JPS6375558A - Analysis of glycohemoglobin - Google Patents
Analysis of glycohemoglobinInfo
- Publication number
- JPS6375558A JPS6375558A JP61221590A JP22159086A JPS6375558A JP S6375558 A JPS6375558 A JP S6375558A JP 61221590 A JP61221590 A JP 61221590A JP 22159086 A JP22159086 A JP 22159086A JP S6375558 A JPS6375558 A JP S6375558A
- Authority
- JP
- Japan
- Prior art keywords
- column
- glycohemoglobin
- silica
- contg
- ion exchange
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000004458 analytical method Methods 0.000 title claims abstract description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 8
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims abstract description 5
- 238000005342 ion exchange Methods 0.000 claims abstract description 5
- 239000000463 material Substances 0.000 claims abstract description 5
- 102000017011 Glycated Hemoglobin A Human genes 0.000 claims description 6
- 108091005995 glycated hemoglobin Proteins 0.000 claims description 6
- 239000003480 eluent Substances 0.000 claims description 3
- 239000007853 buffer solution Substances 0.000 claims 1
- 210000004369 blood Anatomy 0.000 abstract description 6
- 239000008280 blood Substances 0.000 abstract description 6
- 239000000872 buffer Substances 0.000 abstract description 4
- 238000010828 elution Methods 0.000 abstract description 3
- 206010012601 diabetes mellitus Diseases 0.000 abstract description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 abstract 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 abstract 1
- 238000003745 diagnosis Methods 0.000 abstract 1
- 239000007788 liquid Substances 0.000 description 9
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 239000008057 potassium phosphate buffer Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 206010008631 Cholera Diseases 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- HVVWZTWDBSEWIH-UHFFFAOYSA-N [2-(hydroxymethyl)-3-prop-2-enoyloxy-2-(prop-2-enoyloxymethyl)propyl] prop-2-enoate Chemical compound C=CC(=O)OCC(CO)(COC(=O)C=C)COC(=O)C=C HVVWZTWDBSEWIH-UHFFFAOYSA-N 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 239000003219 hemolytic agent Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 description 1
- 229910052939 potassium sulfate Inorganic materials 0.000 description 1
- 235000011151 potassium sulphates Nutrition 0.000 description 1
- UWHCKJMYHZGTIT-UHFFFAOYSA-N tetraethylene glycol Chemical compound OCCOCCOCCOCCO UWHCKJMYHZGTIT-UHFFFAOYSA-N 0.000 description 1
Abstract
Description
【発明の詳細な説明】
（イ）産業上の利用分野
本発明は試料（血液）中のグリコヘモグロビン（Ａ、ｅ
ａ ｌ ＡＺ ｂｌ Ｆ Ｉ Ｌ ＡＺ’ Ｃ＊ ｓ
−ＡＪ ｃ ）の分析法に関する。DETAILED DESCRIPTION OF THE INVENTION (a) Industrial application field
a l AZ bl F I L AZ' C* s
-AJ c ) analysis method.
（ロ）従来技術
グリコヘモグロビンは、赤血球中のヘモグロビンが体内
全循環している間に非酵素的に血液中のグルコースと反
応して形成される。(b) Prior Art Glycated hemoglobin is formed by non-enzymatic reaction of hemoglobin in red blood cells with glucose in the blood while circulating throughout the body.
そして、このグリコヘモグロビンハ、長期間の血液中グ
ルコースの時間平均的な濃度を表示していることから、
グリコヘモグロビンの分析により糖尿病の診断を行って
いる・この分析法としては１例えば特開昭５６−９８６
５７号、特開昭５７−２６７４６号がある。Since this glycated hemoglobin displays the time-averaged concentration of glucose in the blood over a long period of time,
Diabetes is diagnosed by analyzing glycated hemoglobin.This analytical method is 1, for example, JP-A-56-986.
No. 57 and Japanese Unexamined Patent Publication No. 57-26746.
コレラは、いずれもテトラエチレングリコール、テトラ
メチロールメタントリアクリレート等の単量体を基材と
する多異性ポリマーを充てんしたカラムを用いて分析し
ている・（／最発明が解決しようとする問題点
上記物質を基材とするカラムは、シリカを基材とするも
のに比べ耐圧性９分離能共に劣り、特に短いカラムで長
時間安定な分析を行うとき問題となった。Cholera is analyzed using columns packed with polyisomer polymers based on monomers such as tetraethylene glycol and tetramethylolmethane triacrylate. Columns based on the above-mentioned substances are inferior to those based on silica in terms of pressure resistance and resolution, which poses a problem particularly when performing stable analysis over a long period of time with a short column.
