WaterMicrobiology水中微生物微生物学实用教案

1、Pharmaceutical Microbiology PM221These lectures Types of water used in pharmaceutical manufacturing Standards Components of water purification system Water treatment: Deionisation, reverse osmosis and distillation Water storage and distribution Water system design Water testing第1页/共28页第一页，共29页。Pharm

2、aceutical Microbiology PM222Pharmaceutical Water There are various types of water used in pharma industry. May be used in: formulation, processing operations (e.g. cooling, cleaning, sterilisation) Final rinsing of product contact surface. Different grades required depending on use and type of produ

3、ct Water classified based on its source, quality (chemical and microbiological purity), treatment and use. Note that water also required for microbial growth.第2页/共28页第二页，共29页。Pharmaceutical Microbiology PM223Water types 3 main types used in pharma industry: Potable water: Ordinary drinking water fro

4、m mains (tap water) Fit to drink but not usually suitable for pharma applications, especially manufacture of sterile products. Contains wide range of dissolved solids and gases harmless if swallowed, but potentially harmful if injected, could also interact with product. Has variable degree of hardne

5、ss and added Cl for microbial control.第3页/共28页第三页，共29页。Pharmaceutical Microbiology PM224Water Types (contd) Purified water: Specifications for purified water contained in British Pharmacopoeia (BP), EP and USP that describe chemical purity limits. Not necessarily sterile, therefore not suitable for

6、manufacture of parenterals. Used for non-parenteral product formulation Can also be used as initial cleaning agent for some processes. Usually prepared from potable water by deionisation, which removes dissolved substances. May also be produced by distillation or reverse osmosis第4页/共28页第四页，共29页。Phar

7、maceutical Microbiology PM225Water types (contd): Water for Injections (WFI): Standards contained in BP, EP and USP Prepared from potable or purified water by distillation. Water normally deionsied before distillation (stops still “furring up” like kettle). Higher the quality of water fed to still,

8、the greater the quality assurance that water produced by the still will be of high quality. High chemical purity and free from pyrogens. Bulk WFI has low bioburden (sterilised afterwards) Commercially supplied WFI is sterile.第5页/共28页第五页，共29页。Pharmaceutical Microbiology PM226Standards: US Pharmacopoe

9、ia defines water for pharmaceutical use as follows: Water for Injection: Water purified by distillation (or other process) to remove chemicals and microorganisms Prepared from water complying with suitable standard Contains no added substance Purified water: Water obtainable by a suitable process, p

10、repared from water complying with suitable standard, contains no added substance.第6页/共28页第六页，共29页。Pharmaceutical Microbiology PM227USP 23 standardsPurified WaterWFIOrganics0.5ppm TOC0.5ppm TOCConductivity1.3mS/cm at 25oC in-line measurement1.3mS/cm at 25oC in-line measurementEndotoxin by LALNo speci

11、fication0.25 EU/mlBacteria100 cfu/ml80oC Multi-cartridge filters RO unit Deionisation unit 0.5m mm pre-filter - reduce particle and microbial content and extends life of final filter UV sanitizer - (254nm) reduces microbial flora, ozone and chlorine content (but can generate endotoxins) 0.2 m mm fin

12、al filter Storage tank, distribution loop (“dead legs” and low flow should be avoided) and pumps. Distillation still (for WFI production)第8页/共28页第八页，共29页。Pharmaceutical Microbiology PM229Deionisation Usually consists of 1 to 4 cylindrical cartridges Good at removing dissolved solids and gases Poor a

13、t removing other impurities. Stagnant water in cartridges may increase bioburden of water! Needs to be combined with other purification systems for high grade water.第9页/共28页第九页，共29页。Pharmaceutical Microbiology PM2210Deionisation (contd) Deionisation works by: Water passed through deionisation resins

14、 (tiny spherical plastic beads) cationic contaminants in water exchanged with hydrogen ions in resin anionic contaminants in water exchanged for hydroxyl ions in resin. Impurities eventually replace hydrogen and hydroxy ions in resin and resin has to be replaced.第10页/共28页第十页，共29页。Pharmaceutical Micr

15、obiology PM2211第11页/共28页第十一页，共29页。Pharmaceutical Microbiology PM2212Reverse Osmosis Osmosis = movement of water across a semipermeable membrane from an area of high concentration to an area of low concentration. In Reverse osmosis the process is reversed, due to presence of a high pressure pump. Pum

16、p applies pressure to the concentrated side (i.e. impure water), forcing molecules over to the pure side, leaving impurities behind to be removed as waste. Often combined with deionisers第12页/共28页第十二页，共29页。Pharmaceutical Microbiology PM2213Reverse Osmosis Reverse osmosis is a very efficient process.

