Endotoxin is heat resistant and stable in most environmental conditions. For example, it can only be destroyed at high temperatures (185°C for 3 hours or 250°C for 30 min.); by aggressive chemicals such as NaOH or H₂O_2; orremovedbyexpensive chromatographic processes. Consequently, it can be a major problem source in contamination control.

Endotoxin control is a critical quality requirement in the manufacture of parenterals, biologicals, medical devices, and in the operation of dialysis centers, blood donor centers, and in other areas specifically regulated for endotoxin content and the prevention of endotoxin contamination.

For several decades the determination of endotoxin levels were regulated by national & international agencies. In 2001, the International Conference on Harmonization (ICH) issued a guideline for endotoxin testing which has been adopted by the major pharmacopoeias, such as the USP (United States Pharmacopoeia), EP (European Pharmacopoeia) BP (British Pharmacopoeia), and JP (Japanese Pharmacopoeia). The techniques that were accepted include:

Gel-clot limit test
Gel-clot assay
Endpoint Turbidimetric
Kinetic Turbidimetric
Endpoint Chromogenic
Kinetic Chromogenic

From a legal view-point, Gel-clot tests are defined as the standard reference method, except in cases of approved applications such as NDAs, ANDAs, etc., where other techniques can be specified as being legally valid.

Endotoxin standards set the foundation for any of the testing methodologies. The current reference standard (RSE) has been developed through international collaboration, and may be purchased from the USP as EC-6.

Secondary standards such as Control Standard Endotoxin (CSE) may be established and used in determinations provided that the standardization procedure relative to RSE is in compliance with the regulation.

Endotoxin results are expressed as EU/unit or IU/unit. These will be; EU/ml, or EU/ng. IU/ml, or IU/ng; 1 EU = 1 IU.

Special Definitions, Meanings, and Utilities

Endotograph^TM – Definition: Endotographs are transformed Gaussian functions of LAL-endotoxin kinetic reaction data,

where,

C is the concentration,
m is the x-coordinate of the inflection points (averaged over the original replicate curves),
s represents the standard deviation of the x-coordinates from the original replicate curves,
K =1 - INT(log(C_MIN)-1), if INT(log(C_MIN)) = log(C_MIN) and C_MIN < 1, or
K =1 - INT(log(C_MIN)), where C_MIN is the least concentration value during the test.
The relationship between the maximum of the transformed Gaussian-function (y_MAX) and the concentration (C) will be:

Endotograph^TM – Meaning: Endotograph means acquiring valuable new information on the specificity of the LAL-endotoxin kinetic reaction curves. For example, it can differentiate between signal drift and the reaction signals of LAL-endotoxin. This is of particular importance for the lower concentrations of endotoxin standards and samples where the on-set reaction times due to signal drift will give false results.
Endotograph^TM – Utility: Endotograph can be generated from LAL-endotoxin kinetic reaction data by the PyroSolution program. If OOS (out-of-specification) or OOT (out-of-trend) occurs during measurement, Endotographs can then be utilized to review specificity characteristics of the run, and will assist in determination if specificity requirements have been met.
Endotograph^TM – Competition: Can you generate Endotographs from competitor products? NO! Should I invest in purchasing PyroSolution? YES! Consider the following benefits: evaluate & improve specificity, reduce OOS & OOT. An equally important feature of PyroSolution is its full compliance with 21 CFR Part 11; while remaining less expensive than other products offered for LAL-endotoxin measurements.
Endotogram^TM – Definition: Endotograms are transformed derivatives of the LAL-endotoxin kinetic reaction data, such that the relationship between the area under the curve and endotoxin concentration is defined by: C = 10^A-K, where,

C is the endotoxin concentration,
A is the area under the curve,
C_MIN is the least Concentration, and
K = 1 – INT(log(C_MIN) – 1), if INT(log(C_MIN) = log (C_MIN) and C_MIN < 1, or
K = 1 – INT(log(C_MIN)

