Liquid-liquid extraction followed by solid-phase extraction for the determination of lipophilic pesticides in beeswax by gas chromatography-electron-capture detection and matrix-matched calibration

Analytical methods for the simultaneous analysis of lindane, chlorpyriphos, z-chlorfenvinphos, endosulfan A and B, 4,4'-DDE, 4,4'-TDE, acrinathrine, bromopropylate, tetradifon, coumaphos and fluvalinate in pure beeswax samples are studied. For the analysis of bleached beeswaxes, a liquid-liquid extraction with acetonitrile followed by a clean-up on polymeric cartridges is the best option in terms of recovery and precision. However, some interferences that hinder the identification and quantification of important varroacides are found when non-bleached beeswaxes are analyzed. The analysis of all compounds in the latter samples require a clean-up by coupling an ODS cartridge before the polymeric cartridge. Considerations about the influence of the matrix in the quantitative analysis by a classical external standard calibration are also made and the use of a matrix-matched calibration is advised. Recoveries resulted to be about 100% with coefficients of variation between 10% and 20% (n = 5) for concentrations of 0.5 and 5 mg/kg.     

Investigating the history of prehistoric glues by gas chromatography-mass spectrometry

Although organic materials are very sensitive to biochemical alteration processes, they may be preserved for millennia in various archaeological contexts. Remains of adhesives made during prehistory were discovered at different sites, in the form of residues adhering to flint tools and ceramic vessels or as free lumps in sediment. To characterise the natural substances exploited for adhesive production during late prehistory, we undertook GC and GC/MS analysis of 90 samples from 8 sites dating from the Neolithic to Iron Age periods. This paper discusses our approach to the study of organic adhesives preserved in archaeological contexts, with a particular focus on the presentation of the various categories of organic adhesives that we analysed and the choice of chromatographic conditions adapted to the specificity of such samples. The results obtained show that birch bark tar, a triterpenoid adhesive made by destructive distillation of white birch bark, was predominantly used during the neolithic period even though other materials such as various barks or organic fossil substance were also used. During the Bronze and Iron ages, which follow the Neolithic period, adhesive production is evolving through the expansion of the range of the natural substances used (identification of diterpenoid pine resin) and the addition of beeswax as a plasticiser to birch bark tar. By combining chromatographic analysis and archaeological data, it was thus possible to follow the evolution of adhesive making at the end of prehistory, testifying to the inventiveness of the craftsmen whatever the period considered.     

Ultrasonics as a tool for development of pine-needle extract loaded bee wax edible packaging for value addition of Himalayan cheese

In the present study, Himalayan cheese, kradi was coated with beeswax loaded with pine needle extract (PNE) to increase its shelf life and nutraceutical potential. PNE was extracted via ultrasonication and incorporated into beeswax at concentrations, 2:1, 1:1, and 2:3 (grams of beeswax to mL of PNE). The dispersion of PNE in the coatings was carried out using an ultrasonic probe at a frequency of 20 kHz for 15 min and at power rating of 500 W. The coatings were characterised using scanning electron microscopy, light microscopy, dynamic light scattering (DLS), fourier transmission infrared spectroscopy. DLS revealed a hydrodynamic diameter and zeta potential of 12.11 ± 0.41 µm and −19.32 ± 0.61 mV for coating loaded with highest concentration of PNE. The bioactivities of the coating including antioxidant, antidiabetic and antibacterial assays revealed significantly higher values with the increase in PNE concentration. Shelf life and sensory evaluation study including microbiological and sensory analysis revealed inhibition of mould growth and good score of texture and appearance with the increase in concentration of PNE. The study provides a future perspective for application of beeswax loaded PNE coatings in cheese industry.     

Py-GC/MS, GC/MS and FTIR investigations on LATE Roman-Egyptian adhesives from opus sectile: New insights into ancient recipes and technologies

An analytical protocol based on optical microscopy, Fourier transforms infrared spectroscopy (FTIR), analytical pyrolysis in the presence of hexamethyldisilazane followed by gas chromatographic/mass spectrometric analysis (Py-GC/MS) and gas chromatography/mass spectrometry after alkaline hydrolysis, solvent extraction and trimethylsilylation (GC/MS) was used in the chemical characterisation of the original adhesives used to fix monochrome and mosaic glass and stone plaques coming from the Late Roman archaeological site of Antinoopolis (Egypt).FTIR analysis demonstrated the presence of calcite fragments, and Py-GC/MS and GC/MS analyses provided detailed molecular compositions, highlighting the presence of a wide range of compound classes including diterpenoid acids, tricyclic abietanes with a high degree of aromatisation, mid- and long-chain monocarboxylic fatty acids, mono- and di-hydroxy acids, α,ω-dicaboxylic fatty acids, n-alkanols, and n-alkanes. Characteristic biomarkers and their distribution patterns indicated the presence of pine pitch in all the adhesives, which in some cases was admixed with beeswax and brassicaceae seed oil.The results provided new insights into the complex recipes used by artisans in ancient Egypt in the production of adhesives and in the sophisticated manufacture of opus sectile decorations.     

Solid phase microextraction and gas chromatography with ion trap detector (GC-ITD) analysis of amitraz residues in beeswax after hydrolysis to 2,4-dimethylaniline

An analytical method for the determination of amitraz residues in beeswax after hydrolysis to 2,4-dimethylaniline is reported. It consists of wax extraction with an acid buffer solution, head space solid phase microextraction and GC-ITD analysis. The limit of determination is 1 ng g⁻¹. Wax samples from beekepers and commercial foundations were analysed, content of residues varied from <1 to 20.5 ng g⁻¹.     

