Cell Rupture Nanoparticles Emulsions Liposomes

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Cell Rupture Nanoparticles Emulsions Liposomes High Pressure Homogenizers AVESTIN Inc., Ottawa, Canada AVESTIN Europe GmbH, Germany Tel: +1-613-736-0019 Tel: +49-621-724-5980 E-mail: avestin@avestin.com E-mail: avestin@avestin.net

Inside schematic of dynamic homogenizing valve EmulsiFlex Principle of Operation A high pressure pump (C) pushes the product through dynamic homogenizing valve (D). The product can also be passed through a membrane (E). It can be collected (G) or recycled to the reservoir (A) via tubing/pipes or heat exchanger (F). EmulsiFlex equipment can be delivered as: 1-High pressure homogenizers (A, C, D, F, G) 2- High pressure filter/extruder (A, C, E, G) 3-Homogenizer/filter/extruder combination (A, C, D, E, G)

EmulsiFlex General Information Capacity: AVESTIN manufactures homogenizers in many sizes, from a capacity of 1L/hr up to 1000L/hr. Samples as small as 3mL can be processed using the EmulsiFlex -B15, which has a hold back volume of less than 1mL. Pressure: Adjustable between 500-30,000psi/ 34-2,070bar, which is high enough for virtually every homogenization application. Dynamic homogenizing valve is controlled pneumatically; automatic homogenizing pressure control as well as transducers for precision pressure measurement are available as options. Unique Design: There are no o -rings and no gaskets in the product path of AVESTIN homogenizers. All face seals are precision-machined metal to metal to ceramic. Temperature Control: AVESTIN provides stainless steel heat exchangers to control inlet and outlet temperatures. The entire EmulsiFlex -C5 can be immersed in a water bath for temperature control. Filter/Extruder: Most AVESTIN homogenizers can be equipped with a high-pressure filter/extruder. Applications include the extrusion of emulsions or liposome through membranes and sterile filtration at high pressure. The EmulsiFlex -C1000, AVESTIN s largest high-pressure homogenizer, next to the EmulsiFlex -C5, AVESTIN s smallest high-pressure homogenizer. Homogenizing Valve: AVESTIN homogenizers feature dynamic homogenizing valves. These valves can be easily dissassembled for cleaning or inspection. Submicron particles or droplets with diameters less than 50nm and extremely narrow size distributions can be achieved. Cleaning/Sterilization: Instruments are CIP and are suitable for clean room and GMP manufacturing. All wetted parts are autoclavable and can be easily and quickly disassembled for inspection and validation. All materials within wetted parts path will withstand prolonged exposure to high pressure steam for the purposes of sterilization (SIP). Application: Pharmaceuticals, chemicals, cosmetics, coatings, inks, food, etc.

EmulsiFlex Product Line Model Minimum Sample Volume Maximum Flow Rate Holdback Volume Maximum Homogenizing Pressure Pump Type B15 3mL 15mL batch < 1mL 45,000psi/ 3,000bar Pneumatic intensifier C5 7mL 1~5L/hr < 1mL 30,000psi/ 2,070bar Single-acting air driven pump C3 7mL 3L/hr < 1mL 30,000psi/ 2,070bar Single-acting electric pump C50 25mL 15~50L/hr < 4mL D20 50mL 20L/hr < 4mL 30,000psi/ 2,070bar D20A: 15,000psi/ 1,035bar D20B: 30,000psi/ 2,070bar Double-acting air driven pump Electric driven double-acting pump C55 100mL 55L/hr < 6mL C55A: 20,000psi/ 1,380bar C55B: 30,000psi/ 2,070bar Electric motor driven triplex pump C160 250mL 160L/hr < 8mL C160A: 20,000psi/ 1,380bar C160B: 30,000psi/ 2,070bar Electric motor driven triplex pump C500 5L 500L/hr < 15mL C500A: 20,000psi/ 1,380bar C500B: 30,000psi/ 2,070bar Electric motor driven triplex pump C1000 5L 1000L/hr < 15mL C1000A: 20,000psi/ 1,380bar C1000B: 30,000psi/ 2,070bar Electric motor driven triplex pump

