This document discusses pyrogen testing methods. Pyrogens are fever-inducing substances, mainly lipopolysaccharides of bacterial origin. The rabbit pyrogen test, introduced in 1942, involves measuring temperature increases in rabbits injected with a test solution. If temperature increases exceed thresholds, the sample fails the test. The Limulus amebocyte lysate (LAL) test directly measures endotoxins using a lysate from horseshoe crab blood. Both tests are used to ensure medical products are free of pyrogens.
What is pyrogens?
Sources of pyrogens and its elimination methods
Tests for pyrogens-
1. In Vitro Test / LAL Test
2. In Vivo Test / Rabbit Test.
Objective
Principle
Requirements
Procedure
Observation table
Result and interpretation
What is pyrogens?
Sources of pyrogens and its elimination methods
Tests for pyrogens-
1. In Vitro Test / LAL Test
2. In Vivo Test / Rabbit Test.
Objective
Principle
Requirements
Procedure
Observation table
Result and interpretation
Pyrogens include any substance capable of eliciting a febrile (or fever) response upon injection or infection
Endotoxin is a subset of pyrogens that are strictly of gram- negative bacterial origin; they occur (virtually) nowhere else in nature.
Lipopolysaccharide (LPS)is a part of endotoxin, or, endotoxin is the natural complex of LPS occurring in the outer layer of the bilayered gram-negative bacterial cell
Two general methods are used for microbiological assays
Method A: Cylinder plate method or cup plate method.
Method B: Tube assay method or titrimetric method.
Microbiological assay-Principles and methods of different microbiological assay.someshwar mankar
Principles and methods of different microbiological assay. Methods for standardization of
antibiotics, vitamins and amino acids. Assessment of a new antibiotic.
UNIT 6 Fermentation technology, Fermenters, Study of Media, types of fermenta...Shyam Bass
UNIT-6 6th Sem B.Pharma Pharmaceutical Biotechnology-
Following slides include-
Fermentation technology and biotechnological products :
Fermentation methods and general requirements
Study of media
Equipment
Sterilization methods
Aeration process
Stirring
large scale production fermenter design and its various controls
BY- SHYAM BASS
PREPARATION OF BACTERIAL VACCINES:
Steps involved in killed bacterial vaccine preparation:
1. Selection of an antigen:
The exact strain or strains to be incorporated for preparation of bacterial vaccine.
Eg. Cholera vaccine: smooth strains of the two serological types Inaba and Ogawa
TABC vaccine: O and H antigens in S. typhi and S. paratyphi microorganisms and these organisms also contains Vi antigen.
Each strain is carefully checked for freedom from variation and absence of contaminating organisms.
Blood products:Collection, Processing and Storage of whole human blood, dried...Steffi Thomas
blood and its components, conditions for being a donor, anti-coagulants, whole human blood, changes in composition during storage, red blood cells, concentrated platelets, plasma, dried human plasma, fresh frozen plasma, dried human serum, plasma substitutes, ideal properties of plasma substitute, dextran, gum saline, polyvinylpyrrolidone
A reaction intermediate or an intermediate is a molecular entity that is formed from the reactants (or preceding intermediates) and reacts further to give the directly observed products of a chemical reaction.
Pyrogens include any substance capable of eliciting a febrile (or fever) response upon injection or infection
Endotoxin is a subset of pyrogens that are strictly of gram- negative bacterial origin; they occur (virtually) nowhere else in nature.
Lipopolysaccharide (LPS)is a part of endotoxin, or, endotoxin is the natural complex of LPS occurring in the outer layer of the bilayered gram-negative bacterial cell
Two general methods are used for microbiological assays
Method A: Cylinder plate method or cup plate method.
Method B: Tube assay method or titrimetric method.
Microbiological assay-Principles and methods of different microbiological assay.someshwar mankar
Principles and methods of different microbiological assay. Methods for standardization of
antibiotics, vitamins and amino acids. Assessment of a new antibiotic.
