| Project/Area Number |
08672484
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Physical pharmacy
|
|---|
| Research Institution | HEALTH SCIENCES UNIVERSITY OF HOKKAIDO |
|---|
Principal Investigator |
TOHMA Masahiko HEALTH SCIENCES UNIV.OF HOKKAIDO,FAC.PHARMACEUTICAL SCIENCES,PROFESSOR, 薬学部, 教授 (30001043)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KIMURA Akihiko KURUME UNIVERSITY,SCHOOL OF MEDICINE,DEPT.OF PEDIATRICS & CHILD HEALTH,ASSISTANT, 医学部, 助手 (00211201)
YOSHIMURA Teruki HEALTH SCIENCES UNIV.OF HOKKAIDO,FAC.PHARMACEUTICAL SCIENCES,INSTRUCTOR, 薬学部, 講師 (60220737)
MURAI Tsuyoshi HEALTH SCIENCES UNIV.OF HOKKAIDO,FAC.PHARMACEUTICAL SCIENCES,ASSISTANT, 薬学部, 助手 (50260823)
|
|---|
| Project Period (FY) |
1996 – 1997
|
| Project Status |
Completed (Fiscal Year 1997)
|
| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1997: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
|
|---|
| Keywords | BILE ACID BIOSYNTHESIS / CONGENITAL DISORDER / CHOLESTASIS / GAS CHROMATOGRAPHY-MASS SPECTROMETRY / HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY / ENZYME IMMUNOASSAY / 3-OXO-DELTA^4-BILE ACID / 3-OXO-DELTA^4-STEROID 5beta-REDUCTASE / 3-オキソ-Δ^4-ステロイド5β-還元酵素 / 酵素免疫測定法 / 3-オキソ-Δ^1-ステロイド5β-還元酵素 / 胎児-新生児 / 酵素免疫学的測定法 / ガスクロマトグラフィー質量分析 |
|---|
| Research Abstract |
The estsablishment of the diagnosis for congenital liver disease is urgently required for the remedy of liver disorder in early life. The metabolism of bile acids in fetus and neonate is thought to be different from that in adults. The aim of this reaserch is the developement of quantitative analysis of 3-oxo-DELTA^4-bile acids by GC-MS,HPLC and immunological assay, the elucidation of the bile acid profiles in neonatal cholestatic diseases, and the establishment of the diagnosis for the congenital bile acid disorder.
1.The 3-oxo-DELTA^4-bile acids were synthesized chemically from cholic and chenodeoxycholic acids, and their conjugates were also prepared as the authentic samples by condensation with glycine, taurine and/or sulfuric acid.
2.A highly sensitive determination method of the 3-oxo-DELTA^4-bile acids and related bile acids in neonatal urine has been developed by GC-MS using selected ion monitoring with their characteristic fragment ions. Diagnostic bile acid analysis for neonatal liver disease was performed by GC-MS.
3.An improved quantitative analysis of the conjugated 3-oxo-DELTA^4-bile acids in neonatal urine has been developed by HPLC employing the dual wavelength UV detection at 245 and 280nm for 3-oxo-DELTA^4- and 3-oxo-DELTA^<4,6>-bile acids, respectively. These bile acids were predominantly excreted in urine of patients as the glycine and taurine conjugates.
4.An enzyme immunoassay of conjugated 7alpha-hydroxy-3-oxo-DELTA^4-cholanoic acid has been developed using horse radish peroxidase labeled antigen. The conjugated bile acid in urine from patients were significantly higher than that of normal newborns.
|
