WO2015076284A1 - Stabilisant et procédé de stabilisation des lipoprotéines haute densité contenues dans le sérum sanguin ou le plasma sanguin - Google Patents

Stabilisant et procédé de stabilisation des lipoprotéines haute densité contenues dans le sérum sanguin ou le plasma sanguin Download PDF

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WO2015076284A1
WO2015076284A1 PCT/JP2014/080603 JP2014080603W WO2015076284A1 WO 2015076284 A1 WO2015076284 A1 WO 2015076284A1 JP 2014080603 W JP2014080603 W JP 2014080603W WO 2015076284 A1 WO2015076284 A1 WO 2015076284A1
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serum
plasma
hdl
density lipoprotein
component
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PCT/JP2014/080603
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Japanese (ja)
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健太 金城
拓郎 奥田
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協和メデックス株式会社
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Priority to JP2015549165A priority Critical patent/JP6795887B2/ja
Publication of WO2015076284A1 publication Critical patent/WO2015076284A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/96—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood or serum control standard
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors

Definitions

  • the present invention relates to a stabilizer for high-density lipoprotein in serum or plasma, a method for stabilizing high-density lipoprotein in serum or plasma, a method for preserving high-density lipoprotein in serum or plasma, Standards for quantifying components in high density lipoprotein, methods for producing standard products for quantifying components in high density lipoproteins in serum or plasma, methods for quantifying components in high density lipoproteins in serum or plasma, The present invention also relates to a kit for quantifying components in high-density lipoprotein in serum or plasma.
  • Serum and plasma are one of the specimens often used in clinical tests. Serum and plasma contain lipoproteins such as high density lipoprotein (HDL), low density lipoprotein (LDL), and very low density lipoprotein (VLDL). Each lipoprotein contains cholesterol, neutral fat. Ingredients such as phospholipids are included. These lipoproteins are separated by agarose gel electrophoresis based on the difference in charge and by polyacrylamide gel electrophoresis based on the difference in particle diameter. It is known that lipoproteins in serum and plasma are denatured during storage. Since the denatured lipoprotein changes in charge and particle size from the original lipoprotein, the denatured lipoprotein can be detected by agarose gel electrophoresis or polyacrylamide gel electrophoresis.
  • HDL high density lipoprotein
  • LDL low density lipoprotein
  • VLDL very low density lipoprotein
  • Patent Document 1 As a method for stabilizing lipoprotein for suppressing lipoprotein denaturation, a method using 2-methyl-4-isothiazolin-3-one (see Patent Document 1), a method using guanidine sulfate (Patent Document 2). And a method using a buffer having a sulfo group (see Patent Document 3) and the like are known.
  • HDL-C cholesterol in HDL
  • LDL-C cholesterol in LDL
  • Patent Document 4 Method for measuring HDL-C using ionic surfactant, polyanion and albumin
  • Patent Document 5 Method for measuring HDL-C using quaternary ammonium salt and polyanion
  • Patent Document 6 Method for measuring LDL-C using an ethylene polycyclic surfactant
  • Patent Document 8 Method for measuring LDL-C using an arylsulfonic acid derivative
  • Serum or plasma lipoproteins are easily denatured during storage or by lyophilization. This denaturation of lipoproteins often affects the measurement of components in lipoproteins and often precludes accurate measurement of components in lipoproteins.
  • the object of the present invention is to stabilize HDL in serum or plasma, to stabilize HDL in serum or plasma, to accurately measure components in HDL in serum or plasma, in serum or plasma, Provided are a method for storing HDL, a standard for quantifying components in HDL and a method for producing the same, a method for quantifying components in HDL in serum or plasma, and a kit for quantifying components in HDL in serum or plasma. There is.
  • the present inventors have stabilized HDL in serum or plasma by adding a polyoxyethylene polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative to serum or plasma.
  • the present invention was completed by finding the knowledge that components in HDL such as HDL-C can be accurately quantified. That is, the present invention relates to the following [1] to [18].
  • a stabilizer for HDL in serum or plasma containing a polyoxyethylene polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative as an active ingredient.
  • a method for stabilizing HDL in serum or plasma comprising adding a polyoxyethylene polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative to serum or plasma.