Endotogram^TM – Meaning: Endotogram may be considered as the "raw" form of the Endotograph. It complements Endotograph information, and assists in evaluating the "quality" of the LAL-endotoxin reaction measurement. For example, noisy peaks can indicate an optical density gradient across the wells that may be due to temperature turbulence, or poor homogeneity of the solution.
Endotogram^TM – Utility : Endotograms can be generated from LAL-endotoxin kinetic reaction data by the PyroSolution program. They can provide interpretive support for method development during OOS & OOT investigations.
Endotogram^TM – Competition: Can you generate Endotograms from competitive products? NO! Should I invest in purchasing PyroSolution? YES! Consider the following benefits: evaluate & improve reaction quality, reduce OOS & OOT. An equally important feature of PyroSolution is its full compliance with 21 CFR Part 11; while remaining less expensive than other products offered for LAL-endotoxin measurements.
Robust Kinetics – Definition: Robust kinetic is a kinetic methodology, that includes multiple point – up to 35 – optical density data for a specific LAL-endotoxin kinetic reaction time in each microplate well, which is then coupled with an averaging algorithm to reduce the optical density gradient within each well.
Robust Kinetics – Meaning: Robust kinetics means a far greater reliability and improved quality for kinetic turbidimetric measurements.
Robust Kinetics – Utility: Robust kinetics can be defined in the PyroSolution program, and is recommended during kinetic turbidimetric measurements. You can extend the capability of turbidimetric kinetic tests by improving reproducibility, and by reducing OOS & OOT.
Robust Kinetics – Competition: Can you define Robust Kinetics in competitor products? NO! Should I invest in purchasing PyroSolution? YES! Consider the following benefits: improved reliability of kinetic turbidimetric tests, reduce OOS & OOT. An equally important feature of PyroSolution is its full compliance with 21 CFR Part 11; while remaining less expensive than other products offered for LAL-endotoxin measurements.
Endotoxin Measurement Uncertainty (EMU) – Definition: EMU is defined as the data associated with an endotoxin result, that is a contributor to the dispersion of the measurement that characterizes the determination process.
Endotoxin Measurement Uncertainty (EMU) – Meaning: EMU provides meaningful information on the inherent error load of the analytical process. For example, it helps to evaluate the quality constraint of a measurement and highlight, which parameter(s) must be improved to significantly reduce uncertainty while improving accuracy.
Endotoxin Measurement Uncertainty (EMU) – Utility: EMU can be generated from PyroSolution. In addition to evaluating existing methods & results, it can also greatly improve the method development process, and investigations relative to OOS & OOT.
Endotoxin Measurement Uncertainty (EMU) – Competition: Can you generate Endotoxin Measurement Uncertainty from competitive products? NO!
Pyroburden – Definition: Pyroburden is the endotoxin load for a process Pi or Processes Pi relative to time and space. Pyroburden exists if and only if quantitative endotoxin data exists. Pyroburden is expressed by its characteristic parameters jointly referred to as the Pyroburden Characteristic Set (PCS). PCS includes: Endotoxin Area Under the Curve (EAUC), Maximum Endotoxin Concentration (MEC), Number of Endotoxin Peaks (NEP), Minimum Endotoxin Concentration (NEC), Time-to-Maximum (TOM), Time-to-Minimum (TON).
Pyroburden – Internal – Definition: Pyroburden-Internal is the Pyroburden of materials and equipment used.
Pyroburden – External – Definition: Pyroburden-External is the Pyroburden of the environment and people.
Pyroburden – Meaning: Pyroburden may be considered as the endotoxin contamination risk factor associated with a process. Endotoxin load patterns – based on the Pyroburden characteristics parameters - can be established for each process, and during pyroburden monitoring, statistically significant deviations can be used for predictions.
Pyroburden – Utility: Pyroburden can be generated from the PyroSolution program. Two modules – Sample Tracking & EQU – of PyroSolution provide the basis for pyroburden capability. In addition to evaluating the endotoxin risk that may be inherent during processing, cleaning validation can also be supported by PCS. This technology may also be applied for bioburden analysis.
