Mid-IR and Milk Chemistry Lina Jonsson MSc Animal Science Production Manager Miris AB, Sweden 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 Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Gammarays 1 pm X-rays UV VIS IR Micro- Radio waves 1 nm 1 um NIR 1m MIR 0.7 um NITS 1 mm 2.5 um NIRS 10 50 um Liquid Transmission MIR 2.5 um 700 nm 1100 10 um 2500 nm Fat 3.5 5.7 Protein 6.5 Carbohydrate 9.5 Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se History of Mid-IR • • • • • • Liquid Transmission Spectroscopy J.D.S Goulden 1961 IRMA Interference Filters FTIR Solid-State Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Gammarays 1 pm X-rays UV VIS IR Micro- Radio waves 1 nm 1 um NIR 1m MIR 0.7 um NITS 1 mm 2.5 um NIRS 10 50 um Liquid Transmission MIR 2.5 um 700 nm 1100 10 um 2500 nm Fat 3.5 5.7 Protein 6.5 Carbohydrate 9.5 Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Gammarays 1 pm X-rays UV VIS IR Micro- Radio waves 1 nm 1 um NIR 700 nm 1100 1m MIR 0.7 um NITS 1 mm 2.5 um 10 50 um NIRS 2500 nm Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk Spectra Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Principles of Mid-IR IR source Cuvette WavebandDetector CPU Selector Beräknings enhet Energy Energy Wavelength Energy Wavelength Wavelength Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk Spectra Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk Determination mid-IR Milk Water Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk Determination mid-IR Milk Water Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Spectra Milk Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk vs Milkfat Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Infrared Spectroscopy Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Infrared Spectroscopy Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Spectra of Milk and Skimmilk Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk vs Milkprotein Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Infrared Spectroscopy Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Milk vs Lactose Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Infrared Spectroscopy Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se 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 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 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 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 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 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 Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se ”Oiling off” + BAD MILK GOOD MILK 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 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) 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. 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 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) 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) Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Methods • IDF recommended methods • • • • Röse-Gottlieb Kjeldahl Enzymatic Drying oven 102°C Milk fat Milk protein Lactose Dry matter Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Metods Röse-Gottlieb • • • • • Neutral glycerider FFA Phospholipids Sterols Others (fat soluble vitamins.) % 98.7 0.1 0.8 0.3 0.1 Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Methods Kjeldahl Protein-N NPN-N Total-N Cow % % of milk 95 3.04 5 0.16 100 3.20 Human % 75 25 100 % of milk 0.9 0.3 1.2 Urea-N, Creatine-N, Uric-N, Peptide-N, Ammonia-N Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Analytical Methods Enzymatic • Water accounts about • Lactose monohydrate • Lactose anhydrous 5% 5.00 %-units 4.75 %-units 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, CO2, NH3 ) 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 Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Fatty Acid 4:0 6:0 8:0 10:0 12:0 14:0 15:0 16:0 16:1 17:0 18:0 18:1 18:2 CLA 18:3 Cow 4.0 2.6 1.4 3.1 3.3 10.6 0.9 27.7 0.9 0.5 8.2 15.4 2.4 0.47 0.8 (% wt/wt) Human 1.3 3.1 5.1 0.4 20.2 5.7 Rat 1.1 7.0 7.5 8.2 5.9 36.4 13.0 22.6 1.9 0.3 6.5 26.7 16.3 1.4 0.8 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. Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Protein Casein α-casein β-casein Whey κ-casein βlactoglobulin αlactalbumin Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Protein Casein β-casein Whey κ-casein Lactoferrin αlactalbumin 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 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 Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Questions? Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se Thanks for the attention! Miris AB | Kungsgatan 11, 753 18 Uppsala, Sweden | tel: +46 18 146 907 | www.miris.se