Introduction: The regulatory importance of bisphenols and phthalates
The chemical safety of packaging materials that come into contact with food is strictly regulated by the European Union Packaging Ordinance (PPWR) and a number of specific regulations, above all Regulation (EU) No 10/2011 on plastic materials. Two of the most important components of this regulatory framework are the groups of substances known as bisphenols and B phthalates. Bisphenols are monomers used in particular in polycarbonate plastics and epoxy resin coatings, while phthalates are plasticizers that give PVC-based packaging flexibility. Both groups of substances have undergone intensive toxicological examinations on the basis of their endocrine-effective properties (which negatively affect the hormone system) and for many of them specific migration limits (SML) limiting their migration to foods have been set. Nano-Test Laboratories perform bisphenol and phthalate analyzes in full compliance with GLP principles with validated LC-MS/MS and GC-MS/MS procedures and provide our customers with legal compliance assessments and comprehensive technical advice.
Bisphenols: chemical structure, regulatory status and SML values
Bisphenols are synthetic organic compounds in which two hydroxyphenyl groups are connected to each other via a bridge atom or a bridge group. The best known and most commonly used compound of this group, Bisphenol A (BPA), is used as a monomer in the manufacture of polycarbonate plastics and as a starting material in the synthesis of epoxy resins. However, after the endocrine properties of BPA have been scientifically proven, the restrictions on the use of BPA in the European Union have been increasingly tightened. This has prompted manufacturers to use alternatively structurally similar alternative compounds (BPS, BPF, BPAF, etc.) instead of BPA; However, the fear that these alternatives could also have similar toxicological profiles has resulted in the supervisory authorities having also included these connections in the scope of surveillance.
The main regulated bisphenols and their SML values
The following table summarizes the bisphenol compounds that occur most frequently in packaging materials, their chemical structure, their regulatory status and the current SML values:
| Connection | Abbreviation / CAS No. | SML (mg/kg food or simulant) | Regulatory Status | Typical scope |
|---|---|---|---|---|
| Bisphenol A | BPA / 80-05-7 | 0,05 | subject to SML according to 10/2011/EU; prohibited in baby bottles; in the EU in the process of a complete ban on food contact; limited in REACH Annex XVII; On the SVHC Candidate List (Endocrine Disruptor) | Polycarbonate-Plastics, Epoxy Resin Coatings, Canned Inner Coatings |
| Bisphenol S | BPS / 80-09 -1 | 0,05 | Subject to SML according to 10/2011/EU; is increasingly being used as a BPA alternative; toxicological examination is still ongoing | BPA alternative; Thermal paper coatings, sulfone polymers, epoxy resins |
| Bisphenol f | BPF / 620-92-8 | Under observation | Is not yet subject to any SML according to 10/2011/EU; on the EFSA's surveillance list; BPA alternative | Epoxy resins, coatings, adhesives |
| Bisphenol AF | BPAF / 1478-61-1 | Is subject to surveillance | Used in fluorinated polymers; BPA alternative; official examination is still ongoing | Manufacture of fluoropolymers, special coatings |
Note: The European Commission is currently working on a draft prohibiting the use of BPA in all materials that come into contact with food. Nano-Test closely follows these regulatory developments and proactively informs its customers.
Phthalates: regulatory framework for plasticizers
Phthalates (phthalic acid esters) are plasticizers used in particular in polyvinyl chloride (PVC)-based packaging materials to impart flexibility and processability. Since they are not covalently bound to PVC, they can go to the surface (migration) over time and go into food from there. Some of the phthalates are classified as endocrine-effective substances and as reproductive toxic substances, which is why their use in materials that come into contact with food is strictly restricted. For low molecular weight phthalates (such as DBP, BBP, DEHP), far-reaching restrictions also apply under the REACH regulation.