  • a method for stabilizing HDL in serum or plasma comprising the following steps.
  • a method for preserving HDL in serum or plasma comprising adding a polyoxyethylene polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative to serum or plasma.
  • a method for preserving HDL in serum or plasma comprising the following steps. (1) adding a polyoxyethylene polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative to serum or plasma; and (2) A step of storing the mixture obtained in step (1).
  • a method for preserving HDL in serum or plasma comprising the following steps.
  • a method for producing a standard product for quantitative determination of components in HDL in serum or plasma comprising the following steps. (1) adding a polyoxyethylene polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative to serum or plasma; and (2) A step of pricing the concentration of the component in HDL contained in the mixture obtained in step (1) using a known amount of the component. [13] A method for producing a standard product for quantitative determination of components in HDL in serum or plasma, comprising the following steps.
  • a method for quantifying components in HDL in serum or plasma comprising the following steps. (1) a step of measuring the component using a reagent for measuring the component in HDL in serum or plasma; (2) Calibration using the standard product according to any one of [7] to [10] and the measurement reagent of step (1) to express the relationship between the concentration of the component and the measured value for the component Creating a line; and (3) A step of determining the concentration of the component in serum or plasma from the measurement value obtained in step (1) and the calibration curve prepared in step (2).
  • the quantification method according to [15] wherein the component in HDL is cholesterol.
  • a stabilizer for HDL in serum or plasma a method for stabilizing HDL in serum or plasma, serum or plasma for accurately measuring components in HDL such as HDL-C in serum or plasma, Method for storing HDL in plasma, standard product for quantifying components in HDL and method for producing the same, method for quantifying components in HDL in serum or plasma, and kit for quantifying components in HDL in serum or plasma Is provided.
  • the serum and plasma in the present invention may be any serum and plasma prepared from whole blood collected from humans or animals, and serum prepared from whole blood collected from humans. And plasma are preferred.
  • Serum can be prepared by a method of centrifuging whole blood collected using a blood collection tube not containing an anticoagulant, a method of natural sedimentation, or the like.
  • Plasma can be prepared by a method of centrifuging whole blood collected by an anticoagulant-containing blood collection tube, a method of spontaneous sedimentation, or the like.
  • the anticoagulant include ethylenediaminetetraacetic acid (EDTA) dipotassium salt, heparin, sodium fluoride, sodium citrate and the like.
  • animals include primates such as monkeys, gorillas, orangutans, chimpanzees and baboons.
  • HDL High density lipoprotein in the present invention is a lipoprotein having a specific gravity of 1.063 to 1.21.
  • Examples of components in HDL in the present invention include cholesterol and neutral fat, and cholesterol is preferred.
  • the stabilizer of the present invention stabilizes HDL in serum or plasma.
  • the stabilizer of the present invention contains a polyoxyethylene polycyclic phenyl ether derivative (hereinafter referred to as POE polycyclic phenyl ether derivative) or a naphthalenesulfonic acid derivative as an active ingredient.
  • Examples of the POE polycyclic phenyl ether derivative in the present invention include polyoxyethylene polycyclic phenyl ether (hereinafter referred to as POE polycyclic phenyl ether), polyoxyethylene polycyclic phenyl ether sulfate (hereinafter referred to as POE polycyclic phenyl ether sulfate).
  • polyoxyethylene / polyoxypropylene polycyclic phenyl ether hereinafter referred to as POE / POP polycyclic phenyl ether).
  • POE polycyclic phenyl ether sulfates include, for example, New Coal 707-SF, New Coal 707-SFC, New Coal 707-SN, New Coal 714-SF, New Coal 714-SN, New Coal 723-SF, New Coal 740-SF, New Coal 780-SF, New Coal 2607-SF, New Coal 2614-SF (above, manufactured by Nippon Emulsifier Co., Ltd.), Hightenol NF-08, Hightenol NF-0825, High Tenol NF-13, Haitenol NF-17 (Daiichi Kogyo Seiyaku Co., Ltd.) and the like.
  • POE / POP polycyclic phenyl ether examples include, for example, New Coal 707-F, New Coal 710-F, New Coal 714-F, New Coal 2608-F, New Coal 2600-FB, New Coal 2616-F, New Coal 3612-FA (manufactured by Nippon Emulsifier Co., Ltd.) and the like.
