Quantitative analysis of physical and chemical measurands in honey by mid-infrared spectrometry

Fourier transform infrared spectroscopy (FT-IR) was used to determine 20 different measurands in honey. The reference values for 144 honey samples of different botanical origin were determined by classical physical and chemical methods. Partial least squares regression was used to develop the calibration models for the measurands studied. They were validated using independent samples and proved satisfying accuracies for the determination of water (R 2=0.99), glucose (0.94), fructose (0.84), sucrose (0.91), melezitose (0.98) and monosaccharide content (0.82) as well as fructose/glucose ratio (0.98), glucose/water ratio (0.94), electrical conductivity (0.98), pH-value (0.87) and free acidity (0.96). The prediction accuracy for hydroxymethylfurfural, proline and the minor sugars maltose, turanose, erlose, trehalose, isomaltose and kojibiose was rather poor. The results demonstrate that mid-infrared spectrometry is a valuable, rapid and non-destructive tool for the quantitative analysis of the most important measurands in hone

Authentication of the botanical origin of honey using profiles of classical measurands and discriminant analysis

The potential of physical and chemical measurands for the determination of the botanical origin of honey by using both the classical profiling approach and chemometrics was evaluated for the authentication of ten unifloral (acacia, rhododendron, chestnut, dandelion, heather, lime, rape, fir honeydew, metcalfa honeydew) and polyfloral honey types (in total n = 693 samples). The classical approach using a profile for the determination of the botanical origin of honey revealed that the physical and chemical measurands alone do not allow a reliable determination. Pollen analysis is therefore essential for discrimination between unifloral and polyfloral honeys. However, chemometric evaluation of the physical and chemical data by linear discriminant analysis allowed reliable authentication with neither specialized expertise nor pollen or sensory analysis. The error rates calculated by Bayes' theorem ranged from 1.1% (rape and lime honeys) up to 9.9 % (acacia honey

An international proficiency test to detect, identify and quantify ricin in complex matrices

While natural intoxications with seeds of Ricinus communis have long been known, the toxic protein ricin contained in the seeds is of major concern due to its history of criminal, terrorist and military use. In order to harmonize detection capabilities in expert laboratories an international proficiency test was organized that aimed at identifying good analytical practices (qualitative measurements) and determining a consensus concentration on a highly pure ricin reference material (quantitative measurements). Sample materials included highly pure ricin as well as the related R. communis agglutinin spiked into buffer, milk and meat extract; additionally, an organic fertilizer naturally contaminated with R. communis shred was investigated in the proficiency test. The qualitative results showed that either a suitable combination of immunological, MS-based and functional approaches or sophisticated MS-based approaches alone successfully allowed to detect and identify ricin in all samples. In terms of quantification, it was possible to determine a consensus concentration for the highly pure ricin reference material. The results provide a basis for further steps in quality assurance and improve biopreparedness in expert laboratories worldwide.JRC.D.2-Standards for Innovation and sustainable Developmen

Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test

In the framework of the EU project EQuATox a first international proficiency test (PT) on the detection and quantification of BoNT was conducted. Sample materials included BoNT serotypes A, B and E spiked into buffer, milk, meat extract and serum. A variety of methods was applied by the participants combining different principles of detection, identification and quantification. Based on qualitative assays, 95% of all results reported were correct. Successful strategies for BoNT detection were based on a combination of complementary immunological, MS-based and functional methods or on suitable functional in vivo / in vitro approaches (mouse bioassay, hemidiaphrama assay, Endopep-MS assay). Quantification of BoNT/A, BoNT/B and BoNT/E was performed by 48% of participating laboratories. It turned out that precise quantification of BoNT was difficult resulting in a substantial scatter of quantitative data. This was especially true for results obtained by the mouse bioassay which is currently seen as “gold standard” for BoNT detection. The results clearly demonstrate the urgent need of certified BoNT reference materials and the development of methods replacing animal testing. In this context, the BoNT PT provided the valuable information that both the Endopep-MS assay and the hemidiaphrama assay delivered quantitative results superior to the mouse bioassay.JRC.D.2-Standards for Innovation and sustainable Developmen

Quantitative determination of physical and chemical measurands in honey by near-infrared spectrometry

Fourier transform near-infrared spectroscopy (FT-NIR) was evaluated to quantitatively determine 24 different measurands in honey. The reference values of 421 honey samples of different botanical origins were determined by classical physical and chemical methods. Partial least squares regression was used to develop the calibration models for the measurands studied. These calibrations were then validated using independent samples and proved satisfying accuracies for the determination of water (standard error of prediction: 0.3g/100g), glucose (1.3g/100g), fructose (1.6g/100g), sucrose (0.4g/100g), total monosaccharide content (2.6g/100g) as well as fructose/glucose ratio (0.09) and glucose/water ratio (0.12). The prediction accuracy for hydroxymethylfurfural, proline, pH-value, electrical conductivity, free acidity and the minor sugars maltose, turanose, nigerose, erlose, trehalose, isomaltose, kojibiose, melezitose, raffinose, gentiobiose, melibiose, maltotriose was poor and unreliable. The results demonstrate that near-infrared spectrometry is a valuable, rapid and non-destructive tool for the quantitative analysis of some measurands related to the main components in hone

