WO2006054519A1

WO2006054519A1 - Standard item for fractional determination of component of lipoprotein

Info

Publication number: WO2006054519A1
Application number: PCT/JP2005/020845
Authority: WO
Inventors: Isami Tsuboi; Michihiro Taguchi; Akira Miike
Original assignee: Kyowa Medex Co., Ltd.; Bml, Inc.
Priority date: 2004-11-17
Filing date: 2005-11-14
Publication date: 2006-05-26
Also published as: JPWO2006054519A1; JP4704355B2

Abstract

A lyophilized standard item for use in the fractional determination of a component to be measured of lipoprotein is produced by mixing a sulfonated buffering agent with a serum or plasma, lyophilizing the same and evaluating the content of component to be measured in the lyophilization product by the use of component to be measured whose quantity is known. An analytical curve is created by the use of this standard item and a reagent for fractional measurement of component to be measured contained in a lipoprotein. Any component to be measured of an analyte is measured with the use of the reagent for fractional measurement, and the concentration of component to be measured of the analyte is calculated from the thus obtained measurement value and the above analytical curve. Thus, there is provided a standard item for determination of a component to be measured of lipoprotein that ensures easy handling and, even after long-period storage, gives accurate measurement values, and provided a process for producing the standard item. Further, there is provided a method of fractional determination of a component to be measured of an analyte containing a lipoprotein, in which use is made of the above standard item.

Description

Specification

Standard product for fractional determination of components in lipoproteins

Technical field

[0001] The present invention relates to a standard product used for differential quantification of a component to be measured in lipoprotein, a method for producing the same, a kit for fractional quantification of a component to be measured in a sample containing lipoprotein, and a lipoprotein. The present invention relates to a method for fractional quantification of components to be measured in a sample containing protein.

Background art

[0002] For clinical diagnosis, measurement of lipid items is important for prevention of lifestyle-related diseases such as diabetes and improvement of lifestyle habits. Many methods for measuring lipid items in a specimen have been reported, but the so-called homogenous method, in which components in the lipoprotein are measured without separating the lipoprotein in the specimen, has become common. ing. For example, in a discrete type automatic analyzer, a large amount of sample is continuously quantified by a homogenous method using a surfactant such as dextran sulfate or a homogen method using a surfactant. , Ru In any of these methods, the concentration of a component to be measured in a sample is calculated by measuring a standard product containing a measurement target component at a known concentration at the same time as the sample and analyzing the absorbance obtained from the standard product. This is done by using a calibration curve or correlation equation between the amount of information and the amount of information such as absorbance obtained from the specimen.

[0003] In the standard product, the components to be measured are individually contained at a constant concentration or content, so-called pure type standard products, as well as other components in the sample to be measured. A product form containing a known concentration or a known content together with a component, a so-called serum type standard product, is known. For example, a standard product described in Patent Document 1 is known as a serum type standard product.

[0004] In addition, Patent Document 2 discloses lipoproteins for diagnostic purposes that have a water content in the range of 1 to 10% and can be reconstituted into a clear solution with a maximum turbidity of 70 LSU. A lyophilized control or reference plasma or control or reference serum is disclosed. In the case of a standard product of pure product type, for example, when preparing a standard product by dissolving cholesterol or neutral fat, it is necessary to use a large amount of surfactant. As the force changes, pipetting with an automatic analyzer causes problems such as sucking up a larger amount of solution and not giving accurate measurements.

[0005] Although the above-mentioned problems are solved in the serum type standard product, there is a problem in the long-term stability of the standard product, and it is necessary to store it in a frozen state in order to enhance the long-term stability.

[0006] Accordingly, there is a need for stable standard products that provide accurate measurements even after handling and storage for long periods of time.

