Adulteration of honey

adulteration of honey and its detection methods

1. Honey Apis mellifera L.

2. Adulteration of Honey Presented by S.Keerthana 17FT1D7815

3. CONTENTS Introduction Adulteration of honey Detection test Effects Journals Related articles Video clippings Bibliography

4. Introduction Adulteration: Adulteration  is a legal term meaning that a food product fails to meet federal or state standards.  Honey: Honey means the natural sweet substance produced by honey bees from the nectar of blossoms or from secretions of plants which honey bees collect, transform store in honey combs for ripening.

5. Composition of Honey Sr. No. Component Amount (in %) 1. Water (moisture) 20 2. Fructose (Laevulose) 37 3. Glucose (Dextrose) 34 4. Sucrose (Non Reducing Sugar) 5.0 5. Total Ash (Minerals) 0.25 6. Acids Organic 0.20 7. Proteins, Amino Acids, etc. 1.50 8. Other ingredients including pollen grains 1.05

6. Standards For Honey As Per BIS, India Amendment No.1 April 2011 to Is 4941: 1994 Extracted Honey Specification Sr. No. Characteristics Special Grade A Grade Standard Grade 1. Specific Gravity at 27 C, (min) 1.37 1.37 1.37 2. Moisture , % by mass, (Max) 20 22 25 3. Total Reducing Sugar, % by mass, (min.) 70 65 65 4. Sucrose, % by mass, (max.) 5.0 5.0 5.0 5. Fructose-Glucose ratio, (min.) 1.0 1.0 1.0 6. Ash, % by mass, (max.) 0.5 0.5 0.5 7. Acidity (expressed as Formic Acid ), % by mass (max.) 0.2 0.2 0.2 8. Fiehe’s Test 1 Negative 9. Hydroxymethyl Furfural (HMF), mg/kg, (max.) 80 80 80 10. Total counts of pollens and plant elements/g of honey, (max.) 50,000 50,000 50,000 11. Optical Density at 660 nm, %, (max.) 0.3 0.3 0.3 1 If Fiehe’s test is positive, carry out the determination of hydroxymethyl furfural (HMF) content. If it is more than 80 mg/kg , then fructose glucose ratio should be more than 1.00.

7. Adulteration of Honey methods Honey adulteration can be direct or indirect. Direct adulteration means that a substance is added directly to honey. Adulteration with corn sugar Adulteration with cane sugar Adulteration with rice syrup Adulteration with individual sugars Indirect adulteration happens when honeybees are fed adulterating substance.

8. Detection Tests

9. Adulteration of Honey with sweeteners and sugars

10. Contd..

11. Detection of sugar solution

12. Density of fructose=1.69 g/cm³ Density of sucrose =1.59g/cm³ Detection of sugar solution SUGAR : At room temperature, it is highly water-soluble (1800 grams will dissolve in 1 liter of water).

13. Contd..

14. Detection of invert sugar   Fiehe’s Test: Add 5 ml. Of solvent ether to 5 ml. Of honey. Shake well and decant the ether layer in a petri dish. Evaporate completely by blowing the ether layer. Add 2 to 3 ml. Of resorcinol (1 gm. Of resorcinol resublimed in 5 ml. Of conc. HCl .) Appearance of cherry red colour indicates presence of sugar/ jaggery . Hydroxy Methyl Furfural (HMF) occurs in honey due to acid-catalyzed dehydration of hexose sugars. Its value in natural fresh honey varies from 10 to 14 mg/ kg , but it increases upon storage, depending on the pH of honey and on the storage temperature. HMF content of honey also increases upon its adulteration by invert sugars. Presence of invert sugar in honey is assessed by Fiehe's test where HMF reacts with resorcinol and gives a red coloured complex . According to PFA Act, Fiehe's test should be negative for honey, whereas Codex Standards states that HMF content of honey should not be more than 60 mg/kg.

15. Adulteration using sulphite -ammonia caramel Dark honey is usually referred to as “forest honey” and quite often the dye sulphite ammonia caramel (E150d) is added. This is mainly due to the higher sale price of dark honey. Determination of the content of ammonia sulphite caramel is possible with liquid chromatography tandem mass spectrometry (LC-MS/MS). A level of caramel E150d above the limit of quantification methods (> 5 mg/kg) is considered an infringement of regulations ( Bartošová 2013).

16. Methods of detecting indirect adulteration

17. Analysis methods Gas Chromatography (GC) and Liquid Chromatography (LC) analysis: This method may be considered as a replacement of isotopic analysis , which has some limitations . Near Infrared Transflectance Spectroscopy (NIR): It is a rapid, non-destructive and relatively inexpensive method which may be suitable for use as a screening technique in the quality control of honey. Fourier Transform Infrared (FTIR) spectroscopy with Attenuated Total Reflectance (ATR ): In contrast to the time-consuming carbon isotope ratio analysis techniques, these FTIR spectroscopic procedures can be performed in very short time. Protein characterization: The major proteins in honey have different molecular weights depending upon the honeybee species. Therefore, the measurement of major proteins in honey is a useful method to discriminate the honey that produced from different honeybee species. High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection (HPAEC-PAD): It is an efficient tool for the characterization of the honey floral species . This method is less time consuming and less expensive than other methods .

