Reference Product Characterization
Gene Gillman, PhD
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Introduction
• Purpose of Reference Products: • Method Validation and Qualification
• Certified values
• Known repeatability
• Quality Control Process Monitor • Method quality control, control charts
• Data acceptance
• Collaborative Study Material • Establish r and R values
• Reporting of Results • Quality control check for clients and Regulators
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Ideal Reference Products
• Features • Uniformity
• Repeatable content and yield
• Stability • Stable over time
• Target Compounds • Contain all target compounds at quantifiable levels
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Ideal Reference Products
• Controlling variability • Cigarettes
• Simplified blend design (reduce natural variability)
• Simplified product design (reduce product variability)
• Smokeless Tobacco • Simplified blend design (reduce natural variability)
• Packaging material selected to reduce water loss or uptake
• Product Uniformity
• Avoid pouched or whole leaf products
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Current Reference Products
• Features • Uniformity
• Tobacco is a natural product
• Variability is expected
• Design will impact variability
• Banded paper
• Filter Ventilation
• Stability • Finite stability
• Stability of target compounds might vary
• Target Compounds • Target compounds range from % levels of <LOQ/LOD
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1R6F Reference Product
• Features • Uniformity
• Represents Commercial Blend
• Flue Cured 34%
• Burley 24%
• Oriental 12%
• Reconstituted 20%
• Expanded Flue Cured 7%
• Expanded Burley 3%
• Design • Ventilated Filter
• Banded Paper
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1R6F Reference Product
• Example data from the 2015 Characterization of the 1R6F Reference Product • Data collected under short term repeatability conditions
• Short time frame
• Same Technicians and equipment
• 45 Production lots tested for TNCO and Physical Properties
• Data blinded to remove production codes
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1R6F Reference Product
• Impact of Filter Ventilation • Filter Ventilation
• Portion of the puff is directed through the filter
• Variability in filter ventilation will directly impact smoke yields under ISO smoking conditions
• Data • Collected to support the Certification of the 1R6F
• 45 unique production lots
• Data collected in a short time frame
• Short Term Repeatability (r)
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1R6F Reference Product-% Vent
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22.0
24.0
26.0
28.0
30.0
32.0
34.0
36.0
38.0
40.0
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45
% Vent 1R6F
% Vent
Average
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1R6F Reference Product-NFDPM
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7.50
8.00
8.50
9.00
9.50
10.00
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45
NFDPM (mg/cigt.)
NFDPM
Average
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1R6F Reference Product Summary Data for Physical Properties
Pressure
Drop Pressure
Drop
Length (mm)
Weight (g)
Circum. (mm)
Filter Vent
Open Closed
(%) (mm WG) (mm WG)
Average 83.1 0.9 24.8 30.3 107.4 137.5
SD 0.1 0 0 2.4 2.7 2.3
% RSD 0.1 0.8 0.2 8.1 2.5 1.6
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3R4F TSNAs in Filler
• Samples were obtained from four lots of 3R4F cigarettes.
• 200 cigarette were composited from each lot.
• Ground composites analyzed for TSNA content by LC-MS/MS.
Enthalpy Analytical, Inc. 12
Originally Presented as ST27 CORESTA Congress 2014
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TSNA Results-Ground Filler
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Lot # Lot # Lot # Lot #
V348X61B5 V349Y61B5 V348Z61B5 2013 Composite
NNN NNN NNN NNN
ng/g ng/g ng/g ng/g
Average 2476.1 2593.9 2423.8 2351.1
SD 58.4 25.9 11.4 10.5
%RSD 2.4 1.0 0.5 0.4
% Diff* 5.2 9.8 3.0 NA
Lot # Lot # Lot # Lot #
V348X61B5 V349Y61B5 V348Z61B5 2013 Composite
NAT NAT NAT NAT
ng/g ng/g ng/g ng/g
Average 1615.7 1869.1 1759.5 1713.8
SD 63.3 44.3 30.0 26.3
%RSD 3.9 2.4 1.7 1.5
% Diff* -5.9 8.7 2.6 NA
Lot # Lot # Lot # Lot #
V348X61B5 V349Y61B5 V348Z61B5 2013 Composite
NAB NAB NAB NAB
ng/g ng/g ng/g ng/g
Average 76.8 88.9 76.9 72.9
SD 1.3 3.9 1.3 0.6
%RSD 1.7 4.4 1.6 0.8
% Diff* 5.1 19.7 5.3 NA
Lot # Lot # Lot # Lot #
V348X61B5 V349Y61B5 V348Z61B5 2013 Composite
NNK NNK NNK NNK
ng/g ng/g ng/g ng/g
Average 724.1 787.6 745.5 735.4
SD 13.3 13.0 17.4 7.6
%RSD 1.8 1.6 2.3 1.0
% Diff* -1.5 6.9 1.4 NA
* %difference from 2013 composite sample
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TSNAs in 3R4F- Individual cigarette
• TSNAs were determined from 100 individual cigarettes selected from lot of 5000.
• Filler from one cigarette was extracted and analyzed by LC-MS/MS.
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Individual cigarette results-NNN
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1800
1900
2000
2100
2200
2300
2400
2500
2600
2700
1 6 11 16 21 26 31 36 41 46 51 56 61 66 71 76 81 86 91 96
ng/g
ram
NNN in 3R4 Filler
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Individual cigarette results-NNK
Enthalpy Analytical, Inc. 16
400
600
800
1000
1200
1400
1600
1800
1 6 11 16 21 26 31 36 41 46 51 56 61 66 71 76 81 86 91 96
ng/g
ram
NNK in 3R4F Filler
200% of average
160% of average
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TSNA Summary
Enthalpy Analytical, Inc. 17
NNN NAT NAB NNK ng/g ng/g ng/g ng/g
Average 2277.5 1668.0 81.0 732.9 Stdev 123.5 155.2 5.3 136.4 % RSD 5.4 9.3 6.6 18.6
TSNAs in 100 individual cigarettes
4% of the samples had NNK or NAT
greater than 150% of the average of all
samples
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Conclusion
• Design of Reference products will impact method repeatability (r) and
reproducibility (R) values
• Factors that impact product variably should be reduced and controlled
during the design of Reference products
• Simplified blend or pre-production analysis of blend components
• Particle size (smokeless)
• Filter ventilation
• Long term stability of the product requires on-going analysis
• Stability of target compounds over time
• Reduce product moisture changes
• Commercial versus custom packaging
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