The main regulated phthalates and SML values
The following table summarizes the most common phthalate compounds in packaging materials, their chemical structure and SML values in accordance with Regulation (EU) No 10/2011:
| Connection | Abbreviation / CAS No. | SML (mg/kg food or simulant) | Regulatory Status | Typical scope |
|---|---|---|---|---|
| Di (2-ethylhexyl)phthalate | DEHP / 117-81-7 | 1,5 | subject to SML according to 10/2011/EU; REACH Annex XIV (requiring approval); on the SVHC candidate list; prohibited in toys | PVC plasticizer; Seals, Closures, Flexible Foils, Medical Devices |
| Dibutyl phthalate | DBP / 84-74-2 | 0,3 | Subject to SML according to 10/2011/EU; REACH Annex XIV; on the SVHC candidate list; prohibited in cosmetics | Adhesives, printing inks, PVC films, coatings |
| Butylbenzylphthalate | BBP / 85-68-7 | 30 | Subject to SML according to 10/2011/EU; REACH Annex XIV; On the SVHC candidate list | Vinyl coatings, printing inks, laminating adhesives |
| Di-isononylphthalate | DINP / 28553 -12-0 | 9 (as sum of DINP and DIDP) | Subject to SML according to 10/2011/EU; restricted in children's products | PVC plasticizer; Flexible packaging, seals, closures |
| Di isodesyl phthalate | DIDP / 26761-40-0 | 9 (as sum of DINP and DIDP) | subject to SML according to 10/2011/EU; restricted in children's products | PVC plasticizer; Flexible packaging, seals |
| Di-n-octylphthalate | Dnop / 117-84-0 | 5 (as sum of DNOP and DNP) | 10/ 2011/EU is subject to SML | PVC plasticizer; Special Applications |
Note: In the current consolidated version of Regulation 10/2011/EU, the SML values for DEHP, DBP, BBP and DINP are defined on the basis of the specific substances. In addition, combined conformity criteria for the total of these phthalates (e.g. the sum of DEHP + DBP + BBP) must be taken into account.
Analytical methodology in the nano test laboratory
In the nano-test laboratory, bisphenol and phthalate analyzes are carried out using analytical methods, which are optimized, validated and fully compliant with the GLP principles according to the chemical properties of the target substance.
Selection of the analytical platform and method comparison
The following table provides a comparison of the most important analytical platforms used in bisphenol and phthalate analyses and their areas of application:
| Analytical Platform | Technical properties | Destination connections | Advantages |
|---|---|---|---|
| LC-MS/MS (QQQ) | Triple quadrupole; ESI ionization; MRM mode; High selectivity and sensitivity | Bisphenols (BPA, BPS, BPF, BPAF and other analogs); Polar Phthalate Metabolites | Ideal for polar and moderately polar connections; low LOQ values (in the PPB range); No derivatization required |
| GC-MS/MS (QQQ) | Triple quadrupole; egg ionization; MRM mode; High chromatographic selectivity | Phthalates (DEHP, DBP, BBP, DINP, DIDP, DNOP); Low Polar Connections | Reference method for volatile and semi-volatile connections; extensive library support; isomer separation possible |
| HPLC DAD / UV-VIS | Photodiode array detector; Suitable for compounds with chromophores groups | Specific bisphenols; Samples with high concentration | Simple and fast screening; cost-efficient; Suitable for pre-screening |
Analytical workflow: from migration attempt to quantitative determination
The following table describes in detail the end-to-end workflow for bisphenol and phthalate analyzes in the nano-test laboratory:
| Step | Process | Technical Details |
|---|---|---|
| 1. Test setup | Determination of the simulancy and test conditions | Appropriate food simulants and test conditions are selected in accordance with the application scenario specified by the customer in accordance with Annex III and V of Regulation (EU) No 10/2011. For bisphenols and phthalates, Simulan D1 (50% ethanol) or Simulan D2 (vegetable oil) are usually used. |
| 2. Migration test | Controlled contact | The packaging sample is contacted with the selected simulan at a set temperature and over a specified period using standard contact cells or by means of immersion. The contact conditions are continuously monitored in calibrated drying cabinets. |
