JPS6375558A - Glycohemoglobin analysis method - Google Patents
Glycohemoglobin analysis methodInfo
- 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
- glycohemoglobin
- column
- liquid
- analysis method
- analysis
- 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
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
(57)【要約】本公報は電子出願前の出願データであるた
め要約のデータは記録されません。(57) [Abstract] This bulletin contains application data before electronic filing, so abstract data is not recorded.
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 | Glycohemoglobin analysis method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61221590A JPS6375558A (en) | 1986-09-18 | 1986-09-18 | Glycohemoglobin analysis method |
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 | Glycohemoglobin analysis method |
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 | Liquid chromatograph |
| 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 |
| US5358639A (en) * | 1992-04-01 | 1994-10-25 | Hitachi, Ltd. | Method of analyzing hemoglobins |
| 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 (11)
| 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 | Glycohemoglobin separation method, separation device, and separation column |
| US5348649A (en) * | 1990-07-20 | 1994-09-20 | Hitachi, Ltd. | Apparatus for measuring glycohemoglobin |
| JPH055730A (en) * | 1990-11-30 | 1993-01-14 | Hitachi Ltd | Liquid chromatograph |
| 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 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Allison et al. | Dual electrode liquid chromatography detector for thiols and disulfides | |
| Gebauer et al. | Recent application and developments of capillary isotachophoresis | |
| AU609361B2 (en) | Separation of hemoglobin A2 from hemoglobin mixture | |
| Novotny | Capillary biomolecular separations | |
| Kirkegaard et al. | Ion filtration chromatography: a powerful new technique for enzyme purification applied to E. coli alkaline phosphatase | |
| JP3012685B2 (en) | Method and apparatus for analyzing amino acids in biological fluid | |
| Lam et al. | Stereoselective D-and L-amino acid analysis by high-performance liquid chromatography | |
| JP2920092B2 (en) | Method for quantitative measurement of glycated protein | |
| JPH0477500A (en) | Glycohemoglobin separation method, separation device, and separation column | |
| Mario et al. | Capillary zone electrophoresis for the diagnosis of congenital hemoglobinopathies | |
| Fowler | Amino acid sequence of beta-galactosidase. VII. Isolation of the 24 cyanogen bromide peptides | |
| Vacik et al. | Separation and measurement of isoenzymes and other proteins by high-performance liquid chromatography | |
| Candiano et al. | Negative staining of proteins in polyacrylamide gels with methyl trichloroacetate | |
| Maltsev et al. | Determination of myoglobin in human serum by high-performance liquid chromatography with chemiluminescence detection | |
| Anderson | Affinity chromatography of lactate dehydrogenase: An experiment for the undergraduate biochemistry laboratory. | |
| Guaratini et al. | Cathodic stripping voltammetric detection and determination at a hanging mercury-drop electrode of dye contaminants in purified biomaterials: study of the human serum albumin and reactive dye 120 system | |
| Roush et al. | Preferred high-performance liquid chromatographic anion-exchange chromatographic contact region for recombinant rat cytochrome b5 | |
| Iwamoto et al. | Urinary uric acid determination by reversed-phase high-performance liquid chromatography with electrochemical detection | |
| Williams | Capillary electrophoresis in the analytical and physical chemistry laboratories | |
| Weinstein | Enantiomeric analysis of the common protein amino acids by liquid chromatography | |
| Frank et al. | Automated derivatization and gas chromatographie analysis | |
| Katō | Progress in HPLC.: Gel permeation and ion-exchange chromatography of proteins and peptides | |
| Ianniello | Determination of Suttocide® A in Cosmetic Formulations by Single Column Ion Chromatography with Pulsed Amperometric Detection (SCIC-PAD) | |
| Hnatowich | High performance liquid chromatography in studies of radiolabeled antibodies | |
| Kovář | High performance liquid chromatography and biotechnological application of several oxidoreductases |