1. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Mid-IR and Milk Chemistry Lina Jonsson MSc Animal Science Production Manager Miris AB, Sweden

2. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Content What is Mid-IR? -History about Mid-IR -The principle of Mid-IR -The advantage of Mid-IR How to make an accurate calibration and take a correct sample What is milk, what are we measuring? -Analytical methods for milk Human breast milk

3. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Gamma- X-rays UV IR Micro- Radio rays waves 1 pm 1 nm 1 um 1 mm 1 m NIR MIR 0.7 um 2.5 um 10 50 um NITSNIRS 700 nm 1100 2500 nm Liquid Transmission MIR 2.5 um 10 um Fat 3.5 5.7 Protein 6.5 Carbohydrate 9.5 VIS

4. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Liquid Transmission Spectroscopy J.D.S Goulden 1961 IRMA Interference Filters FTIR Solid-State History of Mid-IR

5. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Gamma- X-rays UV IR Micro- Radio rays waves 1 pm 1 nm 1 um 1 mm 1 m NIR MIR 0.7 um 2.5 um 10 50 um NITSNIRS 700 nm 1100 2500 nm Liquid Transmission MIR 2.5 um 10 um Fat 3.5 5.7 Protein 6.5 Carbohydrate 9.5 VIS

6. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Gamma- X-rays UV IR Micro- Radio rays waves 1 pm 1 nm 1 um 1 mm 1 m NIR MIR 0.7 um 2.5 um 10 50 um NITSNIRS 700 nm 1100 2500 nm VIS

7. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk Spectra

8. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Beräknings enhet IR source Cuvette Waveband Selector DetectorCPU Principles of Mid-IR

9. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk Spectra

10. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk Determination mid-IR MilkWater

11. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk Determination mid-IR Milk Water

12. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Spectra Milk

13. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk vs Milkfat

14. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Infrared Spectroscopy

15. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Infrared Spectroscopy

16. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Spectra of Milk and Skimmilk

17. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk vs Milkprotein

18. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Infrared Spectroscopy

19. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk vs Lactose

20. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Infrared Spectroscopy

21. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se

22. The advantages of Mid-IR Quick analytical result The milk can be used after the composition is known The composition of fat protein and lactose in the same analyse Identify dilution before the milk has entered the manufacture

23. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se How to make an accurate calibration and take a correct sample A certified reference lab High quality of the milk Homogenous milk Good sampling routines imperative to obtain representative samples All result must be review critical, mistakes do happen

24. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Sampling Inhomogenity of milk demands special treatments of milk before sampling and/or analyse -Creaming -Aggregation -Foaming Protein Chain

25. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Heating/Mixing Mixing more effective of warm samples (40-43 C) Fat aggregations are dissolved. Condensed water is mixed into the milk

26. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Ultrasound Treatment Previously frozen milk ought to be treated ultrasound 1 second / ml The efficiency of treatment is increased by temp. 40°C is optimal for milk Protein aggregation are dissolved Oil droplets are emulsified

27. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Chemical/Physical factors that interferes with the “true” value of the milk Non Protein Nitrogen (NPN) Lactic acid Lipolysis Free fatty acids Creaming Oiling off

28. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se + ”Oiling off” GOOD MILK BAD MILK

29. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Fat globule (schematic) Lipophile part Phospholipids + membrane Hydrophile part

30. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se What is milk, what are we measuring? What is milk according to definition??? Milk fat Milk protein Lactose Dry matter (Total solids)

31. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Methods for Milk Cow Milk –Different international standards (ISO/IDF/AOAC) for measuring constituents in milk. Also valid for other species. –(ISO 9622:1999; ISO 1211; ISO 8968-1 etc) Human Milk No international standards for human milk methods.

32. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Methods Fat Gerber, Babcock, Lindström Röse-Gottlieb, Mojonnier Turbimetric (Milko-Tester) MIR (Mid Infrared Transmission) NIR (Near Infrared Reflectance/Transmission ) Ultrasonic high frequency

33. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Methods Protein Kjeldahl MIR Mid Infrared Transmission NIR Near Infrared Transmission/Reflectance (Ultrasonic high frequency)

34. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Methods Lactose Luff-Schoorl HPLC Enzymatic MIR Mid Infrared Transmission (NIR) (Ultrasonic)

35. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Methods IDF recommended methods Röse-GottliebMilk fat KjeldahlMilk protein Enzymatic Lactose Drying oven 102°C Dry matter

36. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Metods Röse-Gottlieb % Neutral glycerider 98.7 FFA 0.1 Phospholipids0.8 Sterols0.3 Others (fat soluble vitamins.) 0.1

37. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Methods Kjeldahl Cow Human % of milk % % of milk Protein-N 95 3.04750.9 NPN-N5 0.16250.3 Total-N100 3.201001.2 Urea-N, Creatine-N, Uric-N, Peptide-N, Ammonia-N

38. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Methods Enzymatic Water accounts about 5% Lactose monohydrate 5.00 %-units Lactose anhydrous 4.75 %-units

39. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Methods Drying oven All that evaporate is measured as water Volatile components (Lactic acid, fatty acids, CO 2, NH 3 )

40. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Human breast milk Unsaturated fat Day to day variation Protein composition Lactose

41. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se (% wt/wt) Fatty AcidCowHumanRat 4:04.0 6:02.6 8:01.41.1 10:03.11.37.0 12:03.33.17.5 14:010.65.18.2 15:00.90.4 16:027.720.222.6 16:10.95.71.9 17:00.50.3 18:08.25.96.5 18:115.436.426.7 18:22.413.016.3 CLA0.47 18:30.81.40.8

42. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Human breast milk Day to day variation S. Polberger and B. Selander, 2007.

43. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Protein Casein α-caseinβ-caseinκ - casein Whey β- lactoglobulin α- lactalbumin

44. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Protein Casein β-caseinκ - casein Whey Lactoferrin α- lactalbumin

45. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Methods Oligosaccharides Three to ten monosaccharides including glucose, galactose, fucose, sialic acid, N-acetylglucoseamine Account for about 0.8-1.2 g/100 ml Complicated analytical methods, but mid-infrared methods seems to measure this carbohydrates due to hydroxyl (-OH) groups

46. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Summary Milk is not a solution, it is a emulsion. The different components can aggregate or separate, which makes sampling difficult. The Mid-IR method only need one sample to give the complete major components of milk This reduce the source of error and makes the result more comparable A small amount of milk is needed, the result comes with in a minute No chemicals is needed Approved by IDF

47. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Questions?

48. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Thanks for the attention!