また、カラムのコストの点でも、上記物質を基材とする
ものは比較的高価であった。Also, in terms of cost, columns based on the above substances were relatively expensive.
に）問題点全解決するための手段及び作用本発明は、ｐ
Ｈ５，０〜６０の緩衝液を溶離液とし。) Means and operations for solving all the problems The present invention provides p
Use H5.0-60 buffer as eluent.
シリカが基材でカルボキシル基全イオン交換基とするカ
ラム金柑いて試料中のグリコヘモグロビン全分析するこ
とを特徴とする。The column uses silica as the base material and all carboxyl groups are ion exchange groups, and is characterized by analyzing all the glycated hemoglobin in the sample.
また９分析に当っては時間と共に溶離液のＰＨ’ｔ”変
化させるグラジェント溶離法により分離を行う。In addition, in the 9 analysis, separation is performed by a gradient elution method in which the pH of the eluent is changed over time.
なお、シリカが基材でカルボキシル基をイオン交換基と
するカラムとしては３ｂｉｍ −ｐａｃｋ ＷＣＸ−
１（４ｍｍ１．Ｄ、Ｘ ５０ ｃｍＬ、 、高滓製作所
製）、緩衝液と１〜てはリン酸緩衝液が最適で、そのｐ
Ｈは分離カラムの劣化、各成分の溶出位置の調整という
観点から５．０〜６．０が最適である。In addition, 3bim-pack WCX- is a column with silica as the base material and carboxyl group as the ion exchange group.
1 (4 mm 1.D,
H is optimally 5.0 to 6.0 from the viewpoint of deterioration of the separation column and adjustment of the elution position of each component.
（ト）実施例
本発明全実施例を用いて説明する。試料（血液）を溶血
剤により処理後１次の分析条件で分析した。(G) Examples The present invention will be explained using all the examples. The sample (blood) was treated with a hemolytic agent and then analyzed under the primary analysis conditions.
カラム： Ｓｈｉｍ−ｐａｃｋ ＷＣＸ−１（４ｍｍＩ
、Ｄ、Ｘ ５ｃｍＬ、高滓製作所製）移動相： Ｃｋ）
８０ｍＭ ！Ｊン酸カリウム緩衝液（ｐＨ５，７８）
（Ｂ） １７０ｍＭリン酸カリウム緩衝液（ｐＨ５，４
幻
５０ｍＭ硫酸カリウム
（Ａ） 、 （Ｂ） ２液を準備し、これ會もとに移動
相全作製する。Column: Shim-pack WCX-1 (4mmI
, D, X 5cmL, manufactured by Takashi Seisakusho) Mobile phase: Ck)
80mM! Potassium phosphate buffer (pH 5,78) (B) 170mM potassium phosphate buffer (pH 5,4
Prepare two solutions of 50mM potassium sulfate (A) and (B), and prepare all the mobile phases based on these.
○第１液：（５）液をそのまま用いる。○First liquid: Use liquid (5) as is.
○第２液：（４）液８５チ （ＢＪ液１５％ｐＨは混合
後５．７４となる。○Second liquid: (4) liquid 85 cm (BJ liquid 15% pH is 5.74 after mixing.
○第３液：（４）液４５％ の）液５５％ｐＨは混合後
５５８となる。○Third liquid: (4) liquid 45%) The liquid 55% pH is 558 after mixing.
カラム温度：室温
グラジェントプログラム：０．０１〜７分第２液移動相
流ｌｉｔ ： １．Ｏｍ１３７ｍ ｉ ｎ検 出 器：
Ｕｖ検出器
上記の分析条件で得られたクロマトグラム全第１図に示
す。Column temperature: room temperature Gradient program: 0.01-7 minutes Second liquid mobile phase flow lit: 1. Om137min detector:
Uv Detector The complete chromatogram obtained under the above analysis conditions is shown in FIG.
１がＡｌａ、２がＡＪｂ、３がＦ、４がＬ−Ａ７ｃ、５
が５−ＡＪＣＩ ６がＡ、Ｏである。1 is Ala, 2 is AJb, 3 is F, 4 is L-A7c, 5
is 5-AJCI 6 is A, O.
（へ）効 果
本発明によれば、耐圧性に優れたシリカを基材とするカ
ラムを用いているので、カラムの長さを短くすることが
でき、しかも分析精度が向上する。(f) Effects According to the present invention, since a column based on silica having excellent pressure resistance is used, the length of the column can be shortened, and analysis accuracy is improved.
第１図は９本発明によってグリコヘモグロビンを分析し
たときのクロマトグラムを示す。
ＥＩＪＪ乃−」−。FIG. 1 shows a chromatogram when glycated hemoglobin was analyzed according to the present invention. EIJJノ-”-.
