GC × GC-HRMS with complementary ionization methods in the suspect screening analysis of fragrance allergens: overwhelming or justified?

Modern gas chromatography-mass spectrometry (GC-MS) allows for the analysis of complex samples, such as fragrances. However, identifying all the constituents in natural fragrance mixtures, especially allergens that need to be listed on product labels, is a significant challenge. This is primarily due to the high complexity of the sample and the fact that electron ionization, the most commonly used ionization method in GC-MS, produces numerous nonspecific fragment ions, often resulting in the absence or very low abundance of the molecular ion. These factors affect confidence in assigning the analyte. In this study, we demonstrate that the combination of GC × GC separation, with high mass resolution and accurate mass measurements, as well as chemical ionization in addition to traditional electron ionization, becomes an efficient tool for reliable qualitative analysis of a mixture containing 100 fragrance allergens, even when many of them are closely related species or isomers. The proposed approach expands the applicability of the comprehensive GC × GC-HRMS method, which includes complementary ionization techniques, from studies on anthropogenic priority pollutants and emerging contaminants to the analysis of natural products. Although targeted qualitative and quantitative analysis of allergens in the modern laboratories is well organized, GC × GC-HRMS, being a useful complement to routine quality control of volatile allergens in fragrances, definitely gives an additional contribution to the analytical cases when conventional 1D-GC-MS faces some problems or uncertainties.

Effect of the Combined Ultrasound with Other Technologies on Food Allergenicity: Ultrasound before, under, and after Other Technologies.

Food allergies are a main public health disease in the world. Ultrasound is an environmentally friendly technology that typically leads to protein unfolding and loss of protein structure, which means it has the potential to be combined with other technologies to achieve a great reduction of allergenicity in foods. This review concludes the effects of the combined ultrasound with other technologies on food allergenicity from three combinations: ultrasound before other technologies, ultrasound under other technologies, and ultrasound after other technologies. Each combination affects food allergenicity through different mechanisms: (1) as for ultrasound before other technologies, ultrasound pretreatment can unfold and lose the protein structure to improve the accessibility of other technologies to epitopes; (2) as for ultrasound under other technologies, ultrasound can continuously affect the accessibility of other technologies to epitopes; (3) as for ultrasound after other technologies, ultrasound further induces structural changes to mask and disrupt the epitopes. The reduction of allergenicity is related to the ultrasound/other technologies conditions and food types/cultivars, etc. The comparison of ultrasound before, under, and after other technologies to decrease food allergenicity should be further investigated in the future. The combination of ultrasound with other technologies is promising to produce hypoallergenic foods.

Microfluidic Chip Coupled with MALDI-TOF MS for Multitarget Detection of Allergens in Crucian Carp (Carassius auratus).

The goal of the present study was to establish a rapid, simple method for simultaneous allergy testing of sera from multiple fish-allergic patients. Sera from fish-allergic patients were pooled and used for capturing allergens in fish muscle of crucian carp (Carassius auratus), which was studied as a fish model. Sarcoplasmic proteins of crucian carp (Carassius auratus) were extracted for the analysis of allergens. Anti-human IgE antibody-functionalized magnetic beads were utilized to collect IgE antibodies from human pooled sera. The isolation of allergenic proteins was immunomagnetically performed in microfluidic channels, and the elution of the captured allergenic proteins was done with 5% (v/v) acetic acid aqueous solution. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and peptide mass fingerprinting were used for the analysis of tryptic digests of eluted proteins. Ten potential allergenic proteins were identified from crucian carp (Carassius auratus). The present protocol provides a rapid, efficient, and simple method for simultaneous detection of multiple allergens, based on multitargeted antibodies from pooled sera of allergic patients. The constructed multiple antibody-modified MBs can be applied for the deallergenicity of food matrices. The efficiency of allergen detection can be greatly improved, with promising application in allergen discovery and filtration for other muscle-based foods.

[Analysis of allergen distribution and indoor factors in allergic rhinitis in Chaoshan area].

