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David Baram from Clifton Enterprises
Sampling in theSampling in the Bioprocessing Industries: Bioprocessing Industries:
Expectations andExpectations andthe True Picturethe True Picture
Presented by:David Baram from Clifton Enterprises
Sheffield, Englandemail: [email protected]
Interphex Japan
בס"ד
David Baram from Clifton Enterprises
IntroductionIntroduction• In the Bioprocessing Industry, sampling is fundamental. However,
since people have been doing it for years, it has become a routine. Unfortunately, when things become routine, people start to overlook the related pitfalls and the possible detrimental effect on their processes as a whole.
• This presentation, presented by Clifton Enterprises, is designed to review the various stages of sampling and the potential negative consequences associated with each one of those stages. This presentation will introduce state of the art technology, developed with the support of the British Government through the Department of Trade and Industry, to eliminate these consequences.
• Before I continue, I would like to take this moment to thank Sei Murakami from Hitachi Industrial System Div. for giving me this opportunity to be here today.
David Baram from Clifton Enterprises
The Purpose of SamplingThe Purpose of Sampling
• Monitor the Process For example, by checking:
» pH
» Growth Rate
» Glucose
• Identify Contamination
• Validation
Importance in Choosing Right“The sampling system is responsible for collecting a representative sample of the process stream and delivering it to the analyzer for analysis.
Obviously, the reliability of the sampling system directly affects the reliability of the overall composition analysis system.
The transport delay associated with the sampling system contributes directly to the overall deadtime for an on-line composition measurement. For example, an improperly designed sampling system can result in a transport time of one hour … Sampling deadtime can have a dramatic effect on the performance …”
Chemical and Bio-Process Control (Third edition)
By James B. Riggs and M. Nazmul Karim
David Baram from Clifton Enterprises
David Baram from Clifton Enterprises
Components in Sterile SamplingComponents in Sterile Sampling• Sterile access to the process fluid
– Normally via a Sampling Valve
• Collection System– Open– Bottle– Bag
• Access to Sample for Analysis– Normally via a Syringe
David Baram from Clifton Enterprises
Issues with Common Sampling Issues with Common Sampling ValvesValves
• Mechanism
• Steam Sterilization
• CIP
• Internal Volume
• Dead Volume
• Maintenance
David Baram from Clifton Enterprises
Common Sampling Valve
Problem Areas
Plug & Seat Elastomeric Material Temp. & Chemical
Limitations
Cleanability
& Sterility
Volume Operating Mechanism
Design of Common Sampling Valves
David Baram from Clifton Enterprises
Issues with Common Collecting Issues with Common Collecting EquipmentEquipment
• Complex operation– Difficult to maintain sterility
• Intricate human interaction process– Prone to Human Error
• Requires Wasteful Sterilization Flush
• Lengthy procedure– Detrimental for Ensuring Viable Sample
David Baram from Clifton Enterprises
A typical 4-valve rig for two bottles sampling
• Autoclave a bagged sample bottle with a filter and a valve (V3) sealing the opening.• Unbag the sample bottle as shown, connecting the closed V3 to the closed Tank Sample Valve V2.• Steam behind the flow paths of V2 and V3 as shown in pink.• Close Steam source (V1) and Condensate Drain (V4). You now have a sterile path between your
process and your sample bottle.• Open the Sample bottle (V3) and the Tank Sample Valve (V2), and take your sample as shown in blue.• Close the Sample Valve (V2), then the Sample receptacle Valve (V3). Drain V4 and remove your
Sample Bottle.
