Introduction: The Regulatory Significance of Bisphenols and Phthalates
The chemical safety of packaging materials that come into contact with food is strictly regulated by the European Union’s Packaging Regulation (PPWR) as well as by a number of specific regulations, most notably 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 primarily in polycarbonate plastics and epoxy resin coatings, while phthalates are plasticizers that impart flexibility to PVC-based packaging. Both groups of substances have been subjected to intensive toxicological studies due to their endocrine-disrupting properties (which negatively affect the endocrine system), and specific migration limits (SMLs) have been established for many of them to restrict their migration into food. Nano-Test Laboratories conduct bisphenol and phthalate analyses in full compliance with GLP principles using validated LC-MS/MS and GC-MS/MS methods, and offer our clients regulatory compliance assessments as well as comprehensive technical consulting.
Bisphenols: Chemical Structure, Regulatory Status, and SML Values
Bisphenols are synthetic organic compounds in which two hydroxyphenyl groups are linked to each other via a bridging atom or group. The best-known and most widely used compound in this group, bisphenol A (BPA), is used as a monomer in the production of polycarbonate plastics and as a starting material in the synthesis of epoxy resins. However, after the endocrine-disrupting properties of BPA were scientifically proven, restrictions on the use of BPA in the European Union have been progressively tightened. This has prompted manufacturers to use structurally similar alternative compounds (BPS, BPF, BPAF, etc.) in place of BPA; however, concerns that these alternatives might also exhibit similar toxicological profiles have led regulatory authorities to include these compounds in their monitoring programs as well.
The Most Important Regulated Bisphenols and Their SML Values
The following table summarizes the bisphenol compounds most commonly found in packaging materials, their chemical structure, their regulatory status, and the current SML values:
| Connection | Abbreviation / CAS No. | SML (mg/kg of food or simulant) | Regulatory Status | Typical Applications |
|---|---|---|---|---|
| Bisphenol A | BPA / 80-05-7 | 0,05 | Subject to the SML pursuant to Regulation (EU) No. 10/2011; prohibited in baby bottles; in the process of being completely banned in the EU for contact with food; Restricted under REACH Annex XVII; on the SVHC Candidate List (endocrine disruptor) | Polycarbonate plastics, epoxy resin coatings, interior coatings for food cans |
| Bisphenol S | BPS / 80-09 -1 | 0,05 | Subject to the SML pursuant to 10/2011/EU; increasingly used as an alternative to BPA; toxicological testing is still ongoing | BPA alternatives; thermal paper coatings, sulfone polymers, epoxy resins |
| Bisphenol F | BPF / 620-92-8 | Under observation | Not yet subject to an MRL under Regulation (EU) No. 10/2011; on the EFSA’s watch list; BPA alternative | Epoxy resins, coatings, adhesives |
| Bisphenol AF | BPAF / 1478-61-1 | Subject to monitoring | Used in fluorinated polymers; BPA alternative; regulatory review is still ongoing | Manufacture of Fluoropolymers, Specialty Coatings |
Note: The European Commission is currently working on a draft proposal to ban the use of BPA in all materials that come into contact with food. Nano-Test is closely monitoring these regulatory developments and proactively keeping its customers informed.
Phthalates: Regulatory Framework for Plasticizers
Phthalates (phthalic acid esters) are plasticizers used primarily in polyvinyl chloride (PVC)-based packaging materials to impart flexibility and processability. Since they are not covalently bound to PVC, they can migrate to the surface over time and transfer from there into food. Some phthalates are classified as endocrine disruptors and as substances toxic to reproduction, which is why their use in materials that come into contact with food is strictly restricted. Low-molecular-weight phthalates (such as DBP, BBP, and DEHP) are also subject to far-reaching restrictions under the REACH Regulation.