Objective:To investigate the distribution of common allergens and indoor factors influencing the severity of allergic rhinitis in patients from the Chaoshan region. Methods:Patients diagnosed with allergic rhinitis from Shantou, Jieyang, and Chaozhou were selected for serum allergen-specific IgE testing. A questionnaire survey was conducted to analyze the distribution of allergens and indoor factors affecting the severity of the disease. Results:A total of 1 800 questionnaires were collected, with 1 646 valid responses, resulting in an effective response rate of 91.4%. Among the 1 646 included patients with allergic rhinitis, there were 1 285 children（≤14 years） ，361 adolescents and adults（>14 years）；of which 999 were males and 647 were females. The top three allergens with the highest positive rates were house dust mites（n=1 457, 88.5%）, milk（n=569, 34.6%）, and crab（n=360, 21.9%）. The proportions of allergen sensitization to house dust mites, house dust, dog dander, egg white, milk, fish, crab, shrimp, and beef showed statistically significant differences between children and adolescents and adults（P<0.01）. There were also statistically significant differences in crab and shrimp sensitization between males and females（P<0.01）. Multivariate analysis revealed that active/passive smoking, religious rituals, air conditioning usage, pet ownership, air purifier usage, and bedding drying were indoor factors influencing the severity of allergic rhinitis. Among them, active/passive smoking, religious rituals, air conditioning usage, and pet ownership were risk factors for exacerbating the disease, while air purifier usage and bedding drying were protective factors. Conclusion:House dust mites are the most common allergen in patients with allergic rhinitis in the Chaoshan region. Active/passive smoking, religious rituals, air conditioning usage, and pet ownership can worsen the condition, while air purifier usage and bedding drying can help control the disease. The results of this study can provide clinical reference.

Frequency of perfume in 398 children's cosmetics

Abstract Objective: Perfume (Parfum) or fragrance is a natural or synthetic cosmetic ingredient added to emit a pleasant aroma or to improve the odor of a cosmetic formula. It is a mixture of substances, not revealed by the manufacturer, which may contain ingredients with allergenic potential, endocrine disruptors, and other possible harmful effects on human health. This study aims to analyze children's cosmetics labels to assess the presence of Perfume. Methods: The researchers randomly visited points of sale in Curitiba, the capital of a southern Brazilian state; in order to catalog the largest possible number of children's cosmetics items. Results: 398 children's cosmetics were analyzed and found Parfum on 295 (74.1 %) of the labels, including 90.4 and 79,1 % of the shampoos and wet wipes, respectively. Conclusion: Exposure of children's skin to fragrances can lead to local side effects such as allergies, but also to systemic effects, and the lack of knowledge of the general population and health professionals about its possible deleterious effects emphasizes the importance of changes in the regulation of cosmetics aiming to reduce the use of this ingredient.

A prospective study evaluating the correlation between local weather conditions, pollen counts and pruritus of dogs with atopic dermatitis.

BACKGROUND: Canine atopic dermatitis (cAD) is a hereditary, generally pruritic and predominantly T-cell-driven inflammatory skin disease, involving an interplay between skin barrier abnormalities, allergen sensitisation and microbial dysbiosis. The individual immunological response is predominantly against environmental allergens, including mite antigens; mould spores; and pollen from grasses, trees and weeds. Airborne pollens show fluctuating patterns during the year. OBJECTIVE: The aim of this prospective study was to evaluate the influence of local pollen concentrations and weather conditions on the clinical signs of atopic dogs, and to investigate any possible correlations with the results of intradermal testing (IDT). MATERIALS AND METHODS: Thirty-seven privately owned atopic dogs in Bavaria were surveyed from 1 April to 30 November 2021. Owners were asked to record pruritus using a validated Visual Analog Scale (PVAS) score and the weekly medication of their dog. Furthermore, weather data, including pollen count, rainfall, relative humidity, hours of sunshine and temperature from the dog's location were collected daily. RESULTS: Of the evaluated parameters, only humidity and medication scores correlated positively with the PVAS scores of the atopic dogs. There was no correlation between specific pollen counts and PVAS scores of dogs with positive IDT reactions to that pollen. CONCLUSION AND CLINICAL RELEVANCE: The outcome of this study highlights the importance of a careful interpretation of positive IDT results in dogs with cAD and questions the validity of airborne pollen trap methodology in representing pollen exposure for dogs at ground level.

IgE-Mediated Sensitization to Blo t 21 and Blo t 5 Is Associated With Asthma in the Tropics: A Case-Control Study.