Patent Document 1: Japanese Patent Laid-Open No. 10-17597

Patent Document 2: JP-A-6-300758

Disclosure of the invention

Problems to be solved by the invention

[0008] An object of the present invention is to provide a standard product for fractional quantification of a component to be measured in a lipoprotein which is easy to handle and gives an accurate measurement value even after long-term storage, a method for producing the same, and a lipoprotein. It is an object of the present invention to provide a kit for fractional quantification of components to be measured in a contained sample and a method for fractional quantification of components to be measured in a sample containing lipoproteins. Means for solving the problem

[0009] The present invention is (1) a standard product obtained by freeze-drying serum or plasma to which a buffer having a sulfo group is added, which is used for differential quantification of components to be measured in lipoproteins. In addition, the content of the component to be measured in the standard product is priced by using a known amount of the component to be measured, or (2) having a sulfo group Buffers are N- (2-hydroxyethyl) N, mono (2-sulfoethyl) piperazine, piperazine mono-N, N, mono-bis (2-ethanesulphonic acid), N- (2-acetamido) 2 —Aminoethanesulphonic acid, 3 morpholino-2-hydroxypropanesulfonic acid, N, N-bis (2 hydroxyethyl) 2 aminoethanesulfonic acid, 3 morpholinopropanesulfonic acid, N— [Tris (hydroxymethyl) methyl] — 2 aminoethanesulfonic acid, 3 — [N, N-Bis (2 Hydrochichetyl) amino] — 2-Hydroxypropanesulfonic acid, N— [Tris (hydroxymethyl) methyl] —2 Hydroxy-3 Aminopropanesulfonic acid, Piperazine I N, N , 1bis (2-hydroxypropanesulfonic acid), 3- [4— (2-hydroxyethyl) 1-piperazyl] -2 hydroxypropanesulfonic acid, 3- [4— (2 hydroxyethyl) —1— Perazyl] propanesulfonic acid, N tris (hydroxymethyl) methyl 3-aminopropanosulfonic acid, N cyclohexyl 2-aminoethanesulfonic acid, N cyclohexyl luo 3 amino-2 hydroxypropane sulfonic acid and N-cycloto Xyl 3 aminopropane sulfonic acid power selected one or more buffer materials selected from the above (1) standard product and (3) buffer power having sulfo group N- (2 hydroxyethyl)- The standard product according to (2) above, which is N ′ (2-sulfoethyl) piperazine, and (4) the above components (1) to (3), wherein the component to be measured in lipoprotein is cholesterol or neutral fat (5) The lipoprotein is a lipoprotein selected from the group consisting of high-density lipoprotein, low-density lipoprotein, ultra-low-density lipoprotein, chylomicron, and remnant lipoprotein. the above( 1)

~ (4) Standard product as described in any of the above, or (6) Serum or plasma mixed with a buffer having a sulfo group, and the resulting mixture is freeze-dried and then measured in a freeze-dried product. A method for producing a lyophilized standard used for differential quantification of a component to be measured in lipoprotein, characterized in that the content of the component to be measured is determined using a known amount of the component to be measured (7) Buffering power with sulfo group N- (2 Hydroxyethyl) N, One (2-sulfethenole) piperazine, Piperazine one N, N, One bis (2-ethanesnorephonic acid) , N— (2-acetamido) 2 aminoethanesulfonic acid, 3 morpholino-2-hydroxypropane sulfonic acid, N, N bis (2 hydroxyethyl) 2 aminoethanesulfonic acid, 3 morpholinopropane sulfonic acid, N— [Tris (hydroxymethyl) methyl] —2— Minoethane sulfonic acid, 3— [N, N bis (2 hydroxyethyl) amino] — 2 Hydroxypropane sulfonic acid, N— [Tris (hydroxymethyl) methyl] —2 Hydroxy-3 Aminopropenosolephonic acid, Piperazine 1 N , N'—Bis (2 hydroxyprononsnorephonic acid), 3— [4— (2 hydroxyethyl) 1-piperazyl] —2 Hydroxypropanesulfonic acid, 3— [4— (2 hydroxyethyl) — 1-piperaduryl] propane sulfonic acid, N tris (hydroxymethyl) methyl 3-aminopropane sulfonic acid, N-cyclohexyl 2-aminoethane sulfonic acid, N-cyclohexyl 3-amino 2-hydroxypropane sulfonic acid and N-Cyclohexyl 3 The production method according to (6) above, which is one or more buffering agents selected from aminopropanesulfonic acid, and (8) Buffer having a host group, N-(2-hydroxy E chill) -N, one (2 Suruhoechiru) above a piperidines Rajin (7) (9) The production method according to any one of (6) to (8) above, wherein (9) the component to be measured in lipoprotein is cholesterol or neutral fat, and (10) The method according to any one of (6) to (9) above, wherein the lipoprotein is selected from the group consisting of high density lipoprotein, low density lipoprotein, very low density lipoprotein, chylomicron, and remnant lipoprotein Or a sample containing lipoprotein, comprising (11) a reagent for fractional measurement of components to be measured in lipoprotein and the standard product described in any one of (1) to (5) above Using a kit for fractional quantification of components to be measured in (12) [1] Reagents for fractional measurement of components to be measured in lipoproteins, the components in a sample containing lipoproteins are measured. Step [2] Calibration curve using the standard product according to any one of (1) to (5) above and the reagent for fractionation measurement And [3] calculating the concentration of the component in the sample from the measurement value obtained in the step [1] and the calibration curve created in the step [2]. The present invention relates to a method for fractional quantification of a component to be measured in a specimen containing a characteristic lipoprotein.

