Study of analytical method for conjugated carbohydrates by high performance liquid chromatography.
| Project/Area Number |
62550542
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
工業分析化学
|
|---|
| Research Institution | University of Tokyo |
|---|
Principal Investigator |
WATANABE Noriyuki Department of Industrial Chemistry, Faculty of Engineering, University of Tokyo, Research Associate,, 工学部工業化学科, 助手 (20011116)
|
|---|
| Project Period (FY) |
1987 – 1988
|
| Project Status |
Completed (Fiscal Year 1988)
|
| Budget Amount *help |
¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1988: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1987: ¥1,100,000 (Direct Cost: ¥1,100,000)
|
|---|
| Keywords | Conjugated carbohydrates / High performance liquid chromatography / High sensitive / Galactose infusion test / Electric dialysis / 電気透析 / イノシトールの新しい検出法 / 電気化学検出 / 酸加水分解 / イノシトール / 高速液クロ / 分離分析 |
|---|
| Research Abstract |
Further improvement in electrochemical detection of carbohydrate(CBP method), which has been developed by us was attained. Influence of oxygen dissolved or dissolving from air through teflon tube in eluent and reagent solution was studied in detail. Combination of fused silica capillary tube as the reaction coil, dry reactor bath and on-line degasser made the operation easier and the sensitivity much better. The CBP method was applied to the determination of galactose in human plasma from patients with hepatic diseases undergoing continuous low level galactose infusion. The CBP method proved to be more sensitive and selective than the fluorometric method which had been the most sensitive method so far.
Various separation methods for carbohydrates were tried intensively. Reversed phase ion-pair chromatography seemed to be plausible for acidic carbohydrates. Further study will be required. Amino and acidic carbohydrates provided different retentions among each other by chromatography using hidroxyapatite. Attempt of reversed phase ion-pair mode for neutral sugars failed. Typical components of conjugated carbohydrates(fucose, galactose, glucose and mannose) could be separated within 60 minutes by anion exchange column(HPIC-AS6, dionex) and water as eluent. This separation mode coupled with CBP detection resulted in highly sensitive analytical method tens to hundreds fmol level. An effective method removing concentrated acid from small sample of acid hydrolysate of conjugated carbohydrates was developed by electric dialysis using new kind of anion exchange membrane.
Novel detection method of inositol-3-phosphate, which attracts a lot of attention as second messenger in bio-transduction system was tried. It was found that a coupling od CBP method with a catalytic column as post-column reactor led to high sensitive detection of inositol. This novel detection is expected to be very attractive.
|
Report
(3 results)
Research Products
(6 results)