BACKGROUND AND OBJECTIVE: Sensitization to Blomia tropicalis is associated with asthma in various tropical and subtropical countries; however, information about the specific molecular components associated with this disease is scarce. Using molecular diagnosis, we sought to identify B tropicalis allergens associated with asthma in Colombia. METHODS: Specific IgE (sIgE) to 8 B tropicalis recombinant allergens (Blo t 2, 5, 7, 8, 10, 12, 13, and 21) was determined using an in-house ELISA system in asthma patients (n=272) and controls (n=298) recruited in a national prevalence study performed in several Colombian cities (Barranquilla, Bogotá, Medellín, Cali, and San Andrés). The study sample included children and adults (mean [SD] age, 28 [17] years). Cross-reactivity between Blo t 5 and Blo t 21 was evaluated using ELISA-inhibition. RESULTS: Specific IgE (sIgE) to 8 B tropicalis recombinant allergens (Blo t 2, 5, 7, 8, 10, 12, 13, and 21) was determined using an in-house ELISA system in asthma patients (n=272) and controls (n=298) recruited in a national prevalence study performed in several Colombian cities (Barranquilla, Bogotá, Medellín, Cali, and San Andrés). The study sample included children and adults (mean [SD] age, 28 [17] years). Cross-reactivity between Blo t 5 and Blo t 21 was evaluated using ELISA-inhibition. CONCLUSION: Although Blo t 5 and Blo t 21 are considered common sensitizers, this is the first report of their association with asthma. Both components should be included in molecular panels for diagnosis of allergy in the tropics.

Worldwide Heterogeneity of Food Allergy: Focus on Peach Allergy in Southern Italy.

Food allergy (FA) has shown an increasing prevalence in the last decades, becoming a major public health problem. However, data on the prevalence of FA across the world are heterogeneous because they are influenced by several factors. Among IgE-mediated FA, an important role is played by FA related to plant-derived food which can result from the sensitization to a single protein (specific FA) or to homologous proteins present in different foods (cross-reactive FA) including non-specific lipid transfer proteins (nsLTPs), profilins, and pathogenesis-related class 10 (PR-10). In addition, the clinical presentation of FA is widely heterogeneous ranging from mild symptoms to severe reactions up to anaphylaxis, most frequently associated with nsLTP-related FA (LTP syndrome). Considering the potential life-threatening nature of nsLTP-related FA, the patient's geographical setting should always be taken into account; thereby, it is highly recommended to build a personalized approach for managing FA across the world in the precision medicine era. For this reason, in this review, we aim to provide an overview of the prevalence of nsLTP-mediated allergies in the Mediterranean area and to point out the potential reasons for the different geographical significance of LTP-driven allergies with a particular focus on the allergenic properties of food allergens and their cross reactivity.

A sensitive CRISPR/Cas12a-assisted fluorescent aptasensor for rapid detection of food allergens.

Food allergens elicit abnormal immune system responses among allergic individuals and sensitive detection for allergenic ingredient is greatly significant. To address this need, a novel fluorescent aptasensor, assisted by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR), have been developed for food allergens. In this study, aptamer offers distinctive recognition capabilities in binding specific targets, while CRISPR-associated-12a protein (Cas12a) holds precise cis-cleavage for cutting fluorescent signal probes. Notably, the utilization of Cas12a cis-cleavage activity, rather than trans-cleavage, eliminates the necessity for additional fluorescent probes, thus reducing interference between substances and enhancing sensitivity. Throughout the process, complementary DNA (cDNA) plays a crucial dual role in target recognition conversion and signal presentation, representing a key challenge and innovative aspect of this study. To evaluate the performance of the aptasensor, lysozyme (LYS) is employed as a representative model target of food allergens. Under optimal conditions, the developed aptasensor could achieve an exceptional low limit of detection (LOD) of 6.10 pM with a dynamic detection range of 10 pM-320 pM. The aptasensor demonstrates high selectivity and great recovery rates. This strategy yields promising outcomes, holding the potential to serve as a valuable reference for various food allergens detection.

Cross-Reactivity of Ragweed Pollen Calcium-Binding Proteins and IgE Sensitization in a Ragweed-Allergic Population from Western Romania.

