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TRANSCRIPT
IDF Symposium “Lactose and its Derivatives”14-16 May 2007, Moscow, Russia
3A Business Consulting3A Business Consulting 11
May 15, 2007
Market developments and industry challenges for lactose and lactose
derivatives
3A Business Consulting3A Business Consulting 22
3A Business model
Strategy and businessdevelopment for all elements of the food value chain
Key competenceswithin functional foods, health& wellness and ingredients
Core structure - global network
3A Business Consulting3A Business Consulting 33
3A multi-client reports
3A Business Consulting
November 2005
Global Market Analysis of Whey and Lactose Products 2004-2009 - From commodities to value
added whey protein fractions and lactose derivatives
Dairy Ingredients in Nutritional Sectors Supply/Demand/Forecasts 2005-2010
USA, EUROPE
Date
GIRACT 3A BUSINESS CONSULTING
Website: www.giract.com
Email: [email protected]
Website: www.3abc.dk
Email: [email protected]
S H A I N W R I G H TCONSULTING AND RESEARCH GROUP PTY LTD
China – dairy opportunities unlimited
- dairy production, consumption & trade, trends, players and outlook 2008
3A Business Consulting
February 2006
Update version available from August 2007
3A Business Consulting3A Business Consulting 44
Lactose- introduction
Lactose- marketoverview
Lactose- lactose
derivatives
Market developments and industry challenges for lactose and lactose derivatives
Lactose- summing
up
3A Business Consulting3A Business Consulting 55
Global whey production figures - 2006
177
167
9,6
0 20 40 60 80 100 120 140 160 180 200
Total whey production
Global whey resultingfrom cheese
Global whey resultingfrom casein
million tonsSource: 3A Business Consulting
~ approx. 6%
~ approx. 94%
Liquid whey supply CAGR approx. 2-3%
2001-2006
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Million tons / year
Total world production
177Industrially
utilized
124
Whey powder, lactose
70
WPC/WPI producing permeate
43
Feed, fertilizer,
waste
53 Demineralized, blends, etc.
11
177
World whey utilization - 2006
70%30%
Source: 3A Business Consulting
124
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Whey and lactose product universe
Source: 3A Business Consulting
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CheesewheyCaseinwhey
LactosePermeatepowder
Lactose –pharma grade
Lactosederivatives
GalactoseLactuloseLactitolLactobionic acidGOSLactosucroseSialyllactoseTagatose
Applications:PharmaNutritionHealth careFoodFeed
Added value level
Lactose products universe
Whey raw
materialLactose IILactose I Lactose III
Source: 3A Business Consulting
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Value-adding in whey processing (illustration)
Value creation
Technology requirements
HighLow
Low
High
Whey protein fractionsBioactive peptides
WPI/WPH
WPC80WPC35
WP feedWhey powder
Source: 3A Business Consulting
Permeate powder
LactitolLactulose GOS LBA
Lactosucrose
Sialyllactose
Lactose-pharma
Galactose
Lactose
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Lactose- introduction
Lactose- marketoverview
Lactose- lactose
derivatives
Market developments and industry challenges for lactose and lactose derivatives
Lactose- summing
up
3A Business Consulting3A Business Consulting 1111
Overview of global lactose production 2002-2006
EU 49%
US36%
NZ12%
ROW3%
EU 46%
US37%
NZ13%
ROW4%
Global production: 724,000 tons
Global production: 870,000 tons
2002 2006
The EU and the US are the major producing countries representing more than 83% of the global production (down from 85% in 2002)
New Zealand is the only other major producing country with 13% of the global production corresponding to 110,000 tons (up marginally from 12% in 2002)
Source: USDEC, University of Wisconsin, 3A Business Consulting
CAGR: approx. 5%
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Commodities still make up the most significant part of the lactose business
68%
32%
High value-added lactose ingredients (lactose derivatives) US$ 217 million
Commodities (lactose, permeate powder) US$ 460 million
Source: 3A Business Consulting
66%
34%
Commodities (lactose, permeate powder) US$ 1265 million
High value-added lactose ingredients (lactose derivatives) US$ 665 million
US$ 677 million US$ 1,930 million
Market value growth 185%
2004 2006
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Cheese wheyCasein whey
LactosePermeatepowder
Lactose – pharma grade
Lactosederivatives
Applications:PharmaNutritionHealth careFoodFeed
Added-value level
Illustration of lactose ingredient producers
Whey raw
materialLactose IILactose I Lactose III
Numerous companies
Campina/DMVFriesland Foods DomoMeggleHilmar IngredientsFonterra
Friesland Foods DomoSolvayDaniscoADMFerro PfanstiehlPuracInalcoFresenius-KabiBiofac
Source: 3A Business Consulting
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World's leading lactose producers
Friesland Foods Domo8%
Leprino Foods6%
All others 50%
Arla Foods3%
Fonterra12%
Meggle 4%Lactalis
3%
Hilmar Ingredients
5%
Campina/DMV10%
In 2006, 7 out of the 8 biggest lactose producers in the world are found in the EU and the US
Fonterra has achieved the position as the world's biggest lactose producer with 12% market share
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Carbery, Anchor AlcoholAlcohol
Purac, Cargill, Hendan Jindan, ADM, GalacticLactic acid
Solvay, Sandoz, US Dairy Ingredient CompanyLactobionic acid
Danisco, Purac, Towa, NikkenLactitol
Morinaga, Milei, Solvay, Inalco, Fresenius-Kabi, Relax, Biofac
Lactulose
Hayashibara, EnsuikoLactosucrose
Friesland Foods Domo, Morinaga, Snow Brand, Yakult, Nissin
GOS
ProducersProducts
Lactose/permeate derivatives and producers
Note: In 2006 Arla Foods Ingredients stopped their production of tagatose due to economics
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Lactose segment break down – 2005
Source: 3A Business Consulting/EWPA/ZMP
EU production distribution/applications
Infant formula 18%
Bakery products 5%Pharmaceuticals
28%
Other processed food incl. meats
30%
All other uses3%
Chocolate confectionery
industry 16%
EU consumption approx. 325,000 MT; not same distribution/applications
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Lactose segment break down –US market applications 2005
All other uses13%
Nutraceuticals, Pharmaceuticals
5%
Confectionery industry
16%Infant formula
66%
Source: 3A Business Consulting/ADPI
Total US consumption approx.
130,000 MT
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Lactose price development from 2004 to date
0
0,1
0,2
0,3
0,4
0,5
0,6
0,7
0,8
0,9
1
2004 2005 2006 2007
$/lb
Lactose prices
Growth: >300%
Source: University of Wisconsin, EWPA, Trade interviews
0
0,5
1
1,5
2
2,5
2004 2005 2006 2007
EUR/kg
Growth: >300%
US market
EU market
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Top 10 lactose exporting countries 2005
0 50 100 150 200
Lithuania
Ireland
Denmark
France
Australia
Italy
New Zealand
Germany
Netherlands
US
1,000 MT
Growth since 2002
+ 54%
- 7%
+ 53%
+ 152%
+ 234%
- 13%
+ 93%
+ 15%
+ 23%
+ 9%
Source: FAOSTATS & 3A Business Consulting
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Top 10 lactose importing countries 2005
0 20 40 60 80 100
Germany
United Kingdom
Korea
Spain
France
Vietnam
Mexico
Netherlands
China
Japan
1,000 MT
Growth since 2002
+ 188%
+ 160%
+ 31%
+ 189%
+ 15%
+ 28%
+ 0%
+ 22%
- 33%
+ 0%
Source: FAOSTATS & 3A Business Consulting
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Lactose- introduction
Lactose- marketoverview
Lactose- lactose
derivatives
Market developments and industry challenges for lactose and lactose derivatives
Lactose- summing
up
3A Business Consulting3A Business Consulting 2222
Permeatepowder
Lactose
Biogas
Nisin
Fertilizer
Animal feed
Lactulose
Effluent plant
GOS
LBA
Mineral powder
Beverages
Protein standardisation
Galactose/glucose syrup
Volume
Innovation and value added
Lactose derivatives –strategic mapping
Commodity Main stream Specialty
Alcohol
Lactic acid
Lactosucrose
Lactitol
Galactose
Sialyllactose
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Market positioning/segments: Bulking agent for sugar free products, low sweetness, high viscosity and low hygroscopicity properties, non-cariogenic, reduced calorie-effect. Also some pharma use
Competing products: Food: maltitol, isomalt and erythritol. Possibly isomaltulose, trehalose and tagatose. Pharma: lactulose
Competition: Danisco (DK), Purac (NL), Towa (J), Nikken (J)
Market size: Approx. 10,000 MT and US$ 50 milllionCAGR: 2-4%
Market data - Lactitol
Overall evaluation/market potential/future prospects
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New products with Lactitol
Total number of new products: 865
Source: Mintel, 2001-2007
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Product examples with Lactitol
Nutrabien Chocolate Chip Cookie is sugar-free, low in sodium and high in fiber. The product is free from colorants and preservatives. Also available is an Oats variant.
