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INTERNATIONAL ISO
STANDARD 24381
First edition
2023-11
Bee propolis — Specifications
Propolis d’abeille — Spécifications
Reference number
© ISO 2023
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ii
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Requirements . 3
4.1 Classification of raw propolis types . 3
4.1.1 Temperate, Mediterranean and boreal, brown, Populus spp. propolis . 3
4.1.2 Tropical, green, Baccharis dracunculifolia propolis . 3
4.1.3 Tropical, red, Dalbergia and Clusia propolis . 3
4.1.4 Other types of propolis . 3
4.2 Physical and chemical requirements . 3
4.3 Traceability requirements . 4
5 Test methods . 4
5.1 Reagents . 4
5.2 Sample collection . . . 5
5.3 Sample preparation . 5
5.4 Test methods for physical and chemical requirements . 5
6 Packaging, marking, labelling, storage and transportation . 5
6.1 Packaging . 5
6.2 Marking (label and/or certificate) . 5
6.3 Labelling . 5
6.4 Storage and transportation . 6
Annex A (normative) Ethanol extractables of raw propolis (as dry matter) .7
Annex B (normative) Loss on drying determination . 9
Annex C (normative) Ash content in raw propolis .11
Annex D (normative) Petroleum ether extractables of raw propolis (as dry matter) .13
Annex E (normative) Total phenolic content .15
Annex F (Normative) Total flavonoids content (aluminium chloride method) .18
Annex G (Normative) Total flavonoids content (polyamide method) .20
Annex H (normative) Chemical characterization of polyphenols in poplar propolis —
HPLC/MS .24
Annex I (normative) Chemical characterization of polyphenols in green propolis — HPLC/
PDA . .38
Annex J (normative) Chemical characterization of polyphenols in red propolis by HPLC-
PDA and HPLC-MS .43
Annex K (normative) Determination of antioxidant capacity by DPPH .47
iii
Foreword
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This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 19,
Bee products.
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iv
Introduction
Propolis is a resinous substance produced by worker bees combining plant resins and/or fragments of
newly formed buds with their salivary and wax gland secretions.
The chemical composition of propolis is quite complex. Hundreds of natural compounds such as
flavonoids and phenolic acids have been identified in propolis. Different geographical and plant sources,
bee species, production methods, etc., have a significant influence on the chemical composition of
propolis.
For the purposes of this document, propolis is divided into Populus, Baccharis and Dalbergia types (the
primary sources) and opens the opportunity in the future to cover other types, such as Araucaria spp.,
Betula spp., Castanea spp., Clusia spp., Cupressus spp., Eucalyptus spp., Macaranga spp., Symphonia spp.,
and the mixed plant source propolis (this list is not exhaustive). Only propolis produced by Apis mellifera
bees is covered in this document.
Scientific literature predominantly relates to three main propolis types (brown, green and red) of
which brown (Populus) and green (Baccharis) propolis are the main types traded internationally. This
document considers the complex chemical composition of propolis, and the influences that geographical
and plant species variation, and honey bee sub-species have on the proximate, flavonoid and phenolic
composition of propolis. Propolis is rich in polyphenols, in particular flavonoids, phenolic acids and
derivatives, which can be involved in the biological activities. The decisions made about the types,
methodologies and requirements included in this document were based on the scientific literature
available at the time.
This document sets out the terms, definitions, classification, quality requirements, authenticity
requirements, test method procedures, transportation, storage conditions and packing marks. It aims
to provide a document for the classification and quality control for the international trade of raw
propolis.
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INTERNATIONAL STANDARD ISO 24381:2023(E)
Bee propolis — Specifications
1 Scope
This document specifies the quality requirements, analytical methods, and packaging, marking,
labelling, storage and transportation conditions for bee propolis.
This document is applicable to propolis collected from beehives of Apis mellifera colonies, i.e. raw
propolis.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies. For
undated references, the latest edition of the referenced document (including any amendments) applies.
ISO 22005, Traceability in the feed and food chain — General principles and basic requirements for system
design and implementation
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
3.1
antioxidant capacity
ability of a substance that retards deterioration of oxidation
3.2
ash content
incombustible component remaining after a sample of raw propolis (3.17) is completely burnt
3.3
authenticity requirement
requirement that the addition of resins, extracts or any compounds, and/or bioactive substances in raw
propolis is not allowed
3.4
balsamic
relating to or containing balsam
3.5
beeswax
honey bee secretions including mixtures of substituted long-chain aliphatic hydrocarbons, containing
alkanes, alkyl esters, fatty acids, primary and secondary alcohols, diols, ketones and aldehydes
3.6
contaminant
substance not intentionally added to propolis, which is present as a result of a process such as production
to harvesting, manufacture, processing, preparation, treatment, packing, packaging, transport or
holding of the product, or because of environmental contamination
3.7
ethanol extractables of propolis (as dry matter)
content obtained after an exhaustive extraction process using ethanol (80 % volume/volume) as the
extractor solvent
Note 1 to entry: Also known as genuine, crude, balsamic and dry extract of propolis.
3.8
petroleum ether extractables of propolis (as dry matter)
content obtained after an exhaustive extraction process using petroleum ether as an extraction solvent
3.9
flavonoids
class of plant and fungus secondary metabolites that have the general structure of a 15-carbon skeleton
(abbreviated C6-C3-C6), consisting of two phenyl rings
...