Ragweed pollen allergy is the most common seasonal allergy in western Romania. Prolonged exposure to ragweed pollen may induce sensitization to pan-allergens such as calcium-binding proteins (polcalcins) and progression to more severe symptoms. We aimed to detect IgE sensitization to recombinant Amb a 9 and Amb a 10 in a Romanian population, to assess their potential clinical relevance and cross-reactivity, as well as to investigate the relation with clinical symptoms. rAmb a 9 and rAmb a 10 produced in Escherichia coli were used to detect specific IgE in sera from 87 clinically characterized ragweed-allergic patients in ELISA, for basophil activation experiments and rabbit immunization. Rabbit rAmb a 9- and rAmb a 10-specific sera were used to detect possible cross-reactivity with rArt v 5 and reactivity towards ragweed and mugwort pollen extracts. The results showed an IgE reactivity of 25% to rAmb a 9 and 35% to rAmb a 10. rAmb a 10 induced basophil degranulation in three out of four patients tested. Moreover, polcalcin-negative patients reported significantly more skin symptoms, whereas polcalcin-positive patients tended to report more respiratory symptoms. Furthermore, both rabbit antisera showed low reactivity towards extracts and showed high reactivity to rArt v 5, suggesting strong cross-reactivity. Our study indicated that recombinant ragweed polcalcins might be considered for molecular diagnosis.

Long-term pollen season trends of Fraxinus (ash), Quercus (oak) and Ambrosia artemisiifolia (ragweed) as indicators of anthropogenic climate change impact.

The ongoing climatic change, together with atmospheric pollution, influences the timing, duration and intensity of pollen seasons of some allergenic plant taxa. To study these influences, we correlated the trends in the pollen season characteristics of both woody (Fraxinus, Quercus) and herbaceous (Ambrosia) taxa from two pollen monitoring stations in Slovakia with the trends in meteorological factors and air pollutants during the last two decades. In woody species, the increased temperature during the formation of flower buds in summer and autumn led to an earlier onset and intensification of next year's pollen season, especially in Quercus. The increase of relative air humidity and precipitation during this time also had a positive influence on the intensity of the pollen season of trees. The pollen season of the invasive herbaceous species Ambrosia artemisiifolia was prolonged by increased temperature and humidity during the summer and autumn of the same year, which extended the blooming period and delayed the end of the pollen season. From the studied air pollutants, only three were found to correlate with the intensity of the pollen season of the studied taxa, CO - positively and SO2 and NO2 - negatively. It is important to study these long-term trends since they not only give us valuable insight into the response of plants to changing conditions but also enable the prognosis of the exacerbations of pollen-related allergenic diseases.

[Declaration of allergens on food labels: overview of Latin AmericaDeclaração de alergênicos na rotulagem de alimentos: panorama da América Latina]. / Declaración de alergenos en el etiquetado de alimentos: panorama latinoamericano.

Objective: Identify countries that have legislation on mandatory declarations of food allergens, irradiated foods, and transgenic foods on the nutritional labels of packaged foods. Methods: Exploratory study reviewing the health regulations and technical standards for foods in Latin American countries in order to gather information on declarations of allergens, trace allergens, irradiated foods, and transgenic foods. The information search was carried out through the countries' government web pages. Presentation of the results is descriptive and narrative. Results: Of the 19 countries reviewed, 89% require a declaration of allergens on their nutrition labeling, 76% have legislation that explicitly require a statement on trace allergens, and 82% follow Codex Alimentarius recommendations with some modifications of food categories. Conclusions: Three pending challenges in the Region are: requiring statements on allergens as a food safety measure; making progress toward improved labeling of trace allergens; and ensuring universal availability of epinephrine.

Objetivo: Identificar os países que têm legislação relacionada à declaração obrigatória de alimentos alergênicos, irradiados e transgênicos na rotulagem nutricional de alimentos embalados. Métodos: Estudo exploratório com revisão dos regulamentos sanitários ou normas técnicas de alimentos dos países da América Latina, a fim de coletar informações sobre a declaração de alimentos alergênicos, traços de alergênicos, alimentos irradiados e transgênicos na rotulagem nutricional. A busca de informações foi realizada por meio dos sites governamentais dos países. Os resultados são apresentados de forma descritiva e narrativa. Resultados: Dos 19 países analisados, 89% declaram alergênicos na rotulagem nutricional, 76% incorporam explicitamente a declaração de traços na legislação e 82% seguem as recomendações do Codex Alimentarius, com algumas modificações nas categorias de alimentos. Conclusões: Entre os desafios pendentes na Região estão a implementação da declaração de alergênicos como medida de segurança alimentar e a melhoria da rotulagem de traços de alergênicos e da disponibilidade universal de epinefrina.

Impact of Aeroallergen Sensitization on Chronic Rhinosinusitis.