Alimentos NutraBien –Nutrabien
Chocolate Chip CookieYakult – Yakult Bifiene
Fermented Drink
Yakult Bifiene is a milk-based fermented drink that contains bifidobacterium breve Yakult, which is said to help promote good intestinal function.
Source: Mintel
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Market data - Galactooligosaccharides
Market positioning/segments: Prebiotic, non-cariogenic, positive effect on constipation. Main potential segments include beverages, dairy products, infant formula
Competing products: Other prebiotics/oligosaccharides
Competition: Friesland Foods Domo (NL), Morinaga (J), Snow Brand (J), Yakult (J), Nissin (J); possibly new players
Market size: Approx. 20,000 MT and US$ 120 milllionCAGR: 10-20%
Overall evaluation/market potential/future prospects
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New products with galactooligosaccharides
Total number of new products: 40
Source: Mintel, 2001-2007
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Product examples with galactooligosaccharides
Source: Mintel
Kasdorf – NutriciaBagó Nutrilon Premium – Infant Milk powder
Nutricia Bagó Nutrilon Premium Después de la Primera Infancia Infant Milk Powder is a modified milk powder, with prebiotics, made from partially skimmed milk, maltodextrin, vegetable oil, vitamins and minerals. It is said to help reinforce the immune system of infants.
Coca-Cola – Ooo –Muscat Au Lait
A milky drink with 1% muscat grape juice and oligosaccharide.
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Market data - Lactulose
Market positioning/segments: Pharma: mainly treatment of constipation, Food: as a nutraceutical ingredient in infant formula, diabetic foods, soft drinks etc.
Competing products: Other oligosaccharides in food, lactitol in pharma
Competition: Morinaga (J), Solvay (B), Inalco (I), Fresenius-Kabi (AT), Relax (SA), Biofac (DK)
Market size: Approx. 25,000 MT and US$ 300 milllionCAGR: 2-4%
Overall evaluation/market potential/future prospects
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New products with Lactulose
Total number of new products: 34
Source: Mintel, 2001-2007
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Product examples with Lactulose
Lactulose are prune flavouredlaxatives which are said to relieve constipation. It is suitable for adults and children aged seven years and older.
Merck Génériques –Merck GénériquesLactulose – Prune
Flavoured Laxatives
Source: Mintel
Pasteur has launched Regular Motions Yogurt Drink, which contains synbiotics for regular motions, complex dietary fibre, complex oligosaccharides and prune juice.
Pasteur – Pasteur –Regular Motions
Yoghurt Drink
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Lactose/permeate derivatives overview 2006
Market overview
870
300
35 21 100
100
200
300
400
500
600
700
800
900
1000
Lactose Permeatepowder
Lactulose GOS Lactitol
000´ tons
2-4% 10-20% 0-5% 10-20% 0-5%
Source: 3A Business ConsultingMarket growth rates
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Lactose- introduction
Lactose- marketoverview
Lactose- lactose
derivatives
Market developments and industry challenges for lactose and lactose derivatives
Lactose- summing
up
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Expected market development for lactose
Source: 3A Business Consulting
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Growth opportunities ahead for lactose and derivatives for the industry 2006-2010
Marketgrowth
Value(US$/kg)
β
HighLow
Low
High
Source: 3A Business Consulting
Sialyllactose
LBA
Lactosucrose
Lactose-pharma
GOS
Permeate powder
Lactose
Lactulose
Lactitol
Commodities
High-end derivatives
Emerging
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Regulatory challenges for new dairy ingredients/lactose derivatives
Ingredient approval EU – Novel Foods regulation and the US –FDA Food additive petition/GRAS approval
Documentation of health benefits
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A number of strategic alliances exist within dairy ingredient processing
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Examples of strategic alliances within lactose processing
Lactose-pharma
LactuloseGOS
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Whey ingredients set for growth
The market for industrially utilised whey ingredients will continue to grow particularly in food and nutrition applications
High-value added lactose ingredients will also see moderate growth rates due to increasing demand from the nutritional and pharmaceutical segments, however the pharma market face competition from other excipients e.g. MCC and starch
Lactose derivatives such as lactulose, lactitol alongside galactooligosaccharides are showing interesting new application opportunities and significant annual growth rates
New lactose derivatives such as LBA and sialyllactose are emerging and will develop into commercial products
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Market dynamics for lactose
Global market demand for dairy products and dairy ingredients approx. 2.8% CAGR exceeds current global milk supply growth of less than 1%
The dramatic increase in the price of SMP due to lower production and elimination of EU and US stocks have had a knock-on effect on the price of WP and lactose
A growing demand for lactose for pharma and nutrition usage as well as for lactose derivatives and protein standardization of SMP has further pushed up lactose prices
High end applications for lactose will continue to be locked into lactose, whereas low end applications will try to reformulate product compositions
Increase in lactose supply will not likely keep up with demand
Dairy commodities/ingredients including lactose will remain highpriced well into 2008 and the foreseeable future
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Thank you for your [email protected]
IDF Symposium “Lactose and its Derivatives”14-16 May 2007, Moscow, Russia
University of Alberta
Paul Jelen Dept. of Agricultural, Food and Nutritional
ScienceUniversity of Alberta
Edmonton, Alberta, Canada
Properties of lactose as determinants of crystallization behaviour and of
industrial applications
Lactose
Milk of (almost) all mammalsPrimary source of energy for the neonateHighest concentration (7%) in human milkPrincipal component of cow’s milk (4.5 - 5%; 53% of non-fat solids)Cheapest reducing CHO on the market
Concentration of lactose in milk of different mammals
Human
Cow
Buffalo
Goat
Sheep
7.1
4.6
4.8
4.3
4.8
H2O content (%)H2O content (%)SpeciesSpecies Lactose content (%)Lactose content (%)
87.1
87.3
82.8
86.7
82.0
Schematic representation of the lactose disaccharide molecule
Important basic properties of lactose
Disaccharide (glu-gal)Two stereoisomers (α- and β) > mutarotationReducing sugar >>> browning reactionLow sweetness (20% of sucrose at 5% conc.)Low solubility (18% w/w at 20°C)
> low osmotic pressure> slow crystallization
Forms of lactose
α - lactose, β – lactose In solution the ratio of α : β is about 1 : 2 Effect of temperature and mutarotation on
> solubility> sweetness> crystallization behaviour
In crystalline state> α – lactose contains one H2O molecule> α – lactose anhydride (heating above 130oC)> β – lactose (above 95oC) contains no H2O
Properties of lactose affecting the dairy industry
> Fermentable by lactic acid bacteria > Crystallizes in highly concentrated dairy foods
(sweetened condensed milk, ice cream, whey cheese mysost)
> Low sweetness – unsuitable as a sweetening agent, can be improved by hydrolysis
> Lactose malabsorption limits consumption of dairy foods by lactose intolerant consumers
> Fermentable by aquatic bacteria - high BOD of whey
> Unique crystallization behaviour
Focus of this presentation
Crystallization phenomena Model systems – experimental methodsSolubility Real life dairy foods
Sweetness phenomenasensory impact in dairy foodsuse for protein standardization
Lactose hydrolysisLactose derivatives and pharma-lactose
Centuries of dairy science:lactose
1633 – Bartoletto isolated and described lactose as “essential salt without nitrogen”
1688 - Ettmueller isolated lactose from whey and purified by recrystallization
1814 – 1820 analytical work by Berzelius 1902 – 1942 fundamental work by Hudson1936 – 2007 lifetime achievements of Prof.