| 3. Extraction and Sample Preparation | Isolation of the target analytes | Liquid-liquid extraction or SPE (solid phase extraction) for aqueous simulants; Appropriate solvent extraction and purification for the oily Simulan D2. Matrix effects are corrected by adding internal standards (such as isotope-labeled BPA-D16, DEHP-D4). |
| 4. Instrumental Analysis | Quantitative determination using LC-MS/MS or GC-MS/MS | Bisphenols are analyzed by LC-MS/MS; Phthalates using GC-MS/MS. Two MRM transits (quantitative and confirmation) are tracked for each target connection. The LOQ values were validated to be at least one tenth of the SML limits. |
| 5. Evaluation and Reporting | Comparison with the SML and declaration of conformity | The quantitative results obtained are compared with the legal SML value of the respective substance. In cases where combined limit values apply (e.g. sum of DINP and DIDP), the total values are also calculated. A GLP compliant, comprehensive analysis report is created. |
The integrated service model of nano-test for bisphenol and phthalate analyses
As Nano-Test Labor Services, we do not offer bisphenol and phthalate analyzes as individual examinations, but as an integral part of the PPWR and 10/2011/EU conformity process. Our service model is based on the following four pillars:
| Service pillars | Scope | Benefits for the customer |
|---|---|---|
| Regulatory consulting | Determining which bisphenol and phthalate compounds must be analyzed according to the product composition and the conditions of use; Selecting the right simulation and test conditions | Cost optimization through the right test strategy; Avoidance of missing or unnecessary analyses |
| Analytical Testing Services | Quantitative determination using GLP-compliant validated LC-MS/MS and GC-MS/MS methods | Responsible and traceable analytical data accepted by the supervisory authorities |
| Conformity assessment | Preparation of a declaration of conformity or non-conformity by comparing the results with SML values and combined limit values | A clear and binding assessment of the legal status of the product |
| Support with technical documentation | Provision of comprehensive analysis reports suitable for government requests and the data required for the Declaration of Conformity (DOC) | Support for the customer in the complete and correct creation of their own declaration of conformity |
In the event of an exceeding of the limit value or non-conformity, the specialist team at Nano-Test also offers its customers technical advice on the cause analysis to determine the source of the problem and the planning of corrective measures. As bisphenol and phthalate contaminations are usually due to a specific raw material or additive in the supply chain, the correct identification of the source is crucial for a permanent solution to the problem.
In order to ensure the bisphenol and phthalate compliance of your packaging that you export to the European market, according to PPWR and 10/2011/EU and to carry out your analyzes with a reliable and experienced partner, you can contact our expert team for detailed Get information about the accredited laboratory services and technical advice from Nano-Test.
Introduction: The NIAS concept and the regulatory context
Ensuring the chemical safety of packaging materials coming into contact with foods is not limited to the control of substances intentionally added during the manufacturing process (intentionally added substances - ias). Article 3 of the Framework Regulation (EC) No 1935/2004 of the European Union and Regulation (EU) No 10/2011 on plastics materials oblige the manufacturers to unintentionally added substances (non-intentionally added Substances – NIAS) and conduct a risk assessment. Nias can come from contaminants in the starting materials, from reaction by-products, degradation products or environmental contaminants produced during production. With the introduction of the PPWR, this obligation has become even clearer, so that packaging manufacturers have to tackle the NIAS assessment with a systematic approach. Nano-Test Labor Services offer a comprehensive and GLP-compliant service portfolio in the area of NIAS assessment, ranging from screening and identification studies with advanced analysis techniques to toxicological risk assessment.
