Micromethod for the determination of endotoxins
First Claim
1. A micromethod for the quantitative determination of endotoxins with Limulus Amebocyte Lysate (L.A.L.) reagent, which comprises the steps of:
- (a) admixing the sample containing the endotoxins with a microamount of L.A.L. reagent;
(b) introducing the mixture into a capillary tube;
(c) incubating the capillary tube at a temperature range of 20°
to 40°
C.;
(d) measuring the hydrostatic pressure required to cause the formed viscous mass to flow from said capillary tube; and
(e) determining the quantity of endotoxin in the sample by comparing the measured hydrostatic pressure to a previously determined standard relating hydrostatic pressure to endotoxin content.
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Accused Products
Abstract
An improved micromethod for the quantitative determination of endotoxins with Limulus Amebocyte Lysate (L.A.L.) reagent is based on the discovery that if L.A.L. reagent and a sample containing microquantities of endotoxin are incubated in a capillary tube, a clot will be formed in the plug preventing any outflow of liquids when low hydrostatic pressure is applied. Increasing the pressure will eventually force out the clot plugging the capillary and the liquid will suddenly erupt giving a clear yes-no indication of the presence of endotoxin. The hydrostatic pressure indicated to cause this eruption is a relatively precise indication of the extent of clotting which, in turn, is indicative of the concentration of endotoxin in the liquid. A kit which can be used to perform this test is also disclosed.
35 Citations
8 Claims
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1. A micromethod for the quantitative determination of endotoxins with Limulus Amebocyte Lysate (L.A.L.) reagent, which comprises the steps of:
- (a) admixing the sample containing the endotoxins with a microamount of L.A.L. reagent;
(b) introducing the mixture into a capillary tube;
(c) incubating the capillary tube at a temperature range of 20°
to 40°
C.;
(d) measuring the hydrostatic pressure required to cause the formed viscous mass to flow from said capillary tube; and
(e) determining the quantity of endotoxin in the sample by comparing the measured hydrostatic pressure to a previously determined standard relating hydrostatic pressure to endotoxin content. - View Dependent Claims (2, 3, 4, 5, 6)
- (a) admixing the sample containing the endotoxins with a microamount of L.A.L. reagent;
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7. A kit for carrying out a micromethod for the quantitative determination of endotoxins with Limulus Amebocyte Lysate (L.A.L.) reagent, comprising:
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a plurality of capillary size reaction chamber means for receiving a mixture of a sample containing endotoxin and a microamount of L.A.L. reagent, said reaction chamber means comprising capillary tubes; hydrostatic pressure measuring means for determining the hydrostatic pressure required to cause the endotoxin-L.A.L. reaction product to flow from said capillary tube, said pressure measuring means comprising a watertight vessel having an outlet tube at or near the bottom thereof, said outlet tube including a length of flexible tubing connectable to one of said capillary tubes; and an empirical graph correlating the hydrostatic pressure as measured by said pressure measuring means with the quantitative content of endotoxin in the sample introduced into said reaction chamber means. - View Dependent Claims (8)
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Specification
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- Resources
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Current AssigneeTecgbuib Researcg Abd Devekionebt
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Original AssigneeTECHNION RESEARCH AND DEVELOPMENT FOUNDATION, LTD.
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InventorsNeeman, Ishak, Gaffin, Stephen L.
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Primary Examiner(s)Warden, Robert J.
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Application NumberUS06/235,153Time in Patent Office707 DaysField of Search435/7, 435/29, 435/34, 435/39, 435/287, 435/291, 23/230 B, 422/61US Class Current435/18CPC Class CodesG01N 33/579 involving limulus lysateY10S 435/807 Gas detection apparatusY10S 435/81 Packaged device or kit