The invention's effect

[0010] According to the present invention, there is provided a standard product for fractional quantification of components to be measured in a lipoprotein that is easy to handle and gives an accurate measurement value even after long-term storage. By using the reagent for fractional measurement of the component to be measured, it is possible to accurately perform the fractional quantification of the component to be measured in the sample containing lipoprotein.

BEST MODE FOR CARRYING OUT THE INVENTION

[0011] (Sample)

The specimen containing the component to be measured in the present invention is not particularly limited as long as it contains at least a plurality of various lipoproteins. For example, whole blood, plasma, serum, joint fluid, spinal fluid, saliva, amniotic fluid Urine, sweat, spleen, synovial fluid, etc., but whole blood, plasma, and serum are preferred.

[0012] (Lipoprotein)

Examples of the lipoprotein in the present invention include high density lipoprotein (hereinafter referred to as HDL), low density lipoprotein (hereinafter referred to as LDL), very low density lipoprotein (hereinafter referred to as VLDL), and chylomicron (hereinafter referred to as HDL). , CM)) and remnant lipoproteins.

[0013] (component to be measured in lipoprotein) The components to be measured in the lipoprotein in the present invention include, for example, cholesterol in HDL (HDL cholesterol), neutral fat in HDL, cholesterol in LDL (LDL cholesterol), neutral fat in LDL, Examples include cholesterol in VLDL, neutral fat in VLDL, cholesterol in CM, neutral fat in CM, cholesterol in remnant lipoprotein, and neutral fat in remnant lipoprotein.

[0014] (Serum or plasma)

In the present invention! The term “serum or plasma” used here means the normal meaning of serum or plasma obtained by removing blood cell components and the like from blood. As blood, blood derived from animals such as humans is used.

[0015] (buffering agent)

The buffer having a sulfo group in the present invention is preferably a buffer that gives a pH at which lipoproteins in serum or plasma do not precipitate. For example, pH 6-9 is preferable. Examples of the buffer containing a sulfo group include N— (2-hydroxyethyl) N, one (2—snolechechinole) piperazine (HEPES), piperazine one N, N, one bis (2-ethanesulphonic acid). (PIPES), N— (2acetamido) 2 aminoethanesulfonic acid (ACES), 3 —morpholino 2-hydroxypropanesulfonic acid (MOPSO), N, N-bis (2-hydroxychetetyl) 2 aminoethanesulfonic acid ( BES), 3 morpholinopropanesulfonic acid (MOPS), N— [Tris (hydroxymethyl) methyl] -2-aminoethanesulfonic acid (TES), 3 -— [N, N-bis (2hydroxyethyl) amino] — 2 Hydroxypropane sulfonic acid (DIPSO), N- [Tris (hydroxymethyl) methyl] -2-hydroxy 3-amino-propanosulfonic acid (TAPSO), Piperazine 1 N, N, 1 bis (2-hydroxypropane sulfonate) Acid) (POPSO), 3- [4- (2 Hydroxyethyl) 1-piperazyl] -2 Hydroxypropane sulfonic acid (HEPPSO), 3- [4- (2-Hydroxyethyl) -1-pipera Di-l] propanesulfonic acid [(H) EPPS], N-tris (hydroxymethyl) methyl-3-aminopropanesulfonic acid (TAPS), N cyclohexyl 2-aminoethanesulfonic acid (CHES), N- Examples include cyclohexyl 3-amino 2-hydroxypropane sulfonic acid (CAP SO) and N cyclohexyl 3-aminopropane sulfonic acid (CAPS). Among them, N- (2-hydroxyethyl) N, (2-sulfoethyl) piperazi (HEPES) is particularly preferred.