Andrei Georgievich KHRAMTSOV
Lactose crystallization
Supersaturated solutionConcentrated dairy systems (condensed milk, ice-cream, whey cheese)Whey or whey permeate >> evaporation and cooling for production of lactose commodityCrystalline habit and crystal growth mechanism (α-lactose)
Single crystal method to study the crystallization phenomena
Method described by Professor J.A. Kucharenko from Polytechnical Institute, Kiev, for study of sucrose crystallization
Series of 12 articles published in “The Planter and Sugar Manufacturer” (New Orleans), from May 12 to July 28, 1928 (volumes 80 and 81)
Crystallization velocity, density, effects of impurities, solubility, supersaturation
Single crystal method adapted to study lactose crystallization
Lactose crystal growth: single crystal experiments
Van Kreveld (1966 – 1969): crystals grow from the apex of a pyramid down
Jelen (1971 – 74): effect of supersaturationand growth promotion by mineral impurities
Visser (1980 – 1983): crystal growth retarders, structure of lactose crystal
Jelen (1996) – crystallization velocity with assumption of bottom plane growth
Lactose solubility
Temperature oC Concentration of saturated solution (g / 100 g H2O)
___________________________________________
30 oC 24.050 oC 44.070 oC 77.8 80 oC 98.9
Effect of minerals on lactose solubility at 30oC
Salt added Estimated solubility(5 g / 100 g H2O) (g / 100 g H2O)
__________________________________________________
Control (no salt) 24.4Calcium chloride 24.3Magnesium sulphate 23.0 Lithium chloride 21.9Potassium phosphate 26.5
Lactose crystallization velocity
Average growth rates of lactose crystals in model lactose solutions with or without the addition of salts.
Lactose crystals under microscope
Scanning electron microscopy of spray dried demineralized permeate powder (courtesy Dr. Kalab, Ottawa).
Light microscopy of demineralizedwhey permeate powder (courtesy Dr. Kalab, Ottawa).
Growth rates of lactose crystals (30oC, supersaturation 9 g)
Exposed face
AllSidesTop (truncated)Bottom
Crystallization rate (mg m-2 min-1)
160.9 ± 12.12.5 ± 3.0
-0.6 ± 3.4211.8 ± 69.3
Growth rates of lactose crystals(30oC, supersaturation 10g)
Assumptions
Growth on all sides
Growth on bottom side only______________________________________Sucrose
Approximate Crystallization rate (mg m-2 min-1)
80 (Jelen 1972)
350 (Bhargava/Jelen, 1996)
320 (Smythe, 1971)
Promoters and inhibitors of crystal development
Promoters
Lithium chloride
Calcium chloride
(at low concentrations)
Some phosphates
Other electrolytes (?)
β-lactose
Riboflavin
Galactose
Gelatin
Potassium salts
Inhibitors
Lactose solubility and crystallization in whey, UF permeate or dairy foods
Nucleation vs crystal growthComponents affecting solubility and crystal
growth Very high supersaturation – nucleation
favored Ice cream, frozen desserts, Sw. Cond. MilkNorwegian whey cheese Mysost
Lactose crystals in whey cheese Mysost
Lactose crystals in whey cheese Mysost
Lactose crystals in whey cheese Mysost
Lactose crystals in whey cheese Mysost
Effect of mineral impurities on lactose crystals
Sweetness of lactose
Much lower than sucrose (20 % at 5% conc.)Effect of temperature and concentrationAddition of less that 1% lactose to milk
clearly noticeable (Jelen & Michel, 1999)Lactose used for down standardization of
protein in dry milkSweetness can be increased by hydrolysis
Lactose hydrolysis
Low solubility (sandiness)
Low sweetness
Lactose intolerance
Oligosaccharides
50 Mpersons market in USA alone
Acid + heat
Free enzyme
Immobilized enzyme
Disrupted bacteria
MethodsReasonsReasons
Characteristics of lactose hydrolysis methods
• Acid-catalysed hydrolysis – technological and material problems
• Immobilised enzyme technology– rarely succesful (Valio)
• Membrane - based enzyme reactors– theoretically interesting, rarely used
• Free (soluble) purified enzymes – used in industrial practice– Tetra-lacta process– home use in milk or as a dietary aid (pill)
How widespread is lactoseintolerance in the world?
Lactose hydrolysis by disrupted lactic acid bacteria
• framework: Canadian research network on lactic acid bacteria for applications in dairy industry (University of Alberta).
• objective: to investigate a “simple” approach to the lactose hydrolysis problem using lactase enzyme produced by common dairy bacteria after their disruption as crude enzyme extracts (CEE) obtained by microfluidization
Components of the proposed lactose hydrolysis process
well defined (GRAS) enzyme source (Lactobacillus bulgaricus 11842)
well defined (GRAS) medium (skim milk, whey)culture production, separation and disruption (bead
mill, high pressure homogenizer, Microfluidizer)use of the “dirty” CEE for lactose hydrolysis without
additional purification
Schematic representation of the proposed process
culture propagation (skim milk)
centrifugation or MF
culture disruption milk processing
(NF, HPUF, UF, DF)
viable cell deactivation (MF) lactate, salts
reprocessed milk
addition to milk or whey UF permeate
downstream processing
downstream processing
SEM images of Disrupted L. bulgaricus 11842
Control 1 pass (homogenizer)
Sonication (6 min) Bead Mill (6 min)
Lactose derivatives: GALACTO - OLIGOSACCHARIDES
Di-, tri-, tetra- or higher -saccharides
Intermediate sweetness
Highly heat and acid stable
Bifidogenic factor
Non-digestible
Probiotic foods
Nutraceutical (FOSHU) foods (anticarcinogenic)
Non-cariogenic foods
Competing against inulin
PropertiesProperties Applications
Lactose derivatives as value added products
Lactulose: 16 kt/yearLactitol: 10 kt/year in 1 plantLactobionic acid: potential for 1kt/year in 1
German plantOligosaccharides:
Galactooligosaccharides: n/aLactosucrose: 1.6 kt/year
Pharmaceutical lactose products
α-lactose:100 mesh (>125 µm)Agglomerated / granulatedSpray dried (80% crystals, 20% amorphous)
Anhydrous lactose:α -lactose (heated >130°C)β-lactose (crystallized >93°C)
Pharmaceutical lactose processes
Whey or UF permeate (crystallization)
Crude lactose
Refining recrystallization
Refined α-lactose
Heating >130°C
Anhydrous α -lactose
Spray drying
Spray dried α-lactose
Crystallization on roller driers >93 °C
β-lactose
Industrial processors of pharma-lactose
Manufacturer Product trade name
Lubricant
Meggle
BASF
DMV
TablettoseCellactose
Ludipress
Pharmatose
-Cellulose
PVP
Lactitol
New aspects of lactose science and industrial applications
Use of microcrystalline lactose as a flavourcarrierLactose effects in microencapsulation of fat by WPCCaking in bulk lactoseUse of lactose for protein standardization in non-fat-dry-milk and fluid milk
Traditional uses of isolated lactose
Food(1996 total 425 kt)
Infant foodsConfectioneryOther (bakery, dry mixes)Dairy (protein standardization)
Other(1996 total 175 kt)
PharmaceuticalsFermentationFeedsDerivatives
THANK YOU!