David Baram from Clifton Enterprises
Issues with Accessing Issues with Accessing Samples in Bottles and Common BagsSamples in Bottles and Common Bags
• Requires Safety Cabinet
• Requires replacing a Bottle with a Septum– Risks contamination– Time Consuming
• Sample withdrawal requires syringe with needle– Needle-stick risk to personnel– Insurance factor
• Time required compromises sample viability– pH change– Contamination
David Baram from Clifton Enterprises
• No Maintenance
• Cleanability assured (No Cross Contamination)
• Continuous Steam Sterilization, if required
• No Dead Volume
• Minimal Internal Volume
• 3-Position Valve
David Baram from Clifton Enterprises
Simple 3-Position Mechanism
Sterilize
Shutoff
Sample
David Baram from Clifton Enterprises
Vanasyl Sampling Valve with Vanasyl Sampling Valve with Bottles SamplingBottles Sampling
David Baram from Clifton Enterprises
Vanasyl Sampling Valve with Bottles Sampling
David Baram from Clifton Enterprises
Areas for Bag Technology Applications
• Bioreactor sampling
• Bioreactor additions (antifoam etc.)
• Media/buffer prep & storage
• Product storage & transfer
• Lab & pilot scale bioreactors
• Aseptic connections
David Baram from Clifton Enterprises
Examples of Common Sampling Bags
David Baram from Clifton Enterprises
Example Sampling Bags Assembly
David Baram from Clifton Enterprises
Technical Data for Ethylene-Vinyl Acetate (EVA)(Film Gauge 350 μm @ 232)
Physical Properties
Dimension Value Procedure
Water Vapour Transmission
Rate
g/(m2.day) 4 ASTM F-1249
Oxygen Permeability
cm2/(m2.day.bar) 800 ASTM D-3985
Carbon Dioxide Permeability
cm2/(m2.day.bar) 7000 Mocon Permatran C-IV
David Baram from Clifton Enterprises
Technical Data for Multilayer Ethylene-vinylacetate (EVA) with Barrier(Film Gauge 400 μm @ 232)
EVA Without a Barrier
Physical Properties
Dimension Value Procedure
Water Vapour Transmission
Rate
g/(m2.day) 1.644
ASTM F-1249
Oxygen Permeability cm2/(m2.day.bar) 2.2800
ASTM D-3985
Carbon Dioxide Permeability
cm2/(m2.day.bar) 67000
Mocon Permatran C-IV
UV Transmission
David Baram from Clifton Enterprises
Comparison Between Bottles and the Vanasyl Sampling BagBag vs. Bottle Samples
0
5
10
15
20
25
0 5 10 15 20 25 30 35 40 45 50
Run time
Glu
cose o
r pC
O2 (%
)
BagPCO2
BagGluc.
BottlePC02
BottleGluc.
Bag vs. Bottle pH
7.06
7.08
7.1
7.12
7.14
7.16
7.18
7.2
7.22
7.24
0 5 10 15 20 25 30 35 40 45 50
Time (hrs.)
pH BagpH
BottlepH
David Baram from Clifton Enterprises
The Vanasyl Sampling BagThe Vanasyl Sampling Bag
David Baram from Clifton Enterprises
Sampling Process Flow - Bottle vs. BagCOMMON SAMPLING BAGS SYSTEM VANASYL SAMPLING BAGS SYSTEM
1
2 Operator connects the Sampling Bags to the Operator connects the Sampling Bag to thespecific fermentor specific fermentor and draws a sample
3 Eliminated Eliminated
4 Eliminated
5 Eliminated
6 Eliminated
7
8
9 Eliminated
10 Eliminated
Operator collects 2 Sampling Bag which is kept close by. Or a few are connected to the vessel.
Process material (in the majority of Bag Sampling systems) is run into first bag to purge the bags' tube connections &
Second (or first) bag is filled with sample material
Bag with the sample is placed in a Safety Cabinet where the sample is transferred to a bottle which is then sealed with septum cap.
Process material is run into the first bottle to purge the tree.
Bottles are sterilized, capped, and stored or redistributed to various departments.
Bottles are sterilized, capped, and stored or redistributed to various departments.
Bottle is disassembled, prepped and cleaned.Bottles are disassembled, prepped and cleaned.
Bottle with the sample is placed in a Safety Cabinet where the bottle valve cap is replaced with a septum cap.
Operator collects 1 Sampling Bag which is kept close by
Second bottle is filled with sample material
Operator connects the Sampling Bottles to a specialized sample "tree" attached to the Sample Valve for a specific fermentor.