  • naphthalene sulfonic acid derivatives examples include naphthalene sulfonic acid formaldehyde condensates.
  • Specific examples (commercial products) of naphthalene sulfonic acid formaldehyde condensates include, for example, Disrol SH, Escort (30) (manufactured by Nippon Emulsifier Co., Ltd.), Demol N, Demol NL, Demol RN, Demol RN-L, Demol T , Demol T-45, Demol MS, Demol SN-B (above, manufactured by Kao Corporation) and the like.
  • the content of the POE polycyclic phenyl ether derivative or naphthalenesulfonic acid derivative contained in the stabilizer of the present invention is not particularly limited as long as it is a content capable of stabilizing HDL in serum or plasma.
  • the stabilizer of the present invention is added to mL, the content is 0.01-5 g, and the content is preferably 0.05-1 g.
  • lyophilization is used in the usual sense used in the art, and means that a sample is frozen and depressurized in a frozen state to remove moisture from the sample and dry.
  • the lyophilization conditions are not particularly limited, but are usually -80 to 35 ° C, preferably -80 to 30 ° C, 0.667 to 1333 Pa, preferably 13.3 to 133.3 Pa, and 6 to 120 hours, more preferably For 12 to 120 hours.
  • the water content of the freeze-dried product is usually 10% by weight or less, preferably 1% by weight or less.
  • the method for stabilizing HDL of the present invention includes a step of adding a POE polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative to serum or plasma.
  • the amount of the POE polycyclic phenyl ether derivative or naphthalenesulfonic acid derivative added to serum or plasma is not particularly limited as long as HDL is stabilized in serum or plasma, and is usually per 100 mL of serum or plasma. 0.01 to 5 g, preferably 0.05 to 1 g.
  • the POE polycyclic phenyl ether derivative When a POE polycyclic phenyl ether derivative is added to serum or plasma, the POE polycyclic phenyl ether derivative may be added as the POE polycyclic phenyl ether derivative itself, or as a POE polycyclic phenyl ether derivative aqueous solution. Good.
  • the naphthalenesulfonic acid derivative when the naphthalenesulfonic acid derivative is added to serum or plasma, the naphthalenesulfonic acid derivative may be added as the naphthalenesulfonic acid derivative itself, or may be added as a naphthalenesulfonic acid derivative aqueous solution.
  • an aqueous medium such as deionized water, distilled water, or a buffer solution
  • a buffering agent for preparing the buffer solution include a buffering agent described later.
  • the pH of these aqueous solutions is not particularly limited as long as the lipoprotein in serum or plasma does not precipitate after the aqueous solution is added to serum or plasma, and is preferably pH 6-9.
  • Another aspect of the HDL stabilization method of the present invention includes the following steps. (1) A step of adding a POE polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative to serum or plasma; (2) freeze-drying the mixture obtained in step (1); and (3) A step of dissolving the lyophilized product obtained in step (2) with an aqueous medium.
  • the aqueous medium in step (3) is not particularly limited as long as it dissolves HDL in serum or plasma and can stably maintain HDL in an aqueous solution of HDL, and examples thereof include the above-mentioned aqueous medium. It is done.
  • the method for storing HDL of the present invention includes a step of adding a POE polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative to serum or plasma.
  • Another aspect of the HDL storage method of the present invention includes the following steps. (1) adding a POE polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative to serum or plasma; and (2) A step of storing the mixture obtained in step (1).
  • another aspect of the HDL storage method of the present invention includes the following steps. (1) A step of adding a POE polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative to serum or plasma; (2) a step of freeze-drying the mixture obtained in step (1); (3) dissolving the lyophilized product obtained in step (2) with an aqueous medium; and (4) A step of storing the aqueous solution obtained in step (3).
  • Storing conditions in the HDL storage method of the present invention are not particularly limited as long as HDL is stored stably.
  • the storage temperature in the HDL storage method is not particularly limited as long as the HDL is stored stably, and is usually -80 to 45 ° C, preferably -5 to 30 ° C, and particularly preferably 2 to 10 ° C. .
  • the storage period in the HDL storage method is not particularly limited as long as HDL is stably stored, and is usually 1 day to 2 years.