Optimization of Sample Preparation for the Identification and Quantification of Saxitoxin in Proficiency Test Mussel Sample using Liquid Chromatography-Tandem Mass Spectrometry

Saxitoxin (STX) and some selected paralytic shellfish poisoning (PSP) analogues in mussel samples were identified and quantified with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Sample extraction and purification methods of mussel sample were optimized for LC-MS/MS analysis. The developed method was applied to the analysis of the homogenized mussel samples in the proficiency test (PT) within the EQuATox project (Establishment of Quality Assurance for the Detection of Biological Toxins of Potential Bioterrorism Risk). Ten laboratories from eight countries participated in the STX PT. Identification of PSP toxins in naturally contaminated mussel samples was performed by comparison of product ion spectra and retention times with those of reference standards. The quantitative results were obtained with LC-MS/MS by spiking reference standards in toxic mussel extracts. The results were within the z-score of +/- 1 when compared to the results measured with the official AOAC (Association of Official Analytical Chemists) method 2005.06, pre-column oxidation high-performance liquid chromatography with fluorescence detection (HPLC-FLD).Peer reviewe

Results of a Saxitoxin Proficiency Test Including Characterization of Reference Material and Stability Studies

A saxitoxin (STX) proficiency test (PT) was organized as part of the Establishment of Quality Assurance for the Detection of Biological Toxins of Potential Bioterrorism Risk (EQuATox) project. The aim of this PT was to provide an evaluation of existing methods and the European laboratories' capabilities for the analysis of STX and some of its analogues in real samples. Homogenized mussel material and algal cell materials containing paralytic shellfish poisoning (PSP) toxins were produced as reference sample matrices. The reference material was characterized using various analytical methods. Acidified algal extract samples at two concentration levels were prepared from a bulk culture of PSP toxins producing dinoflagellate Alexandrium ostenfeldii. The homogeneity and stability of the prepared PT samples were studied and found to be fit-for-purpose. Thereafter, eight STX PT samples were sent to ten participating laboratories from eight countries. The PT offered the participating laboratories the possibility to assess their performance regarding the qualitative and quantitative detection of PSP toxins. Various techniques such as official Association of Official Analytical Chemists (AOAC) methods, immunoassays, and liquid chromatography-mass spectrometry were used for sample analyses.Peer reviewe

Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test

In the framework of the EU project EQuATox, a first international proficiency test (PT) on the detection and quantification of botulinum neurotoxins (BoNT) was conducted. Sample materials included BoNT serotypes A, B and E spiked into buffer, milk, meat extract and serum. Different methods were applied by the participants combining different principles of detection, identification and quantification. Based on qualitative assays, 95% of all results reported were correct. Successful strategies for BoNT detection were based on a combination of complementary immunological, MS-based and functional methods or on suitable functional in vivo/in vitro approaches (mouse bioassay, hemidiaphragm assay and Endopep-MS assay). Quantification of BoNT/A, BoNT/B and BoNT/E was performed by 48% of participating laboratories. It turned out that precise quantification of BoNT was difficult, resulting in a substantial scatter of quantitative data. This was especially true for results obtained by the mouse bioassay which is currently considered as “gold standard” for BoNT detection. The results clearly demonstrate the urgent need for certified BoNT reference materials and the development of methods replacing animal testing. In this context, the BoNT PT provided the valuable information that both the Endopep-MS assay and the hemidiaphragm assay delivered quantitative results superior to the mouse bioassay

Authentication of the botanical origin of honey using profiles of classical measurands and discriminant analysis

The potential of physical and chemical measurands for the determination of the botanical origin of honey by using both the classical profiling approach and chemometrics was evaluated for the authentication of ten unifloral (acacia, rhododendron, chestnut, dandelion, heather, lime, rape, fir honeydew, metcalfa honeydew) and polyfloral honey types (in total n = 693 samples). The classical approach using a profile for the determination of the botanical origin of honey revealed that the physical and chemical measurands alone do not allow a reliable determination. Pollen analysis is therefore essential for discrimination between unifloral and polyfloral honeys. However, chemometric evaluation of the physical and chemical data by linear discriminant analysis allowed reliable authentication with neither specialized expertise nor pollen or sensory analysis. The error rates calculated by Bayes' theorem ranged from 1.1% (rape and lime honeys) up to 9.9 % (acacia honey)