LCMS - NMR - HPLC (Polyphenols Measuring Methods)
To see for ourselves any differences between NMR, LC-MS and HPLC, we put them all to the test: We sent a sample from the exact same bottle from olive oil lot Δ25 (produced on the 25th of September 2025), to be analyzed independently by:
- Faculty of Pharmacy at the University of Athens (measuring with NMR 2924 mg/kg total polyphenols)
- Department of Chemistry at the University of Crete (measuring with NMR 3009 mg/kg of polyphenols)
- Department of Analytical Chemistry at the University of Cordoba (measuring with LC-MS 3107 mg/kg total polyphenols)
- Chem Solutions, Certified EU Lab (measuring with HPLC 4412 mg/kg total polyphenols)
For convenience, the comparison of the above results is presented side-by-side as:
POLYPHENOLS MEASUREMENT METHOD FACULTY UNIVERSITY | NMR PHARMACY FACULTY UNIVERSITY OF ATHENS | NMR CHEMISTRY DEPT UNIVERSITY OF CRETE | LC-MS ANALYTICAL CHEMISTRY DEPT. UNIVERSITY OF CORDOBA | HPLC CHEM SOLUTIONS CERTIFIED EU LAB |
Lot / Date of Production | Δ25 / 25th of September 2025 | |||
Date of Analysis | 26 September 2025 | 30 September 2025 | 12 November 2025 | 30 December 2025 |
Location of Analysis | Athens - Greece | Heraclion - Greece | Cordoba - Spain | Athens - Greece |
mg/kg | ||||
Oleocanthal | 1836 | 2075 | 1557 | - |
Oleacein | 816 | 797 | 1238 | - |
Ligstroside Aglycon (monoaldehyde form) | 118 | 109 | 171 | 230.8 |
Ligstroside Aglycon (dialdehyde form) | 72 | 1 | 2694.2 | |
Oleuropein Aglycon (monoaldehyde form) | 68 | 27 | 135 | 164.2 |
Oleuropein Aglycon (dialdehyde form) | 5 | 0 | 1198.8 | |
Free Tyrosol | 10 | - | 4.4 | 7 |
Hydroxytyrosol | - | - | 3 | 2.5 |
Oleocanthalic Acid | - | - | <LOQ | <LOQ |
Apigenin | - | - | <LOQ | 0.22 |
Luteolin | - | - | <LOQ | 0.37 |
Other | - | - | - | 114.31 |
Total Polyphenols | 2924 | 3009 | 3107 | 4412.4 |
CONCLUSION
- Comparing the above analyses in "absolute numbers" (NMR Athens: 2924 mg/kg of total polyphenols - NMR Crete: 3009 mg/kg of total polyphenols - LC-MS: 3107 mg/kg of total polyphenols - HPLC: 4412 mg/kg of total polyphenols), even though each chemical compound may differ slightly, total polyphenols with NMR & LC-MS methods are very similar (3% to 6% higher in LCMS). HPLC demonstrated a significantly higher total polyphenols than the other two methods (around 50% more), while in HPLC, Oleocanthal & Oleacein were mostly identified as DDLA (Dialdehydic Ligstroside Aglycone) & DDOA (Dialdehydic Oleuropein Aglycone)
- IT ALSO HAS TO BE TAKEN INTO ACCOUNT THAT THE DISTANCE FROM OUR ESTATE TO THE UNIVERSITY OF ATHENS IS 270 km (4 HOURS DRIVE), THE DISTANCE FROM OUR ESTATE TO THE UNIVERSITY OF CRETE IS 500 km (2 DAYS EXPRESS SHIPPING), THE DISTANCE FROM OUR ESTATE TO THE UNIVERSITY OF CORDOBA IS +4000 km (5 DAYS EXPRESS SHIPPING) & THE DISTANCE FROM OUR ESTATE TO THE CHEM SOLUTIONS LAB IS 270 km (4 HOURS DRIVE).
- THE MEASUREMENT WITH NMR (ATHENS) TOOK PLACE 1 DAY AFTER PRODUCTION OF THE OLIVE OIL, THE MEASUREMENT WITH NMR (CRETE) TOOK PLACE 5 DAYS AFTER PRODUCTION OF THE OLIVE OIL, THE MEASUREMENT WITH LC-MS (CORDOBA) TOOK PLACE 48 DAYS AFTER PRODUCTION OF THE OLIVE OIL & THE MEASUREMENT WITH HPLC (ATHENS) TOOK PLACE 97 DAYS AFTER PRODUCTION OF THE OLIVE OIL
=> LC-MS & NMR ARE VERY SIMILAR IN RESULTS, LCMS SHOWS SLIGHTLY MORE TOTAL OLIVE OIL POLYPHENOLS (3% TO 6% HIGHER) THAN THE NMR
=> HPLC BY FAR EXCEEDS LC-MS & NMR IN RESULTS, SHOWING +50% HIGHER TOTAL POLYPHENOLS THAN LC-MS OR NMR
IMPORTANT NOTICE: Allegations such as:
- "only HPLC shows the real polyphenols since it is recognized by the International Olive Oil Council"
- "LC-MS is the gold standard"
- "NMR inflates polyphenols by 66% compared to LC-MS and 70% compared to HPLC"
- "to convert NMR to LC-MS, multiply the NMR number by 0.6 to find the LCMS number"
- "to convert NMR to HPLC, multiply the NMR number by 0.66 to find the HPLC number"
THAT WOULD ALSO MEAN BY "COUNTER REVERSING" THAT:
- OUR ABOVE LCMS ANALYSIS 3107 mg/kg WOULD THEORETICALLY CORRESPOND TO, 3107 x 1.66 = 5176 mg/kg , 'NMR CONVERTED'.
- OUR ABOVE HPLC ANALYSIS 4412 mg/kg WOULD THEORETICALLY CORRESPOND TO, 4412 x 1.50 = 6618 mg/kg , 'NMR CONVERTED'.
IN REALITY THE ABOVE EXPERIMENT (ANALYSIS FROM THE SAME BOTTLE WITH NMR, LC-MS & HPLC) DEMONSTRATED THAT ALL THREE METHODS AND RESULTS ARE VALID.
POLYPHENOLS MEASUREMENT METHODS ARE NOT 'CONVERTIBLE'. NMR, LC-MS AND HPLC ARE SCIENTIFICALLY RECOGNIZED WITHIN THE SCIENTIFIC COMMUNITY AND EACH HAS ITS OWN PROVEN BACKGROUND
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