EmulsiFlex -B15 Capacity: 3-15mL batch Pressure: Up to 45,000psi/ 3,000bar Minimum Sample Volume: 3mL Power: Electric power not required Air: 80-150psi/ 5-10bar Dimensions: mm: 335(L) x 280(W) x 650(H) in: 13.3(L) x 11(W) x 25.5(H) Weight: 37kg/ 81lbs

EmulsiFlex -C5 Capacity: 1~5L/hr, depending on pressure and air supply. Pressure: Up to 30,000psi/ 2,070bar Minimum Sample Volume: 7mL Power: electrical power is not required Air: 7.0CFM@100psi (7bar) Dimensions: mm: 430(L) x 160(W) x 320(H) in: 17(L) x 6.3(W) x 12.6(H) Weight: 25kg/ 55lbs

EmulsiFlex -C3 Capacity: 3L/hr (at 60Hz) Pressure: Up to 30,000psi/ 2,070bar Minimum Sample Volume: 7mL Power: 110~220V, 50/60Hz, 1hp/0.75kW Air: 60-120psi, virtually no air consumption Dimensions: mm: 530(L) x 460(W) x 760(H) in: 21(L) x 18(W) x 30(H) Weight: 95kg/ 210lbs

EmulsiFlex -C50 Capacity: 15~50L/hr, depending on pressure and air supply Pressure: Up to 30,000psi/ 2,070bar Minimum Sample Volume: 25mL Power: 110~230V, 50/60Hz (for control panel) Air: 45CFM@100psi (7bar) Dimensions: mm: 610(L) x 530(W) x 260(H) ln: 24(L) x 20.8(W) x 10.2(H) Weight: 80kg/ 176lbs

EmulsiFlex -D20 Capacity: 20L/hr, independent of pressure. Pressure: Two models available A - Fully adjustable up to 15,000psi/ 1,035bar B - Fully adjustable up to 30,000psi/ 2,070bar Minimum Sample Volume: 50mL Power: Electrical motor driven duplex pump. 3-phase, 50/60Hz, 230/400VAC. Dynamic, fully ceramic homogenizing valve with pneumatic control. Virtually no air consumption. Wetted parts are autoclavable. Dimensions: mm:730(l) x 600(W) x 640(H) In: 29(L) x 24(W) x 25(H) Weight: 120kg/ 250lbs

Example of a production scale EmulsiFlex highpressure homogenizer.

EmulsiFlex -C55 Capacity: 55L/hr (at 60Hz) Pressure: Up to 30,000psi/ 2,070bar Minimum Sample Volume: 100mL Power: 208/230/460V, 50/60Hz, 3-phase, 7.5hp/5.5kW Air: 60-120psi, virtually no air consumption Dimensions: mm: 480(L) x 530(W) x 780(H) in: 18.9(L) x 20.8(W) x 30.7(H) Weight: 220kg/ 485lbs

EmulsiFlex -C160 Capacity: 160L/hr (at 60Hz) Pressure: Up to 30,000psi/ 2,070bar Minimum Sample Volume: 250mL Power: 208/230/380/460V, 50/60Hz, 3-phase, 15hp/11kW Air: 60-120psi, virtually no air consumption Dimensions: mm: 1250(L) x 750(W) x 820(H) in: 49(L) x 30(W) x 32.2(H) Weight: 500kg/ 1,100lbs

EmulsiFlex -C500 & C1000 Capacity: 500 or 1000L/hr (at 60Hz) Pressure: Up to 30,000psi/ 2,070bar Minimum Sample Volume: 5L Power: 208/230/380/460V, 50/60Hz, 3-phase, 60hp/75kW Air: 60-120psi, virtually no air consumption Dimensions: mm: 1110(L) x 838 (W) x 1550(H) in: 44(L) x 33(W) x 61(H) Weight: 1,000kg/ 2,200lbs

EmulsiFlex Accessories Heat Exchanger Avestin provides a range of heat exchangers which can be used to control product temperatures. Heat transfer fluids in the range of -25 to 200 O C/ -13 to 400 O F can be used. Simple cooling coils can be fitted where less stringent temperature control is acceptable. All heat exchangers are made of stainless steel to a high quality standard. Heat exchangers are easily and rapidly removed or replaced as required.

