the result. In food microbiology the cold chain record is therefore an inseparable part of the test report.

On the microbiological side there are two different questions and they must not be confused. DETECTION of a pathogen is a qualitative question: is Salmonella present in the sample or not? ENUMERATION of indicator organisms is a quantitative question and an indicator of production hygiene. The absence of a pathogen in a product does not show that hygiene is good; if the total mesophilic aerobic count is high, there is a problem in the process.

On the chemical side, pesticide residues, mycotoxins, heavy metals, additives, veterinary drug residues and process contaminants (acrylamide, furan, PAH) are measured. Multi-residue methods in this area (QuEChERS-based LC-MS/MS and GC-MS/MS) can screen hundreds of active substances in a single analysis; limits are defined separately for each product–substance pair.

On the composition side, protein, fat, moisture, ash, fibre, sugar profile, fatty acid composition, vitamin and mineral content are determined. These values are the basis of the nutrition declaration on the label and the tolerance ranges are defined in legislation — the difference between the declared value and the measured value must stay within that tolerance.

There is NO CE marking on food, and this shapes how the field works. Conformity is demonstrated through operating a HACCP-based food safety system, complying with labelling rules and not exceeding contaminant, residue and microbiological limits. A test result produces evidence not for a certificate but for a functioning system.

Allergen management carries particular weight. The presence of an undeclared allergen in a product is one of the most frequent causes of food recalls. Analysis here runs together with cleaning validation: it is shown that the line has been cleaned adequately after a production run containing an allergen and that the risk of cross-contamination is under control.

Voluntary schemes have become effectively mandatory in this field. Retail chains require their suppliers to hold a certificate under a GFSI-recognised scheme (BRCGS, IFS, FSSC 22000); without it, getting onto the shelf is practically impossible.

Method groups in this field

Pathogen detection
Detection of Salmonella, Listeria monocytogenes, Campylobacter, Cronobacter and Shiga toxin-producing E. coli; culture-based and molecular methods.
Indicator counts
Total mesophilic aerobic count, Enterobacteriaceae, coliforms and E. coli, coagulase-positive staphylococci, yeasts and moulds, lactic acid bacteria, sulphite-reducing Clostridia.
Pesticide and veterinary residues
Multi-residue screening by LC-MS/MS and GC-MS/MS with QuEChERS extraction.
Mycotoxins and contaminants
Aflatoxins, ochratoxin A, deoxynivalenol, patulin; heavy metals (ICP-MS), acrylamide and PAH compounds.
Nutritional composition
Kjeldahl protein, fat and fatty acid profile, moisture, ash, dietary fibre, sugar profile, vitamin and mineral analyses.
Allergens and authenticity
Gluten, milk, egg, nut and soy allergen analyses; molecular methods for species identification and adulteration detection.

Commonly confused in this field

Sampling is half the result. A sample transported at the wrong temperature or delayed gives an invalid result even if it is analysed correctly in the laboratory.
The absence of a pathogen does not show that hygiene is good; indicator counts give separate information and the two are read together.
A result from a sample taken from a lot can be generalised to the whole lot only if the sampling plan was set up statistically.
Limits are defined in separate regulations by product group and are updated regularly; a conformity decision based on an outdated limit table may be wrong.
For exports, the limits of the destination country may differ from EU limits; the same product may be compliant in one market and non-compliant in another.

Which decisions these results support

Verification of the HACCP plan and process monitoring
The basis for the nutrition declaration and allergen information on the label
Health certificates before export and compliance with destination market limits
Supplier and lot-based acceptance control
Shelf-life studies and determining storage conditions
Producing evidence for audits under GFSI-recognised schemes

Related Tests

Frequently Asked Questions About Sweeteners

Are steviol glycosides free because they are natural?

It is not. Steviol glycosides are also permitted as food additives and have maximum use levels by category. Just because it is of natural origin does not mean that it is illegal.