Milk Analysis Laboratory
FAO workshop: Training on the development of good identification and
milk recording systems
Mart Kuresoo, Head of Analysing
Laboratory Department
November 12th, 2013
Topics
•History of the performance tests of milk
samples in Estonia
• Legislation
•General requirements for the laboratory
•Structure of the Laboratory
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Laboratory
Topics
•Equipment and their selection
•Maintenance costs
•Chemicals used and their cost
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Laboratory
Topics
•Logistics of milk samples
•Identification of raw milk components
•Volume of analyses
•Quality system of the laboratory
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Laboratory
History
Gerber Method – 1891 Niklaus Gerber
Kjeldahl Method – 1883 Johan Kjeldahl
Photometric Method – 1963 Foss Electric
Spectrometry – 1975 Foss Electric
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Laboratory
History
1924
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Laboratory
History
1958
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Laboratory
History
1970
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Laboratory
History
1982
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Laboratory
History
1984
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Laboratory
History
1996
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Laboratory
Legislation
•Farm Animals Breeding Act
•Performance Testing Handbook
•ICAR Performance Testing Guide (2012)
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Laboratory
ICAR Performance Testing Guide
(2012)
ICAR's instructions for the laboratory
There are no direct technical requirements
There are requirements for the procedures used:
•Formal reference methods are used;
•Recognised routine methods are used;
•Recommendations for the quality of samples are
presented;
•Recommendations for quality control of analysing
are presented.
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Laboratory
Reference Methods (ICAR)
Has to be internationally recognised
Milk analysed by these methods is used for:
•Calibrating and controlling routine equipment
•Setting control instruments
ISO 1211 – identification of fat content
gravimetrically
ISO 8968 – identification of protein content
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Laboratory
Routine Methods (ICAR)
Instrumental methods should be
standardised or have formal national
recognition:
•ISO 9622/IDF141 – fat, protein, lactose
•ISO 13366 – somatic cells
Methods of national recognition must also
be recognised by ICAR under the protocol of
ICAR
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Laboratory
Requirements for Milk Samples
(ICAR)
•Quality of milk sample is essential
•Sample bottles have to be suitable
•Quantity of sample in the bottle is very
important
•Preservative of milk sample and its usage
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Laboratory
Quality Control in Laboratory (ICAR)
Quality control of reference methods
• Systematic errors in reference analysis transfer to
routine analysis and all comparative tests
External control:
• Routine laboratory has to take part in comparative
tests that can be organised by the national reference
laboratory which itself takes part in international
comparative tests
• Can also take part in international comparative
tests, if needed
• Minimum frequency of comparative tests is 4
times per year
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Laboratory
Quality Control in Laboratory (ICAR)
Internal control:
•In international comparative tests, certified
comparative materials can be used
• Control samples prepared by the
laboratory itself can also be used
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Laboratory
Quality Control in Laboratory (ICAR)
Quality control of routine methods is
performed according to Part II of ISO 8196/
IDF 128
External minimum control 4 times per
year:
•By national expert laboratory
•On the basis of reference samples of the reference
laboratory
•By comparative tests between the laboratories
•Objects of control are at least linearity and
repeatability
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Laboratory
Quality Control in Laboratory (ICAR)
Internal control:
•Performed continuously by routine analysis with
control milk samples (fat, protein, lactose, urea –
ISO9622/IDF141, somatic cells – ISO
13366/IDF148)
•And with certified control material: somatic
cells, bacteria
•Used for:
Calibration, repeatability, transmission, linearity,
transmission factors, homogenisation, zero values
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Laboratory
General Requirements for the
Laboratory
Work area has to be suitable:
•Location
•Size
•Division
•Floor
•Cardinal points
•Possibility to install communications
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Laboratory
General Requirements for the
Laboratory
Environment that is suitable for people and
equipment:
•Ventilation
•Cooling
•Heating
•Recreation area
•Sanitary installations
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Laboratory
Structure of the Laboratory
Depends on the obligations of the
laboratory:
•Analysis of performance testing samples only
•Analysis of dairy samples only
•Analysis of both aforementioned samples together
•Analysis of additional samples
•Control laboratory
•Level and organisation of quality system of the
laboratory
•National requirements
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Laboratory
Laboratory Equipment
Selection of equipment is the matter of free
choice
Compared to the vehicle market, the choice
of analysers is relatively small
The choice of auxiliary equipment is much
wider
It is not wise to use postal services and
Internet sales services
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Laboratory
Laboratory Equipment
Analysers:
•Foss Analytical AS (1956, DK)
•Anritsu Corporation (1890, Japan)
•Bentley Instruments Inc. (1982, USA)
•Delta Instruments B.V. (1988, Netherlands)
•Advanced Instruments Inc.
