Simultaneous qualitative and quantitative analyses of volatile components in Chinese honey of six botanical origins using headspace solid-phase microextraction and gas chromatography-mass spectrometry.

BACKGROUND: Honey aroma is one of its most important properties and it depends on the qualitative and quantitative composition of the volatile compounds. The volatile profile of honey could reveal its botanical origin to avoid a false characterization. Thus, it is of great significance to honey authentication. This study developed and validated a headspace solid-phase microextraction and gas chromatography-mass spectrometry (HS-SPME-GC-MS) method for simultaneous qualitative and quantitative analyses of 34 volatile components in honey. The developed method was applied to 86 honey samples from six different botanical origins, including linden honey, rape honey, jujube honey, vitex honey, lavender honey and acacia honey. RESULTS: The volatile fingerprints and quantitative results were simultaneously obtained by using the full scan and selected ion monitoring (SCAN+SIM) MS scanning mode. The limits of quantification (LOQs) and limits of detection (LODs) of 34 volatile compounds were in the ranges of 1-10 ng/g and 0.3-3 ng/g, respectively. And the spiked recoveries ranged between 70.6% and 126.2%, with the relative standard deviations (RSDs) not higher than 45.4%. A total of 98 volatile compounds were found with relative contents determined, and the 34 volatile compounds were determined with absolute concentrations. Based on the volatile fingerprints and the contents of volatile compounds, honey samples from six botanical origins were well classified by principal component analysis and orthogonal partial least-squares discrimination analysis. CONCLUSIONS: The HS-SPME-GC-MS method was successfully applied to achieve the volatile fingerprints of six types of honey and to quantitatively analyze 34 volatile compounds with satisfying sensitivity and accuracy. Chemometrics analysis showed significant correlations between honey types and volatiles. These results reveal the characteristics of volatile compounds in six types of unifloral honey and provide some supports for honey authentication. © 2023 Society of Chemical Industry.

Comprehensive investigation of the content and the origin of matrine-type alkaloids in Chinese honeys.

A HPLC-MS/MS method for simultaneous determination of four matrine-type alkaloids (matrine, oxymatrine, sophocarpine and sophoridine) in honey was established and was applied to 567 Chinese honey samples. The overall detection rate was 61.0 % and all eight Sophora viciifolia Hance honey samples were detected with the average content of 1183.3 µg/kg. Among 383 Acacia honey samples, matrine-type alkaloids have the highest detection rates in Gansu (98.8 %) and Shaanxi (86.5 %) provinces, which were significantly correlated with the distribution of S. viciifolia Hance. Moreover, matrine-type alkaloids in fruits and flowers of S. viciifolia Hance were as high as 7.06 g/kg and 2.65 g/kg respectively. The melissopalynological analysis showed that the concentration of oxymatrine was positively correlated with the pollen frequency of S. viciifolia Hance (R2 = 0.737) in representative Acacia honey samples. It can be concluded that the matrine-type alkaloids in Chinese honeys come from the nectariferous plants S. viciifolia Hance.

Hemolymph Metabolism Analysis of Honey Bee (Apis mellifera L.) Response to Different Bee Pollens.

Pollen is essential to the development of honey bees. The nutrients in bee pollen vary greatly among plant species. Here, we analyzed the differences in the amino acid compositions of pear (Pyrus bretschneideri), rape (Brassica napus), and apricot (Armeniaca sibirica) pollens and investigated the variation in hemolymph metabolites and metabolic pathways through untargeted metabolomics in caged adult bees at days 7 and 14. The results showed that the levels of five essential amino acids (isoleucine, phenylalanine, lysine, methionine, and histidine) were the highest in pear pollen, and the levels of four amino acids (isoleucine: 50.75 ± 1.93 mg/kg, phenylalanine: 87.25 ± 2.66 mg/kg, methionine: 16.00 ± 0.71 mg/kg and histidine: 647.50 ± 24.80 mg/kg) were significantly higher in pear pollen than in the other two kinds of bee pollen (p < 0.05). The number of metabolites in bee hemolymph on day 14 (615) was significantly lower than that on day 7 (1466). The key metabolic pathways of bees, namely, "sphingolipid metabolism (p = 0.0091)", "tryptophan metabolism (p = 0.0245)", and "cysteine and methionine metabolism (p = 0.0277)", were significantly affected on day 7. There was no meaningful pathway enrichment on day 14. In conclusion, pear pollen had higher nutritional value among the three bee pollens in terms of amino acid level, followed by rape and apricot pollen, and the difference in amino acid composition among bee pollens was reflected in the lipid and amino acid metabolism pathways of early adult honey bee hemolymph. This study provides new insights into the physiological and metabolic functions of different bee pollens in bees.

