Bacterial Endotoxin Test (LAL)

The Bacterial Endotoxin Test (LAL) test is in vitro assay for detection and quantitation of bacterial endotoxin. Validated test methods performed at Nelson Laboratories includes the gel clot technique and the kinetic turbidimetric and chromogenic (colorimetric) assays.

Optional Tests for Bacterial Endotoxin Test (LAL)

Study Outline

SOLID SAMPLE EXTRACTIONS: Extraction of the devices are performed using non-pyrogenic water.  The extraction process involves flushing and/or washing the devices with an appropriate amount of extract liquid that completely immerses or flushes the device.  USP <161> Extraction Parameters are followed for fluid pathway extractions.  Immersion extractions are performed by washing the devices at 37-40°C for 40-60 minutes with intermitten swirling on an orbital shaker.

TEST PROCEDURE: A BET Test (LAL) test involves testing the liquid sample or the sample extract with Limulus Amebocyte Lysate (LAL).  LAL is an aqueous extract of the blood cells of horseshoe crabs which forms a clot or change in color, depending on the technique, in the presence of bacterial endotoxin.  The test sample is compared to a standard series of Control Standard Endotoxin (CSE) dilutions.  The endpoint or reaction times of these dilutions are used to calculate the amount of endotoxin present in the sample. All tests are performed in at least duplicate.  A positive control of the sample and negative control using non-pyrogenic water are also performed.  An initial validation on 3 lots must be performed to validate this method of testing for a given product.

SCORE: The endotoxin limit for devices used intrathecally (cerebrospinal fluid contact) is 2.15 EU/device (Endotoxin Units/device).  The limit for other medical devices is 20.0 EU/device.  For drugs and liquids, refer to the BET Guideline published by US FDA (December 1987).

REFERENCES:

United States Pharmacopeia 28 & National Formulary 23, 2005.  Bacterial endotoxins Test <85>, and Transfusion and Infusion Assemblies and Simliar Medical Devices <161>.  United States Pharmacopeial Convention, Inc. Rockville, MD.

U.S. Food & Drug Administration, U.S. Department of Health & Human Services, U.S. Public Health Service, December, 1987.  Guideline on Validation of the Limulus Amebocyte Lysate Test as an End Product Endotoxin Test for Human and Animal Parenteral Drugs, Biological Products, and Medical Devices.

Interim Guidance for Human and Veterinary Drug Products and Biologicals, U.S. Dept of Health & Human Service, FDA, July, 1991.

European Pharmacopeia, 4th edition.  2002.  2.6.14 Bacterial Endotoxins.  Council of Europe, 67075 Strasbourg.

ANSI/AAMI ST72:2002, Bacterial endotoxins- Test methodologies, routine monitioring, and alternatives to batch testing. Association for the Advancement of Medical Instrumentation, Arlington, VA.

Sample Requirements

USP bacterial endotoxins testing requires 3% of the production lot with a minimum of 3 and maximum of 10 devices to be pooled and tested.  The FDA guideline allows the testing of 2 devices for lot sizes under 3.

SOLID DEVICES: 3% of the production lot (Minimum of 3, Maximum of 10 Devices)

LIQUID SAMPLES: 1 ml

POWDER SAMPLES: Enough to reconstitute to a minimum of 1 ml

Devices may be sterile or non-sterile, depending on the validated test method. Sterile products are recommended for lot release testing.

Liquid or powder materials should be shipped in sterile containers that are free of detectable endotoxin.  Containers for liquids should be leak-proof and sealed with Parafilm®.  To minimize endotoxin adsorption, avoid using polypropylene plastics.  Polystyrene plastics are recommended.  If samples are non-sterile, consider shipping the sample on ice with overnight delivery.

Nelson Experts:

• Amy Jo Karren 801-290-7834, 800-826-2088
• Quinton Inglet 801-290-7890, 800-826-2088
• Zachary Anderson 801-290-7888, 800-826-2088

Sample Requirements

USP bacterial endotoxins testing requires 3% of the production lot with a minimum of 3 and maximum of 10 devices to be pooled and tested.  The FDA guideline allows the testing of 2 devices for lot sizes under 3.

SOLID DEVICES: 3% of the production lot (Minimum of 3, Maximum of 10 Devices)

LIQUID SAMPLES: 1 ml

POWDER SAMPLES: Enough to reconstitute to a minimum of 1 ml

Devices may be sterile or non-sterile, depending on the validated test method. Sterile products are recommended for lot release testing.

Liquid or powder materials should be shipped in sterile containers that are free of detectable endotoxin.  Containers for liquids should be leak-proof and sealed with Parafilm®.  To minimize endotoxin adsorption, avoid using polypropylene plastics.  Polystyrene plastics are recommended.  If samples are non-sterile, consider shipping the sample on ice with overnight delivery.

Bacterial Endotoxin Test (LAL) Tests

test code	pricing options	price	primary uom
LAL110	Bacterial Endotoxin Test (LAL): Kinetic Turbidimetric Method
	Per test (device immersion, device flush, or liquid/powder)	$120.00	each
LAL140	Bacterial Endotoxin Test (LAL) Water, kinetic turbidimetric method
	Per test (water)	$90.00	each
LAL150	Bacterial Endotoxin Test (LAL): Kinetic Chromogenic Method
	Per test	$120.00	each
LAL210	Bacterial Endotoxin Test (LAL): Gel Clot Method
	Per test	$145.00	each
LAL310	Gel clot validation, inhibition and enhancement, 3 lots
	Validation testing 3 lots (recommended)	$475.00	each
LAL320	Gel clot validation, inhibition and enhancement, 1 lot
	Validation testing 1 lot	$265.00	each
LAL330	Endotoxin quantitation, gel clot method
	Gel clot quantitation, each dilution	$30.00	each
LAL710	Bacterial Endotoxin Test (LAL): Additional dilutions, kinetic method
	Kinetic method, each dilution	$30.00	each
LAL730	Bacterial Endotoxin Test (LAL): Large device/kit or manipulation
	Per sample	$40.00	per sample