UNIT 6 Fermentation technology, Fermenters, Study of Media, types of fermenta...Shyam Bass
UNIT-6 6th Sem B.Pharma Pharmaceutical Biotechnology-
Following slides include-
Fermentation technology and biotechnological products :
Fermentation methods and general requirements
Study of media
Equipment
Sterilization methods
Aeration process
Stirring
large scale production fermenter design and its various controls
BY- SHYAM BASS
PREPARATION OF BACTERIAL VACCINES:
Steps involved in killed bacterial vaccine preparation:
1. Selection of an antigen:
The exact strain or strains to be incorporated for preparation of bacterial vaccine.
Eg. Cholera vaccine: smooth strains of the two serological types Inaba and Ogawa
TABC vaccine: O and H antigens in S. typhi and S. paratyphi microorganisms and these organisms also contains Vi antigen.
Each strain is carefully checked for freedom from variation and absence of contaminating organisms.
Blood products:Collection, Processing and Storage of whole human blood, dried...Steffi Thomas
blood and its components, conditions for being a donor, anti-coagulants, whole human blood, changes in composition during storage, red blood cells, concentrated platelets, plasma, dried human plasma, fresh frozen plasma, dried human serum, plasma substitutes, ideal properties of plasma substitute, dextran, gum saline, polyvinylpyrrolidone
A reaction intermediate or an intermediate is a molecular entity that is formed from the reactants (or preceding intermediates) and reacts further to give the directly observed products of a chemical reaction.
To avoid contamination, the aseptic technique is the method of reducing or removing contaminants from entering the operative field in surgery or medicine.
University Institute of Pharmaceutical Sciences is a flag bearer of excellence in Pharmaceutical education and research in the country. Here is another initiative to make study material available to everyone worldwide. Based on the new PCI guidelines and syllabus here we have a presentation dealing with the quality control tests of parenteral as referred in the pharmacopoeia.
Thank you for reading. Hope it was of help to you.
UIPS,PU team
We all have good and bad thoughts from time to time and situation to situation. We are bombarded daily with spiraling thoughts(both negative and positive) creating all-consuming feel , making us difficult to manage with associated suffering. Good thoughts are like our Mob Signal (Positive thought) amidst noise(negative thought) in the atmosphere. Negative thoughts like noise outweigh positive thoughts. These thoughts often create unwanted confusion, trouble, stress and frustration in our mind as well as chaos in our physical world. Negative thoughts are also known as “distorted thinking”.
How to Make a Field invisible in Odoo 17Celine George
It is possible to hide or invisible some fields in odoo. Commonly using “invisible” attribute in the field definition to invisible the fields. This slide will show how to make a field invisible in odoo 17.
Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
Model Attribute Check Company Auto PropertyCeline George
In Odoo, the multi-company feature allows you to manage multiple companies within a single Odoo database instance. Each company can have its own configurations while still sharing common resources such as products, customers, and suppliers.
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
How to Create Map Views in the Odoo 17 ERPCeline George
The map views are useful for providing a geographical representation of data. They allow users to visualize and analyze the data in a more intuitive manner.
2. Pyrogens
Pyrogens - fever inducing organic substances
Responsible for many febrile reaction
These are Endotoxin.
Having nature
Endogenous (inside body)
Exogenous (outside body)
Exogenous pyrogens –
mainly lipopolysaccharides
bacterial origin, but not necessary
3. Endotoxin characteristic
thermostable
water-soluble
unaffected by the common bactericides
non-volatile
These are the reasons why pyrogens are
difficult to destroy once produced in a
product
4. Test for pyrogens = Rabbit test
the development of the test for pyrogens reach
in 1920
a pyrogen test was introduced into the USP
XII (1942)
The test consists of measuring the rise in body
temperature in healthy rabbits by the
intravenous injection of a sterile solution of
the substance under the test.
5. Why the Rabbit?
Reproducible pyrogenic response
Other species not predictable
Similar threshold pyrogenic response to
humans
Rabbit chosen for economic purposes
6. Rabbit Pyrogen Test
Rabbits must be healthy and mature
New Zealand or Belgian Whites used mostly
Either sex may be used
Must be individually housed between 20 and
23°C
Not varies more than ± 3º c.