IDF Symposium “Lactose and its Derivatives”14-16 May 2007, Moscow, Russia
LACTOSE CRYSTALLIZATION
FROM SATURATED SOLUTIONS
Aram Galstyan
GNU All-Russia Dairy Research Institute, Moscow, RussiaDepartment of gerodietical and special products
Тел./Факс: +7 (495) 236-02-36www:vnimi.org
E-mail: [email protected]
Product qualitative index*
Consistence
HomogeneousNot less than 400.000 crystals in 1 mm3 of product
Crystals average size ≤ 10 µm
Technological operations
Crystallization FermentationGerms
Vacuum
Isohydric Lactose preparation
Immobilizing
Consumed
Small dispersionlactose (3-4 µm )
Powder Suspension
* Радаева И.А., Гордезиани В.С., Шулькина С.П. Технология молочных консервов и ЗЦМ: Справочник– М.:Агропромиздат, 1986.Чекулаева Л.В., Чекулаев Н.М Сгущенные молочные консервы. – М.: Легкая и пищевая пром-ть, 1982. Чекулаева Л.В., Полянский К.К., Голубева Л.В. Технология продуктов консервирования молока и молочного сырья.– Воронеж: Изд. ВГУ, 1996.
Introduction
Technological requirements
Crystallization
Saturation level
To control energy exchange processes
Prime material availability
To control mass distribution processes
Fermentation
To have a ferment
To know the activity, optimal conditions
To know the properties, decide the adding time
Introduction
Crystallization
Homogeneous Heterogeneous
Heterogeneous crystallization nucleus
Homogeneouscrystallization nucleus
The composition of exuded crystalsdoes not correspond to
composition of crystallization centers
The composition of exuded crystalscorresponds to
composition of crystallization centers
1) Shabalin V.N., Shatoxina S.N. Morphology of biological fluids. – M.:Chrizostom, 20012) Rosenberger F. Fundamentals of Crystal Growth.— Berlin, 19793) Ohtaki H. Crystalization Processes. – Wiley, 20014) Hartel R.W. Crystallization In Foods.- Kluwer Academic Publishers, 20015) Веригин А. Н., Щупляк И. А., Михалев М. Ф. Кристаллизация в дисперсных системах: Инж. мет. расчета.- Л.: Химия, 1986 6) Портнов В.Н., Чупрунов Е.В. Возникновение и рост кристаллов.-М.:Изд. Физ.-мат.лит., 2006
http://www.wikipedia.org http://www.xumuk.ru http://www.paceka.ru http://www.cheresources.com http://www.geo.web.ru и др.
Introduction
Condensed milk with sugar Boiled condensed milk with sugar
Raw materials acceptance, storage and preparation
Dry milk products reduction at
water temperature 40-60 °C Lactose fermentative hydrolyses (39-41 °C, for 3h)1
Dispergration of the fatty
component
Heating (75-77 °C)
Sugar dissolution
Heating and pasteurization
(90-95 °C for 2-3 min) Heating
Cooling (32-37 °C)
Crystallization 3
Homogenization
(15-16 Pa) at T ≥ 30 °C Heat treatment (cooking) (95 °C and more for 1-3 hours) 2
Cooling (20 ± 2 °C) Cooling to 80-85 °C and homogenization (15-16 MPa)
Packaging Packaging
Marking Marking
Storage and sale
1 Employed with a purpose of prevention the uncontrolled crystallization & speeding-up cooking time 2 Duration depends on the temperature & fermentation process 3 The adding of priming material in vacuum meant. In manufacture of sugar- containing condensed milk “cooking” is not employed.
Principle technological schemes of manufacture of sugar containing condensed milk products
Materials and Methods
Model product’s normalized indices
≤ 15Allowed size of lactose crystals, µm ,
≤ 3-615
Viscosity, Pa*сvalid within 2 monthsvalid within 2 to 12 months
≤ 45Acidity,0Т
≥ 28,5≥8,5
Total mass proportion of milk dry materials, %also fat, %
≥ 43,5Mass proportion of saccharose, %≤ 26,5Mass proportion of moisture, %
StandardsIndex appellation
Materials and Methods
Х51000,0Total productХ4255,0WaterХ3450,0Granulated sugar (dry materials proportion 99,8%)Х282,0Milk fat (fat proportion 99,8%)
Х1213,0Dry skimmed milk (dry materials proportion 95%, fat proportion 1,5%)
CodingRecipesComponents
Recipes of sugar-containing condensed milk productsand established conventional coding
Х5=Х4 + Х1 + Х2 + Х3
Y14StoringY7Cooling
Y13MarkingY6Pasteurization
Y12MixingY5Homogenization
Y11PackingY4Heating
Y10CookingY3Dispergration
Y9CrystallizationY2Germs bringing in
Y8VacuumingY1Dissolution
CodingAppellation ofthe operation
CodingAppellation ofthe operation
Materials and Methods
Y15 X5---ТО15
Y13 X5Y14X5Y14X5Y14X5ТО14
Y7 X5Y13X5Y13X5Y13X5ТО13
Y10 X5Y11 X5Y11 X5Y11 X5ТО12
Y11 X5Y9X4123Y9X4123Y9X4123ТО11
Y7X4123Y7X4123Y7X4123Y7X4123ТО10
Y8X4123← ZY8X4123← ZY8X4123Y8X4123ТО9
Y6X4123Y6X4123Y6X4123Y6X4123ТО8
Y1X3Y1X3Y1X3Y1X3ТО7
Y4X412Y4X412Y4X412← ZY4X412ТО6
Y5X412Y5X412Y5X412Y5X412ТО5
Y3X2Y3X2Y3X2Y3X2ТО4
Y4X41Y4X41Y4X41Y4X41ТО3
Y1X1Y1X1Y1X1Y1X1ТО2
Y4X4Y4X4Y4X4Y4X4← ZТО1
C/CBА
Possible modification of germs adding timeOperationssequence
ТО – technological operation; Z – time point of bringing in of germs
Operative models: condensed milk with sugar (A,B,C) andcondensed milk with sugar “cooked” (С/)
Materials and Methods
12015 302
0 Time (Days)
9060 7545 105
Time point of sampling for microscoping
0 – end of technological process and beginning of analysis120 – end of analysis
Storage temperature
6-100C
Materials and Methods
14-15CaCO3Calcium carbonate3
not presentTiO2Titanium dioxide2
Not present
≥ 98
SiO2Silicon dioxide1
Solubility in water at рН≤7,0, mg/l
Properties, %
Molecular formula
Products№
2 60040175 00010
7 00030220 0009
21 00020270 0008
50 00015500 0007
98 00012770 0006
Lactose crystals likely amount in 1mm3 of
product, M
Crystals average size, µm
Lactose crystals likely amount in 1mm3 of
product, M
Crystals average size, µm
∑ ∑∆= )2/( 2nvnаUU – homogeneity coefficient; ∆а – crystal size limit, µm; n – crystals frequency; v – given and mean sizes divergence, µm
D – mean value of crystals size, µm ; n – crystals frequency; а – crystals linear size, µm
Coefficient of crystals homogeneity was calculated by the means of N. Figurovskii’s formula
Crystals average size was calculated by formula:
D = ∑na / ∑n
Materials and Methods
Distribution of samples with no crystallization effect when germ size is 3 µm and its corresponding dosage 0,02…0,10% of product mass
14%
16%
22%
31%
46%
0
10
20
30
40
50
60
700.10
0.09
0.080.05