Definition, sources and classification of NIAS
NIAS is a broad term that encompasses all chemical substances that are not intentionally added to the formulation of a packaging material, but may be present in the final product. The sources of these substances are diverse and can arise throughout the entire lifecycle of the packaging. A correct understanding of NIAS is crucial for the development of an effective analysis strategy and the performance of a risk assessment.
Classification of NIAS sources
The following table systematically summarizes the most important mechanisms of formation, sources and typical examples for each source:
| NIAS source | Mechanism of formation | Typical NIAS examples |
|---|---|---|
| Contaminations in the starting materials | Residues from the manufacturing process of monomers, additives or residues of solvents originating from the production process | Monomer isomers, solvent residues, catalyst residues, contaminations in chemicals of technical quality |
| Reaction by-products | Undesired compounds formed during polymerization, cross-linking or curing reactions undesired compounds | Oligomers (dimers, trimers), polymerization by-products, degradation products |
| Decomposition products | Thermal, photochemical or hydrolytic degradation of the polymer or additives | Degradation products of antioxidants, degradation products of UV stabilizers, depolymerization oligomers |
| Contaminations from recycling</ strong> | Foreign substances that enter the recycled raw material or residues from previous use | Printing ink components, adhesive residues, residues of previous products, MOSH/MOAH |
| Environmental contaminants | Substances that occur during production, storage or transport from the environment | PAH (polycyclic aromatic hydrocarbons), PCB, pesticide residues |
Note: The scope of the NIAS assessment is determined on a case-by-case basis based on the composition of the packaging material, the manufacturing process and the intended conditions of use. The guidelines published by the EFSA provide the basic methodological framework for the NIAS assessment.
Analytical strategy in NIAS assessment
The NIAS assessment represents one of the most complex areas of analytical chemistry due to the need to screen and identify a variety of known and unknown substances simultaneously. In the Nano-Test laboratory, the NIAS assessment is carried out using the following multi-stage and systematic approach.
Step-by-step NIAS assessment strategy
The NIAS assessment process applied by Nano-Test consists of four main phases, which are summarized in the following table:
| Phase | Step | Explanation |
|---|---|---|
| 1. Pre-evaluation and data collection | Review of information on formulation and manufacturing process | Theoretical determination of all raw materials, additives, production parameters, and potential NIAS sources (impurities, reaction by-products, decomposition products). In this phase, supplier declarations and safety data sheets (SDS) provided by the customer are also evaluated. |
| 2. Non-targeted screening (non-targeted screening) | Comprehensive screening using high-resolution mass spectrometry (HRMS) | The extract obtained after a suitable extraction and/or migration test is analyzed with LC-QTOF-MS and GC-QTOF-MS systems. The obtained data are evaluated with advanced data processing software to perform a library match and identify unknown peaks. |
| 3. Identification and semiquantitative determination | Structural elucidation of identified NIAS candidates | The structural characterization of unknown compounds is based on the exact mass obtained from the HRMS data, the isotope distribution and the MS/MS fragmentation pattern. As far as possible, validation is based on reference standards. The semi-quantitative estimate is based on an approach based on internal standards or structural analogues. |
| 4. Toxicological Risk Assessment | Evaluation of the identified NIAS with regard to food safety | For each identified NIAS, a risk assessment is performed using the EFSA's Threshold of Toxicological Concern (TTC) approach or substance-specific toxicological data. The Cramer classification and genotoxicity warnings (QSAR) are taken into account. |
Nano Test NIAS Analysis Platforms
The main analytical platforms used in the Nano Test Laboratory for the NIAS assessment, as well as their specific roles in NIAS analysis, are shown in the following table:
| Analytical Platform | Technical properties | Role in NIAS analysis | Target network profile |
|---|---|---|---|
| LC-QTOF-MS | High Resolution (‚30,000 FWHM), High Mass Accuracy (±3 ppm), MS/MS Fragmentation | Target-free screening and identification of polar and moderately polar NIAS | Oligomers, degradation products, antioxidants and their metabolites, plasticizers, lubricants |
| GC-QTOF-MS | High Resolution, Egg and Ci Ionization, Extensive Library (NIST, Wiley) | Targets Free Screening and Identification of Volatile and Semi-Flecked NIAS | Volatile organic compounds (VOCs), low molecular weight oligomers, solvent residues |
| LC-MS/MS (QQQ) | Triple-quadrupole, MRM mode, high selectivity and sensitivity | Targeted quantitative determination and validation of identified NIAS | Specific NIAS compounds (BPA, phthalates, PAA, etc.) |
| Headspace GC-MS | Headspace sampling, optimized for volatile connections | Screening and Quantification of Volatile NIAS | Residual monomers (styrene, butadiene), solvents, volatile degradation products |
Toxicological risk assessment and TTC approach
The ultimate goal of the NIAS assessment is to determine whether the identified substances pose a risk to consumer health. The “Threshold of Toxicological Concern” (TTC) approach adopted by EFSA offers a scientifically based risk assessment tool for substances whose structure is known but for which only limited toxicological data are available.
