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US3986930A
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Lipase activity determining method and reagent
Assignee: Sumitomo Pharma Co Ltd
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Filing: 1975-05-22
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Granted: 1976-10-19
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Expires: 1995-05-22
Abstract
Abstract A reagent for determining lipase activity, which comprises as the active ingredient one of the S-acyl compounds of the formula: ##EQU1## WHEREIN N IS AN INTEGER OF 2 TO 4, INCLUSIVE; N' IS AN INTEGER OF 2 TO 4, INCLUSIVE OR OF 9 TO 13, INCLUSIVE; M IS AN INTEGER OF 3, 7 OR 11; M' IS AN INTEGER OF 4 OR 12; Y IS AN INTEGER OF 1 OR 2; X is sulfur atom or oxygen atom; Y is sulfur atom, oxygen atom or ethylene group; provided that when y is 1, m' is 4 and when y is 2, m' is 12, and a method for determining lipase activity by reacting the S-acyl compound with lipase in a buffer solution containing a chromogenic sulfhydryl reagent and then subjecting the resultant to a colorimetry.
Inventors
- 👤 Shigeru Kurooka
- 👤 Masahisa Hashimoto
- 👤 Masatsugu Tomita
- 👤 Akio Maki
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 10:40
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/34; C12Q1/44; C07; C07C; C07C327/00
Claims Analysis
Total Claims: 44
Claim 1
Independent Claim
120 words
A reagent for determining lipase activity comprising a chromogenic sulfhyldryl reagent and, as an active ingredient one of the S-acyl compounds of the formula: ##EQU2## wherein n is an integer of 2 to 4, inclusive; n' is an integer of 2 to 4, inclusive or of 9 to 13, inclusive; m is an integer of 3, 7 or 11; m' is an integer of 4 or 12, y is an integer of 1 or 2; X is sulfur atom or oxygen atom; Y is sulfur atom, oxygen atom or ethylene group; provided that when y is 1, m' oxygen atom or ethylene group; provided that when y is 1, m' is 4 and when y is 2, m' is 12.
Claim 2
Dependent Claim
120 words
A reagent for determining lipase activity comprising a chromogenic sulfhyldryl reagent and, as an active ingredient one of the S-acyl compounds of the formula: ##EQU2## wherein n is an integer of 2 to 4, inclusive; n' is an integer of 2 to 4, inclusive or of 9 to 13, inclusive; m is an integer of 3, 7 or 11; m' is an integer of 4 or 12, y is an integer of 1 or 2; X is sulfur atom or oxygen atom; Y is sulfur atom, oxygen atom or ethylene group; provided that when y is 1, m' oxygen atom or ethylene group; provided that when y is 1, m' is 4 and when y is 2, m' is 12.
Claim 3
Dependent Claim
52 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR42## wherein n' is an integer of 3 or 11; m is an integer of 3, 7 or 11; X is sulfur atom or oxygen atom; and Y is sulfur atom, oxygen atom or ethylene group.
Claim 4
Dependent Claim
50 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR43## wherein n is an integer of 2 to 4, inclusive; n' is an integer of 2 to 4, inclusive or of 9 to 13, inclusive; and X is sulfur atom or oxygen atom.
Claim 5
Dependent Claim
41 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR44## wherein n' is an integer of 2 to 4, inclusive or of 9 to 13, inclusive; and X is sulfur atom or oxygen atom.
Claim 6
Dependent Claim
25 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR45## wherein X is sulfur atom or oxygen atom.
Claim 7
Dependent Claim
17 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR46##
Claim 8
Dependent Claim
17 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR47##
Claim 9
Dependent Claim
17 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR48##
Claim 10
Dependent Claim
37 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR49## wherein m is an integer of 3, 7 or 11; and Y is sulfur atom, oxygen atom or ethylene group.
Claim 11
Dependent Claim
27 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR50## wherein Y is sulfur atom, oxygen atom or ethylene group.