Claims (1)
が基材でカルボキシル基をイオン交換基とするカラムを
用いて試料中のグリコヘモグロビンを分析することを特
徴とするグリコヘモグロビン分析法。1. Glycohemoglobin analysis, which is characterized by analyzing glycated hemoglobin in a sample using a buffer solution with a pH of 5.0 to 6.0 as an eluent and a column with silica as a base material and carboxyl groups as ion exchange groups. Law.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61221590A JPS6375558A (en) | 1986-09-18 | 1986-09-18 | Analysis of glycohemoglobin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61221590A JPS6375558A (en) | 1986-09-18 | 1986-09-18 | Analysis of glycohemoglobin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS6375558A true JPS6375558A (en) | 1988-04-05 |
Family
ID=16769134
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61221590A Pending JPS6375558A (en) | 1986-09-18 | 1986-09-18 | Analysis of glycohemoglobin |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6375558A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4124058A1 (en) * | 1990-07-20 | 1992-01-23 | Hitachi Ltd | Glyco haemoglobin measurement by HPLC - by simultaneously sepg. glyco haemoglobin, haemoglobin and haemoglobin derivs. and removing labile haemoglobin to measure stable haemoglobin |
| JPH055730A (en) * | 1990-11-30 | 1993-01-14 | Hitachi Ltd | Apparatus for liquid chromatography |
| EP0563865A2 (en) * | 1992-04-01 | 1993-10-06 | Hitachi, Ltd. | Method and apparatus for analyzing hemoglobins and solution for suppressing column deterioration for use therein |
| US5292818A (en) * | 1982-07-20 | 1994-03-08 | Sekisui Kagaku Kogyo Kabushiki Kaisha | Method for producing a carrier for cation exchange liquid chromatography and a method for determining glycosylated hemoglobins using the carrier |
| US5294336A (en) * | 1989-09-18 | 1994-03-15 | Hitachi, Ltd. | Apparatus for liquid chromatography for separating AIC components from hemoglobin in blood |
| US5407568A (en) * | 1992-09-17 | 1995-04-18 | Hitachi, Ltd. | Separation column containing S-carboxyalkylcysteine |
| US5730867A (en) * | 1988-06-10 | 1998-03-24 | Drew; Keith Raymond | Method and apparatus for low pressure liquid chromatography |
-
1986
- 1986-09-18 JP JP61221590A patent/JPS6375558A/en active Pending
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5292818A (en) * | 1982-07-20 | 1994-03-08 | Sekisui Kagaku Kogyo Kabushiki Kaisha | Method for producing a carrier for cation exchange liquid chromatography and a method for determining glycosylated hemoglobins using the carrier |
| US5730867A (en) * | 1988-06-10 | 1998-03-24 | Drew; Keith Raymond | Method and apparatus for low pressure liquid chromatography |
| US5294336A (en) * | 1989-09-18 | 1994-03-15 | Hitachi, Ltd. | Apparatus for liquid chromatography for separating AIC components from hemoglobin in blood |
| DE4124058A1 (en) * | 1990-07-20 | 1992-01-23 | Hitachi Ltd | Glyco haemoglobin measurement by HPLC - by simultaneously sepg. glyco haemoglobin, haemoglobin and haemoglobin derivs. and removing labile haemoglobin to measure stable haemoglobin |
| JPH0477500A (en) * | 1990-07-20 | 1992-03-11 | Hitachi Ltd | Method for separating glycohemoglobin and separation device therefor |
| US5348649A (en) * | 1990-07-20 | 1994-09-20 | Hitachi, Ltd. | Apparatus for measuring glycohemoglobin |
| JPH055730A (en) * | 1990-11-30 | 1993-01-14 | Hitachi Ltd | Apparatus for liquid chromatography |
| EP0563865A2 (en) * | 1992-04-01 | 1993-10-06 | Hitachi, Ltd. | Method and apparatus for analyzing hemoglobins and solution for suppressing column deterioration for use therein |
| US5358639A (en) * | 1992-04-01 | 1994-10-25 | Hitachi, Ltd. | Method of analyzing hemoglobins |
| US5468379A (en) * | 1992-04-01 | 1995-11-21 | Hitachi, Ltd. | Solution for suppressing deterioration of a separation column for analyzing hemoglobins |
| EP0563865A3 (en) * | 1992-04-01 | 1997-10-15 | Hitachi Ltd | Method and apparatus for analyzing hemoglobins and solution for suppressing column deterioration for use therein |
| US5407568A (en) * | 1992-09-17 | 1995-04-18 | Hitachi, Ltd. | Separation column containing S-carboxyalkylcysteine |
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