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Tomorrows Results Today

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D- Count Applications for the Food and Cosmetic Industry Elke Kohler, Chemunex SA, Paris D-Count : Non-Filterable Product Testing D-Count Technology ... – PowerPoint PPT presentation

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Title: Tomorrows Results Today


1
D- CountApplicationsfor the Food and Cosmetic
IndustryElke Kohler, Chemunex SA, Paris
2
D-Count Non-Filterable Product Testing
Real-time Microbiology Analysis in Food and
Cosmetics
3
D-Count Technology
Viability substrate
Bacteria
Enzyme
Cell Labelling 12 minutes
Free fluorochrome
Flow Cell
Detector
Cell Counting 1 minutes
Laser
Detector
4
FluorassureÒ Viability Markers
  • Criteria of Viability
  • Enzyme activity
  • Membrane integrity
  • Applicable for
  • Total Viable Count
  • Yeast Moulds

5
Cell Labelling Technology
Fluorescent Signal
Fluorescence Based Cell Labels
515 nm
IncidentLight
488 nm
6
D-Count Counting Principle
  • Flow Cell
  • Linear fluid stream
  • Cells separated into
  • single entities
  • Sensitive detectors

7
D-Count Optical Bench
Sample
  • Green emission
  • Blue light

Detectors
Flow cell
Objective Lens
Red emission
Dichroic filter
Beam splitter
Laser beam
8
Data Processing
PMT Detector
Data Processing
Digital Signal Processor collects blocks of data
  • Colour fingerprint
  • Light intensity
  • Signal shape

Rejected Background
Results
9
D-Count Main Menu
10
Sample identification
11
Presentation of results
12
Presentation of results
13
Dedicated Automation
  • Capacity
  • 50 samples / batch
  • High Throughput Automate
  • up to 60 samples per hour
  • Simple sample pre-treatment
  • Product sample directly loaded onto automate
  • samples containing big particles are
    centrifiltrated before loading

14
D-Count
  • Combining Speed with Sensitivity
  • Designed for non filterable products
  • Direct Counting in less than 30 minutes
  • Automated system
  • Automate capacity 50 -64 samples
  • High throughput
  • Sensitivity down to 5x102/ml sample
  • Linearity between 5x102 to 5x105/ml

15
D-CountBenefits from speed and sensitivity
  • Final products testing in hours not days
  • Fast detection of problems (Raw materials ? Final
    product)
  • Immediate response to contamination incidents
  • Rapid confirmation of corrective action
    effectiveness
  • Real time trending of the production process
  • Decrease inventory holding and warehouse costs
  • Decrease product losses
  • Minimise production disruption
  • Increase production flexibility

16
B a c t i F l o w
Compact Microbial Analyser
Real-time microbiology analysis in Food,
beverages cosmetics
17
Based on Proven Technologies
  • Fluorescent cell labelling (Fluorassure)
  • Same reagent technologies as used on D-Count
    ChemScan RDI
  • Validated on more than 500 strains
  • Bench Top Microbial Analyser
  • Compact Flow cytometer specifically designed for
    microbial detection
  • Laser based detection
  • Simple to operate
  • Data acquisition, analysis and real time result
    display
  • Automatic data interpretation
  • Quantitative results in minutes

18
FluorassureÒ Viability Markers
Viability substrate
  • Criteria of Viability
  • Enzyme activity
  • Membrane integrity

Enzyme
Bacteria
Free fluorochrome
  • Applicable for
  • Total Viable Count
  • Total Count
  • Yeast Moulds
  • Specifics (on going)

19
Quantitative Microbial Counting
  • Flow Cell
  • Linear fluid stream
  • Cells Separated into
  • single entities
  • Sensitive detectors

Labelled Sample
Detector 1
Laser
Detector 2
B a c t i F l o w
Sample
20
R a p i d
  • Time to result
  • 10 minutes labelling prior to analysis
  • 1 minute analysis per sample
  • Simple sample preparation adapted to product
    matrices

No cell multiplication required
Labelling 10 min.
Counting 1 min.
21
S e n s i t i v e
  • Combining Speed with Sensitivity
  • Quantitative counting cell by cell
  • Sensitivity of detection less than 100 cells/ml
    labelled solution
  • Linear response up to 100,000 micro-organisms
  • Direct Counting in less than 15 minutes
  • Protocol available for Presence / Absence within
    24 hours

