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Life Sciences

Pall Laboratory for the Quality Control
Brice MONTEIL MENA Area Manager
Filtration. Separation. Solution. SM

 Pallchek presentation  HPLC Analytical Sample Prep Mobile phase  Microbiology

Analytical Filtration 1. Filter Media Properties 4. How do we make membranes? 5. Types of Media 3. Chromatography and HPLC 6. What is Filtration 2. Filtration and Sample Preparation Choosing the best device .

What is Filtration? • The process of removing particles from a liquid or gaseous stream as it passes through a porous medium. .

Why Use Filters? • Collect and/or Concentrate a Sample for Analysis • Clarify a Solution or Gaseous Stream • Sterilize a Solution or Gaseous Stream • Protect Critical Instrumentation from Particles or Liquids .

Types of Media • Depth Media • Membranes • Combination Filters .

What is Depth Media • A Depth Filter is a filter consisting of either multiple layers or a single layer of a medium having depth. . which captures contaminants within its structure. as opposed to on the surface.

.Membrane Filtration • A Membrane Filter typically traps contaminants larger than the pore size on the addressed surface of the membrane.

A Combination Filter • A Combination Filter combines different membrane pore sizes. . They can offer an economical alternative to using individual prefilters and final filters. or combines depth media and a membrane filter to create selfcontained serial filter units.

Important Filter Media Properties • • • • • • Pore Size Rating Hydrophilic or Hydrophobic Porosity (Flow rate) Chemical Compatibility Extractables Surface Area .

2µm rated membranes must retain 100% of the microorganism Brevundimonas diminuta according to ASTM F838-83 methodology .Pore Size Ratings • Absolute Rated • Specifies the pore size at which a challenge organism of a particular size will be retained with 100% efficiency under specific test conditions • Example: 0.

Nominally rated filters vary significantly in the filtration industry .Pore Size Ratings • Nominal Rated • • Specifies the pore size at which a challenge particle of a particular size will be retained with an efficiency of 60-98%.

Possessing an Affinity for Water (likes water) • Hydrophobic . such as organic solvents .Lacking an Affinity for or Repels Water (dislikes water) • Will Not wet in water but will wet in low surface tension liquids.Hydrophilic or Hydrophobic • Hydrophilic .

higher flow) • Not pore size.Porosity • The measure of quantity of all the open spaces (pores) in the membrane • Membranes are generally 50% to 90% open space • Directly proportional to the flow rate of the membrane (more pores. but more a reflection of number of pores .

Chemical Compatibility • The filter media’s resistance to select chemicals • Pore structure should not be adversely effected by chemical exposure Least Resistant FP Vericel Supor HT-Tuffryn Nylon GN Metricel Most Resistant PTFE GHP Metricel PP .

Inhibits growth / effects recovery Environmental Analysis .Kills cells Micro Analysis .Add extraneous peaks Cell Culture .Metals contaminants .Extractables • Contaminants eluting from the filter media or device which may adversely effect effluent quality • • • • HPLC Analysis .

• Contaminants may be:
• • •

Wetting agents in filter media Sterilization residuals (EtO) Additives in polymer or housing components (colorants, mold release agents, etc..)
• Remember – its not just about the filter, consider the housing

Surface Area
Doubling surface area will yield at least two times more service life and as much as four times more.

How do we make membranes?
Membranes can be made in the 4 following ways

• • • •

Phase-Inversion Stretching Track-Etching Sintering

Phase-Inversion • Used to produce most of BioSciences membranes • Most versatile method for producing membrane • Three major components are combined • • • Polymer Solvent Pore former .

Phase-Inversion • Polymer • Makes up the body of the membrane • Solvent • Dissolves the polymer into a liquid state Inability to dissolve the polymer Polymer forms around the pore former droplets • Pore Former • • .

Stretching • Commonly used to produce PTFE membrane • A dense plastic film is carefully stretched in all directions • Pores are formed under the conditions of the stretching .

or neutral • • • • • • charge) HT Tuffryn (polysulfone) FP Vericel (polyvinylidene fluoride-PVDF) GN-4 & GN-6 Metricel (mixed cellulose esters) GHP (hydrophilic polypropylene) MetricelPolypropylene Zylon PTFE or “Teflon” (polyterafluoroethylene) .Some Pall BioScience Membranes • Supor(polyethersulfone) • Biodyne (nylon 6.-.6 with +.