[Simultaneous determination of delta13C values of glycerol and ethanol in wine by liquid chromatography coupled with isotope ratio mass spectrometry].

A novel procedure was established for the characterization of delta13C values of glycerol and ethanol in wine by liquid chromatography-isotope ratio mass spectrometry (LC-IRMS). Several parameters influencing the separation of glycerol and ethanol from wine matrix were optimized. The precision and accuracy of the proposed method were 0.15 per thousand to 0.26 per thousand and 0.11 per thousand to 0.28 per thousand, respectively. The results obtained for 40 wine samples displayed that the delta13C value of glycerol ranged from--26.87 per thousand to--32.96 per thousand and that of ethanol ranged from--24.06 per thousand to--28.29 per thousand. Close correlations (R = 0.82) were obtained between the delta13C values of glycerol and ethanol. The proposed method didn't need complex sample treatment, and the delta13C values of glycerol and ethanol in wine can be simultaneously determined, thus improving the method in terms of simplicity and speed compared with traditional methods.

Analysis method study on 839 pesticide and chemical contaminant multiresidues in animal muscles by gel permeation chromatography cleanup, GC/MS, and LC/MS/MS.

The paper reports the study of gel permeation chromatography (GPC), gas chromatography/mass spectrometry (GS/MS), and column chromatography tandem MS (LC/MS/MS) for 839 pesticides and chemical contaminants, through which a GPC data bank has been established for 744 pesticides, a GC/MS data bank for 541 pesticides, and an LC/MS/MS data bank for 464 pesticides. On the basis of this study, a new method for quantitative determination of 587 pesticide residues in animal muscles such as beef, mutton, pork, chicken, and rabbit has been established using GPC cleanup followed by GC/MS and LC/MS/MS. In the method, 10 g animal samples were mixed with 20 g sodium sulfate and extracted twice with 35 mL cyclohexane-ethyl acetate (1 + 1) by blender homogenization followed by centrifugation, filtration, and concentration. An equivalent of 5 g sample was injected into a 400 x 25 mm S-X3 GPC column, with cyclohexane-ethyl acetate (1 + 1) as the mobile phase at a flow rate of 5 mL/min. The 22-40 min fraction was collected for subsequent analysis. For the 478 pesticides determined by GC/MS, the portions collected from GPC were concentrated to 0.5 mL and exchanged twice with 5 mL hexane. For the 379 pesticides determined by LC/MS/MS, the portions collected from GPC were dissolved with acetonitrile-water (60 + 40) after taking the extract to dryness with nitrogen gas. At the limit of quantification (LOQ) and 10 LOQ fortification levels of 0.1-16 000 microg/kg, recoveries were within 40-130%, among which 563 pesticide recoveries were between 60 and 130%, accounting for 96% of the compounds; 24 analytes were recovered between 40 and 60%, accounting for 4% of the compounds. The relative standard deviation was below 30% for all 587 pesticides. The limits of detection for the method were 0.1-1600 microg/kg. In comparison with GC/MS, LC/MS/MS increased the detection sensitivity 2-1000 times for 236 pesticides.

Validation study on 660 pesticide residues in animal tissues by gel permeation chromatography cleanup/gas chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry.