EmulsiFlex Accessories (Continued) Peak Reset Meter & Transducer AVESTIN manufactures a unique, electronic peak reset meter to process the signals generated by an autoclavable pressure transducer for sterile appliations. The peak pressure is displayed and reset every three seconds. Provisions are made to output pressure measurements to a recorder for monitoring or automating processing. Automatic Pressure Control Simple system. Attaches to any lab scale Avestin High Pressure Homogenizer without modification. User sets the pressure once and the pressure will be maintained during the entire process time. The set pressure maintains even if the product property changes during the homogenization. Filter/Extruder For most Avestin homogenizers Diameter: 47mm or 90mm Pressure: Up to 6,500psi/ 450bar

Applications Cell Rupture Strain: Machine: Pressure: Schizosaccharomyces pombe EmulsiFlex-C160 28,000psi/ 193MPa Sample cfu/ml % viable % NOT viable control 5.1x10 7 ~100 ~0 1 pass 5.0x10 3 0.01 99.99 Strain: Machine: Pressure: E. coli (BL21) EmulsiFlex-C160 20,000psi/ 138MPa A control sample of unruptured Schizosaccharomyces pombe yeast. Schizosaccharomyces pombe after one pass at 28000psi/ 193MPa through the Emulsi- Flex-C160. Only cell fragments remain. Sample cfu/ml % viable % NOT viable control 4.06x10 9 ~100 ~0 1 pass 1.77x10 7 0.44 99.56 2 pass 1.70x10 6 0.04 99.96 3 pass 2.4x10 4 5.9x10-4 ~100 Emulsions Oil in water emulsion Sample Pressure (psi) Diameter (nm) unprocessed - 656 1 pass 16,000 77 3 passes 16,000 59 5 passes 16,000 42 Pharmaceutical Emulsion Sample Homogenizing Pressure (psi) Mean Droplet Diameter (nm) control - 3,500 1 pass 20,000 590 3 passes 20,000 185 5 passes 20,000 140 Liposomes AVESTIN s high pressure homogenizers are used all over the world for liposome research and production. Only EmulsiFlex homogenizers offer the combination of high pressure homogenization, high pressure membrane filtration or a combined, homogenization/filtration process. Dispersions A whole range of dispersions can be run through EmulsiFlex high pressure homogenizers to reduce particle size as well as make the size distribution uniform. EmulsiFlex homogenizers are involved in the processing of dispersions such as drug dispersions, herb extracts, industrial coatings and pigments. Liposomes prepared using the EmulsiFlex high pressure homogenizer, extruder or both in combination. Combined homogenization yields 113.7nm in one pass and 89.9nm in two passes.

LiposoFast -Basic Extrusion of unilamellar, homogeneous liposomes in minutes Capacity: 0.2mL to 1.0mL Over 6000 instruments sold worldwide Available for a FREE trial. Visit for more information LiposoFast -Stabilizer Facilitates the repetive use and extrusion of highly concentrated membranes. Available for a FREE trial. Visit for more information.

LiposoFast -50 Extrusion as simple one-step procedure Lipid suspensions are forced through a membrane with a fixed pore size Produces homogeneous unilamellar liposomes Temperature control jacket 5-50 ml capacity

Production Facility 16,000ft 2 / 1500m 2 of floor space Located in Ottawa, a 15 minute drive from the Parliament of Canada Avestin, Inc. 2450 Don Reid Drive Ottawa, Canada K1H 1E1 Tel: +1-613-736-0019 Fax: +1-613-736-8086 Avestin Europe GmbH Weinheimer Str. 64b 68309 Mannheim, Germany Tel: +49-621-724-5980 Fax: +49-621-724-5813 avestin@avestin.com