. or organic resin.The basic HPLC Mobile phase. refractive index or fluorescence. usually packed with small particles of porous silica. Can be ultraviolet. This is where the “multistage” separation Process occurs. The detector contains a “flow cell” mobile phase carries sample though HPLC Eluent Reservoir Pump Sample Injector Column Detector Fraction Collector Pump is used to force eluent (mobile phase) through the column and detector. Recorder The Column. The pump should provide constant Flow. Detector. Usually organic solvent mixture. alumina. Needs to be of high grade and filtered – use the SolVac.

. • This will reduce system downtime and maintenance. • Protect columns. • Reduce incorrect results. noisy baselines caused by contamination and noise in the detector due to release of dissolved gases in the mobile phase.Sample Prep • Both “samples” and “mobile phase” should be filtered.

Selecting membrane pore size • The filter pore size should be determined based on the column packing size. This space is labelled the Flow Path. Ideally. . you would not want contamination to fit into the space between the particles of packing. As you can see the column packing particles touch each other.

you should use a 0. the flow path is 0. . • When an HPLC column has a packing size of 3 µm or smaller.2 µm filter because a 0.• We can use basic mathematics to determine the size of the flow path If the column packing is 3 µm in diameter.43 µm.45 µm filter may let particles through that will plug the column.

. • Aqueous Samples • Choose a hydrophilic membrane. • Aqueous and Organic-solvent Solutions • Pall Life Sciences patented hydrophilic polypropylene (GHP) membrane is a universal membrane for both aqueous and organic applications.• When filtering solvents and chemicals selecting a chemically compatible membrane is important. or PVDF membranes. Selecting the correct membrane • Aggressive Organic Solvents • Choose a chemically resistant membrane. for example. Pall GHP. PES. for example PTFE. Nylon.

HPLC certified device • Contaminants eluting from the filter media or device which may effect effluent quality .Extractables It is important to use a clean.

to spin filters and syringe filters • Devices are also available with a single layer or combination filter (with a prefilter) • By increasing surface area and using a prefilter we will increase throughput • If sample size is small we must consider holdup volume (the amount of sample that will be left in the device) Selecting the best device .• Filters come in a variety of sizes ranging from the area within a single well of a 96-well plate.

® Acrodisc PSF Syringe Filter Three Powerful Benefits in One .

GxF multi-layered prefilter provides maximum throughput • The Acrodisc® PSF syringe filter has a serial prefilter to allow for two to four times the throughput and faster flow rates than standard devices. 1 . • The multi-layered prefilter traps particulate rated from > 40 to 1 µm.

Automation certification • Caliper (Zymark) certifies Pall syringe filters for worry-free performance in their automated workstations. 2 .

cause high column back-pressure. • Pall Acrodisc PSF syringe filters with GHP (hydrophilic polypropylene) membrane offer the most efficient removal of particulate and prolong the life of HPLC system components. 3 . and shorten the normal lifetime of the column. • Injection of samples containing particulates will eventually block the column inlet.Best protection for HPLC columns Extending column life up to 46 times • Plugging is the most frequently encountered cause of High Performance Liquid Chromatography (HPLC) column failure.

Retention Efficiency / Column Life Study • Examine how much longer an HPLC column might last when samples are filtered with Pall Acrodisc premium syringe filters • Inject UNFILTERED mixture of latex spheres • Repeat injections until column plugs • Inject FILTERED mixture into new column • Repeat injections until column plugs • Plot pressure vs. injection number .

Results .

Your experience… • Have you ever had a column plug unexpectedly? • Have you ever had unexpected peaks in your chromatograms? • Has your analyte concentration change after filtration? • Do you consider analyte adsorption in your filter equivalency testing? .

9 mL well • HPLC certified for low extractables.Other Pall Sample Prep Products • AcroPrep 24 • 24 x 1. • 500 µL sample volume. • GHP Nanosep • Various pore in a centrifugal size and device. • AcroPrep 96 well filter plates • 1ml size wells . for low extractables. membrane • HPLC certified configurations.

Filtering Mobile Phase • Traditionally labs will use a 47mm glass filter funnel to filter • Glass filter funnels are easy to break • They are open systems. therefore it is easy to spill aggressive solvents .

• Durable plastic construction. .SolVac Filter Holder • Draws directly from HPLC solvent bottle. Less likely to spill aggressive solvents than glass funnels or disposable cups. Less likely to break than glass funnels or assemblies.

Using the Solvac – its safe and easy .

Which membrane should I use? • Pall supplies a whole range of 47mm membranes which will fit into the SolVac filter holder • We would recommend our GHP PP membrane for the majority of requirements. .