[0016] In addition, two or more of these sulfo group-containing buffering agents may be used in combination, but the amount of the buffering agent added when mixing the buffering agent with serum or plasma is such that serum or plasma lOOmL is added. 10 to 500 mmol is preferred, and 20 to 200 mmol is more preferred.

In addition to the buffer with sulfo group, lactate buffer, citrate buffer, acetate buffer, succinate buffer, phthalate buffer, phosphate buffer, triethanolamine buffer, Diethanolamine buffer, lysine buffer, barbitur buffer, imidazole buffer, malate buffer, oxalate buffer, glycine buffer, borate buffer, carbonate buffer, dalysin buffer, Good buffer Agents and the like. Good buffers include, for example, MES (2-morpholinoethanesulfonic acid) buffer, Bis-Tris [bis (2-hydroxyethyl) iminotris (hydroxymethyl) methane] buffer, Tris [tris (hydroxymethyl) aminomethane ] Buffer, ADA [N- (2-acetamido) iminoniacetic acid] buffer, etc. can be used in combination.

[0017] (Additive)

The standard product of the present invention may contain additives such as an aqueous medium, metal ions, salts, surfactants, preservatives, sugar compounds and the like as necessary. Examples of the aqueous medium include deionized water and distilled water. Examples of the metal ion include magnesium ion, calcium ion, manganese ion, zinc ion and the like. Examples of the salts include sodium salt sodium and potassium salt. Examples of the surfactant include nonionic surfactants, cationic surfactants, anionic surfactants, and amphoteric surfactants. Examples of antiseptics include antibiotics such as sodium azizuma, streptomycin, gentamicin, Bioace, Procrine 300 (trademark), Proxel GXL (trademark), and the like.

[0018] Examples of the sugar compound include monosaccharides and disaccharides, with disaccharides being preferred. Examples of monosaccharides include glucose, fructose, and fucose. Examples of the disaccharide include saccharose, trehalose, maltose and the like. Two or more monosaccharides and disaccharides can be used in combination. In addition, the amount of sugar compound added when sugar compounds are mixed with serum or plasma is preferably 0.5 to 20 g, more preferably 1 to 15 g, relative to serum or plasma lOOmL. 25-12. 5 g is particularly preferred. [0019] (Reagent for differential measurement of components to be measured in lipoprotein)

Reagents for fractionation measurement of components to be measured in lipoproteins include multiple types of lipoproteins that can fractionate specific lipoproteins such as HDL, LDL, VLDL, CM, and remnant lipoproteins by means such as centrifugation. Reagents that measure fractions in specific lipoproteins in the presence of the same reaction solution, so-called homogenous measurement reagents are not particularly limited. Cholesterol measurement reagent in HD L, neutral fat in HDL Measuring reagent, cholesterol measuring reagent in LDL, neutral fat measuring reagent in LDL, cholesterol measuring reagent in VLDL, neutral fat measuring reagent in VLDL, cholesterol measuring reagent in CM, neutral fat in CM Examples include a measuring reagent, a cholesterol measuring reagent in remnant lipoprotein, and a neutral fat measuring reagent in remnant lipoprotein. As a separate measurement reagent for a component to be measured in a strong lipoprotein, for example, not only a reagent prepared in-house but also a commercially available measurement reagent can be used. Commercially available measuring reagents include, for example, HDL cholesterol measurement reagent Detamina I HDL (Kyowa Medettas), Detamina I HDL (Kyowa Medex), Colles Test N HDL (Daiichi Kagaku) And Detamina L LDL (manufactured by Kyowa Medettas), which is an LDL cholesterol measurement reagent.

