The Limulus amebocyte lysate (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 Limulus Amebocyte Lysate (LAL)
Additional dilutions required to overcome interference or quantify bacterial endotoxin in a sample for the Limulus Amebocyte Lysate (LAL) test.
Additional dilutions required to quantify bacterial endotoxin in a sample for the Limulus Amebocyte Lysate (LAL) test by the gel-clot method
Inhibition/Enhancement testing on one lot of product to demonstrate validity of the Limulus Amebocyte Lysate (LAL) test by the gel-clot test.
Inhibition/Enhancement testing on three lots of product to demonstrate validity of the Limulus Amebocyte Lysate (LAL) test by the gel-clot test, and perform a complete validation.
Assay to detect bacterial endotoxin in a water sample by the Limulus Amebocyte Lysate (LAL) test utilizing the gel-clot technique.
Assay to detect bacterial endotoxin in a liquid or powder sample by the Limulus Amebocyte Lysate (LAL) test utilizing the gel-clot technique.
Assay to detect bacterial endotoxin on the extract of the fluid pathway of a device by the Limulus Amebocyte Lysate (LAL) test utilizing the gel-clot technique.
Assay to detect bacterial endotoxin on an entire device that is extracted by immersion by the Limulus Amebocyte Lysate (LAL) test utilizing the gel-clot technique.
LAL Water, kinetic method
USP 85, USP 161, USP 797, AAMI ST72:2002, EP 2.6.14
Assay to detect bacterial endotoxin in a water sample by the Limulus Amebocyte Lysate (LAL) test utilizing the kinetic technique.
Assay to detect bacterial endotoxin in a liquid or powder sample by the Limulus Amebocyte Lysate (LAL) test utilizing the kinetic technique.
Assay to detect bacterial endotoxin on the extract of the fluid pathway of a device by the Limulus Amebocyte Lysate (LAL) test utilizing the gel-clot technique.
Assay to detect bacterial endotoxin on an entire device that is extracted by immersion by the Limulus Amebocyte Lysate (LAL) test utilizing the kinetic technique.
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 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 LAL 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
- Mark Christensen 801-290-7888, 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.
Limulus Amebocyte Lysate (LAL) Tests
| test code |
pricing options |
price |
primary uom |
| LAL110 |
Device immersion, kinetic method |
| |
Per test |
$110.00 |
each |
| LAL120 |
Device flush, kinetic method |
| |
Per test |
$110.00 |
each |
| LAL130 |
Liquid/powder, kinetic method |
| |
Per test |
$110.00 |
each |
| LAL140 |
LAL Water, kinetic method |
| |
Per test |
$85.00 |
each |
| LAL210 |
Device immersion, gel clot method |
| |
Per test |
$140.00 |
each |
| LAL220 |
Device flush, gel clot method |
| |
Per test |
$140.00 |
each |
| LAL230 |
Liquid/powder, gel clot method |
| |
Per test |
$140.00 |
each |
| LAL240 |
LAL water, gel clot method |
| |
Per test |
$95.00 |
each |
| LAL310 |
Gel clot validation, inhibition and enhancement, 3 lots |
| |
Validation testing 3 lots (recommended) |
$450.00 |
each |
| LAL320 |
Gel clot validation, inhibition and enhancement, 1 lot |
| |
Validation testing 1 lot |
$250.00 |
each |
| LAL330 |
Endotoxin quantitation, gel clot method |
| |
Gel clot quantitation, each dilution |
$25.00 |
each |
| LAL710 |
LAL Test: Additional dilutions, kinetic method |
| |
Kinetic method, each dilution |
$25.00 |
each |
| LAL730 |
LAL Test: Large device/kit or manipulation |
| |
Per sample |
$40.00 |
per sample |