Summary • Filtration should be used in chromatography • Samples and mobile phase should be • • • filtered Choose membrane based on compatibility Choose EFA based on volume to be filtered Choose pore size based on column bead size Not all 0.45µm filters are equal Filtration does extend column life • • .

Microbiology .

. The accurate and reliable detection and identification of micro-organisms is critical. • Pall Corporation began providing membranes for microbial analysis over 50 years ago.Microbiology • Micro-organisms can halt the production of pharmaceuticals and cause disease.

The MF Technique .

food and beverage industries.Advantages of MF technique • Permits usage of large sample volumes. • Used in water. pharmaceutical. • Reduced preparation time. cosmetics. • Effective and accepted technique. . • Provides presence or absence information within few days.

What do we need to do the MF technique? • • • • • • • • Vacuum Pressure Pump with VacuShield Filtering Flask Stainless Steel Forceps-smooth tip Petri Dishes-50mm Media Tubing Bunsen Burner Catch bottle .

. the mixed cellulose ester GN Metricel membranes set the standard worldwide.Microbiology • For microbial analysis. where culturing of sensitive organisms can be difficult and identification is critical for process control and public safety. • These membranes provide uniform and consistent growth of organisms to ensure quality in pharmaceutical products.

.Why use a Pall membrane? • The GN6 mixed cellulose esters membrane is most accepted filter media for microbiological analysis. • The unique dot grid pattern provides easy quantification of bacterial colonies without growth inhibition or enhancement.

45µm Pore Size White Membrane Black Gridlines E. Total Coliform. coli. Total Bacteria .GN-6 Metricel Membrane • Mixed Cellulose • • • • • Ester > 90% Recoveries 0.

Supor 200 Membrane • • • • • • Polyethersulfone > 90% Recoveries 0. analysis .2µm White Membrane Black Gridlines Pseudomonas sp.

Light Colored Bacteria .Metricel Black Membrane • • • • • • • Polyethersulfone > 85% Recoveries 0.45µm and 0.8µm Black Membrane White Gridline Yeast. Mold.


MicroFunnel™ Filter Funnels .

• Ideal for quality control analysis of aqueous fluids used in pharmaceutical production. Individually labelled for lot traceability.MicroFunnel™ Filter Funnels • Increase laboratory efficiency with convenient. ready-to-use disposable filter funnels • Test any aqueous solution for microbial contamination using the principles of the Membrane Filter (MF) Technique. .

or Metricel Black membrane 0. 100 mL and 300 mL size GN-6.45 µm gridded Removable membrane . Supor.Culture in place or remove filter for culturing on agar plate.2 gridded or plain Supor 0.MicroFunnel™ Filter Funnels • • • • • • 47 mm. Gamma irradiated .

MicroFunnel™ Filter Funnels • Easy snap apart design • Fits directly on Manifold • Individually Bagged • LP Version has additional Petri Dish with Dry Absorbent Pad • Extensive Literature .

vacushield. tubing.What do I need to use with a funnel? Pump. catch vessel Forceps Manifold (this is a single place) Petri dishes (depending on the funnel) Media .


The Supor MF Plus has been validated with samples hotter than 90ºC. .MicroFunnel™ Plus Filter Funnel • Combines sample cup and filter funnel into one product to prevent contamination of the sample. • Optimized for use in hot loop water applications.

• Patented design • Current method required collection of sample in separate cup. . The filter has been validated with samples hotter than 90ºC.MicroFunnel™ Plus Filter Funnel • Innovative design • No other product like it on the market. • MicroFunnel Plus filter funnel eliminates the need for a separate sample cup.

. • The high temperature and constant circulation helps control any growth of microorganisms.Hot water loop • Water that is constantly circulating at elevated temperature of 70 . • Water is used to produce Sterile WFI or simply purified water.90ºC or higher.

Design Features .

How to use it ? .

MicroFunnel™ Benefits • Less material to • order • inventory • dispose of • Less potential for contamination • no sample transfer • filtered air drawn in .

• Provides added contamination control for sampling fluids for microbial testing. .• Fluid is sampled MicroFunnel Plus AP filter funnel into the port on the lid so it never comes in contact with the environment.


MicroFunnel Plus AP filter funnel Vented Lid Supor Membrane Seal at Base Vent Tab Benefits : • Contamination Control – Aseptic sampling port – No sample transfer – Filtered air drawn in • Time Saving – Less handling .