Nano Test uses the following systematic approach in this crucial phase of NIAS assessment:
Cramer Classification and TTC Thresholds
The following table summarizes the toxicological potential of the substances according to the Cramer classification system and the corresponding TTC thresholds:
| Cramer class | Toxicity potential | TTC threshold (μg/kg body weight/day) | Typical chemical structures |
|---|---|---|---|
| Class I | Low toxicity potential | 30 | Simple aliphatic hydrocarbons, normal metabolic products |
| Class II | Mean toxicity potential | 9 | Compounds with functional groups without aromatic rings, esters, ethers |
| Class III | High toxicity potential | 1, 5 | Aromatic compounds, heterocyclic structures, conjugated systems |
| Genotoxic substances | Suspected DNA-reactive genotoxicity | 0.0025 (for a lifetime risk of cancer of 1:10⁵) | Aflatoxin-like structures, N-nitroso compounds, DNA intercalators |
Note: EFSA does not recommend using the TTC approach for genotoxic carcinogens. Minimizing exposure (Alara principle) is of fundamental importance for such substances. In addition, the TTC approach may not be suitable for substance groups with potentially neurotoxic effects, such as organophosphates and carbamates.
Nano-Test's integrated NIAS assessment service
As Nano-Test Labor Services, we offer the NIAS assessment with a multidisciplinary and integrated approach. Our service model consists of the components summarized in the following table:
The NIAS assessment is not just an analytical test, but a multi-layered process that requires well-founded chemical and toxicological expertise. As a nano test, at every stage of this process, we act in accordance with the GLP principles, scientific care and a customer-oriented approach to consulting. PPWR and the relevant EU regulations and conduct your screening and risk assessment processes with a reliable partner, you can contact our team of experts for detailed information on Nano-Test's NIAS Valuation Services get.
| Performance component | Scope | Benefit for the customer |
|---|---|---|
| Formulation and Process Analysis | Detailed investigation of the raw materials used, additives and the production process; Theoretical mapping of potential NIAS sources | Correct elaboration of the analysis strategy, avoidance of unnecessary analyses |
| Untargeted Screening (HRMS) | Comprehensive, target-free chemical screening using LC-QTOF-MS and GC-QTOF-MS | Detection of unexpected contaminants and unknown NIAS |
| Structural Enlightenment and Half-Quantification | Structural characterization based on HRMS data and semi-quantitative estimate based on internal standards | Reliable identification of unknown peaks and determination of their approximate concentrations |
| Toxicological risk assessment | Comprehensive Risk Assessment Using TTC Approach, Cramer Classification and QSAR Toxicity Prediction (in Silico) | Scientifically based assessment of the food safety risk of identified NIAS |
| Compliance with regulatory requirements | Comprehensive report for the DOC (Declaration of Conformity), which contains all analytical results and the risk assessment | Complete documentation for official applications and customer audits |