[0020] (Standard product manufacturing method)

In the production method of the standard product of the present invention, a sulfo group-containing buffer and serum or plasma are mixed, and the resulting mixture is freeze-dried, and the content of the component to be measured in the freeze-dried product is known. The method is not particularly limited as long as it is a method that uses a component to be measured, and when mixing serum or plasma with a buffer having a sulfo group, the buffer having a sulfo group is an aqueous solution (buffer). The pH of the aqueous solution that can be mixed is selected so that the protein in the serum or plasma does not precipitate after mixing, and pH 6-9 is preferred.

In the present invention, lyophilization is used in the usual meaning used in the art, and the sample is frozen, depressurized in the frozen state, and dried by removing moisture from the sample vessel. Means. The lyophilization conditions are not particularly limited, but are usually −80 to 20 ° C., preferably −80 to 15. It is carried out at a temperature of C at a pressure of 0.6667 to 1333 Pa, preferably 13.3 to 133.3 Pa for 6 to 72 hours, more preferably 12 to 72 hours. The water content of the freeze-dried product is usually 10% by weight or less, preferably 1% by weight or less. One aspect of the standard product manufacturing method is described below. The

[0022] Serum or plasma is cooled to 2-8 ° C, and an aqueous solution containing the above-mentioned buffer having a sulfo group is added thereto. After adjusting the pH with about ImolZL sodium hydroxide or about ImolZL hydrochloric acid solution, the pH-adjusted serum or plasma-containing aqueous solution is filtered using a filtration membrane of about 0.2 to 0.4 microns. To do. Dispense the resulting solution into vials for freeze-drying and half-clamp. The solution dispensed in vials is freeze-dried at a low temperature of, for example, -30 ° C or lower, for example, at a vacuum of 133.3 Pa, and finally dried up at 0-20 ° C while gradually warming. Prepare a standard product in a lyophilized state.

[0023] (Standard product pricing)

Pricing of a lyophilized standard means that the content of the component to be measured in the lipoprotein contained in the manufactured standard is determined using a known amount of the component to be measured. . The measurement components in the lyophilized standard product obtained are standardized by, for example, using multiple homogenous methods such as the recommendation of the US Department of Health's Disease Control and Prevention Center (CDC) recommendation for multiple serum or plasma samples from healthy individuals. When measuring a plurality of sera or plasma and standard samples at the same time using a known amount of a component to be measured by the method and using an automated analyzer, etc., the measured value in the serum or plasma is the CDC recommended method, etc. This can be done by determining the content of the component to be measured in the standard product so that it matches the measured value obtained by the standard method. (Kit for fractional determination of components to be measured in samples containing lipoproteins)

The kit for differential quantification of the component to be measured in the sample containing the lipoprotein of the present invention includes the standard product of the present invention and the reagent for differential measurement of the component to be measured in the lipoprotein. There is no particular limitation, and when the kit of the present invention is used, fractional measurement of components to be measured in a sample containing lipoprotein can be easily performed.

[0024] (Method for fractional determination of components to be measured in samples containing lipoproteins)

As a method for differential measurement of a component to be measured in a sample containing the lipoprotein of the present invention, (1) a component for measurement of a component to be measured in a lipoprotein is used to determine the component to be measured in a sample. A step of measuring, (2) a step of preparing a calibration curve using the standard product in which the amount of the component to be measured according to the present invention is priced and the fractionation measuring reagent, and (3) the step of (1) above. From the measured values obtained and the calibration curve created in step (2), the concentration of the component to be measured in the sample is calculated. In the fractional measurement step of the component to be measured in the above step (1), there is no particular limitation as long as it is a quantification method having a process for producing a specific step such as HDL, LDL, VLDL, CM, and remnant lipoprotein. A method of fractionating specific components of lipoproteins in a state where multiple types of lipoproteins coexist in the same reaction solution without fractionating lipoproteins by means such as centrifugation, so-called homogenous measurement method is used. As a powerful homogenous measurement method, specifically, cholesterol measurement method in HDL, neutral fat measurement method in HDL, cholesterol measurement method in LDL, neutral fat measurement method in LDL, cholesterol measurement in VLDL Method, Method for measuring triglyceride in VLDL, Method for measuring cholesterol in CM, Method for measuring triglyceride in CM, Method for measuring cholesterol in remnant lipoprotein, Remnan Neutral fat measuring method in lipoproteins and the like. Thus, the method for measuring the components to be measured for lipoproteins in a specimen by the homogenous method is that the lipoproteins in the specimen are not separated physically, and in the presence of multiple types of lipoproteins, Means a method of measuring the components of

[0025] 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 a standard product and a specimen are automatically weighed and measured.

