Compendial Methods

visual inspection to ensure products are free of visible particulates

a liquid is considered clear if its clarity is the same as that of water - Ph.Eur. 2.2.1

USP <785>

USP <791>

What are Bacterial Endotoxins?

A bacterial endotoxin is a pyrogen and is found in the cell wall of Gram-negative bacteria. Beyond a certain limit, endotoxin introduced into bloodstream or spinal fluid can cause fever, organ failure, septic shock and even death. As per US Pharmacopeia, 2011, Chapter 85, the Bacterial Endotoxin Test (BET) is a method to detect and/or quantify endotoxins from Gram-negative bacteria using Limulus amoebocyte lysate (LAL) from the horseshoe crab (Limulus polyphemus or Tachypleus tridentatus). FDA Guidance for Industry: Pyrogen and Endotoxins 2012, recommends all injectable and implantable drug products and devices to be tested for (below limit of specified range) endotoxin content before release to any patient.

How is Bacterial Endotoxin detected?

Bacterial endotoxins, when present, activate the proteolytic activity of proenzyme Factor C found in circulating amebocytes of horseshoe crab (Limulus polyphermus). Endotoxin levels are determined by measuring the activity of Factor C in the presence of a synthetic peptide substrate that releases p-nitroaniline (pNA) after proteolysis, producing a yellow color that can be measured at an absorbance of 405 nm.

What we provide?

We provide a wide range of analytical services to support the biopharmaceutical product development, including essential release assays like the BET. Our laboratories offer state-of-the-art equipment for endotoxin testing which can be either cartridge-based or plate-based, adhering to cGLP, cGMP and other regulatory guidelines.

What is Bioburden Testing?

A Bioburden test is used to assess the total number of viable microorganisms on any surface like a medical device, a container or a component. This test is generally conducted for any substance which needs to be controlled or monitored to give a proper assessment of manufacturing procedures before sterilization. This test may also be used as an indicator for the number of rounds (or doses) of sterilization (for example: gamma radiation) required to achieve desired sterility in a medical device, material or sample.

How is a Bioburden test conducted?

Bioburden tests can be conducted in two ways- membrane filtration method and pour plate method. Membrane filtration method is used for aqueous solutions >2 mL and involves vacuum manifold along with Tryptic Soy Agar cassettes. Pour plate method using Tryptic Soy Agar plates, is meant for aqueous samples between 0.1 mL and 2 mL. The Tryptic Soy agar cassettes or plates are incubated at 20 °C – 35 °C for 2-7 days, as per USP <61> and <62>. Release of Drug Products with different microbes is decided as per acceptance criteria in USP<1111>.

What we provide?

We provide a wide range of analytical services to support the biopharmaceutical product development, including essential release assays like the Bioburden testing for Drug Products. Our laboratories offer both methods- Membrane filtration and Pour plate method, adhering to USP, cGLP, cGMP and other regulatory guidelines.