PURPOSE: This study investigated the impact of aeroallergens on the development and progression of chronic rhinosinusitis (CRS), with a focus on the specific associations between aeroallergens and CRS according to allergen type, number, and extent of sensitization. METHODS: The medical records of 256 CRS patients were retrospectively analyzed. All were divided into nonallergic, house dust mite (HDM)-allergic, pollen-allergic, and double allergic groups via specific immunoglobulin E (IgE) testing. Clinical characteristics, computed tomography (CT) scores, olfactory functions, and demographic data were compared. Correlation analysis was performed to explore the relationships between the extent of allergen sensitization and CRS severity. Binary logistic regression analysis was used to identify risk factors for hyposmia and anosmia. RESULTS: The allergic group exhibited higher total CT scores than the nonallergic group (P = 0.001). Sensitivity to HDM or pollen allergens alone was not significantly associated with increased CRS severity. No significant differences were observed between the effects of HDM and pollen allergens on CRS severity. However, the double allergic group exhibited significantly higher CT scores (P < 0.001, < 0.001, and 0.003) than the other groups. Although the prevalence rates of anosmia and hyposmia were notably higher in the double allergic group, the difference was not statistically significant. The maximum specific IgE levels to HDM and pollen allergens positively correlated with the CT scores (P = 0.001 and 0.001, respectively). CONCLUSIONS: Allergen sensitization, particularly to multiple common allergens, contributed to CRS severity. CRS patients sensitized to both HDM and pollen allergens tended to experience the diminished olfactory function. These findings underscore the importance of considering the allergen sensitization pattern when assessing CRS severity and its potential progression.

Lipidomics reveals the serum profiles of pediatric allergic rhinitis and its severity.

Allergic rhinitis (AR) is a prevalent upper airway chronic inflammatory disease in children worldwide. The role of bioactive lipids in the regulation of AR has been recognized, but the underlying serum lipidomic basis of its pathology remains unclear. We utilized ultra-performance liquid chromatography (UPLC)-Q-Exactive Orbitrap/mass spectrometry (MS) to investigate the serum lipidomic profiles of children with AR. The lipidomic analysis identified 42 lipids that were differentially expressed (p < 0.05, fold change > 2) between the AR (n = 75) and normal control groups (n = 44). Specifically, the serum levels of diacylglycerol (DG), triacylglycerol (TG), fatty acid (FA), lysophosphatidylcholine (LPC), lysophosphatidylethanolamine, phosphatidyl-ethanolamine, and cardiolipins were significantly higher in the AR group. The diagnostic potential of the identified lipids was further evaluated using receiver operating characteristic curve analysis. The analysis revealed that five lipids, including FA 30:7, LPC O-18:1, LPC 18:0, LPC 16:0, and DG 34:0, had area under the curve values greater than 0.9 (p < 0.05). Furthermore, serum levels of IgE and IL-33, markers of AR severity, were found to have a significant positive correlation (p < 0.05) with DGs, LPCs, TGs, and FAs in AR patients. This study revealed the lipid disorders associated with AR and its severity, providing new insights into the pathological process of AR.

Algae-derived compounds: Bioactivity, allergenicity and technologies enhancing their values.

As a rapidly growing source of human nutrients, algae biosynthesize diverse metabolites which have promising bioactivities. However, the potential allergenicity of algal components hinder their widespread adoption. This review provides a comprehensive review of various macro and micronutrients derived from algal biomass, with particular focus on bioactive compounds, including peptides, polyphenols, carotenoids, omega-3 fatty acids and phycocyanins. The approaches used to produce algal bioactive compounds and their health benefits (antioxidant, antidiabetic, cardioprotective, anti-inflammatory and immunomodulatory) are summarised. This review particularly focuses on the state-of-the-art of precision fermentation, encapsulation, cold plasma, high-pressure processing, pulsed electric field, and subcritical water to reduce the allergenicity of algal compounds while increasing their bioactivity and bioavailability. By providing insights into current challenges of algae-derived compounds and opportunities for advancement, this review contributes to the ongoing discourse on maximizing their application potential in the food nutraceuticals, and pharmaceuticals industries.

Efficient experimental method for purifying allergens from aqueous extracts.

