Membrane filters sterilizing

The earliest commercially available filters were manufactured in two pore sizes 0.45 and 0.8 pm. The 0.45 pm-rated membranes were considered to be stefilizing-grade filters and were successfully used in the sterile filtration of pharmaceuticals and parenterals. The membrane filters were qualified using Serratia marcescens a standard bacterium, having dimensions of 0.6 x 1 pm. However, in the late 1960s it became apparent that the matrix of the 0.45 pm-rated filters could be penetrated by some pseudomonad-like organisms (1). For sterile filtration apphcations in the 1990s, 0.2 pm-rated membranes are the industry standard in the manufacture of sterile parenterals and pharmaceuticals. 

Verification of the microbial retention efficiency of the membrane filters may be undertaken using either Hquid or aerosol challenge tests. A Hquid challenge test is more stringent. Furthermore, this test can provide retention information for process conditions such as extreme moisture after sterilization or air entrained with water drops. A Hquid challenge is performed using a protocol similar to that described for Hquid filtration. 

Another standard test, which is much simpler and more convenient, is the membrane filter technique. A suitable volume of sample is filtered through a sterile, 0.45-p.m membrane filter. The filter is placed in a petri dish containing a specific growth medium (M-Endo nutrient broth, M-Endo medium) and incubated for 24 h at 35°C. If after this time the colonies show the characteristic green sheen, this is taken as positive evidence for the presence of the coliform group (see Water, sewage). 

As mentioned in Chapter 3, DI water has now replaced distilled water for most laboratory purposes. PVC pipe, which is both inexpensive and easy to install, is normally employed to carry the water to points of usage. It is very important that the lines do not have dead legs where water is allowed to stand, since this could encourage bacterial growth. In cases where sterile water is needed, special steps must be taken. The water may be boiled, which also drives out dissolved gases, or ultraviolet radiation may be employed, a common method in the cosmetics industry. Membrane filters will also effectively remove bacteria. Laboratories needing sterile water often use two bacteria removal systems of... 

They may also be required in industrial applications where they become part of venting systems on fermenters, centrifuges, autoclaves and freeze-dryers. Certain types of filter (membrane filters) also have an important role in sterility testing, where they can be employed to trap and concentrate contaminating organisms from solutions under... 

Other applications of filters include sterilization of venting or displacement air in tissue and microbiological culture (carbon filters and hydrophobic membrane filters) decontamination of air in mechanical ventilators (glass fibre filters) treatment of exhausted air ftom microbiological safety cabinets (HEPA filters) and the clarification and sterilization of medical gases (glass wool depth filters and hydrophobic membrane filters). 

Filtration =s0.22,um pore size, sterile membrane filter... 

Articles which are to he discharged from the clean room (or elsewhere) to the aseptic area must he sterilized. To achieve this they should be transferred via a double-ended sterilizer (i.e. with a door at each end). If it is not possible, or required, that they be discharged directly to the aseptic area, they should be (i) double-wrapped before sterilization (ii) transferred immediately after sterilization to a clean environment until required and (iii) transferred from this clean environment via a double-doored hatch (where the outer wrapping is removed) to the aseptic area (where the inner wrapper is removed at the workbench). Hatchways and sterilizers should be arranged so that only one side of the entry into an aseptic area may be opened at any one time. Solutions manufactured in the clean room may be brought into the aseptic area through a sterile 0.22-/im bacteria-proof membrane filter. 

Membrane filtration is the technique reeommended by most pharmacopoeias and involves filtration of fluids through a sterile membrane filter (pore size = 0.45 im), any microorganism present being retained on the surfaee of the filter. After washing in situ, the filter is divided aseptically and portions transferred to suitable culture media which are then incubated at the appropriate temperature for the required period of time. Water-soluble solids can be dissolved in a suitable diluent and processed in this way. 

Injections and infusion fluids must be manufactured in a manner that will minimize or eliminate extraneous particulate matter. Parenteral solutions are generally filtered through 0.22 pm membrane filters to achieve sterility and remove particulate matter. Prefiltration through a coarser filter is often necessary to maintain adequate flow rates, or to prevent clogging of the filters during large-scale manufacturing. A talc or carbon filtration aid (or other filter aids) may also be necessary. If talc is used, it should be pretreated with a dilute acid solution to remove surface alkali and metals. 

In the first example, procaine penicillin, an aqueous vehicle containing the soluble components (such as lecithin, sodium citrate, povidone, and polyoxyethylene sorbitan monooleate) is filtered through a 0.22 pm membrane filter, heat sterilized, and transferred into a presterilized mixing-filling tank. The sterile antibiotic powder, which has previously been produced by freeze-drying, sterile crystallization, or spray-drying, is aseptically added to the sterile solution while mixing. After all tests have been completed on the bulk formulation, it is aseptically filled. 

In general, aqueous ophthalmic solutions are manufactured by methods that call for the dissolution of the active ingredient and all or a portion of the excipients into all or a portion of the water and the sterilization of this solution by heat or by sterilizing filtration through sterile depth or membrane filter media into a sterile receptacle. If incomplete at this point, this sterile solution is then mixed with the additional required sterile components, such as previously sterilized solutions of viscosity-imparting agents, preservatives, and so on, and the batch is brought to final volume with additional sterile water. 

Because membrane filtration is the only currently acceptable method of sterilizing protein pharmaceuticals, the adsorption and inactivation of proteins on membranes is of particular concern during formulation development. Pitt [56] examined nonspecific protein binding of polymeric microporous membranes typically used in sterilization by membrane filtration. Nitrocellulose and nylon membranes had extremely high protein adsorption, followed by polysulfone, cellulose diacetate, and hydrophilic polyvinylidene fluoride membranes. In a subsequent study by Truskey et al. [46], protein conformational changes after filtration were observed by CD spectroscopy, particularly with nylon and polysulfone membrane filters. The conformational changes were related to the tendency of the membrane to adsorb the protein, although the precise mechanism was unclear. 

Glass-filtration device, or a 500-mL filter-sterilization flask with a 0.45-pm membrane. 

Subcutaneously/IM - Prepare soluble tablets in sterile water and filter through a 0.22 micron membrane filter. 

Sterility of antibiotic membrane filter STM No. Lot No. F9AM31473 is sterile (incubated for 21 days)... 

Using a sterile forceps (flame the tips of the forceps and cool to room temperature before using), take a presterilized 47-mm, 0.45-ltm, mixed esters of cellulose (MFC) tortuous path membrane filter (hereafter referred to as membrane filter) and place filter, grid side up, in the filter holder. 

Aseptically decant the Tween solution, splitting the total volume present equally among each of four sterile membrane filter holders. 

Pour an additional 10 ml (approximately) of sterile 0.1% Tween solution into each filter using aseptic technique. This rinse is intended to wash the interior walls of the filtration units and carry any bacteria adhering to these surfaces onto the surface of the membrane filter. 

Using sterile forceps (or forceps whose tips have been flamed and then cooled to room temperature), carefully remove each membrane filter and place onto prepoured TSA plate. 

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