Study On The Fingerprint Of Organic Acids In Pape Honey

Honey is a traditional, inartificial and nutritional healthy food. In recent years, as the improvement of living standards, the demand for honey is increasingly, and the consumption of honey has increased year by year. But at the same time, the problems of honey market has also become Seriously. Untill now, the methods used to control the quality of honey is still not completely scientific and rational, resulting that the honey export of our country blocked again and again. So a scientific and rational method is necessary to establelished to control honey quality. Studies show that more traces active substances has exist in honey that come from the nectar plants, different single-flower honey has the different characteristics composition. Therefore, different types of trace active substances can be studied, in combination with fingerprinting, establish the fingerprint of different active ingredients and use it to identify the truth of honey. The use of fingerprinting technology to identify the authenticity of honey has irreplaceable advantages to other methods.In this paper, the rape honey of different origins was used for study, and analysisd by HPLC and GC metheods combined with GC-MS techniques, Established the HPLC and GC fingerprinting. through the similarity analysis and clustering analysis to study the similarities and differences between the honey samples. The conclusions were as follows:1. Though the experiment, the best conditions to extract Short-chain organic acids in rape honey was used the Bond Elutes SAX column for sample purification, the system pH = 7, the elution volume was 4 mL. And the optimistic HPLC conditions were as follows:the column was Waters C18-MS-â…¡(4.6 mm×250 mm,5μm); mobile phase was 0.04mol/L solution of potassium dihydrogen phosphate (pH 2.30 to 2.35); flow rate was 0.7 mL/min; injection volume:20μL and the detection wavelength was 210 nm. Under these conditions,9 standard organic acids were separated and the honey samples too, The detection limits ranged from 0.06 to 9.4 mg/kg, the relative standard deviations of precision, stability, repeatability were controlled within 5%.2. Established the HPLC fingerprint of Short-chain organic acids in rape honey, Through the similarity analysis and clustering analysis,12 batches of rape honey samples from different areas were divided into three categories:No.1,4,5 samples Clustered into one group, No.10 samples into a category separately, and the other eight samples into a category.The study can be used to classification and identification the property of honey. 3. The best conditions to extract Long-chain organic acids in rape honey was used ether as extraction solvent and by the method of ultrasonic assisted extraction, Long-chain organic acids were esterfied with 3 mL 1% sulfuric acid-methanol, esterification temperature is 60℃, reaction time was 50 min. Best GC conditions were using HP-5 (30 m×0.25 mm,0.25μm) capillary column for separation, temperature program was 60℃(2min)â†'8℃/min; 180℃(1min)â†'3℃/min; 200℃(2min)â†'3℃/min; 240℃(4min), split ratio was 20:1; under these conditions, the relative standard deviations of precision, stability, repeatability were controlled within5%4. Studied the different resources of honey samples by GC-MS. We found that the type and the content of fatty acids were different in the four types of honey. For exemple, in the rape honey, Eicosanoids was identified, but not found in other three plants sources honey, so it maybe was the marker substances of rape honey, but it needs large number statistical experiments to be certain.5. Established the GC fingerprint of Long-chain organic acids in rape honey, Through the similarity analysis, clustering analysis and principal component analysis,55 batches of rape honey samples from different areas were divided into four categories:No. 33,34,50 samples Clustered into one group, the No.9 sample into a category separately, the No.12,14,15,16 sample s Clustered into a category and the other samples into a category,the basic results are consistent with the similarity analysis. This study can be used to classification and identification the property of honey.6. Compared the result analysis by HPLC and GC, we found that both methods were clustered 8 rape honey producd in bubei together, the two methods has coincided, it can be used to identification of variety and quality of honey.7. Application of the established fingerprints to commercial rape honey, we found that the commercial honey has fewer peak compered to standard sample, as the standard rape honey added, the peaks increased, indicating that the purity of commercial honey samples is not enough, and the result of GC-MS also revealed that the commercial honey also contains the Tricosane acid and octadecanoic acid, that were not detected in standard samples, It’s demonstrate the practicality of the established fingerprints.