0.02
Y14X5
Y13X5
Y11 X5
Y9X4123
Y7X4123
Y8X4123← Z
Y6X4123
Y1X3
Y4X412
Y5X412
Y3X2
Y4X41
Y1X1
Y4X4
Model C
Results
51%
68% 21%
28%
40%
0
10
20
30
40
50
60
700.10
0.09
0.080.05
0.02
Distribution of samples with no crystallization effect when germ size is 4 µm and its corresponding dosage 0,02…0,10% of product mass
Y14X5
Y13X5
Y11 X5
Y9X4123
Y7X4123
Y8X4123← Z
Y6X4123
Y1X3
Y4X412
Y5X412
Y3X2
Y4X41
Y1X1
Y4X4
Model C
Results
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4C
oeff
icie
nt o
f hom
ogen
eity
, U
3 4
Germ size, µm
Silicon dioxideTitanium dioxideCalcium carbonate
Y14X5
Y13X5
Y11 X5
Y9X4123
Y7X4123
Y8X4123← Z
Y6X4123
Y1X3
Y4X412
Y5X412
Y3X2
Y4X41
Y1X1
Y4X4
Model C
Lactose crystals homogeneity coefficient when priming material size is 3-4 µm
Results
Silic
on d
ioxi
de
Tita
nium
dio
xide
Cal
cium
car
bona
te
0
2
4
6
8
Lact
ose
crys
tals
size
, µm
0.082 0.089 0.098
Dosage of priming material, %Y14X5
Y13X5
Y11 X5
Y9X4123
Y7X4123
Y8X4123
Y6X4123
Y1X3
Y4X412
Y5X412
Y3X2
Y4X41
Y1X1
Y4X4← Z
Model А
Rational dosage of priming material for lactose crystallization by model A
Results
Silic
on d
ioxi
de
Titan
ium
dio
xide
Calci
um ca
rbon
ate
0
2
4
6
8
Lact
ose
crys
tals
size,
µm
0.051 0.061 0.078
Dosage of priming material, %Y14X5
Y13X5
Y11 X5
Y9X4123
Y7X4123
Y8X4123
Y6X4123
Y1X3
Y4X412← Z
Y5X412
Y3X2
Y4X41
Y1X1
Y4X4
Model B
Rational dosage of priming material for lactose crystallization by model B
Results
Silic
on d
ioxi
de
Tita
nium
dio
xide
Cal
cium
car
bona
te
0
2
4
6
8
Lact
ose
crys
tals
size
, µm
0.022 0.029 0.031
Dosage of priming material, %Y14X5
Y13X5
Y11 X5
Y9X4123
Y7X4123
Y8X4123← Z
Y6X4123
Y1X3
Y4X412
Y5X412
Y3X2
Y4X41
Y1X1
Y4X4
Model C
Rational dosage of priming material for lactose crystallization by model C
Results
Y15 X5
Y13 X5
Y7 X5
Y10 X5
Y11 X5
Y7X4123
Y8X4123← Z
Y6X4123
Y1X3
Y4X412
Y5X412
Y3X2
Y4X41
Y1X1
Y4X4
Model C/
Thermal treatment at 117 0С during 1 hour
Uncontrolled cooling at room temperature
Germs adding0,03% to products mass
Packing in metal cans №7 (0,4 kg)
Operative model of packaged product lactose crystallization after its thermal treatment
Results
U = 0.64М = ∑na / ∑n = 6.98Y15 X5
Y13 X5
∑nv2=484.1--∑an=698∑n=100∆a = 2Y7 X5
Y10 X5
----0≥ 25IVY11 X5
----024Y7X4123
----022Y8X4123← Z
----020Y6X4123
----018Y1X3
----016
III
Y4X412
49.349.287.0214114Y5X412
100.825.205.0248412II
Y3X2
36.59.123.0240410Y4X41
53.11.041.02408518Y1X1
13.50.960.9884146Y4X4
230.98.882.98104264
I
C/
nv2v2Discrepancy, v
Productna
Crystals frequency, n
Crystals size, а, µm
Crystals groupModel
Results of packaged product lactose crystallization after its thermal treatment
Results
----500.0000,827,090,031CaCO3
----≥770.0000,855,670,029TiO2
500.0000.646.980,022≥770.0000,884,480,022SiO2
M, in 1mm3UD, µmК, %M, in 1mm3UD, µmК, %
C/C
Variant of modification of germs adding timePriming material type
(1-2µm)
270.0000,597,680,078220.0000,419,130,098CaCO3
500.0000,636,860,061270.0000,548,260,089TiO2
500.0000,826,650,051770.0000,766,410,082SiO2
M, in 1mm3UD, µmК, %M, in 1mm3UD,µmК, %
BА
Variant of modification of germs adding timePriming material type
(1-2µm)
Results of lactose crystallization depending on the operating model and priming material type
SiO2>TiO2>CaCO3Efficacy gradation
Results
Farther investigations algorithm Priming material choice
Physico-chemicalproperties
Security
Technologicalpeculiarities
Modeling systems
Development of product technology
Availability
Investigation methodsPRODUCT
Products indices
Preservation stability
Security
Technological parameters
Technological regulations
Economical expediency
Methods of control
Crystallography
Methods of control
Formationconformity
Growth conformity
Efficacy
Production approval
Thank you for attention!Спасибо за внимание!
Aram Galstyan
GNU All-Russia Dairy Research Institute, Moscow, RussiaDepartment of gerodietical and special products
Tel/Fax: +7 (495) 236-02-36www.vnimi.org
E-mail: [email protected]
Acknowledgements
Academician Kharitonov V.DProfessor Radaeva I. A.Chief of the laboratory Petrov A. N.
All colleagues and staff at the Department of Gerodietical
and special products
IDF Symposium “Lactose and its Derivatives”14-16 May 2007, Moscow, Russia
IDF International Symposium -Lactose and its Derivatives
Analytical Methods for Lactose Quantification in Milk and Milk Products
Rachid Kouaouci, Ing., M.Sc., ChemistValacta, Ste-Anne de Bellevue, Québec, Canada
Centre d’expertise en production laitière du Québec
IDF International Symposium -Lactose and its Derivatives
METHODS
Mid-infraredPolarimetryGravimetryEnzymatic essayDifferential pHChromatography ( HPLC )
IDF International Symposium -Lactose and its Derivatives
MID-INFRAREDIDF 141C:2000; AOAC 972.16
Principle: Measurement of the absorption of the hydroxyl group (OH) at 9.6 µm.
Advantages:Very fast (400 samples/hour)No sample preparationWidely used
Disadvantages:Indirect methodNo differentiation between carbohydratesLimitation to fluid samples
IDF International Symposium -Lactose and its Derivatives
POLARIMETRYAOAC 896.01
Principle:1-Precipatation of fat and protein2-Measurement of the specific rotation of the polarized
light due to the asymmetric carbon of lactose
Advantages:Costs
Disadvantages:Interference with optically active components No differentiation between carbohydratesEmpirical calculation
IDF International Symposium -Lactose and its Derivatives
POLARIMETRYAOAC 896.01
Calculation:C [g/100ml] = a[Vm + Vr – 0.01145Vm ( F+P) ]
Vm (b)a: observed readingVm: Volume of milk usedVr: Volume of reagent usedb: rotation in a 40 cm tube of 100 ml solution
containing 1 gram of lactose at t 0C ( b=2.096at 25 0C)
F: %fat P: % protein
IDF International Symposium -Lactose and its Derivatives
GRAVIMETRYAOAC 930.28
Principle: Lactose + CuSO4
Advantages:Very simple procedureCosts
Disadvantages:Empirical calculationInterference with all reducing carbohydratesNo differentiation between carbohydrates
Cu2O
IDF International Symposium -Lactose and its Derivatives
GRAVIMETRYAOAC 930.28
Hammond table for calculating Lactose
7.78.5.
122.2.