Studying the molecular and immunological basis of allergic diseases often requires purified native allergens. The methodologies for protein purification are usually difficult and may not be completely successful. The objective of this work was to describe a methodology to purify allergens from their natural source, while maintaining their native form. The purification strategy consists of a three-step protocol and was used for purifying five specific allergens, Ole e 1, Amb a 1, Alt a 1, Bet v 1 and Cup a 1. Total proteins were extracted in PBS (pH 7.2). Then, the target allergens were pre-purified and enriched by salting-out using increasing concentrations of ammonium sulfate. The allergens were further purified by anion exchange chromatography. Purification of Amb a 1 required an extra step of cation exchange chromatography. The detection of the allergens in the fractions obtained were screened by SDS-PAGE, and Western blot when needed. Further characterization of purified Amb a 1 was performed by mass spectrometry. Ole e 1, Alt a 1, Bet v 1 and Cup a 1 were obtained at > 90 % purity. Amb a 1 was obtained at > 85 % purity. Overall, we propose an easy-to-perform purification approach that allows obtaining highly pure allergens. Since it does not involve neither chaotropic nor organic reagents, we anticipate that the structural and biological functions of the purified molecule remain intact. This method provides a basis for native allergen purification that can be tailored according to specific needs.

Comparative Seed Proteome Profile Reveals No Alternation of Major Allergens in High-Yielding Mung Bean Cultivars.

Mung bean contains up to 32.6% protein and is one of the great sources of plant-based protein. Because many allergens also function as defense-related proteins, it is important to determine their abundance levels in the high-yielding, disease-resistant cultivars. In this study, for the first time, we compared the seed proteome of high-yielding mung bean cultivars developed by a conventional breeding approach. Using a label-free quantitative proteomic platform, we successfully identified and quantified a total of 1373 proteins. Comparative analysis between the high-yielding disease-resistant cultivar (MC5) and the other three cultivars showed that a total of 69 common proteins were significantly altered in their abundances across all cultivars. Bioinformatic analysis of these altered proteins demonstrated that PDF1 (a defensin-like protein) exhibited high sequence similarity and epitope matching with the established peanut allergens, indicating a potential mung bean allergen that showed a cultivar-specific response. Conversely, known mung bean allergen proteins such as PR-2/PR-10 (Vig r 1), Vig r 2, Vig r 4, LTP1, ß-conglycinin, and glycinin G4 showed no alternation in the MC5 compared to other cultivars. Taken together, our findings suggest that the known allergen profiles may not be impacted by the conventional plant breeding method to develop improved mung bean cultivars.

Performance and overview of clinically relevant areas of application of saliva testing in the cat.

Introduction: The cat represents an important model in order to investigate basic physiological knowledge of salivary secretion as well as pharmacokinetics of active substances. Objective: The aim of the study was to review in which diagnostic application areas saliva testing is routinely used and in which areas it could be further explored in the future. Materials and methods: Literature relevant to the research question was collected in March 2022 using the Pubmed database. Results: The diagnosis of infectious diseases in cat saliva is one of the most important fields of application. Saliva diagnostics may also indicate dental diseases, allergies or kidney and other metabolic diseases. Sexual and stress hormones can also be measured in cat saliva. A number of clinically relevant allergens in cat saliva that may cause allergies in humans has been investigated and described, in addition to infectious agents that can be transmitted from cats to humans. Conclusions: Saliva testing in cats can be useful in many areas, including the detection of infectious diseases, allergies and dental disease. However, it is far from being used to its full potential within veterinary medicine.

Diagnostics of IgE-mediated occupational allergies: Between reality, requirements, and opportunities.

Occupational skin and respiratory allergies are among the most common occupational diseases in Germany. The identification of the allergy trigger is essential for the recognition of an occupational allergy as well as for effective individual prevention. However, occupational type I allergens are among the "rare" allergens and the possibilities of guideline-compliant diagnosis using quality-tested skin test solutions is becoming increasingly difficult due to the reduction in commercially available test allergens. In order to guarantee meaningful diagnostic workup for all affected insured persons with suspected occupational type I allergies and to ensure this in the future, a durable optimization, standardization, and availability of allergy tests for occupational allergic diseases is urgently required. The need for action has been recognized by the German Social Accident Insurance (DGUV), and steps to eliminate the diagnostic gaps have been initiated by a joint research project at the Institute for Prevention and Occupational Medicine of the DGUV (IPA) and the Paul Ehrlich Institut (PEI). The evaluation of alternative methods for the production of standardized test allergen solutions can also be used for newly emerging allergens in the workplace. New allergen sources at workplaces and thus also sensitization and allergies among employees can be expected as a result of changes in work processes and the introduction of new technologies and/or working materials, which are also introduced in connection with climate change and the concept of sustainability.