Claim 12
Dependent Claim
61 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR51## wherein m is an integer of 3, 7 or 11; m' is an integer of 4 or 12; and y is an integer of 1 or 2; provided that when y is 1, m' is 4 and when y is 2, m' is 12.
Claim 13
Dependent Claim
27 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR52## wherein m is an integer of 3, 7 or 11.
Claim 14
Dependent Claim
43 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR53## wherein n is an integer of 2 to 4, inclusive; and n' is an integer of 2 to 4, inclusive or of 9 to 13, inclusive.
Claim 15
Dependent Claim
33 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR54## wherein n' is an integer of 2 to 4, inclusive or of 9 to 13, inclusive.
Claim 16
Dependent Claim
17 words
The reagent according to claim 1, wherein the S-acyl compound is the one of the formula: ##STR55##
Claim 17
Independent Claim
139 words
A method for determining lipase activity, which comprises subjecting one of the S-acyl compounds of the formula: ##STR56## wherein n is an integer of 2 to 2, inclusive; n' is an integer of 2 to 4, inclusive or of 9 to 13, inclusive; m is an integer of 3, 7 or 11; m' is an integer of 4 or 12; y is an integer of 1 or 2; X is sulfur atom or oxygen atom; Y is sulfur atom, oxygen atom or ethylene group; provided that when y is 1, m' is 4 and when y is 2, m' is 12, to the lipolytic action of lipase in a buffer solution containing a chromogenic sulfhydryl reagent and then subjecting the resulting SH-compound obtained by the hydrolysis of the above S-acyl compound to a colorimetry with the chromogenic sulfhydryl reagent.
Claim 18
Independent Claim
139 words
A method for determining lipase activity, which comprises subjecting one of the S-acyl compounds of the formula: ##STR56## wherein n is an integer of 2 to 2, inclusive; n' is an integer of 2 to 4, inclusive or of 9 to 13, inclusive; m is an integer of 3, 7 or 11; m' is an integer of 4 or 12; y is an integer of 1 or 2; X is sulfur atom or oxygen atom; Y is sulfur atom, oxygen atom or ethylene group; provided that when y is 1, m' is 4 and when y is 2, m' is 12, to the lipolytic action of lipase in a buffer solution containing a chromogenic sulfhydryl reagent and then subjecting the resulting SH-compound obtained by the hydrolysis of the above S-acyl compound to a colorimetry with the chromogenic sulfhydryl reagent.
Claim 19
Dependent Claim
12 words
The method according to claim 16, wherein the buffer is Tris-HCl buffer.
Claim 20
Dependent Claim
27 words
The method according to claim 16, wherein a carboxylesterase and/or arylesterase inhibitor is added to the reaction system prior to the reaction of S-acyl compound and lipase.
Claim 21
Dependent Claim
23 words
The method according to claim 18, wherein the enzyme inhibitor is a member selected from the group consisting of phenylmethylsulfonyl fluoride and diisopropylfluorophosphate.
Claim 22
Dependent Claim
13 words
The method according to claim 18, wherein the enzyme inhibitor is phenylmethylsulfonyl fluoride.
Claim 23
Dependent Claim
20 words
The method according to claim 20, wherein the phenylmethylsulfonyl fluoride is used in a concentration of 0.03 to 3 mM.
Claim 24
Dependent Claim
20 words
The method according to claim 20, wherein the phenylmethylsulfonyl fluoride is used in a concentration of 0.1 to 1.5 mM.
Claim 25
Dependent Claim
20 words
The method according to claim 20, wherein the phenylmethylsulfonyl fluoride is used in a concentration of 0.2 to 0.8 mM.
Claim 26
Dependent Claim
20 words
The method according to claim 20, wherein the phenylmethylsulfonyl fluoride is used in a concentration of 0.3 to 0.5 mM.
Claim 27
Dependent Claim
24 words
The method according to claim 16, wherein acetone is added to the reaction mixture of the S-acyl compound and lipase to stop the reaction.