  • the standard for quantifying components in HDL in serum or plasma of the present invention may be in solution or lyophilized.
  • the standard product of the present invention contains HDL in serum or plasma and a POE polycyclic phenyl ether derivative or naphthalene sulfonic acid derivative, and the content of the component in the HDL is a value. It is attached.
  • the standard product is in a lyophilized state, contains HDL in freeze-dried serum or plasma, and POE polycyclic phenyl ether derivative or naphthalenesulfonic acid derivative, and the HDL The content of the components in it is priced.
  • the standard product of the present invention can be used not only as a standard product for quantifying components in HDL in serum or plasma, but also as a quality control substance in quantifying components in HDL in serum or plasma.
  • the method for producing the standard product of the present invention includes the following steps. (1) adding a POE polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative to serum or plasma; and (2) A step of pricing the concentration of the component in HDL contained in the mixture obtained in step (1) using a known amount of the component.
  • the method for producing the standard product of the present invention includes the following steps. (1) A step of adding a POE polycyclic phenyl ether derivative or a naphthalenesulfonic acid derivative to serum or plasma; (2) freeze-drying the mixture obtained in step (1); and (3) A step of pricing the content of the component in HDL contained in the freeze-dried product obtained in step (2) using a known amount of the component.
  • the amount of the POE polycyclic phenyl ether derivative or naphthalenesulfonic acid derivative added to serum or plasma is not particularly limited as long as HDL is stabilized in serum or plasma. In general, it is 0.01 to 5 g, preferably 0.05 to 1 g per 100 mL of serum or plasma.
  • Serum or plasma is cooled to 2-8 ° C., and an aqueous solution of POE polycyclic phenyl ether derivative or naphthalenesulfonic acid derivative is added to the cooled serum or plasma.
  • an aqueous solution of POE polycyclic phenyl ether derivative or naphthalenesulfonic acid derivative is added to the cooled serum or plasma.
  • the pH-adjusted aqueous solution is filtered to about 0.2-0.4 microns. Filter using a membrane to obtain a standard product.
  • Serum or plasma is cooled to 2-8 ° C., and an aqueous solution of POE polycyclic phenyl ether derivative or naphthalenesulfonic acid derivative is added to the cooled serum or plasma.
  • an aqueous solution of POE polycyclic phenyl ether derivative or naphthalenesulfonic acid derivative is added to the cooled serum or plasma.
  • the pH-adjusted aqueous solution is filtered to about 0.2-0.4 microns. Filter using a membrane. The resulting solution is dispensed into lyophilized vials and half-capped.
  • the solution dispensed in the vial is frozen at a low temperature of -30 ° C or lower, then lyophilized under vacuum conditions of 133.3 Pa, and finally dried at 0 to 20 ° C with gradual warming.
  • the standard product in a lyophilized state is used after dissolving in a known amount of an aqueous medium in preparing a calibration curve.
  • the aqueous medium for dissolving the lyophilized standard is not particularly limited as long as it is an aqueous medium that enables the method of quantifying the components in HDL of the present invention.
  • deionized water, distilled water, buffer solution Etc examples of the buffering agent for preparing the buffer solution include a buffering agent described later.
  • pricing of a standard product means determining the content of a component in HDL to be measured contained in a manufactured standard product using a standard serum containing a known amount of the component.
  • the standard serum containing a known amount of the component is determined by a method that does not depend on a homogeneous method such as a standard analysis method of the US Center for Disease Control and Prevention (CDC). Serum.
  • CDC US Center for Disease Control and Prevention
  • Serum Using this standard serum and the manufactured standard as a specimen, measurement is performed by a homogeneous method using an actual automatic analyzer, and a measurement value (for example, absorbance) of a standard serum containing a known amount of the component is manufactured.
  • the measured value of the standard product (for example, absorbance) is compared to determine the content of the component in the manufactured standard product.
  • the manufactured standard product is used as a standard serum in the laboratory to determine the measurement value of the specimen.
  • the content of the POE polycyclic phenyl ether derivative and naphthalene sulfonic acid derivative contained in the standard product of the present invention should be such that the HDL in the standard product is stably maintained and the components in the HDL in the standard product can be accurately measured.