The Most Important Regulated Phthalates and SML Values
The following table summarizes the phthalate compounds most commonly found in packaging materials, their chemical structures, and the SML values in accordance with Regulation (EU) No. 10/2011:
| Connection | Abbreviation / CAS No. | SML (mg/kg of food or simulant) | Regulatory Status | Typical Applications |
|---|---|---|---|---|
| (2-Ethylhexyl)phthalate | DEHP / 117-81-7 | 1,5 | Subject to the SML pursuant to 10/2011/EU; REACH Annex XIV (subject to authorization); on the SVHC Candidate List; prohibited in toys | PVC plasticizers; seals, fasteners, flexible films, medical devices |
| Dibutyl phthalate | DBP / 84-74-2 | 0,3 | Subject to the SML pursuant to 10/2011/EU; REACH Annex XIV; on the SVHC Candidate List; prohibited in cosmetics | Adhesives, printing inks, PVC films, coatings |
| Butylbenzyl phthalate | BBP / 85-68-7 | 30 | Subject to the SML pursuant to 10/2011/EU; REACH Annex XIV; on the SVHC Candidate List | Vinyl coatings, printing inks, laminating adhesives |
| Diisononyl phthalate | DINP / 28553-12-0 | 9 (as the sum of DINP and DIDP) | Subject to the SML pursuant to 10/2011/EU; restricted in children's products | PVC plasticizers; flexible packaging, gaskets, closures |
| Di-isodecyl phthalate | DIDP / 26761-40-0 | 9 (as the sum of DINP and DIDP) | Subject to the SML pursuant to 10/2011/EU; restricted in children's products | PVC plasticizers; flexible packaging, seals |
| Di-n-octyl phthalate | DNOP / 117-84-0 | 5 (as the sum of DNOP and DNP) | 10/2011/EU is subject to the SML | PVC plasticizers; special applications |
Note: In the current consolidated version of Regulation (EU) No. 10/2011, the SML values for DEHP, DBP, BBP, and DINP are specified on the basis of the respective individual substances. In addition, combined compliance criteria for the total sum of these phthalates (e.g., the sum of DEHP + DBP + BBP) must be taken into account.
Analytical Methods in the Nano-Test Laboratory
In the Nano-Test laboratory, bisphenol and phthalate analyses are performed using analytical methods that have been optimized and validated in accordance with the chemical properties of the target substance and are fully compliant with GLP principles.
Selection of the Analytical Platform and Comparison of Methods
The following table provides a comparison of the most important analytical platforms used in bisphenol and phthalate analyses, as well as their areas of application:
| Analytical Platform | Technical Specifications | 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 compounds; low LOQ values (in the ppb range); no derivatization required |
| GC-MS/MS (QQQ) | Triple quadrupole; EI ionization; MRM mode; high chromatographic resolution | Phthalates (DEHP, DBP, BBP, DINP, DIDP, DNOP); low-polarity compounds | Reference method for volatile and semi-volatile compounds; extensive library support; isomer separation possible |
| HPLC-DAD / UV-Vis | Photodiode array detector; suitable for compounds containing chromophore groups | Specific bisphenols; samples with high concentrations | Simple and rapid screening; cost-effective; suitable for prescreening |
Analytical Workflow: From Migration Testing to Quantitative Determination
The following table provides a detailed description of the end-to-end workflow for bisphenol and phthalate analyses at the Nano-Test laboratory:
| Step | Process | Technical Details |
|---|---|---|
| 1. Experimental Setup | Determination of the simulation and test conditions | Based on the application scenario specified by the customer, suitable food simulants and test conditions are selected in accordance with Annexes III and V of Regulation (EU) No. 10/2011. For bisphenols and phthalates, Simulan D1 (50% ethanol) or Simulan D2 (vegetable oil) are generally used. |
| 2. Migration Attempt | Controlled Contact | The packaging sample is brought into contact with the selected simulant using standard contact cells or by immersion at a specified temperature and for a specified period of time. The contact conditions are continuously monitored in calibrated drying ovens. |
| 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 simulant D2. Matrix effects are corrected by adding internal standards (such as isotope-labeled BPA-d16 and DEHP-d4). |
| 4. Instrumental Analysis | Quantitative analysis using LC-MS/MS or GC-MS/MS | Bisphenols are analyzed using LC-MS/MS; phthalates using GC-MS/MS. For each target compound, two MRM runs (quantitative and confirmatory) are performed. The LOQ values were validated to ensure that they are at least one-tenth of the SML limits. |
| 5. Analysis and Reporting | Comparison with the SML and Declaration of Conformity | The quantitative results obtained are compared with the statutory SML value for the respective substance. In cases where combined limit values apply (e.g., the sum of DINP and DIDP), the total values are also calculated. A comprehensive analysis report that complies with GLP is prepared. |
Nano-Test's Integrated Service Model for Bisphenol and Phthalate Analysis
At Nano-Test Labor Services, we do not offer bisphenol and phthalate analyses as standalone tests, but rather as an integral part of the PPWR and 10/2011/EU compliance 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 based on the product composition and conditions of use; selecting the appropriate simulation and test conditions | Cost Optimization Through the Right Testing Strategy; Avoiding Missing or Unnecessary Analyses |
| Analytical Testing Services | Quantitative determination using GLP-compliant, validated LC-MS/MS and GC-MS/MS methods | Analytical data that is accepted, reasonable, and traceable by regulatory authorities |
| Conformity Assessment | Preparation of a declaration of conformity or nonconformity by comparing the results with SML values and combined limit values | A clear and definitive assessment of the product's legal status |
| Support for Technical Documentation | Provision of comprehensive analytical reports suitable for regulatory submissions, as well as the data required for the Declaration of Conformity (DoC) | Assisting the customer in preparing their own declaration of conformity completely and correctly |
In the event that a limit value is exceeded or nonconformity is detected, Nano-Test’s team of experts also offers its customers technical consulting to analyze the causes, identify the source of the problem, and plan corrective actions. Since bisphenol and phthalate contamination can usually be traced back to a specific raw material or additive in the supply chain, correctly identifying the source is crucial for finding a lasting solution to the problem.