342.0
11.312.4
.179.0
.489.7
Lactose.H2O(mg)
Cu2O(mg)
IDF International Symposium -Lactose and its Derivatives
ENZYMATIC ESSAYIDF 79-1,2/ISO 5765-1,2 (2002); AOAC 930.28
Principle: 1- Lactose
2- β-Galactose + β-Galactose dehydrogenase
+NAD+ (nicotinamide Adenine-dinucleotide)
NADH
3-Measurment of the amount of NADH by absorbance at 340 nm
+ β-GalactosidaseGlucose + β-Galactose
IDF International Symposium -Lactose and its Derivatives
ENZYMATIC ESSAYIDF 79-1,2/ISO 5765-1,2 (2002); AOAC 930.28
Advantages:CostsSolid samples can be used
Disadvantages:More elaborate procedureNo differentiation between carbohydrates
IDF International Symposium -Lactose and its Derivatives
Differential pHIDF/ISO draft
Principle:1- Lactose Glucose + β-Galactose2- Glucose + ATP + Glucokinase
Glucose-6P + ADP + H+
3- Measurement of the pH
+ β-Galactosidase
IDF International Symposium -Lactose and its Derivatives
HPLCIDF 198 │ISO 22622 (FDIS)
• Principle: 1-Sample + Internal Std1
2-Filtration of the sample3-Injection through an analytical column4-Detection by a differential refractometer5-Quantification
+ Biggs Solution2Precipitation of fat
and protein
2Zinc acetate + Glacial acetic acid+ Phosphotungstic acid1Internal Std: Melezitose ( Glu-Fru-Fru )
IDF International Symposium -Lactose and its Derivatives
HPLCIDF 198 │ISO 22622 (FDIS)
Chromatographic conditions:Mobile phase: degassed HPLC grade waterFlow rate: 0.6ml / minInternal detector temperature: 350 CColumn temperature: 850 CVolume to be injected: 20 µlRun time: 15 minColumn type: Styrene divinylbenzene resin
IDF International Symposium -Lactose and its Derivatives
HPLCIDF 198 │ISO 22622 (FDIS)
Chromatogram of raw milk sample
IDF International Symposium -Lactose and its Derivatives
HPLCIDF 198 │ISO 22622 (FDIS)
Chromatogram of raw milk sample with added lactulose
IDF International Symposium -Lactose and its Derivatives
HPLCIDF 198 │ISO 22622 (FDIS)
Chromatogram of lactose-reduced milk sample
IDF International Symposium -Lactose and its Derivatives
HPLCIDF 198 │ISO 22622 (FDIS)
Chromatogram of raw milk sample with added sucrose and fructose
IDF International Symposium -Lactose and its Derivatives
HPLCIDF 198 │ISO 22622 (FDIS)
Advantages:Direct methodDifferentiation between carbohydratesAutomated methodReference methodFlexibility
Disadvantages:Costs
IDF International Symposium -Lactose and its Derivatives
HPLCIDF 198 │ISO 22622 (DIS)
IDF International Symposium -Lactose and its Derivatives
HPLC
IDF 198 │ISO 22622 (DIS)
IDF International Symposium -Lactose and its Derivatives
HPLCIDF 198 │ISO 22622 (DIS)
IDF International Symposium -Lactose and its Derivatives
HPLCIDF 198 │ISO 22622 (DIS)
IDF International Symposium -Lactose and its Derivatives
HPLCIDF 198 │ISO 22622 (DIS)
IDF International Symposium -Lactose and its Derivatives
HPLCIDF 198 │ISO 22622 (DIS)
IDF International Symposium -Lactose and its Derivatives
REFERENCES
• Brons, C, and Olieman, C, 1983. Study of the high performance liquid chromatography separation of reducing sugars applied to the determination of lactose in milk, J. Chromato., 259, 79.
• IDF 141C:2000 -Whole Milk- Determination of milkfat, protein and lactose content, Guidance on the operation of Mid-infrared instruments.
• IDF 79-1:2002│ISO 5765-1- Dried milk, dried ice-mixes and processed cheese- Determination of lactose content- Part 1: Enzymatic method utilizing the glucose moiety of the lactose.
• IDF 79-2:2002│ISO 5765-2- Dried milk, dried ice-mixes and processed cheese- Determination of lactose content- Part 2: Enzymatic method utilizing the galactose moiety of the lactose.
• IDF 106:2004│ISO 5548- Caseins and caseinates- Determination of lactose content- Photometric method.
• Official Methods of Analysis, 16th Ed., 4th Revision, 1998, AOAC INTERNATIONAL, Gaithersburg, MD, methods 896.01, 984.15, 930.28 and 972.16.
• Nickerson, T. A., Vujicic, I. F., and Lin, A. Y, 1976. Colorimetric estimation of lactose and its hydrolytic products. J. Dairy Sci., 59, 386.
IDF International Symposium -Lactose and its Derivatives
Many thanks for your attention!
Hi, I need to measure my lactose level!
IDF Symposium “Lactose and its Derivatives”14-16 May 2007, Moscow, Russia
Primary Nucleation of Alpha Lactose Monohydrate: The Effect of Supersaturation
and Temperature
J.S McLeod, A.H.J Paterson, JR Jones and JE Bronlund
Nucleation – The effect on crystal size
Nucleation Mechanisms
Primary Nucleation Mechanisms
Primary nucleation equations
3 2
3 2
16exp3( ) (ln )
mHom
K R
VJ AkT S
πσ⎛ ⎞= −⎜ ⎟
⎝ ⎠
3 2
3 2
16 ( )exp3( ) (ln )
mHen
K R
V qJ AkT S
πσ⎛ ⎞= −⎜ ⎟
⎝ ⎠
T Hom HenJ J J= +
Lactose Nucleation – Previous Work
“In the manufacture of lactose it is desirable to secure a maximum yield of crystals in a minimum time, and to secure crystals which may be readily washed with a minimum of loss.” Herrington, (1934)
Effect of Supersaturation on Nucleation Shi, (1990), Griffith, (1982), Kauter, (2003), and Butler (1998)
Effect of temperature on nucleation (Shi, 1990)
Effect of temperature on nucleation (Kauter, 2003)
Experimental Setup
Relating induction time to the nucleation rate
NN
T
NtJ
∝
3 2
3 2
16exp3( ) (ln )
N N mN
K R
N N VtJ A kT S
πσ⎛ ⎞= = −⎜ ⎟
⎝ ⎠
Typical absorbance curve for nucleation
Time to reach critical absorbance vs. supersaturation
The effect of temperature on nucleation rates at different absolute supersaturations
Primary Nucleation Mechanisms
-2 -2Hen R Hom R(-B [lnS ] ) (-B [lnS ] )
N Hen Homt = A exp +A exp× ×× ×
Nucleation Results 25 Degrees
Nucleation Results 40 Degrees
Nucleation Results 60 Degrees
Nucleation Results 50 Degrees
Effect of temperature on the point where the homogenous nucleation begins to appear
20.671.972.176020.102.271.485020.122.711.014020.203.690.5925
Absolute SupersaturationCα-Cαs (g /100 g water)
Relative Supersaturation(Cα/Cαs)
(lnSR)-2Temperature °C
Conclusions
Increasing the supersaturation increases the nucleation rate of alpha lactose monohydrate
The effect of temperature on the nucleation rate at high supersaturations is negligible – temperature becomes more important as supersaturation is decreased
The point at which homogenous nucleation appears as an importantmechanism is independent of temperature (when viewed in absolutesupersaturation).
THANK YOU
Funding and Support
New Zealand Foundation for Research Science and Technology
Fonterra
Massey University
NZIFST (Dairy Division)
IDF Symposium “Lactose and its Derivatives”14-16 May 2007, Moscow, Russia
Analysis of a sticky impurity:lactose phosphate: a contaminant of
lactose
Dr. Rob Sleigh,Food Science Australia.
Estelle Lifran, Dr. Jim Hourigan, Dr. Rosalie Durham, Dr. Linh Vu,
Centre for Plant and Food Sciences, UWS.
16th May 2007
IDF symposium
Lactose & its derivatives
Role of lactose-phosphate in lactose crystallisation
Lactose phosphate: strong inhibition effect on lactose crystal growth(Visser 1980, 1984 and 1988)
Pharmaceutical grade lactose contaminated by 270 to 400ppm lactose phosphate; preferentially integrated in the crystals; can not be washed off.
→ Impact for industrial crystallisation:Poor control of particle size, size distributions and yield.
New IEL lactose is free of lactose phosphate and other impurities; used as the control in this work.
Study also relevant to pre-crystallisation of spray-dried lactose and dairy powders.