22
E a s y t o U s e
  • Ready to use Reagents
  • Simple preparation for sample products
  • Validated protocols for a full range of
    applications
  • Fully automated detection and counting
  • No data interpretation required
  • Full data traceability
  • PC with Windows XP interface

23
B a c t i F l o w A p p l i c a t i o n s
Dairy Products
Cosmetics
Fruit based Products
Water
Pure cultures
24
D-COUNT and BACTIFLOW DISCRIMINANTS
25
Fingerprint Colour
Fluorochrome Emission Spectrum
Green channel
Transmission
Red channel
Wavelength (nm)
26
Ratio at Primary Peak
Intensity
H2
Green channel

 Peak ratio 
H1
H1
Intensity
 Samples
Red channel
H2
Samples
27
Signal
Fluorescence Intensity
Fluorescence Intensity
Baseline
Samples
(Signal) - (Baseline)
 Samples
28
Signalauswahl
Ausgewählte Signale
Schwellenwert
Hintergrund
29
D-COUNT and BACTIFLOW APPLICATIONS
30
Applications
  • Beverages
  • Fruit Juices
  • Soft drinks concentrates
  • Food
  • Milk products
  • Fruit products
  • Frozen vegetables
  • Salads

31
Applicable to Range of Products
  • Non sterile pharmaceuticals
  • Syrups
  • Ointments
  • Tablets
  • Lotions
  • Cosmetic products
  • Shampoos and hair conditioner
  • Creams
  • Make-up and Foundation
  • Toothpaste
  • Lotions

32
Applications
  • Final product
  • Microbial Limit Test
  • Presence / Absence test
  • In Process
  • Raw Materials
  • Bioburden
  • Cleaning
  • validation
  • Trending

T V C Yeast Moulds E. coli (on going) Other
specifics (on going)
D-Count
  • Environmental
  • Surface
  • Personnel

33
Applications
  • Presence / Absence Test
  • 24 hours
  • TVC - Direct Counting
  • 90 minutes
  • Yeast / Moulds - Direct Counting
  • 90 minutes
  • E. coli and coliform tests (availabilty
    sept. - oct. 01)
  • 20 hours

34
Food Beverages
  • Applications
  • Yeast detection in Fermented milk products
  • TVC or Yeast counts in Juice concentrates
  • Sterility test in Desserts
  • TVC or Yeast mould detection in Fruit juices
    beverages
  • TVC in Process Water

35
D-COUNT Sample Background Spiking Experiments
36
D-Count Evaluation
  • Pure culture correlation studies
  • Non contaminated product analysis
  • Inoculated product analysis

37
Pure Cultures Studies
  • Range of micro-organisms tested
  • Staphylococcus aureus
  • Escherichia coli
  • Bacillus circulans
  • Bacillus licheniformis
  • Enterobacter cloacae
  • Candida albicans
  • Aspergillus niger
  • Penicillium decumbeus
  • Methods
  • Micro-organisms isolated from plates
  • Selected colony incubated for 24 hours at 30C
    in TSB
  • Serial dilution in physiological water
  • Analysis by D-Count plate

38
D-Count Evaluation
  • Pure culture correlation studies
  • Non contaminated product analysis
  • Inoculated product analysis

39
Pure Culture Correlation
Enterobacter cloacae Candida albicans ATCC
10231
40
Pure Culture Correlation
Staph. aureus ATCC 6538P Bacillus circulans
41
Pure Culture Correlation
E. coli ATCC 8739 Bacillus licheniformis
42
Pure Culture Correlation
Aspergillus niger Penicillium decumbeus
43
Pure Culture Correlation
Combined plot of all micro-organisms analysed
Staphylococcus aureus Escherichia coli Bacillus
circulans Bacillus licheniformis Enterobacter
cloacae Candida albicans
44
Conclusions
  • Good correlation with all micro-organisms tested
  • Sensitivity of detection less than 100 cells per
    ml
  • Linear response up to 5 x 10e5 micro-organisms
    per ml
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