A new method using gel permeation chromatography (GPC) cleanup followed by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS-MS) has been established for quantitative determination of 437 pesticide residues in animal tissues such as beef, mutton, pork, chicken, and rabbit. Based on an appraisal of the characteristics of both GC-MS and LC-MS-MS, validation experiments were conducted for 660 pesticides. In the method, 10 g animal samples were mixed with 20 g sodium sulfate and extracted with 35 mL of cyclohexane+ethyl acetate (1+1) twice by blender homogenization, centrifugation, and filtration. Evaporation was conducted and an equivalent of 5 g sample was injected into a 400 mm x 25 mm S-X3 GPC column, with cyclohexane+ethyl acetate (1+1) as the mobile phase at a flow rate of 5 mL/min. The 22-40 min fraction was collected for subsequent analysis. For the 368 pesticides determined by GC-MS, the portions collected from GPC were concentrated to 0.5 mL and exchanged with 5 mL hexane twice. For the 69 pesticides by LC-MS-MS, the portions collected from GPC were dissolved with acetonitrile+water (60+40) after taking the extract to dryness with nitrogen gas. In the linear range of each pesticide, the correlation coefficient was r > or = 0.98, exceptions being dinobuton, linuron, and fenamiphos sulfoxide. At the low, medium and high three fortification levels of 0.2-4800 microg/kg, recoveries fell within 40-120%, among which 417 pesticides recoveries between 60% and 120%, accounting for 95%, 20 analytes between 40% and 60%, accounting for 5%. The relative standard deviation was below 28% for all 437 pesticides. The limits of detection for the method were 0.2-600 microg/kg, depending on each pesticide.

Simultaneous determination of 16 sulfonamides in honey by liquid chromatography/tandem mass spectrometry.

A method is described for the determination of 16 sulfonamides in honey. Samples are dissolved in phosphoric acid solution (pH2), cleaned up with 2 solid-phase extraction (SPE) cartridges, an aromatic sulfonic cation-exchange cartridge and an Oasis HLB SPE cartridge, and analyzed both qualitatively and quantitatively by liquid chromatography/tandem mass spectrometry (LC/MS/MS) under the selected conditions. Without exception, calibration curves were linear (r = > 0.995), when sulfamethizole was between 1.0 and 25.0 microg/kg; sulfacetamide, sulfapyridine, sulfadiazine, sulfachloropyridazine, sulfamethoxazole, sulfamerazine, sulfisoxazole, sulfamonomethoxine, and sulfadoxine were between 2.0 and 50.0 microg/kg; sulfamethoxypyridazine, sulfadimethoxine, and sulfathiazole were between 4.0 and 100.0 microg/kg; sulfamethazine and sulfameter were between 8.0 and 200.0 microg/kg; and sulfaphenazole was between 12.0 and 300.0 microg/kg. Average recoveries at 4 fortification levels in the range of 1.0-300 microg/kg in honey were 70.9-102.5%, and relative standard deviations were 2.02-11.52%. The limits of quantitation for the 16 sulfonamides were between 1.0 and 12.0 microg/kg, with the LC/MS/MS method.

Liquid chromatography-fluorescence detection for simultaneous analysis of sulfonamide residues in honey.

A rapid, accurate LC analytical method has been developed for determination of eight sulfonamides (sulfacetamide, sulfapyridine, sulfamerazine, sulfamethoxypyridazine, sulfameter, sulfachloropyridazine, sulfamethoxazole and sulfadimethoxine) in honey. The sample was dissolved in phosphoric acid solution (pH 2). After filtration, the sample solution was cleaned by use of two solid-phase extraction (SPE) cartridges-an aromatic sulfonic cation-exchange cartridge and an Oasis HLB cartridge. The eight sulfonamides were then derivatized with fluorescamine and the derivatives were determined by LC with fluorescence detection at excitation and emission wavelengths of 405 and 495 nm, respectively. Average recoveries at three fortification levels in the range 0.02-0.50 mg kg(-1) in twelve different kinds of honey were 73.5-94.1% with coefficients of variation of 4.35-16.60%. The limit of detection (LOD) was 0.002 mg kg(-1) for sulfacetamide, sulfapyridine, sulfamerazine, and sulfamethoxypyridazine; that for sulfameter, sulfachloropyridazine, sulfamethoxazole and sulfadimethoxine was 0.005 mg kg(-1). The limit of quantitation (LOQ) was 0.005 mg kg(-1) for sulfacetamide, sulfapyridine, sulfamerazine, and sulfamethoxypyridazine; that for sulfameter, sulfachloropyridazine, sulfamethoxazole, and sulfadimethoxine was 0.010 mg kg(-1). The method is suitable for determination of multiresidue sulfonamides in the various kinds of honey.