Pharmaceutical Microbiology Insights

How to Perform the Water BET (Bacterial Endotoxin Test) – Step-by-Step Guide | Pharmaceutical Microbiology Insights Introduction The Bacterial Endotoxin Test (BET) — often performed using Limulus Amebocyte Lysate (LAL) reagents — is a critical quality control assay for pharmaceutical water systems (Purified Water, Water For Injection). This guide explains how to perform a Water BET in a laboratory-appropriate SOP format and provides tips for reliable results and regulatory compliance. Quick note: Regulatory endotoxin limits and specific acceptance criteria should be taken from your applicable pharmacopeia (USP / BP / EP) or product specifications. Always follow your lab's approved SOPs and safety rules. Principle LAL reagents react with bacterial endotoxin (lipopolysaccharide) and produce a measurable response. There are three common LAL methods: ...

How to Calibrate a Heating Block: Step-by-Step Guide for Accurate Lab Results In laboratories, heating blocks are essential equipment used for maintaining precise temperatures during experiments. Over time, the heating block’s accuracy can drift due to wear, environmental factors, or electrical variations. Proper heating block calibration ensures that your lab results remain reliable and reproducible. In this detailed guide, we will explain the process of calibrating a heating block step by step, the tools required, and tips to maintain accuracy. Why Heating Block Calibration is Important Calibration is the process of adjusting and verifying the accuracy of an instrument. For heating blocks, calibration is crucial because: It ensures experimental consistency and reproducibility. Prevents overheating or underheating of samples. Maintains compliance with lab standards and regulatory requirements. Extends the life of the heating block by preventing temperature-related dam...

·          "Log reduction" is a mathematical term (as is "log increase") used to show the relative number of live microbes eliminated from a surface by disinfecting or cleaning. ·          For example, a "5-log reduction" means lowering the number of microorganisms by 100,000-fold, that is, if a surface has 100,000 pathogenic microbes on it, a 5-log reduction would reduce the number of microorganisms to one. Log Reductions 1 log reduction means the number of germs is 10 times smaller (10 1 ) 2 log reduction means the number of germs is 100 times smaller (10 2 ) 3 log reduction means the number of germs is 1000 times smaller(10 3 ) 4 log reduction means the number of germs is 10,000 times smaller(10 4 ) 5 log reduction means the number of germs is 100,000 times smaller(10 5 ) 6 log reduction means the number of germs is 1,000,000 times smaller(10 6 ) 7 log reduction means the number of ger...

Ø   1912 : pyrogen test was done by Rabbit Test method. Ø   USP XII –(Rabbit test ) approved method (1942 to 1980 ) Pyrogen : Ø   A pyrogen is a foreign substance that causes a fever (temperature elevation) in an animal’s body. Typically, pyrogenic substances include endotoxin and other bacterial by products. Vaccines and other injectable drugs must be confirmed to be pyrogen free according to regulatory requirements of 21CFR, USP, and EP. Pyrogen Testing : 1.        Rabbit test method (Sham test). 2.        LAL test  ( Bacterial Endotoxin Test). Rabbit Test Method:  3 rabbits is selected  Endo toxin sample is inoculated (10mL/kg per body weight)  3 days rabbit temperature is observed  Rabbit temperature is with in 37 o C.(sample does not have endotoxins or toxic)  Rabbit temperature out off 37 o C.(sample have endotoxins or toxic)  37 o C =healthy human temp...

Name of the Sterilization Process Microorganisms Used 1.     Steam sterilization Geo bacillus Stearothermophilus (7953) 2.     Hydrogen peroxide sterilization 3.     Formaldehyde Strilization 4.     Propylene Oxide Sterilization 5.     Ozone Sterilization 1.     Ethylene Oxide sterilization Bacillus Atrophaeus (9372) 2.     Dry heat sterilization 3.     Chloroxidene sterilization 4.     Ozone Sterilization 1.     Low temperature or steam sterilization Bacillus Subtilis (6633) 1.     Ionizing radiation Bacillus pumpilus (27142) 2.     Ultra violet sterilization 1.     Foo...

·          Culture media contains nutrients and  physical growth parameters necessary for microbial growth. All microorganisms cannot grow in a single culture medium and in fact many can’t grow in any known culture medium. ·          Organisms that cannot grow in artificial culture medium are known as obligate parasites. Mycobacterium  and  Chlamydias are obligate parasites. Bacterial culture media can be distinguished on the basis of composition, consistency and purpose. 1.     Solid medium (Agar medium) : solid medium contains agar at a concentration of 1.5-2.0% or some other, mostly inert solidifying agent. Solid medium has physical structure and  allows bacteria to grow in physically informative or useful ways (e.g. as colonies or in streaks). Solid medium is useful for isolating bacteria or for determining the colony cha...

·          Bacterial Generation Time (or) Doubling Time 1. Lag Phase 2. Exponential (or) logarithomic Phase (or) Log phase 3. Stationary phase 4. Decline Phase 1. Lag Phase: ·          When microorganism is introduced in to the first medium. ·          It take some time to adjust which the new environment. ·          Cellular metabolism is accelerated. ·          Cells are increasing in size. ·          By bacteria replicate. 2. Exponential (or) Log phase: ·          The microorganism rapidly growing and divided state. ·          Organism DNA replicate begin by Binary fission at a constant rate. ·        ...

·          WHO(World Health Organization) & EU (European Union) viable monitoring : Class Settling plates cfu/m 2 /4 hours Active Air sampling  Cfu/m3 Surface Monitoring Cfu/plate (25 cm 2 ) Personal monitoring Cfu/glove 5 A <1 < 1 < 1 < 1 6 B 5 10 5 5 7 C 50 100 25 -- 8 D 100 200 50 -- ·          Schedule-M (Drug and Cosmotic Act 1940) Limits : Class Settling plates cfu/ 2 hours Active Air sampling  Cfu/m3 Surface Monitoring Cfu/plate (25 cm 2 ) Personal monitoring Cfu/glove 5 A < 1 < 1 < 1 < 1 6 B 5 10 5 5 7 C 50 100 25 -- 8 D ...