•Anadis Instruments
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Laboratory
Laboratory Equipment
Comby Foss FT+ (470,000 €)
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Laboratory
Laboratory Equipment
The operating principle of MilkoScan™ FT+
is based on Fourier Infrared Transmission
(FTIR) and it operates in the central area of
infrared of 3-10 µm.
Component Measuring range Performance range Repeatability
Accuracy* Bulk
Accuracy Single cow
Fat
0-15%
Protein
0-10%
Lactose
0-10%
Solids
0-20%
Urea Patented¹
2-15%
2-10%
2-10%
2-20%
10-100mg/dl
Citric Acid
0.1-0.5%
FPD(Screening)
0.1-0.5%
Sd< 0.005% Sd < 0.01% Sd < 0.015%
400-600m°C 450-550 m°C Sd <0.5 m°C Sd <4 m°C N/A
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Cv < 0.5%
Cv < 0.5%
Cv < 0.5%
Cv < 0.5%
10-100mg/dl
Cv < 1.0% Cv < 1.5%
Cv < 0.9% Cv < 1.5%
Cv < 0.9% Cv < 1.5%
Cv < 1.0% Cv < 1.5%
Sd <1.5mg/dlSd < 3mg/dl Sd < 3.5mg/dl
Laboratory
Laboratory Equipment
The operating principle of Fossomatic FC is
based on flow cytometry where the DNA of
somatic cells' nuclei is stained and counted
in flow.
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Laboratory
Laboratory Equipment
Auxiliary equipment:
•Water purification equipment
•Washing equipment
•Drying equipment
•Refrigerating equipment
•Ventilating equipment
•Compressed air equipment, driers
•Environment control equipment
•Hydrobaths
•Tables
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Laboratory
Laboratory Equipment
Control laboratory:
•Scales – analytical and larger
•Drying ovens
•Hydrobaths
•Magnetic stirrer
•Block-digestion apparatus – Büchi K435
•Distillation apparatus – Büchi B-324
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Laboratory
Laboratory Equipment
Control laboratory:
•Automatic and manual titrators
•Membrane technology – protein separation
•CL-10 Micro – urea reference
•Cryoscope – freezing spot – and its purpose
•Thermometres – classic, electronic –
calibrated
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Laboratory
Laboratory Equipment
Maintenance and maintenance costs
Analysers:
•Competence of manufacturer's representatives,
contract?