Aim: to control lactose processing in a way relevant to the factory scale; to understand the impact of seeding and impurities on lactose crystallisation kinetics.
alpha-lactose
lactose-phosphate
O
OHOH
HH
H
H
HOH
OH
O
O
OH
HH
OH
H H
OHH
OH
O
OHOH
HH
H
H
HOH
OP
OH
OHO
O
O
OH
HH
OH
H H
OHH
OH
Crystallisation inhibition by lactose phosphate
alpha-lactose
lactose-phosphate
O
OHOH
HH
H
H
HOH
OH
O
O
OH
HH
OH
H H
OHH
OH
O
OHOH
HH
H
H
HOH
OP
OH
OHO
O
O
OH
HH
OH
H H
OHH
OH
0 5 10 15 20 25
50
100
150
200
250
300
Median size (µm)
0 5 10 15 20 25
50
100
150
200
250
300
Median size (µm)
Time (h)
(1) Commercial Pharmaceutical grade(2) Ultra-pure lactose + 60 ppm LP and (3) Ultra-pure lactose (control)
(1)
(2)
(3)
Fast cooling
Fast cooling (1) with 0.4% seeds
(1) with 1% seeds
0 5 10 15 20 250
0.1
0.2
0.3
Solid content (g/g)
0 5 10 15 20 250
0.1
0.2
0.3
Solid content (g/g)
Time (h)
(1)
(2)
(3)
(1) Commercial pharmaceutical grade(2) Ultra-pure lactose + 60 ppm LP and (3) Ultra-pure lactose (control)
(1) with 1% seeds
(1) with 0.4% seeds Fast cooling
Fast cooling
Analysis of the different forms of phosphorus in lactose
Total phosphorus (P):
- ICP-AES
- Spectrophotometric method (FIL-IDF 42B, 1990)
Inorganic phosphorus (Pi):
- Spectrophotometric method (Chen, 1956; FIL-IDF 42B, 1990)
- Same spectrophotometric method under milder conditions (Lowry et al., 1953; Chen, 1956)
- Flow Injection Analysis (FIA) (automated version)
Organic phosphorus/lactose phosphate:
- Indirect: difference between P and Pi
- Direct: CE
Total P content of dairy powders measured by ICP-AES
Concentrations in mg/kg powder (ppm), n= 2
Mineral Skim milk Whey Edible grade Pharma grade Pharma grade Pharma grade Pharma grade Ultra-pure(ppm) powder powder lactose β-lactose 1_1 1_2 2 lactoseCa 11917 6046 1000 22 76 1 12 4K 17334 14107 124 72 18 34 4 0Mg 1047 1058 129 4 3 2 1 0Na 3611 4534 138 72 2 21 3.1 0P 10834 6550 438 37 35 26 19 0Sulphated ash (%) 9.54 7.12 0.320 0.0500 0.0300 0.0200 0.0100 0
Similar results found with spectrophotometric method (FIL-IDF 42B, 1990)
Large variations of total phosphorus content depending on manufacturer and batch of lactose
Determination of the inorganic phosphorus content of lactose powders
Reference methods: Fiske & Subbarow, 1925. Modified by Lowry, 1946, 1953, and Chen et al., 1956, FIL-IDF42B, 1990.
Principle: sulfuric acid + ascorbic acid + ammonium molybdate added to sample, incubated at 38°C for 2 hours, develops blue colour.
The inorganic phosphate ions and molybdate ions form molybdophosphoric acid. This acid is reduced to phosphomolybdenumblue, the concentration of which is measured by spectrophotometry.
The blue colour produced is proportional to the inorganic phosphorus concentration.
Inorganic phosphorus content of dairy powders (n = 10)
No blue colour formationUltra-pure lactose
1.10 ± 01.10 ± 0.21.10 ± 0.1Pharmaceutical grade α-lactose 2
1.77 ± 0.051.50 ± 0.11.78 ± 0.1Pharmaceutical grade α-lactose 1 Batch 1
2.05 ± 0.071.85 ± 0.12.01 ± 0.1Pharmaceutical grade α-lactose 1 Batch 2
2.25 ± 0.21.95 ± 0.42.40 ± 0.3Pharmaceutical grade β-lactose
137 ± 3.5103 ± 4.1145 ± 4.8Edible grade lactose
1840 ± 281833 ± 421850 ± 45Whey powder
2195 ± 642012 ± 732226 ± 98Skim milk powder
FIA(ppm)
pH 4(ppm)
IDF(ppm)
Sample
ppm: mg phosphorus per kg powder
IDF: FIL-IDF42B, 1990, pH = 0.85
pH 4: Lowry et al., 1953
FIA: Flow Injection Analysis, automatic spectrophotometric method
Direct analysis of lactose phosphate in dairy products by capillary electrophoresis (CE)
Method for analysis of anionic compounds (Soga and Imaizumi, 2001; Izco et al., 2003).
Direct detection of lactose-phosphate, Pi and carbohydrates present in samples in one analysis.
Method applied to lactose, cheese, whey and milk
Development of a direct method of analysis of dairy powders by CE
Uses indirect UV detection mode with 20mM PDC + 0.5mM CTAB asbackground electrolyte, optimisation of temperature, pH and injection pressure
Can analyse lactose phosphate in the presence of lactose and other organic compounds without pre-treatment or risk of overloading
Calibration and validation
Lactose phosphate from lactose
Lactose-1-phosphate standards
y = 9755.1x - 51.816R2 = 0.9979
0
2000
4000
6000
8000
10000
12000
0 0.5 1 1.5
Lactose-1-phosphate concentration (mM)
AU day 1
day 2day 3
Lactose-1-phosphate standards (0.9 to 2.4mM)
Small intra-day (0.5%) and day to day (2.7%)variations
Linearity, precision and accuracy all within the range of similar studies
(Castro et al. 1989; Izco et al., 2003)
Analysis of lactose phosphate in lactose powders
Lactose phosphateLactose-1-phosphate
lactose
Citrates
Inorganic phosphates
Commercial pharmaceutical grade lactose powders
IEL lactose
1. IEL lactose free of lactose phosphate
2. Quantification of lactose phosphate in lactose samples using CE
Determination of the lactose phosphate content of lactose powders by CE
Sample Lactose phosphate
(ppm)
Edible grade lactose 368 ± 6
Pharmaceutical grade β-lactose 250 ± 4.8
Pharmaceutical grade α-lactose 1 Batch 2 252 ± 3.9
Pharmaceutical grade α-lactose 1 Batch 1 204 ± 3.7
Pharmaceutical grade α-lactose 2 144 ± 2.5
Ultra-pure lactose No detectable peak
Results expressed as means ± standard deviations, n = 10
ppm: mg lactose phosphate per kg lactose powder
Results are lower than in Visser (1988) estimation of lactose phosphate in batches of pharmaceutical grade lactose (: 270 to 400 ppm)
Determination of the organic phosphorus content of lactose powders by difference
Sample Pa
(ppm) Pi
a (ppm)
Poa
(ppm)
Lactose phosphateb
(ppm)
Edible grade lactose 438 103 335 4556
Pharmaceutical grade β-lactose 37 1.9 35 476
Pharmaceutical grade α-lactose 1 Batch 2 35 1.8 33 449
Pharmaceutical grade α-lactose 1 Batch 1 26 1.5 24 326
Pharmaceutical grade α-lactose 2 19 1.1 18 245
Ultra-pure lactose No blue colour formation
a expressed in mg of phosphorus per kg of lactose
b expressed in mg of lactose phosphate per kg of lactose
- =
To be compared to the value found by the direct method368
250
252
204
144
All the organic phosphorus present in lactose samples is not lactose phosphate as was previously assumed
Other sources of organic phosphorus exist
Analysis of lactose phosphate extracted from lactose by HPAEC-PAD
1,2 and 3:
Unknown peaks
4: Lactose phosphate
5: N-acetylneuraminic acid
6: KDN
Both sialic acid analogues
Showing pharmaceutical grade lactose contains other acidic components, especially GOS
Impurities other than lactose phosphate could also be phosphorylated
High performance anion exchange chromatography with pulsed amperometricdetection
Very sensitive, down to picomoles (1x10-12
moles)
Need extensive pre-treatment of lactose samples before analysis
Causing loss of lactose phosphate therefore no quantification possible for impure samples
Summary
Analysis of the different forms of phosphorus in lactose powders
1. Suitability of the spectrophotometric method to measure totaland inorganic phosphorus, in the absence of organic phosphates
2. Lactose phosphate can not be quantified indirectly
3. Lactose phosphate can be analysed in lactose and other dairy products by CE
4. Lactose phosphate is only one of several acidic contaminants present in traces (picomole order) in pharmaceutical grade lactose
CE was used as a tool to monitor lactose phosphate integration
During seeded batch isothermal and cooling crystallisations (Lifran et al.,Powder technology (2006), doi:10.1016/j.powtec.2006.11.010).