Comparative evaluation of UV-vis-IR Nd:YAG laser cleaning of beeswax layers on granite substrates

The beeswax treatment applied in the sixties to prevent rain water from penetrating the outer stone surface of valuable granitic Galician monuments is contributing to the acceleration of the superficial degradation process of these monuments. At present, the northern sector of the renaissance frieze in the Cloister of the Cathedral of Santiago de Compostela is one of the most representative examples.Conventional wax removal methods (water, chemical and mechanical cleaning) can possibly destruct important details of the relief. Therefore laser removal is considered as a good alternative.In this work, we report systematic investigations of the effect of laser cleaning at different Nd:YAG laser wavelengths (266, 355, 532 and 1064 nm) on representative samples of the real historical surfaces.Laser removal of beeswax on granite at neither of the four wavelengths of the Nd:YAG laser is not a layer by layer removal process. For each irradiance and wavelength there is a maximum thickness that can be completely removed by a single pulse. Above this thickness the waxy material is not removed, although it undergoes thermal modifications; since the fraction of radiation that reaches the granite substrate is not enough to trigger the ejection of material.Our results show that the wax-granite interface plays a fundamental role in granite cleaning, and when the wax is weakened by absorption of radiation at 266 nm, the removal process becomes more efficient.     

Analysis of wax ester molecular species by high performance liquid chromatography/atmospheric pressure chemical ionisation mass spectrometry

High chromatographic resolution of wax esters (WEs) was achieved by non-aqueous reversed-phase liquid chromatography on a Nova-Pak C18 column by optimising the acetonitrile/ethyl acetate mobile phase gradient. The retention behaviour of WEs was studied in this chromatographic system. The WEs eluted according to their equivalent carbon number (ECN) values; within the group of WEs with the identical ECN, the most unsaturated species tended to elute first. The isobaric WEs with different positions of the ester moiety were separated from each other whenever the lengths of the chains were sufficiently different. The methyl-branched esters eluted at shorter retention times than the straight-chained analogues, and the resolution among methyl-branched WEs depended on the position of the branching. The analytes were detected by atmospheric pressure chemical ionisation mass spectrometry (APCI-MS) using data-dependent scanning. WEs provided simple full-scan spectra with abundant protonated molecules and low-intensity fragments. Collision-induced dissociation (CID) promoted identification of the WE molecular species. The responses of WEs were found to be dependent on the number of double bonds and on the alkyl-chain length; the limits of the detection ranged from 20 μmol/L to 200 nmol/L. The HPLC/APCI-MS was applied for the analysis of the WEs isolated from honeycomb beeswax, jojoba oil and human hair. Good agreement between reported results and the literature data was achieved, with several novel polyunsaturated WEs also being found.     

气相色谱-质谱法测定蜂蜡净油中限用成分

提出了气相色谱-质谱法测定蜂蜡净油中限用成分含量的方法。样品经二氯甲烷-三氯甲烷(1+1)混合溶剂溶解,采用不同极性的两根色谱柱分离后,选择离子监测模式下质谱法进行测定。共鉴定出96种化合物,其中香豆素和马索亚内酯是禁用成分。苯甲醇、肉桂醇、香豆素、马索亚内酯、苯甲酸苄酯、水杨酸苄酯和肉桂酸苄酯等7种限用物质的质量浓度在0.08～80mg.L-1范围内呈线性关系,检出限(3S/N)在0.01～0.23mg.kg-1之间。样品加标回收率在86%～101%之间,相对标准偏差(n=6)小于4.0%。     

Determination of synthetic acaricides residues in beeswax by high-performance liquid chromatography with photodiode array detector

A multiresidue HPLC method for identification and quantification of the synthetic acaricides fluvalinate, coumaphos, bromopropylate and its metabolite 4,4′-dibromobenzophenone in beeswax has been developed. Different techniques were tested and modified. The method consists of a sample preparation with isooctane followed by solid phase extraction using Florisil columns. Determination of the synthetic acaricides is achieved by HPLC with a photodiode array detector. Analytical performance of the proposed method, including sensitivity, accuracy and precision was satisfactory. The LOD for the analytes varied between 0.1 and 0.2 μg g⁻¹ wax and the recoveries between 70 and 110%. Relative standard deviation of the repeatability of the method is <15% and reproducibility is <31%.     

Development and validation of a liquid chromatography with mass spectrometry method to determine resveratrol and piceid isomers in beeswax

This paper represents the first report of a liquid chromatography coupled to electrospray ionization mass spectrometry method for simultaneously analyzing resveratrol and piceid isomers (cis and trans) in beeswax. An efficient extraction procedure has been proposed (average analyte recoveries were between 89 and 95%); this involved a solid–liquid extraction using a mixture of ethanol and water (80:20, v/v) and a concentration step in a rotary evaporator. The separation of all the compounds was achieved using a C₁₈ column and a mobile phase composed of ammonium formate 0.03 M in water and acetonitrile in gradient elution mode at a flow rate of 1 mL/min. The method was fully validated in terms of selectivity, limits of detection and quantification, linearity, precision, and accuracy. The limits of detection and quantification ranged from 1.0 to 1.7 and 3.5 to 5.5 μg/kg, respectively. Finally, the proposed method was applied to analyze beeswax samples collected from experimental and organic apiaries.