•Competence of maintenance staff of laboratory
equipment
•Cost of physical and moral ageing of the equipment
•New equipment periodically available in the
market
•Original spare parts – according to pricelist
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Laboratory
Laboratory Equipment
Maintenance and maintenance costs
Analysers:
•Analogue spare parts from other
manufacturers
•Spare parts prepared on-site
•Impact of the quality of milk sample on
maintenance costs and operation speed of
analyser
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Laboratory
Laboratory Equipment
Maintenance and maintenance costs
Other equipment:
•Periodic verification and calibration of
scales – according to pricelist
•Other equipment according to the
recommendations of the manufacturer and
nature of work – 20-60 € per hour, spare
parts, used materials
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Laboratory
Chemicals
We use only the products of recognised
manufacturers and suppliers, for which the list has
been compiled
Chemical kits for analysers:
MSC FT+: S 6060 - zero concentrate - 69 €
MSC FT+: concentrate of washing solution - 94 €
MSC 6000: S470 - concentrate of washing solution - 81 €
Fossomatic 5000:
reagent E Buffer - 250 €
reagent B Colour - 1359 € - for 25,000 samples
reagent D Washing - 128 €
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Laboratory
Chemicals
Analytical chemicals:
Chemical registry has been created which contains
name and location in store book and physical
location in laboratory
Store book contains precise data related to
chemical:
•Where it was bought from and on the basis of
which document
•Manufacturer
•Quantity
•Usage period
•Shelf-life
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Laboratory
Chemicals
Analytical chemicals
Quantities used:
•Users keep constantly account of this
•Quality manager conducts inspections and
submits orders, if needed
In case of quality deviations, the reason is
determined and manufacturer or supplier is
replaced, if needed
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Laboratory
Milk Sample Collection
•Sample collection is not part of laboratory's
competence, but all successive phases
depend on this
•Samples are collected from approximately
300 points
•Milk samples are collected from each point
once a month
•There are 17 sample collection rounds
•There are two cars and two drivers
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Laboratory
Handling of Milk Samples
Conservation:
•Physical and chemical properties of milk
have to be conserved from collecting the
sample to analysing
•Must not affect reference samples
•Must not affect reference analyses
•Can affect routine analysis to an
insignificant extent, the impact of which is
eliminated upon calibration
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Laboratory
Handling of Milk Samples
Preservative:
•Has to conform to national health protection
requirements
•Has to conform to environmental requirements
•Historically, potassium dichromate was used
•Bronopol tablets are the best
•Samples have to be shaken half an hour after
collecting them
•Samples can be stored up to 72 hours
•Sample bottle 30-70 ml; price 0.16 €
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Laboratory
Handling of Milk Samples
Transportation:
•Warming has to be avoided
•Freezing has to be avoided
•Thermo-insulated van according to quantity of
samples and possible additional services
•Taking road conditions into account upon making
the choice
•Acceptable conditions for the driver
•Navigation device
•Availability of maintenance
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Laboratory
Handling of Milk Samples
Transportation:
•Sample boxes
•Importance of material
•Size
•Separator
•Suitability for storage
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Laboratory
Handling of Milk Samples
Storage:
•In barn – in cool room
•In laboratory's refrigerator – 0-4 °C
•Preanalysis period in laboratory
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Laboratory
Identified Milk Components
Minimal necessity for performance testing:
•Fat
•Protein
•Somatic cells
In addition, it is possible to offer:
•Urea
•Ketone bodies – acetone and BHB
•Fractions of fatty acids
•Mastitis pathogens
•etc.
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Laboratory
Identified Milk Components
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Laboratory
Number of Analyses Conducted in ARC Laboratory
In 1971
In 1988
In 1993
In 2010
In 2012
31,000
2,561,000
1781000
877,314
890,756
Laboris analüüsitud proovide arv
(sh piimatööstuse ja jk proovid kuid v.a dna-proovid)
90000
88000
86000
84000
2008
82000
2009
80000
2010
78000
2011
2012
76000
74000
72000
70000
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VEB
MÄR
JAN
APR
MAI
JUN
JUL
AUG
SEP
OKT
NOV
Laboratory
DET
Work Volume in 2012
1,350,000 prepared sample bottles
890,756 milk samples for
performance testing
78,648 milk samples from dairies
16,875 completed sample boxes
2,271 mastitis samples
364 times control milk to dairies
52 control milks for analysers
1 CECA LAIT for practice
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Laboratory
Number of Samples in 2012
ANALÜÜSITUD PIIMAPROOVE tk.
JAN
VEB
MÄR
APR
MAI JUN
JUL
AUG
SEP
OKT
NOV
73178 73398 74775 75300 74242 74189 75057 75181 76048 75808 71323
JK
89,82 92,03 92,04 91,98 90,25 91,95 91,33 92,49 92,73 91,19 92,52
%
jk lisapr.
