Correct Ways to Use LAL accessories

The Limulus Amebocyte Lysate (LAL) assay is an efficient method for the detection of bacterial endotoxin in Gram-negative bacteria. It was developed in 1960 based on the clotting reaction of haemolymph from the Atlantic horseshoe crab. There are a number of LAL accessories available for bacterial endotoxin testing, for instance, the Toxinometer® Analysis System with Toximaster® software program, BioClean® plates, lysate reagent water, reaction and dilution tubes.   All of these accessories are manufactured to bring ease and accuracy to the LAL test.

The Toxinometer® is a kinetic incubating tube reader, which can be used for endotoxin testing in a wide range of fields and sample quantities. The rate of endotoxin reaction is directly dependent on the concentration of endotoxin present in a standard or sample. For example, the smaller the amount of endotoxin present the slower the endotoxin mixture reacts and the greater the amount of endotoxin present the faster the mixture reacts.  The amount of time required for the reaction to take place is called the reaction time.

The Toxinometer® Analysis System can be used for kinetic turbidimetric, kinetic chromogenic and Gel-clot assays.  The kinetic turbidimetric and Gel-clot results can be obtained for the same sample, which allows for the direct correlation between the two methodologies. The Toxinometer® uses the measured reaction time and the reaction time of known endotoxin standards to calculate the endotoxin concentrations in a sample. It utilizes the rate of turbidity or the increase in colour intensity to calculate reaction times depending whether the assay ran is turbidimetric or chromogenic. It is designed to achieve testing with ease. The Toxinometer® Analysis System is computer operated, can be easily expanded (extension modules available to extend number of samples) and has exceptional user-friendliness.  Usage of the Toxinometer® instead of a microplate reader decreases the risk of contamination by a huge factor.  The Toxinometer® can test using both multi test and single test reagents and it also has the ability to run a new assay while one test is still running giving the user the ease to begin analysing immediately. As an additional feature, it countenances archiving of standard curves, as well as, allows variable temperature settings and measurement of up to 16 samples simultaneously.

The current Toxinometer® Analysis System, known as the ET-6000, comes with a dedicated laptop that is pre-loaded with the Toximaster® QC7 software package along with power cables, extra fuses, USB connectors, a cover for its photometric zone, reaction tube stand, validation documents and instruction manual. The QC 7 user management tool is designed on an individual login basis to control access within the Toximaster® program.

At the start of the program a window appears with four options (1. arrange sample and then start measurement, 2. start measurement now, 3. load protocol and then start measurement and 4. browse measurement data) to bring ease in setting your protocol. Edit, load and save a protocol keys can be easily seen in the toolbar. There are four test group options with multiple sample type options to choose from. The information of the sample can also be customized according to the requirements of the test. The program also gives the option to set parameters, generate plots for single or multiple samples, draw standard curves, generate group reports and allow for print outs. The print out includes all relevant information. The sample ID, activating time, elapsed time, light difference signal and light transmission rate can be seen for all the samples at any time. The samples that do not gel during the cycle show a blank led next to the sample position, while an orange light is visible next to a sample position signifying that the sample continues to gel. The samples, which have gelled during the time cycle will beep and show a red light next to the sample position signifying the assay is complete.

FUJIFILM Wako Chemicals also offers endotoxin-free pipette tips, test tubes and aluminium caps, PYROSTAR™ ES-F reagents in single and multi-test configurations, Control Standard Endotoxin, Limulus Color KY in single and multi-test configurations (for chromogenic testing), and lysate reagent water to accompany usage of the Toxinometer® Analysis System.  In addition, the Toximaster® software program is 21CFR, part 11 compliant. The Toxinometer ET-7000 Series is expected be released in 2019.

LAL REAGENT KIT:

PYROSTAR ES-F Plate with CSE	Kit of 100 PYROSTAR™ ES-F (2 ML) without CSE	Endotoxin Extracting Solution for LAL Test