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Method for measuring activated partial thromboplastin time and reagent kit for activated partial thromboplastin time measurement
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A thromboplastin time and compound technology, which is applied in the field of reagents and kits for the determination of activated partial thromboplastin time, and can solve the problems of increasing environmental burden, strict restrictions on the use of phenol and the like
Pending Publication Date: 2021-04-09
SYSMEX CORP
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However, phenol is a substance that imposes an environmental burden, and the use of phenol is increasingly restricted
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Embodiment 1
[0064] Example 1: Study on the Inhibitory Effect of Precipitation of Ellagic Acid Compounds
[0065] In the APTT reagent containing the metal ion complex of ellagic acid as the activator, when the salt concentration increased, the particle size of the complex increased due to the salting-out effect, and finally precipitated. Utilizing this salting-out principle, the precipitation-inhibiting effects of various additives in ellagic acid metal ion complex solutions were confirmed.
[0066] (1) Preparation of ellagic acid metal ion complex solution
[0067] 50 mM HEPES, 8 mM TAPS, and 1% by mass glycine as buffer components, 100 μM ellagic acid as a metal ion complex of ellagic acid, 60 μM zinc chloride, and 50 μM aluminum chloride were prepared as preservatives. 0.05% ProClin (trademark) 300 (Sigma-Aldrich) in metal ion complex solution of ellagic acid. This solution is mixed with at least one selected from gallic acid, pyrogallol, hydroquinone, 5-methylpyrogallol, resorcinol, ...
Embodiment 2
[0072] Example 2: Study on the Inhibition Effect of Abnormal Initial Reactions of Solidification
[0073] Whether or not initial abnormalities can be suppressed was investigated by using an APTT reagent containing pyrogallol or gallic acid.
[0074] (1) Preparation of APTT reagent
[0075] 50 mM HEPES, 8 mM TAPS, and 1% by mass glycine as buffer components, 100 μM ellagic acid as a metal ion complex of ellagic acid, 60 μM zinc chloride, and 50 μM aluminum chloride were prepared as preservatives. 0.05% ProClin (trademark) 300 (Sigma-Aldrich) in metal ion complex solution of ellagic acid. The complex solution was mixed with gallic acid or pyrogallol as an additive to obtain a complex solution containing gallic acid or pyrogallol at a concentration of 100 μM. As a control, an ellagic acid metal ion complex solution without additives was used. As phospholipid components, PE (L-α-phosphatidylethanolamine), PC (L-α-phosphatidylcholine) and PS (L-α-phosphatidylserine) were adjuste...
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Abstract
The invention relates to a method for measuring activated partial thromboplastin time, and a reagent kit for activated partial thromboplastin time measurement. The method comprises: mixing: a blood sample; a first reagent comprising an ellagic acid compound and a compound represented by Formula (I); and a second reagent comprising a calcium ion; and measuring activated partial thromboplastin time using the mixture prepared in the mixing step, wherein the compound represented by Formula (I) is shown below wherein X is a hydrogen atom, -(C=O)-Y, or -(C=O)-OZ, Y is a hydroxyl group, a hydrogen atom, a linear or branched alkyl group having a carbon number of 1 or more and 6 or less, or a phenyl group, and Z is an alkali metal.
Description
technical field [0001] The present invention relates to a reagent and kit for measuring activated partial thromboplastin time. In addition, the present invention relates to a method for measuring an activated partial thromboplastin time, and a method for inhibiting precipitation of an ellagic acid compound. Background technique [0002] Ellagic acid has the function of activating the contact factor system, so ellagic acid is often used as an activator of coagulation for activating a reagent for measuring partial thromboplastin time (APTT). However, ellagic acid tends to precipitate in an aqueous solvent such as that used in the APTT measurement reagent. Once the precipitation of ellagic acid occurs, the ellagic acid necessary for the coagulation reaction is insufficient, and the coagulation time cannot be accurately measured. Conventionally, phenol was added to a reagent for APTT measurement containing ellagic acid as a substance that inhibits the precipitation of ellagic ...
Claims
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