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Study Outline

The Bacterial Endotoxins test can be performed by the kinetic

turbidimetric, kinetic chromogenic, or gel-clot test methods. However, the kinetic test

methods have significant advantages over the gel-clot test.

Sample extractions: Device

extractions are performed using water free of detectable endotoxins. The extraction

process involves flushing and/or washing the devices with the minimum amount of liquid

to properly extract the device. Devices with a non-pyrogenic fluid pathway are

extracted by flushing the relevant fluid pathway with extraction liquid that has been

heated to 37 ± 1°C and maintaining contact for not less than one hour. Device

immersions are performed by washing the devices in extraction liquid at 37 – 40°C for

>60 minutes with intermittent swirling on an orbital shaker.

Test procedure: A BET

involves analyzing the liquid sample or sample extract using Limulus Amebocyte Lysate

(LAL). LAL is a reagent made from the blood of the horseshoe crab. In the presence of

bacterial endotoxins, the lysate reacts to form a clot or cause a color change

depending on the technique. The test sample is compared to a standard curve made from

known endotoxin concentrations. All tests are performed in at least duplicate. A

positive product control and negative control are included as part of each

assay.

Validation: It is required to demonstrate that the test sample does not

interfere with the ability to detect endotoxins. This is accomplished with the positive

product control (also called the spike recovery) for the kinetic test methods, and with

a separate inhibition and enhancement assay for the gel-clot method.

Samples that

require a treatment such as reconstitution in a solvent other than water, heat

denaturing, centrifugation, or filtration should be validated to demonstrate that the

treatment does not result in the loss of endotoxins. This is accomplished by

inoculating the sample with endotoxins and subjecting the inoculated sample to the

chosen treatment.

The endotoxin limit for medical devices is not more than 20.0

EU/device unless the device will have contact with cerebrospinal fluid, in which case

the limit is 2.15 EU/device. Ophthalmic implants and OVDs may be required to meet a

lower limit of 0.2 EU/device or 0.2 EU/mL. Endotoxin limits for pharmaceutical products

are based on the maximum dose that can be administered in a 1-hour time period.

Testing

Locations

Salt Lake City, UT, USA

Itasca, IL, USA

Ontario, CA, USA

Mexico City,

Mexico

Petit-Rechain, Belgium

Leuven, Belgium

Somercotes, England

Rantigny,

France

Wiesbaden, Germany

Shanghai, China

Learn more about our locations and their

certifications.

Contact Us

If you have additional questions about Bacterial Endotoxins

Test (BET) services, or would like to consult with the experts at Nelson Labs, just

send us a request or call us at +1 (801) 290-7500.

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