Author details

10.5 Blocking the reaction

Growing and Handling of Bacterial Cultures

By diazo coupling reagent

By acetic acid

taken at 405 nm.

and is stirred to mix.

10.6 Measurement at 545 nm

11. Results from software

1.51" to collect the results.

Conflict of interest

104

After completion of the incubation period, place tube rack in ice water bath, then

blocks the reaction immediately from one of the two methods mentioned below:

Absorbance reading (using spectrophotometer) measurement at 405 nm If 0.8 M acetic acid is used to block the reaction, then absorbance reading is

Note: The readings. Glass photocell is used for reading at 405 nm. Because the volume of the tube content is not sufficient, the distilled water is added to each tube

When Diazo coupling reagent is used for blockage of the reaction then the

All the absorbance readings are fed in the "Software reader for window version

Endotoxin=ml in product <sup>¼</sup> conc:of std:in solution � sample ABS � blank ABS

MVC ¼ M=K Where the lowest sensitivity of lysate, M is the maximum dose/kg body weight

> MVD ¼ concentration of product in 1 ml=MVC ELC ¼ X MVD

The author(s) confirm that this chapter content has no conflict of interest.

Std:ABS � blank ABS � DF

Add 0.4 ml of 0.8 M acetic acid into each tube and stir to mix.

Add 0.5 ml from bottle no 8 s to each tube and stir to mix. Add 0.5 ml from bottle no 9 s to each tube and stir to mix. Add 0.5 ml from bottle no 10s to each tube and stir to mix.

Three bottles of the reagent are used as under;

reading is taken at 545 nm. Note all the readings.

11.1 Manual measurement of endotoxin

Calculations of MVC, MVD and ELC.

and K is constant having value equal to 5.

Following Formula is used to calculate the results

Note: Distilled water is used for reference in both cases.

Yasir Mehmood Faculty of Pharmaceutical Sciences, Government College University Faisalabad, Pakistan

\*Address all correspondence to: yasir\_dpharm@hotmail.com

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