To ensure that your packaging, which you export to the European market, complies with bisphenol and phthalate regulations in accordance with the PPWR and Regulation (EU) No. 10/2011, and to have your analyses conducted by a reliable and experienced partner, you can contact our team of experts to receive detailed information about Nano-Test’s accredited laboratory services and technical consulting.
Introduction: The NIAS Concept and the Regulatory Context
Ensuring the chemical safety of packaging materials that come into contact with food is not limited to monitoring substances that are intentionally added during the manufacturing process (Intentionally Added Substances—IAS). Article 3 of the European Union’s Framework Regulation (EC) No. 1935/2004 and Regulation (EU) No. 10/2011 on plastic materials require manufacturers to identify unintentionally added substances (NIAS) and conduct a risk assessment. NIAS can originate from impurities in raw materials, by-products of reactions occurring during production, degradation products, or environmental contaminants. With the introduction of the PPWR, this obligation has become even clearer, requiring packaging manufacturers to approach NIAS assessment with a systematic approach. Nano-Test Labor Services offers a comprehensive and GLP-compliant portfolio of services in the field of NIAS assessment, ranging from screening and identification tests using advanced analytical techniques to toxicological risk assessment.
Definition, Sources, and Classification of NIAS
NIAS is a broad term that encompasses all chemical substances that were 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 life cycle of the packaging. A proper understanding of NIAS is crucial for developing an effective analytical strategy and conducting a risk assessment.
Classification of NIAS Sources
The following table systematically summarizes the main formation mechanisms, sources, and typical examples for each source:
| NIAS Source | Mechanism of Formation | Typical NIAS Examples |
|---|---|---|
| Impurities in the raw materials | Residues from the monomer manufacturing process, additives, or solvent residues originating from the production process | Monomer isomers, solvent residues, catalyst residues, impurities in technical-grade chemicals |
| Reaction byproducts | Undesirable compounds formed during polymerization, cross-linking, or curing reactions | Oligomers (dimers, trimers), polymerization byproducts, decomposition products |
| Degradation products | Thermal, photochemical, or hydrolytic degradation of the polymer or additives | Decomposition products of antioxidants, decomposition products of UV stabilizers, depolymerization oligomers |
| Verunreinigungen aus dem Recycling</ strong> | Foreign substances that enter the recycled raw material, or residues from its previous use | Printing ink components, adhesive residues, residues from previous products, MOSH/MOAH |
| Environmental contaminants | Substances that originate from the environment during manufacturing, storage, or transport | PAHs (polycyclic aromatic hydrocarbons), PCBs, pesticide residues |
Note: The scope of the NIAS assessment is determined on a case-by-case basis, taking into account the composition of the packaging material, the manufacturing process, and the intended conditions of use. The guidance documents published by EFSA provide the basic methodological framework for the NIAS assessment.
Analytical Strategy for the NIAS Assessment
NIAS assessment is one of the most complex areas of analytical chemistry due to the need to simultaneously screen and identify a large number of known and unknown substances. At the Nano-Test Laboratory, NIAS assessment is conducted using the following multi-step, systematic approach.
