Testing Methods (DTE 1 - 15 December 2020)

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Testing Methods

  • Isotope Testing (3 delta)
  • EA
  • LC
  • ISOTOPE TESTING (all three delta tests — max, p-h, fru-glu) is done to calculate C4 and C3 sugars. The method used is isotope ratio mass spectrometry (IRMS) to determine the sugar
  • For C4 sugar analysis, laboratories do elemental analysis (EA) and for C3 sugar they use liquid chromatography (LC) . But it is the “nature” of the sugar type that only C4 sugar can be quantified as carbon 12,13 ratios of corn is available and used. FSSAI 2020 standard for checking the purity of honey is based on isotope testing.
  • Rice syrup marker – tests for Specific Marker for Rice (SMR) i.e.. , 2-AFGP and Trace Marker for Rice (TMR) .
  • gold-standard for detecting adulteration through Nuclear Magnetic Resonance Spectroscopy (NMR) . NMR imaging helps to get a full picture of the honey and its constituents. It is then able to identify both the origin of the honey and its authenticity.
Nuclear Magnetic Resonance Spectroscopy (NMR)
  • Column (1) : Quantifies C4 sugars
  • Column (2) to (4) : By applying EA/LC-IRMS (liquid chromatography/elemental analysis-isotope ratio mass spectrometry) for the determination of the — 13 C, adulteration with both C4 and C3 sugars could be detected
  • Column (5) : Foreign oligosaccharides are starch-based sugars such as from rice, corn, and wheat — can be both C4 and C3 sugars
  • Column (6) and (7) : These are markers for rice syrup, thus identify C3 sugars
  • Column (8) : NMR shows addition of sugar syrup — could be either C4 or C3 plant based
Foreign Oligosaccharides Are Starch-Based Sugars


Samples were sent for laboratory test to the Centre for Analysis and Learning in Livestock and Food (CALF) at National Dairy Development Board (NDDB) in Gujarat.

Centre for Analysis and Learning in Livestock and Food (CALF)
National Dairy Development Board (NDDB)
  • 13 top and smaller honey brands were selected
  • Most of the top brands passed the laboratory tests for Indian standards
  • Laboratory in India didn՚t find adulteration of C3 and C4 sugar in these brands
  • However, most of the smaller brands failed the laboratory tests for Indian standards
  • Adulteration with C4 sugar is most common
  • Of the 13 brands only three brands passed
  • Of the 22 samples only five samples passed test. Rest was adulterated

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