Pyroburden – Competition: Can you obtain pyroburden in competitor products? NO! Should I invest in purchasing PyroSolution? YES! Consider the following benefits: improved understanding of endotoxin contamination, control, management, and intervention using knowledge in hand. Reduce OOS & OOT, and improve compliance. An equally important feature of PyroSolution is its full compliance with 21 CFR Part 11; while remaining less expensive than other products offered for LAL-endotoxin measurements.
PyroSolution – is a 21 CFR Part 11 compliant software for the Endotoxin Information Management System. The program includes a control & measurement module for various microplate readers, such as BioTek’s EL808, BioTek’s PowerWave, BioTek’s CERES900, BioTek’s EL340, BioWhittaker’s KQCL, Anthos’ HTIII, Labsystems’ Twinreader, AsysHitech’s DigiScan. PyroSolution is able to control readers in a parallel mode; for example, 2 x BioTek EL808 & 2 x KQCL readers – 4 at the same time – can be used in parallel operation with a single SQL database. Integrated Sample Tracking and Endotoxin Decision Management Support extends the capability of the programs into a fully-fledged Endotoxin Information Management System. PyroSolution utilizes a sophisticated signal processing technology to improve accuracy, reproducibility, sensitivity, specificity, and robustness of raw data. Should I consider buying PyroSolution? YES! In addition to the benefits that you have already read; there are many, many more! But don’t just take our word, order a demo version for trial and see for yourself.
PyroGel – is a 21 CFR Part 11 compliant software for gel-clot determinations of Endotoxin. The program utilizes ICH protocols for the measurements. A sophisticated graphical wizard is used for set up, load, & calculations, while advanced reporting & modeling capabilities extend its utility from routine endotoxin QC to endotoxin QC/QA management. The software controls and monitors the set temperature at 8 different points of the PyroTherm temperature block thereby improving reliability, while providing prompt and full documentation. Included also is single process temperature programming for depyrogenation, cooling, and gel-clot measurements. Data can also be integrated with the quantitative results obtained in PyroSolution. Should I consider buying PyroGel? YES! In addition to the benefits that you have already read; there are many, many more! But don’t just take our word, order a demo version for trial and see for yourself.
PyroDoQ – is a Design Qualification (DQ) & method development software solution product for LAL-endotoxin measurement. DQ may be the most critical, but least understood component of the DQ-IQ-OQ-PQ process. DQ is tightly integrated to PQ! You cannot get more performance specification, or performance quality, or performance stability out of a system, than has been built into the system! What specification & qualities do I need for my measurements? PyroDoQ provides accurate information of the specification & quality characteristics required for a particular application. Should I consider buying PyroDoQ? YES! In addition to the benefits that you have already read; there are many, many more! PyroDoQ reduces the risk of getting the wrong system. You can also synchronize DQ with PQ! Order a demo version for trial and see for yourself.
PyroSuite – is a collection of software solution products developed to cover all of the requirements for endotoxin testing, endotoxin control, endotoxin information management & endotoxin executive management. It includes the PyroSolution, PyroGel, and PyroDoQ software packages with appropriate database & EQU integration. Is PyroSuite more than PyroSolution, PyroGel, and PyroDoQ in a box? YES! PyroSuite offers added database integration and integrated EQU models that will optimize method development, and endotoxin control, to develop an endotoxin knowledge base; and for the execution of effective and efficient prevention of endotoxin contamination while achieving maximum economic benefits.
EQU* – is an SQL query & modeling Application Integration System. It can connect to multiple databases and extract information for an integrated model. It can also be considered as a knowledge engineering tool; for example, to create a new database for extracted information for use in a recursive mode. EQU* also uses a natural language interface with embedded parameters and dynamic sentence creation for SQL-queries. EQUANT and EQUAL are derivatives of EQU*; EQUANT is a real-time extension of EQU, whereas, EQUAL is a library of EQU applications.

Endotoxin Safety & Hazard Measures

It may be fatal if absorbed or introduced through skin,

In the case of contact, immediately remove contaminated clothing and flush the skin area with plenty of water for at least 15 minutes.
Cover open cuts before use.