CURRENT SAMPLE BOTTLE PROCESS
Bottles are steamed for a half hour and allowed to cool with one bottle open to avoid forming a vacuum and to allow condensate to leave sample
Operator collects 2 Sterile Sampling Bottles from the Supply which is kept in cabinets, ready for use
The bag with the sample can either be kept frozen for validation or discarded.
The Sampling Bag is labelled and sent for analysis where the samples are drawn via a syringe (without a needle), which is directly connected to the Vanasyl Sampling Bag.
Bottles are removed and the second bottle is labelled and sent for analysis where the samples are drawn through the septum with a needle attached to a syringe.
Bottle with the sample is labelled and sent for analysis where the samples are drawn through the septum with a needle attached to a syringe.
The bags are discarded. The bottle with sample material that if not required to be kept frozen for validation, is rinsed and placed in a bucket for pickup by Glassware's
First bottle with purge material, and second bottle with sample material that if not required to be kept frozen for validation, are rinsed and placed in a bucket for pickup by Glassware's
David Baram from Clifton Enterprises
A Year’s Cost Comparison Between Bottle Sampling and
Vanasyl Bag Sampling (True in 2013)
Number of Samples/year on 14 Fermenters 11,814
Process Bottles/year 23,628
Sampling Bags/year 11,814
Estimated Bottle Sampling Costs
Req. Material for Maintaining the Valves and Bottle (Replacement Bottles, Filters, Diaphragms, Replacement Valves, etc.) $308,960.00
Maintenance Labour Costs (1,656 hours for the bottles and 504 hours on valves @ $40/hour (technician)) $86,400.00
Labour Costs Associated with Sample Taking (11,814 samples, 30 Minutes per Sample @ $70/hour - scientist) $413,490.00
Bottle Sampling Estimated Total Costs $808,850.00
Single Bottle Sampling Estimated Costs $68.47
Estimated Vanasyl Bag Sampling Costs
Bags Costs (11,814 Sample Bags using 150 ml @ $36 each) $425,304.00
Req. Material for Maintaining the Valves and Bottle (O-Rings for the Ingold, Inlet, and Outlet fittings) $300.00
Maintenance Labour Costs (70 Hours @ $40/hour - Technician) $2,800.00
Labour Costs Associated with Sample Taking (11,814 samples, 7 Minutes per Sample @ $70/hour - scientist) $96,481.00
Bags Sampling Estimated Total Costs $524,885.00Single Bag Sampling Estimated Costs $44.43
ESTIMATED YEARLY SAVINGS - GLASS BOTTLES Vs VANASYL BAGS $283,965.00
David Baram from Clifton Enterprises
Further Advantages of the Vanasyl Sampling System
• The Vanasyl Sampling System offers 4 easy sampling steps instead of 10 complicated sampling steps as well as:
– More than 99% reduction in product loss. (See Table above)
– Eliminates introduction of steam into process and sample.
– Eliminates false samples due to cross contamination.
– Eliminate the dilution of sample with condensate
• No delays between collection and testing ensuring sample integrity.
• No need for a safety cabinet.
• No use of needles or sharps (eliminate needle injuries and disposal issues).
• Sample can be frozen in the bag to -120C.
Bottle Sampling: A Minimum of 250 ml of product flashing per sample.
A yearly product loss from 11,814 samples: 2,953.50 Litres/year
Vanasyl Bag Sampling: A Maximum 2 ml of product flashing per sample.
A yearly product loss from 11,814 samples: 23.60 Litres/year
David Baram from Clifton Enterprises
Vanasyl Bag Sampling
David Baram from Clifton Enterprises
Vanasyl Sampling Bag System
David Baram from Clifton Enterprises
Clave Connection on Vanasyl Sampling Bag
David Baram from Clifton Enterprises
ConclusionConclusion
• This presentation should raised the awareness of sampling and its related issues and hopefully was beneficial for all those who attended.