18. Contd.. Liquid Chromatography Coupled to Isotope Ratio Mass Spectrometry (HPLC-IRMS) : The new procedure has advantages over existing methods in terms of analysis time, sensitivity, lack of sample preparation, reduced consumption of reagents, and simplicity of the operative procedure . In addition , it is the first isotopic method developed that allows beet sugar addition detection. Calorimetric methods (Application of DSC): Application of DSC showed the possibility of using the glass transition temperature to distinguish between honeys and syrups and is a powerful technique for characterizing the thermal behavior of honeys and for detecting the effect of adulteration on physicochemical and structural properties of samples. Stable Carbon Isotope Ratio Analysis (SCIRA): It is determined by the isotope ratio Fourier Transform (FT) Raman spectroscopy: FT-Raman spectroscopy is successfully applicable to detect beet and cane invert syrups. This method can also be used to discriminate between the types of adulterants irrespective of its floral origin. Microscopic detection: Microscopic analysis of adulterated honeys with cane sugar exhibited parenchyma cells, single ring vessels and epidermal cells. Overall the microscopic procedure is a good screening method for the detection of adulteration of honey with cane sugar products.

19. Cordella et al. (2005) describe the development of high performance anion exchange chromatography with Pulsed Amperometric Detection (HPAEC-PAD) for the analysis of honey to detect adulteration combined with chemometric techniques for processing chromatograms for better discrimination of pure and adulterated honey. This method was investigated using honey samples containing between 10% and 40% of different industrial sugar syrups used for the feeding of honey bees. Journal Papers

20. Contd.. Ruiz- Matute et al. (2010a) studied the sugar composition of high-fructose corn syrup (HFCS ) using gas chromatography coupled with mass spectrometry (GC-MS). Sucrose syrups were analysed in parallel as a control. HFCS was shown to contain fructosyl -fructose and other unknown sugars that could be fructosyl -glucose . Guler et al . (2014) investigated the sensitivity of methods for the analysis of carbonisotope ratios. They analysed a total of 100 samples of unadulterated honey, honey made by bees fed with different amounts (5, 20 and 100 litres /colony) of sugar syrups. These syrups included corn syrups with high fructose-85 (HFC-85%), with moderate fructose-55 (HFC-55%), bee-feeding syrups (BFS), glucose syrups (GMS), and sucrose syrups (SS).

21. Health effects Although the adulteration of honey is not injurious to health, problems of honey fraud negatively influence market growth by damaging consumer confidence.

22. Effects Pesticides residues include acaricides , organic acids, insecticides, fungicides, herbicides, and bactericides. Many of these contaminants are banned because of their well-documented health hazards such as carcinogenic effect on humans . Lactam antibiotics cause cutaneous eruptions, dermatitis, gastrointestinal symptoms and anaphylaxis at very low doses Long-term effects of exposure to antibiotic residues include microbiological hazards, carcinogenicity, reproductive effects, and teratogenicity. Microbiological effects are one of the major health problems in human beings. Certain drugs like nitrofurans and nitroimidazoles can cause cancer in human being. Similarly, some drugs can produce reproductive and teratogenic effects at very low doses.

23. Articles Over 150 different pesticides have been found in colony samples  The maximum limits of pesticide residues in honey are not included in the Codex Alimentarius . The European Union legislation has regulated the MRLs for three amitraz , coumaphos , and cyamizole , which are 0.2, 0.1, and 1 mg ·, respectively. The US Environmental Protection Agency has established MRLs for amitraz (1 mg·), coumaphos (0.1 mg·), and fluvalinate (0.05 mg·)

24. In India, a study was carried out to explore the extent of pesticide residue in honey produced in the various parts of Himachal Pradesh. It was found that HCH and its isomers were the most frequently detected followed by dichlor-diphenyl-trichlorethylene (DDT) and its isomers. Malathion’s residue was found exceeding the MRLs (5 ppb) proposed by the Ministry of Commerce, Government of India. The results showed that honey from natural vegetation contained lesser residues. In Turkey, 24 organochlorine pesticide residues in 109 different honey samples collected from stores and open markets in Konya, Turkey, were analyzed by gas chromatography-electron capture detection. Aldrin ,  cis -chlordane, trans-chlordane, oxy-chlordane, 2,4(′)-DDE, and 4,4(′)-DDE were found in all honey samples.  In Spain, residues of thymol were found in honey collected from the beehives, ranged from 0.75 to 8.20  μg ·

25. News articles

26. Bibliography Commission Regulation (EC) No. 2377/90 of 26 June 1990 laying down a Community Procedure for the stablemen of maximum residue limits of veterinary medicinal products in foodstuff of animal origin (as amended by regulations) ECC No.2034/96 (OJ L272 25.10.1996, p 2), No2686/98 (OJ L337 12.12.1998, p 20) No. 1931/99 (OJ L240 10.09.1999, p 3), and No. 239/99(OJ L 290 12.11.1999, p 5). Food and Drug Administration of the United States, Pesticide tolerances, 2003,  http://www.cfsan.fda.gov/ . D. Beyoǧlu and G. Z. Omurtag , “Occurrence of naphthalene in honey consumed in Turkey as determined by high-pressure liquid chromatography,” Journal of Food Protection, vol. 70, no. 7, pp. 1735–1738, 2007.  View at Google Scholar  ·  View at Scopus S. Adamczyk , R. Lázaro , C. Pérez- Arquillué , P. Conchello , and A. Herrera, “Evaluation of residues of essential oil components in honey after different anti- varroa treatments,” Journal of Agricultural and Food Chemistry, vol. 53, no. 26, pp. 10085–10090, 2005.  View at Publisher  ·  View at Google Scholar  ·  View at Scopus Mehryar , Laleh , and Mohsen Esmaiili . "Honey and honey adulteration detection: A Review."  Proceeding 11th International Congress on Engineering and Food. Athens, Greece, Elsevier Procedia . Vol. 3. 2011 . Al- Waili , Noori , et al. "Antibiotic, pesticide, and microbial contaminants of honey: human health hazards."  The Scientific World Journal  2012 (2012).