  • the standard product of the present invention may contain additives such as buffers, metal ions, salts, surfactants, preservatives, sugar compounds and the like as necessary.
  • Buffers include, for example, lactate buffer, citrate buffer, acetate buffer, succinate buffer, phthalate buffer, phosphate buffer, triethanolamine buffer, diethanolamine buffer, lysine buffer, barbitur tool Buffering agents, imidazole buffers, malic acid buffers, oxalic acid buffers, glycine buffers, boric acid buffers, carbonate buffers, glycine buffers, Good buffers, and the like can be mentioned.
  • Examples of the good buffer include 2-morpholinoethanesulfonic acid (MES), bis (2-hydroxyethyl) iminotris (hydroxymethyl) methane (Bis-Tris), N- (2-acetamido) iminodiacetic acid (ADA), Piperazine-N, N'-bis (2-ethanesulfonic acid) (PIPES), N- (2-acetamido) -2-aminoethanesulfonic acid (ACES), 3-morpholino-2-hydroxypropanesulfonic acid (MOPSO) N, N-bis (2-hydroxyethyl) -2-aminoethanesulfonic acid (BES), 3-morpholinopropanesulfonic acid (MOPS), N- [tris (hydroxymethyl) methyl] -2-aminoethanesulfonic acid (TES), 2- [4- (2-hydroxyethyl) -1-piperazinyl] ethanesulfonic acid (HEPES), 3- [N
  • Examples of metal ions include magnesium ions, calcium ions, manganese ions, and zinc ions.
  • Examples of the salts include sodium chloride and potassium chloride.
  • Examples of the surfactant include nonionic surfactants, cationic surfactants, anionic surfactants, and amphoteric surfactants.
  • Examples of the preservative include antibiotics such as sodium azide, streptomycin, gentamicin, Bioace, Procrine 300 (trademark), Proxel GXL (trademark) and the like.
  • sugar compound examples include monosaccharides and disaccharides, and disaccharides are preferable.
  • monosaccharides include glucose, fructose, and fucose.
  • disaccharide examples include saccharose, trehalose, maltose and the like. Two or more monosaccharides, disaccharides and the like can be used in combination.
  • the amount of the sugar compound added when the sugar compound is mixed with serum or plasma is usually 0.5 to 20 ⁇ g, preferably 1 to 15 ⁇ g, more preferably 1.25 to 12.5 ⁇ g with respect to 100 ⁇ mL of serum or plasma. .
  • the method for quantifying components in HDL in serum or plasma of the present invention comprises the following steps. (1) a step of measuring the component using a reagent for measuring the component in HDL in serum or plasma; (2) using the standard product of the present invention and the measurement reagent of step (1) to create a calibration curve representing the relationship between the concentration of the component and the measured value for the component; and (3) A step of determining the concentration of the component in serum or plasma from the measurement value obtained in step (1) and the calibration curve prepared in step (2).
  • a known method for measuring the component in HDL can be used.
  • the method for measuring components in HDL include a method for measuring HDL-C, a method for measuring neutral fat in HDL (hereinafter referred to as HDL-TG), and the like.
  • HDL-TG a method for measuring neutral fat in HDL
  • HDL-TG a method for measuring neutral fat in HDL
  • a method for measuring the components therein that is, a so-called homogeneous method is preferred.
  • a specimen and an enzyme for measuring cholesterol described in WO 2004/035816 are reacted in an aqueous medium containing a nonionic surfactant, a polyanion, and albumin.
  • the enzyme for measuring cholesterol is a combination of (i) cholesterol ester hydrolase and cholesterol oxidase, or (ii) cholesterol ester hydrolase, oxidized coenzyme, and cholesterol dehydrogenase. It is a combination.
  • the kit for quantifying components in HDL in serum or plasma of the present invention contains the standard product of the present invention and a reagent for measuring components in HDL. By using the kit of the present invention, the components in HDL can be easily quantified.
  • a reagent for measuring a component in HDL a known reagent for measuring a component in HDL can be used. Examples of the reagent for measuring components in HDL include HDL-C measuring reagent and HDL-TG measuring reagent.
  • HDL is not fractionated by means of centrifugation or the like, but in a state where a plurality of lipoproteins such as LDL and VLDL coexist in the same reaction solution in addition to HDL.