Meierei 8084 6313 6386 6440 7944 6197 7111 5958 5942 7278 5650
9,92 7,92 7,86 7,86 9,65 7,67 8,64 7,32 7,24 8,74 7,32
%
Tänav
%
Kokku
152
0,26
158
0,21
1278
5
81469 79755 81238 81866 82359 80787 82282 81384 82104 83236 77187 77870
971537
972010
2271
9
DNA proove 149
44
0,05
178
77
0,10
307
126
0,16
448
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73
0,10
169
301
0,38
134
14
0,03
109
145
0,19
198
24
0,03
142
50
0,07
172
114
0,16
Kokku Päevas proove
tööpäevi 254
72257 890756
3507
92,91
1318
5,2
5355 78658
756
6,88
DET
225
40
Laboratory
Quality System of the Laboratory
•Laboratory is accredited since 15 January 1999
•Initial accreditation on the basis of ISO/IEC 25 and
EN 45001
•New accreditation once in every five years
•Accreditation on the basis of EN ISO/IEC 17025
•We use EAC as accreditation institution
•Estonian Accreditation Centre (EAC) is the
member of European co-operation for Accreditation
(EA) since June 2000
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Laboratory
Quality System of the Laboratory
•The objective of the quality system is to issue
reliable analysis results
•This enables to recognise the analysis results
everywhere
•No need for repetitive analyses
•Quality of raw materials and finished products
is guaranteed
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Laboratory
Quality System of the Laboratory
Quality management system:
•Includes all works performed in laboratory
•Guarantees the satisfaction of the needs
and expectations of customers using
laboratory services
•Guarantees confidentiality
unnecessary expenses
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and
avoids
Laboratory
Quality System of the Laboratory
Tools of the quality management system:
•Quality management system
documentation
•Management system audits and inspections
•Implementation of measures on the basis of
these audits and inspections for improving
quality
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Laboratory
Quality System of the Laboratory
Quality manual:
•Describes the laboratory’s quality
management system
• The purpose of the quality manual is to
support and contribute to the
implementation of the laboratory's quality
policy
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Laboratory
Quality System of the Laboratory
Quality manual:
Pursuant to the statutes, includes the
following fields of activity of analyses
laboratory:
•Collection and transportation of milk
samples
•Analysis of milk samples
•Issuing analysis results
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Laboratory
Quality System of the Laboratory
•Quality management system
documentation is prepared by quality
manager and approved by laboratory
manager
•Relevant quality management system
documentation is presented to the staff by
the quality manager and consistently
implemented by the staff
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Laboratory
Competence Tests CECA LAIT and ICAR
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Laboratory
Quality System of the Laboratory
Fat content
Rasvasisalduse hälve
hälve
0,5
0,4
0,3
0,2
0,1
0
-0,1
-0,2
-0,3
-0,4
-0,5
0,41
0,16
0,16
0,13
0,11
0,05
0,09 0,08
0,07 0,08
0,02 0,07 0,07 0,05 0,02 0,09 0,11 0,09 0,07
0,14
1.
2.
-0,02 -0,02
3.
4.
5.
6.
7.
8.
9.
10. 11.
12. 13.
14. 15. 16.
17. 18.
19. 20. 21.
22.
osavõtu kord
standardhälve
Standardhälve
0,3
0,2
0,12
0,11
0,05 0,06 0,06 0,05 0,07 0,04 0,03 0,04 0,04 0,06 0,05 0,06 0,05
0,06
0,1
0,11
0,05 0,05 0,05 0,06
0
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
osavõtu kord
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Laboratory
Quality System of the Laboratory
Nitrogen content
hälve
Lämmastikusisalduse hälve
0,095
0,1
0,08
0,06
0,04
0,014
0,0160,0050,002
0,022
0
-0,002
0,02
-0,002
-0,006
-0,005
-0,012
0
-0,02
-0,04 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23.