17、Removes: 100% suspended solids 99.5% bacteria, viruses, pyrogens. 97-99.5% organics 94-99% inorganics.第13页/共28页第十三页，共29页。Pharmaceutical Microbiology PM2214Reverse Osmosis第14页/共28页第十四页，共29页。Pharmaceutical Microbiology PM2215Distillation Water heated to boiling point and resultant water vapour, or ste

18、am, is condensed. Condensate (or distillate) is then collected. Not an “on-demand” method. Water has to be distilled and then stored in advance. Wasteful process as much as 95% of feedwater may be wasted, leaving only about 5% as pure product. Also requires high energy input. Still design important

19、to avoid carry-over (“entrainment”) of water droplets that contain impurities第15页/共28页第十五页，共29页。Pharmaceutical Microbiology PM2216Distillation第16页/共28页第十六页，共29页。Pharmaceutical Microbiology PM2217Water Storage and Distribution Once water has been distilled, it must be held in appropriate holding and

20、distribution system in order to prevent microbial contamination and growth. This can be achieved as follows: Use water immediately on collection from still Only applicable to small-scale use Collect water in clean vessel, seal and sterilse Only applicable to small-scale use. Collect water, pipe dire

21、ctly from still to insulated holding tank and hold at 80oC. Pump from tank when required. Connect insulated holding tank to ring-piping system, continuously circulate water back to tank at 80oC. Method most commonly used in large-scale sterile manufacturing.第17页/共28页第十七页，共29页。Pharmaceutical Microbio

22、logy PM2218Steam Where steam comes in contact with product or with product-contact surfaces, “clean steam” required. E.g. steam supplied to autoclave chamber SIP systems. Clean steam produced in specially designed “Clean steam generators” or “Pure steam generators” from purified or distilled water第1

23、8页/共28页第十八页，共29页。Pharmaceutical Microbiology PM2219Water System Design Quality What type of water will be used and where? Quantity How much water will be used, flow rate? Time Usage profile: e.g. 8 hrs per day, 24/7? Space Where is water system / loop being placed? Money Overall budget.第19页/共28页第十九页

24、，共29页。Pharmaceutical Microbiology PM2220Water System Design Physical layout Create a usage layout Start with a drawing of the building and mark use points Colour code the use points to indicate usage needs Flow, daily usage, quality, ergonimics.第20页/共28页第二十页，共29页。Pharmaceutical Microbiology PM2221第2

25、1页/共28页第二十一页，共29页。Pharmaceutical Microbiology PM2222Multi-MediaSoftenSoftener5FilterBreakTankUVunit1 FilterCarbon FilterCausticadditionTo main storageChlorination6014 60096006602468076815Pump6804603668736004 6007ReverseOsmosis2nd passReverseOsmosis 1st pass 6808Heat ExchangeDeionisation5FilterWater

26、treatment 第22页/共28页第二十二页，共29页。Pharmaceutical Microbiology PM2223Ozone (O3) Loop Sanitisation User points interlocked Ozone generator activated, UV unit deactivated Ozone concentration monitored Sanitisation timer started when ozone concentration in return rises to 200 ppb When time out period expire

27、s, Ozone generator deactivated, UV unit activated User points unlocked when concentration at UV outlet falls to 10 ppb第23页/共28页第二十三页，共29页。Pharmaceutical Microbiology PM2224Microbiological Water testing Guidelines e.g. USP Standard Methods for the examination of water and wastewater. Pour plate metho

28、d or membrane filtration method used for enumeration of microbes in sample, MPN / serial dilutions and selective agar for detection of coliforms. Endotoxins (WFI) tested by LAL or other method (rabbit test)第24页/共28页第二十四页，共29页。Pharmaceutical Microbiology PM2225Chemical testing TOC can be determined u

29、sing a spectrophotometer Sample placed in cuvette, absorbance read第25页/共28页第二十五页，共29页。Pharmaceutical Microbiology PM2226Routine monitoring Sampling sites and frequencies to be established Establish alert and action limits Worksheets to record test results Trend results over time Carry out investigation/corrective action if action limits exceeded or alert limits exceeded a number of times.第26页/共28页第二十六页，共29页。Pharmaceutical Microbiology PM2227Summary The 3 main types of water used in pharmac