  • a reagent for measuring the components therein that is, a reagent used in a measuring method by the so-called homogeneous method is preferable.
  • Examples of the HDL-C measuring reagent used in the HDL-C measuring method by the homogeneous method include HDL-C containing an enzyme for measuring cholesterol, a nonionic surfactant, a polyanion and albumin described in, for example, WO 2004/035816 pamphlet.
  • Examples include a measurement reagent, an enzyme for measuring cholesterol, a quaternary ammonium salt having a specific structure, and a HDL-C measurement reagent containing a polyanion described in WO 2006/118199 pamphlet.
  • the enzyme for measuring cholesterol is a combination of (i) cholesterol ester hydrolase and cholesterol oxidase, or (ii) cholesterol ester hydrolase, oxidized coenzyme, and cholesterol dehydrogenase.
  • a commercially available measuring reagent can also be used as the HDL-C measuring reagent used in the homogeneous method.
  • Examples of commercially available measuring reagents include Determiner L HDL-C (manufactured by Kyowa Medex), Metabolid HDL-C (manufactured by Kyowa Medex), and “Seiken” HDL-EX N (manufactured by Denka Seiken). ) And the like.
  • the standard product of the present invention and the kit of the present invention are suitably used in a measurement method using an automatic analyzer in which components in serum or plasma in HDL are automatically and continuously measured.
  • the effects of the HDL stabilizer, the stabilization method, and the storage method of the present invention can be verified, for example, by the following method.
  • Pooled serum or pooled plasma is prepared by mixing serum or plasma collected from a plurality of healthy individuals.
  • the prepared pool serum or pool plasma is divided into two, and the HDL stabilizer of the present invention is added to one pool serum or pool plasma, and the pool sera or pool to which the HDL stabilizer of the present invention is added Plasma (group A serum or plasma) and pooled serum or pooled plasma (group B serum or plasma) to which the HDL stabilizer of the present invention is not added are prepared.
  • a certain amount of group A serum or plasma is dispensed into a plurality of containers of the same shape and material, and if necessary, freeze-dried by the method described above.
  • pricing is performed by the above-described method to prepare a standard product (standard product A).
  • a standard product (standard product B) is prepared using serum or plasma of group B.
  • the sample is stored for 7 days at 5 ° C. before storage of standard solution A, and after storage of standard solution A ( 1 after storage of standard product A to N after storage of standard product A).
  • Serum (manufactured by Discovery Life Science), trehalose (manufactured by Hayashibara), New Coal 740 (60) (POE polycyclic phenyl ether derivative; manufactured by Nippon Emulsifier Co., Ltd.), New Coal 740-SF (POE polycyclic phenyl ether derivative; Japan) Emulsifier Co., Ltd.), Demol N (Naphthalenesulfonic acid derivative; Kao Co., Ltd.), Borosilicate glass vial [manufactured by Yamato Special Glass Co., Ltd. (capacity: 2 mL; diameter: 16 mm; brown)], freeze dryer (Tokyo Rika Instruments) Manufactured by EYELA DRC-1100, FDU-2100).
  • the serum 1 before storage (the serum 1 before storage 1 serum 1 before storage 3) a 5 ° C. for 7 days Save and sera obtained 1 after storage (serum 1 after storage 1 serum 1 after storage 3)
  • “Metabolide HDL-C” as an HDL-C measurement reagent, and measuring each sample according to the operation procedure described in the package insert accompanying “Metabolide HDL-C” after the absorbance 1 store (absorbance 1 after storage 1 to the absorbance 1 after storage 3) were measured for each sample, the mean value was calculated absorbance 1 after storage 1 to the absorbance 1 after storage 3 as the mean absorbance 1 after storage.
  • Measure by the same method except using freeze-dried serum 2-4 and freeze-dried serum 0 instead of freeze-dried serum 1, and calculate average absorbance 2 after storage -average absorbance 4 storage and after average absorbance 0 storage did.
  • the residual rate (%) was calculated according to the following formula (II) using the calculated average absorbance before 1 storage and after the average absorbance after 1 storage .
  • the freeze-dried serum 1 was measured three times in accordance with the above procedure, and the residual ratio in each measurement of all three measurements was calculated, and the average value (average residual ratio) of the calculated residual ratio was determined.