-0,007
-0,015
-0,003
-0,033
-0,019 -0,008
-0,008-0,008
-0,06 -0,003
-0,052
-0,08
-0,062
osavõtu kord
standardhälve
Standardhälve
0,05 0,041
0,04
0,03
0,0190,016
0,0180,0240,015
0,014
0,014
0,012
0,02
0,0100,0100,0110,007 0,01 0,01 0,01 0,0070,013
0,008 0,01 0,008
0,008
0,01
0
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22.
osavõtu kord
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Laboratory
Quality System of the Laboratory
Ring tests of bacteria counter
Kokkulangevus %-des
BSC149(84)
(2000-2012 a)
110
100
kokkulangevuse %
90
80
70
60
50
40
30
20
10
0
2.
9. 16. 23. 30. 37. 44. 51. 58. 65. 72. 79. 86. 93. 100. 107.114. 121. 128.135. 142. 149.156. 163.
osavõtu kord
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Laboratory
Quality System of the Laboratory
Results of ring tests (70 to 80 participants)
rasv
CECALAIT
ICAR
lämmastik
CECALAIT
ICAR
laktoos
ICAR
CECALAIT
külmumistäpp
somaatilised rakud
CECALAIT CECALAIT
ICAR
karbamiid
CECALAIT
ICAR
osalejaid koht osalejaid koht osalejaid koht osalejaid koht osalejaid koht osalejaid koht osalejaid koht osalejaid koht osalejaid koht osalejaid koht osalejaid koht
4.03.1998
2.09.1998
10.03.1999
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8.03.2000
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8.03.2002
3.09.2002
6.03.2003
4.09.2003
9.03.2004
6.09.2004
9.03.2005
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6.03.2007
1.03.2008
10.03.2009
7.09.2010
7.03.2011
6.03.2012
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Quality System of the Laboratory
The quality system is audited once a year:
•Performance of previous audit
•Results of control visit by accrediting body
•Results of conducted internal audits
•State of quality manual
•Results of comparative results and their necessity in the
future
•Changes in quality management system; efficiency of
the system
•Results of the handling of complaints; customer
satisfaction
•Staff and training; efficiency of training
•Sufficiency of staff and equipment
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Laboratory
Identification of Mastitis Pathogens
Mastitis 12 - 2010
Mastitis 16 - 2012
1. Staphylococcus aureus
2. Staphylococcus sp.
3. Streptococcus agalactiae
4. Streptococcus dysgalactiae
5. Streptococcus uberis
6. Escherichia coli
7. Enterococcus sp. (incl. E. faecalis and E. faecium)
8. Klebsiella sp. (incl. K. oxytoca and K. pneumoniae)
9. Serratia marcescens
10. Corynebacterium bovis
11. Arcanobacter pyogenes, Peptostreptococcus indolicus
12. Mycoplasma bovis
13. Mycoplasma spp
14. yeast
15. alga Prototheca spp
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Laboratory
Identification of Mastitis Pathogens
• A test Pathoproof Mastitis PCR Assey identifies 15
bacteria causing mastitis. In addition to pathogens,
the beta-lactamase gene is also identified from a milk
sample that shows penicillin resistance
•Identification is conducted by performance testing or
from separately collected samples, incl. general milk
and mammary quarters
•Bottles are labelled with a bar code
•Identified pathogen and its level is reported: +,2+,3+
•Price of samples:
1-4 samples - 30 €
5-9 samples - 27 €
>10 samples - 25 €
www.jkkeskus.ee
Laboratory
Identification of Ketone Bodies
Acetone
BHB
•Ketosis is a metabolic disease that causes
major expenses in some herds
•Ketosis is usually a problem for the whole herd
•Subclinical limit of ketosis is 0.15 mmol/l for
acetone and 0.1 mmol/l for BHB
•Objective is to give a monthly overview of cows
who have begun milking (1 to 2 months)
www.jkkeskus.ee
Laboratory
Thank you and I hope
you have learned
something new!
www.jkkeskus.ee
Kreutzwaldi 48A
Tartu 50094, Estonia
Phone (+372) 738 7700
www.jkkeskus.ee
Laboratory