Claim 28
Dependent Claim
14 words
The method according to claim 16, wherein the chromogenic sulfhydryl reagent is 5,5'-dithiobis(2-nitrobenzoic acid).
Claim 29
Dependent Claim
34 words
The method according to claim 18, wherein the buffer is Tris-HCl buffer, the enzyme inhibitor is phenylmethylsulfonyl fluoride in the range of 0.3 to 0.5 mM and the chromogenic sulfhydryl reagent is 5,5'-dithiobis(2-nitrobenzoic acid).
Claim 30
Dependent Claim
18 words
The method according to claim 18, which is applied to the determination of lipase activity in human serum.
Claim 31
Dependent Claim
17 words
The method according to claim 16, wherein the S-acyl compound is the one of the formula: ##STR57##
Claim 32
Dependent Claim
17 words
The method according to claim 16, wherein the S-acyl compound is the one of the formula: ##STR58##
Claim 33
Dependent Claim
117 words
A kit for the determination of lipase activity comprising a. one of the S-acyl compounds of the formula: ##STR59## wherein n is an integer of 2 to 4, inclusive; n' is an integer of 2 to 4, inclusive or of 9 to 13, inclusive; m is an integer of 3, 7 or 11; m' is an integer of 4 or 12; y is an integer of 1 or 2; X is sulfur atom or oxygen atom; Y is sulfur atom, oxygen atom or ethylene group; provided that when y is 1, m' is 4 and when y is 2, m' is 12, b. a buffer, c. a chromogenic sulfhydryl reagent, and d. a carboxylesterase and/or arylesterase inhibitor
Claim 34
Dependent Claim
117 words
A kit for the determination of lipase activity comprising a. one of the S-acyl compounds of the formula: ##STR59## wherein n is an integer of 2 to 4, inclusive; n' is an integer of 2 to 4, inclusive or of 9 to 13, inclusive; m is an integer of 3, 7 or 11; m' is an integer of 4 or 12; y is an integer of 1 or 2; X is sulfur atom or oxygen atom; Y is sulfur atom, oxygen atom or ethylene group; provided that when y is 1, m' is 4 and when y is 2, m' is 12, b. a buffer, c. a chromogenic sulfhydryl reagent, and d. a carboxylesterase and/or arylesterase inhibitor
Claim 35
Dependent Claim
20 words
The kit according to claim 31, wherein the S-acyl compound is in the form of a solution in an alcohol.
Claim 36
Dependent Claim
13 words
The kit according to claim 31, wherein the buffer (b) is Tris-HCl buffer.
Claim 37
Dependent Claim
17 words
The kit according to claim 31, wherein the buffer (b) is in the form of a powder.
Claim 38
Dependent Claim
15 words
The kit according to claim 31, wherein the chromogenic sulfhydryl reagent (c) is 5,5'-dithiobis(2-nitrobenzoic acid).
Claim 39
Dependent Claim
24 words
The kit according to claim 31, wherein the chromogenic sulfhydryl reagent (c) is a mixed powder of 5,5'-dithiobis (2-nitrobenzoic acid) and an inert powder.
Claim 40
Dependent Claim
12 words
The kit according to claim 36, wherein the inert powder is mannitol.
Claim 41
Dependent Claim
14 words
The kit according to claim 31, wherein the enzyme inhibitor (d) is phenylmethylsulfonyl fluoride.
Claim 42
Dependent Claim
17 words
The kit according to claim 31, wherein the S-acyl compound is the one of the formula: ##STR60##
Claim 43
Dependent Claim
17 words
The kit according to claim 31, wherein the S-acyl compound is the one of the formula: ##STR61##
Claim 44
Dependent Claim
14 words
The reagent according to claim 1 wherein the chromogenic sulfhydryl reagent is 5,5'-dithiobis(2-nitrobenzoic acid).
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1979-10-19 | 1980-04-19 | 1980-10-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1983-10-19 | 1984-04-19 | 1984-10-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1987-10-19 | 1988-04-19 | 1988-10-19 | $7700.00 | Window Closed / Lapsed |