US4798804A — Serum pretreatment in digoxin immunoassay

Claim 1

Independent Claim 78 words

A method for preparing a sample suspected of containing digoxin for determination of digoxin in said sample in an assay, which method comprises: (a) contacting in a medium a sample with β-cyclodextrin in an amount and under conditions sufficient to allow a substantial portion of digoxin in said sample to bind to said β-cyclodextrin, (b) separating β-cyclodextrin with digoxin bound thereto from at least one other component of the medium, and (c) releasing said digoxin from said β-cyclodextrin.

Claim 2

Independent Claim 78 words

A method for preparing a sample suspected of containing digoxin for determination of digoxin in said sample in an assay, which method comprises: (a) contacting in a medium a sample with β-cyclodextrin in an amount and under conditions sufficient to allow a substantial portion of digoxin in said sample to bind to said β-cyclodextrin, (b) separating β-cyclodextrin with digoxin bound thereto from at least one other component of the medium, and (c) releasing said digoxin from said β-cyclodextrin.

Claim 3

Dependent Claim 11 words

The method of claim 1 wherein said sample is whole blood.

Claim 4

Dependent Claim 11 words

The method of claim 1 wherein said sample is blood serum.

Claim 5

Dependent Claim 23 words

The method of claim 1 wherein said β-cyclodextrin is in substantial excess over the amount of digoxin suspected of being in said sample.

Claim 6

Dependent Claim 30 words

The method of claim 1 wherein the amount of said β-cyclodextrin is about from 10 6 to 10 9 times the amount of digoxin suspected of being in said sample.

Claim 7

Dependent Claim 11 words

The method of claim 1 wherein the β-cyclodextrin is water soluble.

Claim 8

Dependent Claim 10 words

The method of claim 1 wherein the β-cyclodextrin is polymeric.

Claim 9

Dependent Claim 23 words

The method of claim 1 wherein said sample is contacted with said β-cyclodextrin for a period of from about 0.5 to 5 minutes.

Claim 10

Dependent Claim 18 words

The method of claim 1 wherein digoxin bound to said β-cyclodextrin is separated from the solution by filtration.

Claim 11

Dependent Claim 16 words

The method of claim 1 wherein digoxin bound to said β-cyclodextrin is separated from the solution.

Claim 12

Dependent Claim 36 words

The method of claim 1 wherein said digoxin is released from said β-cyclodextrin by contacting digoxin bound to said β-cyclodextrin with a releasing agent that is capable of displacing said digoxin by binding to said β-cyclodextrin.

Claim 13

Dependent Claim 95 words

The method of claim 11 wherein said releasing agent is a compound of the formula: Y--R--X wherein R is a saturated or unsaturated six-membered ring unsubstituted at positions 2, 3, 5 and 6 and X and Y are independently substituents at the 1 and 4 positions of R and are independently selected from the group consisting of hydrogen, hydroxy, carboxy, lower alkyl of from 1 to 5 carbon atoms, and lower alkyl of from 1 to 5 carbon atoms substituted with hydroxy, carboxy, amino, and/or sulfonate, with the proviso that said compound is water soluble.

Claim 14

Dependent Claim 23 words

The method of claim 12 wherein said releasing agent is selected from the group consisting of phenols, cyclohexanols and water soluble 1,4-substituted benzenes.

Claim 15

Dependent Claim 92 words

In an assay method for the determination of digoxin in a sample suspected of containing digoxin wherein said sample is combined with reagents for detecting said digoxin, the improvement which comprises: (a) prior to conducting said assay method contacting in a medium a sample with β-cyclodextrin in an amount and under conditions sufficient to allow a substantial portion of digoxin in said sample to bind to said β-cyclodextrin, (b) separating β-cyclodextrin with digoxin bound thereto from at least one other component of the medium, and (c) releasing said digoxin from said β-cyclodextrin.

Claim 16

Dependent Claim 92 words

In an assay method for the determination of digoxin in a sample suspected of containing digoxin wherein said sample is combined with reagents for detecting said digoxin, the improvement which comprises: (a) prior to conducting said assay method contacting in a medium a sample with β-cyclodextrin in an amount and under conditions sufficient to allow a substantial portion of digoxin in said sample to bind to said β-cyclodextrin, (b) separating β-cyclodextrin with digoxin bound thereto from at least one other component of the medium, and (c) releasing said digoxin from said β-cyclodextrin.

Claim 17

Dependent Claim 23 words

The method of claim 14 wherein said β-cyclodextrin is in substantial excess of the amount of digoxin suspected of being in the sample.

Claim 18

Dependent Claim 11 words

The method of claim 14 wherein the β-cyclodextrin is water soluble.

Claim 19

Dependent Claim 10 words

The method of claim 14 wherein the β-cyclodextrin is polymeric.

Claim 20

Dependent Claim 18 words

The method of claim 14 wherein β-cyclodextrin with digoxin bound thereto is separated from the medium by filtration.

Claim 21

Dependent Claim 35 words

The method of claim 14 wherein said digoxin is released from said β-cyclodextrin by contacting said cyclodextrin with digoxin bound thereto with a releasing agent capable of displacing said digoxin and binding to said β-cyclodextrin.

Claim 22

Independent Claim 143 words

A method for preparing a sample suspected of containing digoxin for an assay method to determine digoxin in said sample, which method comprises: (a) contacting in an aqueous medium a sample with an amount of soluble or polymeric β-cyclodextrin in substantial excess over the amount of digoxin suspected of being in said sample, (b) filtering said medium to separate therefrom β-cyclodextrin with digoxin bound thereto, and (c) treating said β-cyclodextrin with an agent selected from compounds of the formula: Y'--R--X' wherein R is a saturated or unsaturated six-membered ring unsubstituted at positions 2, 3, 5 and 6 and Y' and X' are independently substituents at the 1 and 4 positions of R and are independently selected from the group consisting of hydrogen, hydroxy, carboxy, and lower alkyl of from 1 to 5 carbon atoms, with the proviso that said compound is water soluble.

Claim 23

Independent Claim 143 words

A method for preparing a sample suspected of containing digoxin for an assay method to determine digoxin in said sample, which method comprises: (a) contacting in an aqueous medium a sample with an amount of soluble or polymeric β-cyclodextrin in substantial excess over the amount of digoxin suspected of being in said sample, (b) filtering said medium to separate therefrom β-cyclodextrin with digoxin bound thereto, and (c) treating said β-cyclodextrin with an agent selected from compounds of the formula: Y'--R--X' wherein R is a saturated or unsaturated six-membered ring unsubstituted at positions 2, 3, 5 and 6 and Y' and X' are independently substituents at the 1 and 4 positions of R and are independently selected from the group consisting of hydrogen, hydroxy, carboxy, and lower alkyl of from 1 to 5 carbon atoms, with the proviso that said compound is water soluble.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1992-01-17	1992-07-18	1993-01-17	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1996-01-17	1996-07-18	1997-01-17	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	2000-01-17	2000-07-18	2001-01-17	$7700.00	Window Closed / Lapsed

Serum pretreatment in digoxin immunoassay

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