Examples of the present invention are shown below, but the present invention is not limited to these. In this example, the following enzymes, reagents and instruments were used.

[0027] Azizuna sodium (manufactured by Wako Pure Chemical Industries, Ltd.), Bjarbin (product name, CV Y-2YT, manufactured by Sato Ampoule Co., Ltd.), freeze dryer (FDM 05S, manufactured by Sanpec Co., Ltd.).

Example 1

[0028] Manufacture of standard products for quantitative determination of components to be measured in samples containing lipoproteins

10 mL of a 10% sodium azide solution was added to 8 mL of fresh human serum, and 2 mL of ImolZL HEPES solution was added to this and stirred. This solution was aliquoted into vials in lm L, frozen at 40 ° C, dried and lyophilized at 13. 3-133.3 Pa for 20 hours to produce a standard product. In Tables 1 and 2, the standard product is described as HEPESO. 2molZL. This is equivalent to serum lOOmL supplemented with 25 mmol of buffer. A control using PBS instead of the HE PES solution was used as a control.

Example 2 [0029] Quantification of HDL cholesterol in samples using standard products

Each of the freeze-dried products prepared in Example 1 was dissolved in 0.5 mL of purified water, and the concentration of HDL cholesterol in the sample was determined as a commercially available homogenous HDL measuring reagent, Detaminaichi HDL (manufactured by Kyowa Medettas; frozen) Measurement was performed with an automatic analyzer (Hitachi 7170) using a dry reagent product (denoted as reagent DH) and Detamina I L HDL (manufactured by Kyowa Medettas; liquid reagent product (denoted as reagent LH)). Table 1 shows the measurement results.

[0030] [Table 1]

[0031] As is apparent from Table 1, the standard product containing HEPES was stronger than the control product containing no HEPES, with no difference in the measured values between the products.

Example 3

[0032] Quantification of components to be measured in samples containing lipoproteins using standard products

In order to examine the stability of the freeze-dried product obtained in Example 1, each freeze-dried product was stored for 1 week in a heated state at 40 ° C. Two reagent products for HDL cholesterol measurement used in Example 2, and Correstest N HDL [Daiichi Chemical Co., Ltd .; liquid reagent product (denoted as reagent NH)] and LDL cholesterol measurement reagent Detamina L LDL [ Using a liquid reagent product (reagent LL)] manufactured by Kyowa Medex Co., Ltd.], the measured values before and after storage were compared. The results are shown in Table 2. The numerical values in Table 2 show the ratio of the measured values before and after storage, that is, the measured cholesterol value in the standard product (control product) after storage Z) the measured cholesterol value in the standard product (control product) before storage.

[0033] [Table 2]

[0034] As shown in Table 2, for standard products containing HEPES, measured values were measured before and after storage. The fluctuation was kept below 2%, and the effect of adding HEPES was recognized.

Industrial applicability

According to the present invention, a standard product used for differential quantification of components to be measured in lipoprotein, useful for diagnosis of lifestyle-related diseases such as arteriosclerosis, a method for producing the standard product, A kit for fractional quantification of a component to be measured and a method for fractional quantification of the component to be measured in a specimen are provided.

Claims

The scope of the claims

[1] Standard product obtained by freeze-drying serum or plasma to which a sulfo group-containing buffer is added, which is used for differential quantification of components to be measured in lipoproteins. A standard product in a lyophilized state, characterized in that the content of the component to be measured is priced using a known amount of the component to be measured.

