Patent EP0456252A2 successfully refreshed from Google Patents.
EP0456252A2
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Method for determination of pyrogen content
Assignee: Daicel Corp
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Filing: 1991-05-10
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Granted: —
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Expires: 2011-05-10
Abstract
Abstract A method for determination of a pyrogen content by using a centrifugation type filtration unit for centrifugal filtration is disclosed. A sepcimen is brough into contact with an adsorbent having pyrogen absorbability in the filtration unit and any non-adsorbed material is separated and removed by centrifugation so that a pyrogen is specifically adsorbed by the adsorbent and then the pyrogen adsorbed on the adsorbent is reacted with a detection reagent to determine the pyrogen content.
Inventors
- 👤 Tetsuya Tosa
- 👤 Tadashi Sato
- 👤 Taizo Watanabe
- 👤 Satoshi Minobe
- 👤 Masahiro Nawata
- 👤 Shinji Nagamatsu
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-09 03:00
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/34; C12Q1/37; G; G01; G01N; G01N5/00; G01N5/02; G01N33/00; G01N33/48; G01N33/50; G01N33/579; C12Q2337/00; C12Q2337/10; C12Q2337/12; C12Q2337/20; C12Q2337/22
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
113 words
A method for determination of a pyrogen content which comprises using a centrifugation type filtration unit for centrifugal filtration comprising a filtrate cup and a specimen cup removably attached to the opening part of the filtrate cup, said specimen cup being communicated with the filtrate cup through a filtration membrane, bringing a specimen into contact with an adsorbent having pyrogen absorbability in the specimen cup, separating any non-adsorbed material through the filtration membrane under conditions for accelerating permeation through the membrane to remove the non-adsorbed material into the filtrate cup and then reacting a pyrogen adsorbed on the adsorbent retained in the specimen cup with a detecting reagent to determine the pyrogen content.
Claim 2
Dependent Claim
58 words
A method according to claim 1, wherein, after completion of a reaction with Limulus lysate in the presence of a synthetic substrate, the adsorbent is separated from the reaction solution through the filtration membrane by centrifugation to transfer the reaction solution transferred in the filtrate cup and the reaction solution is subjected to determination of the pyrogen content.
Claim 3
Dependent Claim
35 words
A method according to claim 1, wherein the filtration membrane is a ultrafiltration membrane having a cutoff molecular weight of 10,000 to 3,000,000, or a microfiltration membrane a pore opening of 0.01 to 10 µm.
Claim 4
Dependent Claim
185 words
A method according to claim 1, wherein the adsorbent is a heterocyclic type absorbent composed of a nitrogen containing cyclic compound of the formula: R-A-X wherein R is a nitrogen containing heterocyclic group, A is a single bond, an alkylene group having 1 to 12 carbon atoms or an alkenylene group having 2 to 12 carbon atoms, and X is hydrogen amino, hydorxy or carboxyl, when A is a single bond, or X is amino, hydorxy or carboxyl, when A is an alkylene or alkenylene group, said heterocyclic group and alkylene group being optionally sustituted with one or more groups selected from carboxyl, oxo, an alkyl group having 1 to 6 carbon atoms, hydroxy, amino and an alkoxy group having 1 to 6 carbon atoms; and a water-insoluble carrier having hydroxy, amino carboxyl or a halogen atom; said nitrogen containing cyclic compound being bound to said water-insoluble carrier directly or through a spacer which is a compound selected from the group consisting of NH₂(CH₂) n NH₂, NH₂(CH₂) n COOH, NH₂(CH₂) n OH and HOOC(CH₂) n COOH wherein n is an integer of 1 to 12.
Claim 5
Dependent Claim
47 words
A method according to claim 4, wherein the nitrogen containing heterocyclic group R is imidazole nucleus, pyrazole nucleus, pyrimidine nulceus, pyridazine nucleus, pyrazine nucleus, purine nucleus, acridine nucleus, triazole nucleus, oxadiazole nucleus, tetrazole nucleus, indazole nucleus, benzotriazole nucleus, benzopyridazine nucleus, benzopyrimidine nucleus, benzopyrazine nucleus and naphthylizine nucleus.
Claim 6
Dependent Claim
118 words
A method according to claim 1, wherein the adsorbent is an aliphatic type adsorbent composed of an aliphatic nitorgen containing compound having a functional chain containing 3 to 50 chain-constituting atoms, said aliphatic nitorgen atom being that of a primary amino, secondary amino, tertiary amino, quaternary ammonium or imino and bound to an aliphatic carbon atom; and a porous base material having pore size of 50 nm to 1 µm; said aliphatic nitrogen containing compound being bound to said base material directly or through a spacer which is a compound selected from the group consisting of NH₂(CH₂) n NH₂, NH₂(CH₂) n COOH, NH₂(CH₂) n OH and HOOC(CH₂) n COOH wherein n is an integer of 1 to 12..
Claim 7
Dependent Claim
17 words
A method according to claim 1, wherein separation of the non-adsorben material is carried out by centrifugation.
Claim 8
Dependent Claim
41 words
A method according to claim 1, wherein a solution of the specimen is brought into contact with the adsorbent at 4 to 40°C, pH 4 to 8 and an inonic strength of not higher than 0.2 for at least 15 mintues.
Claim 9
Dependent Claim
21 words
A method accroding to claim 2, wherein the reaction is carried out at 25 to 40°C for 10 to 120 minutes.
Claim 10
Dependent Claim
15 words
A method according to claim 2, wherein the synthetic substrate is Ac-Ile-Glu-Ala-Arg-pNA, Boc-Leu-Gly-Arg-pNA or Boc-Leu-Gly-Arg-MCA.
Claim 11
Independent Claim
66 words
A method for determination of a pyrogen content by using a pyrogen detecting reagent which comprises bringing a pyrogen in a solution of a specimen into contact with a solid adsorbent and, after solid-liquid separation, reacting the adsorbent with a detecting reagent to determine the pyrogen content, said solid absorbent comprising a compound containing an aliphatic nitrogen and a base material bound thereto through a spacer.
Claim 12
Dependent Claim
118 words
A method according to claim 11, wherein the adsorbent is an aliphatic type adsorbent composed of an aliphatic nitorgen containing compound having a functional chain containing 3 to 50 chain-constituting atoms, said aliphatic nitorgen atom being that of a primary amino, secondary amino, tertiary amino, quaternary ammonium or imino and bound to an aliphatic carbon atom; and a porous base material having pore size of 50 nm to 1 µm; said aliphatic nitrogen containing compound being bound to said base material directly or through a spacer which is a compound selected from the group consisting of NH₂(CH₂) n NH₂, NH₂(CH₂) n COOH, NH₂(CH₂) n OH and HOOC(CH₂) n COOH wherein n is an integer of 1 to 12.
References Cited (Backward Citations)
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