Free from disturbances likely to excite them.
7. equipment and material used in test (glassware,
syringes, needles etc)
Must be free from Pyrogens by heating at 250º c
for not less then 30 minutes or any other method
retaining boxes (comfortable for rabbits as
possible)
Thermometers or thermistor probe (standardized
position in rectum, precision of ± 0.1°C)
8.
9. Rabbit pyrogen test
Preliminary test (Sham Test)
intravenous injection of sterile pyrogen-free
saline solution
Warm the pyrogen free solution up to 38.5ºc
to exclude any animal showing an unusual
response to the trauma (shock) of injection
any animal showing a temperature variation
greater than 0.6°C is not used in the main test
10. Rabbit pyrogen test -
main test:
group of 3 rabbits
preparation and injection of the product:
warming the product
dissolving or dilution
duration of injection: not more than 4 min
the injected volume: not less than 0.5 ml per 1 kg and not
more than 10 ml per kg of body mass
determination of the initial and maximum temperature
all rabbits should have initial Temperature: from 38.0 to
39.8°C
the differences in initial Temperature should not differ from
one another by more than 1°C
11. Interpretation of the results:
the test is carried out on the first group of 3 rabbits; if
necessary on further groups of 3 rabbits to a total of 4
groups, depending on the results obtained
intervals of passing or failing of products are on the
basis of summed temperature response
12. The result of pyrogen test:
No.of Rabbits Individual
Tempt. rise
(°c)
Tempt.
Rise in
group (°c)
Test
3 rabbits 0.6 1.4 Passes
If above not passes
3+5 = 8 rabbits
0.6 3.7 Passes
If above test not passes the sample is said to pyrogenic.
13. LAL Test
Limulus amebocyte lysate test.
to measure the concentration of endotoxins of
gram-negative bacterial origin
reagent: amoebocyte lysate from horseshoe
crab, Limulus polyphemus
15. Principle
The addition of solution containing endotoxin
to a solution of lysate produce turbidity.
The rate of reaction depends upon
concentration of endotoxin , the pH and the
temperature.
The endotoxin reference standard is the freeze
dried.
The test is based on the primitive blood-
clotting mechanism of the horseshoe crab
16. Commercially derived LAL reagents
bleeding adult crabs into an anticlotting solution
washing and centrifuging to collect the amebocyte
lysing in 3% NaCl
lysate is washed and lyophilized for storage
activity varies on a seasonal basis and
standardization is necessary.
17. Test performance (short)
avoid endotoxin contamination
Before the test:
interfering factors should not be present
equipment should be depyrogenated
the sensitivity of the lysate should be known
Test:
equal volume of LAL reagent and test solution (usually 0.1
ml of each) are mixed in a depyrogenated test-tube
incubation at 37°C, 1 hour
remove the tube - invert in one smooth motion (180°) - read
(observe) the result
18. Endotoxin concentration monitoring
Following method are used to monitor the endotoxin
concentration
Method A: El-clot method: limit test
Method B: semi-quantitative gel-clot method
Method C: kinetic turbidimetric method
Method D: kinetic chromogenic method
Method E: end-point chromomeric method
19. different techniques:
the gel-clot technique - gel formation
the turbidimetric technique - the development
of turbidity after cleavage of an endogenous
substrate
the chromogenic technique - the development
of color after cleavage of a synthetic peptide-
chromogen complex
Monkeys, horses, dogs, cats, and rabbit have reproducible responses
Rats, guinea pigs, mice, hamsters, chicks, etc, are irregular and unpredictable
Rabbit and dog chosen chosen for economic purposes, BUT…
Rabbit has labile thermoregulatory process, and is susceptible to false positives. A negative test is more significant than a positive one. The dog has a more stable thermoregulatory system, and is less sensitive to pyrogen. Therefore a positive is more significant than a negative one.
Similar threshold pyrogenic response to humans. HOWEVER, as dose is increased, humans respond more vigorously.
USP XX had only two requirements for the animals, that they be healthy and mature.
NZ or Belgian Whites are mostly used
Either sex may be used, but kept to a single sex to avoid outside stimuli