[2] Buffer power with sulfo group N— (2-hydroxyethyl) N, 1 (2-sulfoethyl) piperazine, piperazine 1 N, N, 1 bis (2-ethanesolephonic acid), N— (2-acetamido) -2 aminoethanesulfonic acid, 3 morpholino-2-hydroxypropanesulfonic acid, N, N bis (2 hydroxyethyl) 2 aminoethanesulfonic acid, 3 morpholinopropanosulfonic acid, N— [tris (hydroxy Methyl) methyl] -2-aminoethanesulfonic acid, 3- [N, N bis (2hydroxyethyl) amino] -2 hydroxypropanesulfonic acid, N- [tris (hydroxymethyl) methyl] -2 hydroxy-1 3 Minopropanesulfonic acid, piperazine 1 N, N, 1 bis (2 hydroxypropanesulfonic acid), 3- [4- (2 Hydrochichetyl) -1-piperazyl] -2 hydroxypropane Sulfonic acid, 3- [4- (2-hydroxyethyl) 1-piperadul] propanesulfonic acid, N-tris (hydroxymethyl) methyl 3-aminopropanesulfonic acid, N-cyclohexyl 2 aminoethanesulfone 2. The standard product according to claim 1, which is one or more buffering agents selected from acids, N-cyclohexyl 3amino 2 hydroxypropane sulfonic acid and N-cyclohexyl 3 amino propane sulfonic acid.

[3] The standard product according to claim 2, which is N- (2-hydroxyethyl) N, mono (2-sulfoethyl) piperazine having a sulfo group-containing buffer.

4. The standard product according to any one of claims 1 to 3, wherein the component to be measured in the lipoprotein is cholesterol or neutral fat.

5. The lipoprotein is a lipoprotein selected from the group consisting of high-density lipoprotein, low-density lipoprotein, very low-density lipoprotein, chylomicron, and remnant lipoprotein. Standard product.

[6] Serum or plasma and a sulfone-containing buffer are mixed, and the resulting mixture is lyophilized, and then the content of the component to be measured in the lyophilizate is determined to a known amount. Using A method for producing a standard product in a lyophilized state, which is used for differential quantification of components to be measured in lipoprotein, characterized in that

[7] Buffer power with sulfo group N— (2 Hydroxyethyl) N, One (2-Sulfoethyl) Piperazine, Piperazine One N, N, One Bis (2-Ethansnorephonic acid), N— ( 2-acetoamide) -2 aminoethanesulfonic acid, 3 morpholino-2-hydroxypropanesulfonic acid, N, N bis (2 hydroxyethyl) 2 aminoethanesulfonic acid, 3 morpholinopropanosulfonic acid, N— [tris (hydroxymethyl ) Methyl] -2-aminoethanesulfonic acid, 3- [N, N bis (2hydroxyethyl) amino] -2 hydroxypropanesulfonic acid, N- [tris (hydroxymethyl) methyl] -2 hydroxy-1-3amino Propanesulfonic acid, piperazine 1 N, N, 1 bis (2 hydroxypropanesulfonic acid), 3- [4- (2 Hydroxichetyl) -1-piperazyl] -2 hydroxypropane Ruphonic acid, 3- [4- (2-hydroxyethyl) 1-piperadul] propanesulfonic acid, N-tris (hydroxymethyl) methyl 3-aminopropanesulfonic acid, N-cyclohexyl 2 aminoethanesulfone 7. The production method according to claim 6, which is one or more buffering agents selected from acids, N-cyclohexyl 3amino 2 hydroxypropanesulfonic acid and N-cyclohexyl 3 aminopropanesulfonic acid.

8. The production method according to claim 7, wherein the buffering power having a sulfo group is N- (2 hydroxyethyl) N, mono (2-sulfoethyl) piperazine.

[9] The method according to any one of [6] to [8], wherein the component to be measured in the lipoprotein is cholesterol or neutral fat.

[10] The lipoprotein selected from the group consisting of high-density lipoprotein, low-density lipoprotein, ultra-low-density lipoprotein, chylomicron, and remnant lipoprotein strength according to any one of claims 6 to 9. Manufacturing method.

[11] A reagent for fractional measurement of a component to be measured in lipoprotein and the standard product described in any one of claims 1 to 5 should be used to measure in a sample containing lipoprotein Kit for fractional determination of components.

[12] (1) a step of measuring the component in a sample containing lipoprotein using a reagent for fractional measurement of the component to be measured in lipoprotein, (2) any one of claims 1 to 5 Standard products listed A step of preparing a calibration curve using the fractionation measurement reagent, and (3) the component in the sample from the measured value obtained in the step (1) and the calibration curve created in the step (2). A method for differential quantification of a component to be measured in a specimen containing lipoprotein, comprising a step of calculating a concentration.