Acknowledgements
We would like to thank Food Science Australia, the Centre for Plant and Food Sciences of the University of Western Sydney and Dairy Australia for their support of the project.
Special Thanks to Estelle Lifran, Linh Vu, Jim Hourigan, and Rosalie Durham.
Thank You For Your Attention
IDF Symposium “Lactose and its Derivatives”14-16 May 2007, Moscow, Russia
COMPUTER MODELLING COMPUTER MODELLING AND FORECASTING OF AND FORECASTING OF
PHYSICAL AND CHEMICAL PHYSICAL AND CHEMICAL PROPERTIES OF LACTOSEPROPERTIES OF LACTOSE
A.V.SEROVA.V.SEROVNorthNorth--Caucasian State Technical Caucasian State Technical
UniversityUniversityStavropolStavropol, , RUSSIARUSSIA
112,3112,3113,8113,8109,9109,9110,0110,0114,9114,9113,6113,6108,2108,2105,6105,6110,1110,1110,7110,7109,7109,7109,7109,7110,1110,1112,7112,7114,3114,3104,5104,5109,9109,9117,5117,5
113,8113,8110,2110,2112.9112.9112.2112.2120,3120,3117,6117,6102,7102,7109,3109,3110,0110,0111,6111,6111,8111,8113,9113,9113,1113,1112,1112,1116,4116,4112,7112,7115,3115,3117,0117,0
110,9110,9110,9110,9108,9108,9109,0109,0112,2112,2111,2111,2107,0107,0107,7107,7108,8108,8110,5110,5108,1108,1111,4111,4110,0110,0108,4108,4112,0112,0106,8106,8110,3110,3117,1117,1
GalactoseGalactose residueresidueСС(1(1''))--СС(2(2''))--СС(3(3''))СС(2(2''))--СС(3(3''))--СС(4(4''))СС(3(3''))--СС(4(4''))--СС(5(5''))СС(4(4''))--СС(5(5''))--ОО(5(5''))СС(5(5''))--ОО(5(5''))--СС(1(1''))ОО(5(5''))--СС(1(1''))--СС(2(2''))ОО(5(5''))--СС(1(1''))--ОО(1(1''))СС(2(2''))--СС(1(1''))--ОО(1(1''))СС(1(1''))--СС(2(2''))--ОО(2(2''))СС(3(3''))--СС(2(2''))--ОО(2(2''))СС(2(2''))--СС(3(3''))--ОО(3(3''))СС(4(4''))--СС(3(3''))--ОО(3(3''))СС(3(3''))--СС(4(4''))--ОО(4(4''))СС(5(5''))--СС(4(4''))--ОО(4(4''))СС(4(4''))--СС(5(5''))--СС(6(6''))ОО(5(5''))--СС(5(5''))--СС(6(6''))СС(5(5''))--СС(6(6''))--ОО(6(6''))GlycosidicGlycosidic lincagelincageСС(1(1''))--ОО(1(1''))--СС(4)(4)
АМАМ11РМРМ33Experiment byExperiment by KK. . HirotsuHirotsuandandAA. . ShimadaShimada
AngleAngle
Table 1 Table 1 -- Dependence of Dependence of valentvalent angles values in the angles values in the method of optimization of method of optimization of αα--lactose moleculelactose molecule
Table 1 Table 1 -- Dependence of Dependence of valentvalent angles values in the angles values in the method of optimization of method of optimization of αα--lactose moleculelactose molecule
((continue)continue)
108,9108,9108,9108,9109,6109,6112,1112,1114,3114,3112,0112,0103,6103,6110,9110,9112,2112,2110,9110,9112,5112,5109,0109,0112,6112,6105,3105,3111,0111,0106,5106,5111,3111,3
112,2112,2110,5110,5111,2111,2111,9111,9116,3116,3113,0113,0107,9107,9109,3109,3112,3112,3111,6111,6110,1110,1106,3106,3109,6109,6104,4104,4111,7111,7104,6104,6112,2112,2
110,9110,9110,3110,3111,1111,1107,9107,9114,1114,1109,7109,7111,5111,5108,8108,8111,1111,1112,7112,7107,0107,0111,6111,6110,6110,6107,0107,0113,7113,7107,2107,2111,2111,2
Glucose residueGlucose residueСС(1)(1)--СС(2)(2)--СС(3)(3)СС(2)(2)--СС(3)(3)--СС(4)(4)СС(3)(3)--СС(4)(4)--СС(5)(5)СС(4)(4)--СС(5)(5)--ОО(5)(5)СС(5)(5)--ОО(5)(5)--СС(1)(1)ОО(5)(5)--СС(1)(1)--СС(2)(2)ОО(5)(5)--СС(1)(1)--ОО(1)(1)СС(2)(2)--СС(1)(1)--ОО(1)(1)СС(1)(1)--СС(2)(2)--ОО(2)(2)СС(3)(3)--СС(2)(2)--ОО(2)(2)СС(2)(2)--СС(3)(3)--ОО(3)(3)СС(4)(4)--СС(3)(3)--ОО(3)(3)СС(3)(3)--СС(4)(4)--ОО(1(1´́))СС(5)(5)--СС(4)(4)--ОО(1(1´́))СС(4)(4)--СС(5)(5)--СС(6)(6)ОО(5)(5)--СС(5)(5)--СС(6)(6)СС(5)(5)--СС(6)(6)--ОО(6)(6)
АМАМ11РМРМ33Experiment byExperiment by KK. . HirotsuHirotsu andandAA. . ShimadaShimada
AngleAngle
Fig 1 Alpha-lactose molecule after semiempirical method optimization in АМ1 parametrization on Polak-Ribiere algorithm
Fig 2. Values of an effective charge of alphaFig 2. Values of an effective charge of alpha--lactose molecule atoms and its lactose molecule atoms and its formation heat after geometrical optimization by means of formation heat after geometrical optimization by means of semiempiricalsemiempirical
method in method in АМАМ1 1 parametrizationparametrization on on PolakPolak--RibiereRibiere algorithmalgorithm
Fig. 3 Map of distribution of molecular electrostatic potential Fig. 3 Map of distribution of molecular electrostatic potential of of αα--lactose moleculelactose molecule
Fig. 4 Map of threeFig. 4 Map of three--dimensional distribution of dimensional distribution of molecular electrostatic potential of molecular electrostatic potential of αα--lactose moleculelactose molecule
Fig 5 MetalFig 5 Metal--organic complexorganic complex
OH H
H
H
H
C C
O
CO
Ca++
Figure 6 Figure 6 -- Structure of complexStructure of complex tetrahydroxoboratetetrahydroxoborate--lactoselactose
Fig. 7 Fig. 7 -- Possible Possible isomerizationisomerization mechanism of lactose at presence of a mechanism of lactose at presence of a tetrahydroxoboratetetrahydroxoborate--ionion
Figure 8 Figure 8 -- Spectrum of the 13Spectrum of the 13СС nuclear magnetic resonance of nuclear magnetic resonance of anomericanomeric centers in the equilibrium centers in the equilibrium lactuloselactulose solutionsolution
IDF Symposium “Lactose and its Derivatives”14-16 May 2007, Moscow, Russia