Step-by-Step NIAS Assessment Strategy
The NIAS evaluation process used by Nano-Test consists of four main phases, which are summarized in the following table:
| Phase | Step | Explanation |
|---|---|---|
| 1. Preliminary Assessment and Data Collection | Review of information regarding the formulation and manufacturing process | Theoretical identification of all raw materials, additives, production parameters, and potential sources of NIAS (impurities, reaction byproducts, decomposition products) used. During this phase, the supplier declarations and safety data sheets (SDS) provided by the customer are also evaluated. |
| 2. Non-Targeted Screening | Comprehensive screening using high-resolution mass spectrometry (HRMS) | The extract obtained following an appropriate extraction and/or migration test is analyzed using LC-QTOF-MS and GC-QTOF-MS systems. The data obtained are evaluated using advanced data processing software to perform a library search and identify unknown peaks. |
| 3. Identification and semiquantitative determination | Structural characterization of the identified NIAS candidates | The structural characterization of unknown compounds is based on the exact mass, isotopic distribution, and MS/MS fragmentation patterns obtained from the HRMS data. Where possible, validation is performed using reference standards. Semiquantitative estimation is performed using an approach based on internal standards or structural analogs. |
| 4. Toxicological Risk Assessment | Assessment of the identified NIAS with regard to food safety | For each identified NIAS, a risk assessment is conducted using 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 NIAS evaluation, along with their specific roles in NIAS analysis, are compared in the following table:
| Analytical Platform | Technical Specifications | Role in the NIAS analysis | Target Composite 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, EI and CI ionization, extensive library (NIST, Wiley) | Targets: Unbiased screening and identification of volatile and semi-volatile 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 compounds | Screening and Quantification of Volatile NIAS | Residual monomers (styrene, butadiene), solvents, volatile degradation products |
Toxicological Risk Assessment and the 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 provides a scientifically sound tool for risk assessment of substances whose structure is known but for which only limited toxicological data are available.
During this crucial phase of the NIAS evaluation, Nano-Test applies the following systematic approach:
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 | Moderate toxicity potential | 9 | Compounds containing 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 cancer risk of 1 in 10⁵) | Aflatoxin-like structures, N-nitroso compounds, DNA intercalators |
Note: The EFSA does not recommend applying the TTC approach to genotoxic carcinogens. For such substances, minimizing exposure (the ALARA principle) is of fundamental importance. Furthermore, the TTC approach may not be suitable for groups of substances with potentially neurotoxic effects, such as organophosphates and carbamates.
Nano-Test's Integrated NIAS Evaluation Service
At Nano-Test Labor Services, we offer NIAS assessment using a multidisciplinary and integrated approach. Our service model consists of the components summarized in the following table:
The NIAS assessment is not merely an analytical test, but a multifaceted process that requires in-depth chemical and toxicological expertise. As Nano-Test, we operate in accordance with GLP principles, scientific rigor, and a client-focused consulting approach at every stage of this process.To ensure that your packaging, which you export to the European market, complies with NIAS under the PPWR and relevant EU regulations, and to carry out your screening and risk assessment processes with a reliable partner, you can contact our team of experts to receive detailed information about Nano-Test’s NIAS assessment services.
| Service Component | Scope | Benefits for the Customer |
|---|---|---|
| Formulation and Process Analysis | Detailed analysis of the raw materials and additives used, as well as the production process; theoretical mapping of potential NIAS sources | Proper development of the analysis strategy; avoiding unnecessary analyses |
| Non-targeted screening (HRMS) | Comprehensive, non-targeted chemical screening using LC-QTOF-MS and GC-QTOF-MS | Detection of unexpected contaminants and unknown NIAS |
| Structural Analysis and Semi-Quantification | Structural characterization based on HRMS data and semi-quantitative estimation using internal standards | Reliable identification of unknown peaks and determination of their approximate concentrations |
| Toxicological Risk Assessment | Comprehensive risk assessment using the TTC approach, the Cramer classification, and QSAR toxicity prediction (in silico) | Scientifically Based Assessment of the Food Safety Risk Posed by the Identified NIAS |
| Report on Compliance with Regulatory Requirements | Comprehensive report for the DoC (Declaration of Conformity) that includes all analytical results and the risk assessment | Complete documentation for regulatory applications and customer audits |