  • the combination of the average absorbance 1 before storage and the average absorbance 1 after storage instead of the combination of the average absorbance 2 before storage and the average absorbance 2 storage, the combination of the average absorbance 3 before storage and the average absorbance 3 storage , and the average absorbance 4 before storage mean absorbance 4 combination after storage, and, in the same procedure as above except for using the mean absorbance 0 after storage and the average absorbance 0 combination after storage, respectively, each of the freeze-lyophilized serum 2-4 and lyophilized serum 0
  • measurement was performed three times in total, and the residual rate in each measurement of all three measurements was calculated for each freeze-dried serum, and the average value (average residual rate) of the calculated residual rate was calculated. Determined for each lyophilized serum. Further, the calculated residual ratio was subjected to a Dunnett test (Dunnett test) with a significance level of 0.05. The results are shown in Table 2.
  • the lyophilized serum prepared by adding the stabilizer of the present invention has a higher residual rate than the lyophilized serum prepared without adding the stabilizer of the present invention. It was. In the standard product for HDL-C determination, even when the standard product is stored in solution, a very high residual rate of 100% is required for accurate HDL-C measurement. Is done. Therefore, the difference in the residual ratio between the case where the stabilizer of the present invention is added and the case where it is not added in Table 2 is important. In addition, when Dunnett's test was performed with a significance level of 0.05, all the lyophilized sera prepared by adding the stabilizer of the present invention remained in the lyophilized serum prepared by adding the stabilizer of the present invention.
  • a stabilizer for HDL in serum or plasma a method for stabilizing HDL in serum or plasma, a method for storing HDL in serum or plasma, and an accurate measurement of components in HDL in serum or plasma.
  • a standard product and a production method thereof a method for quantifying components in HDL in serum or plasma, and a kit for quantifying components in HDL in serum or plasma.
  • Stabilizer for HDL in serum or plasma of the present invention method for stabilizing HDL in serum or plasma, method for storing HDL in serum or plasma, standard for quantification of components in HDL in serum or plasma, and The production method, the method for quantifying components in HDL in serum or plasma, and the kit for quantifying components in HDL in serum or plasma are useful for diagnosis of metabolic syndrome and the like.

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Abstract

Pour mesurer avec précision les composants des lipoprotéines haute densité (HDL) contenues dans le sérum sanguin ou le plasma sanguin, la solution selon l'invention porte sur un stabilisant pour HDL contenues dans le sérum sanguin ou le plasma sanguin, sur un procédé de stabilisation des HDL contenues dans le sérum sanguin ou le plasma sanguin, sur un procédé de stockage des HDL contenues dans le sérum sanguin ou le plasma sanguin, et sur un standard de référence permettant de déterminer les composants des HDL. Le stabilisant pour les HDL contenues dans le sérum sanguin ou le plasma sanguin contient un dérivé d'éther phénylique polycyclique de polyoxyéthylène ou un dérivé d'acide naphtalènesulfonique à titre de principe actif. Le procédé de stabilisation et le procédé de stockage des HDL contenues dans le sérum sanguin ou le plasma sanguin sont caractérisés en ce qu'un dérivé d'éther phénylique polycyclique de polyoxyéthylène ou en ce qu'un dérivé d'acide naphtalènesulfonique est ajouté au sérum sanguin ou au plasma sanguin. Le standard de référence permettant de déterminer les composants des HDL contenues dans le sérum sanguin ou le plasma sanguin est caractérisé en ce qu'il contient les HDL contenues dans le sérum sanguin ou le plasma sanguin, et un dérivé d'éther phénylique polycyclique de polyoxyéthylène ou un dérivé d'acide naphtalènesulfonique, une valeur étant donnée pour la teneur en composants des HDL.
PCT/JP2014/080603 2013-11-21 2014-11-19 Stabilisant et procédé de stabilisation des lipoprotéines haute densité contenues dans le sérum sanguin ou le plasma sanguin WO2015076284A1 (fr)

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WO2022009973A1 (fr) * 2020-07-10 2022-01-13 積水メディカル株式会社 Composition à teneur en tarc, et procédé d'amélioration de stabilité de conservation de tarc

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