1995 Fiscal Year Final Research Report Summary
Development of HETE Analysis by means of mass Spectrometry
| Project/Area Number |
06672265
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
応用薬理学・医療系薬学
|
|---|
| Research Institution | Tohoku University |
|---|
Principal Investigator |
OHYAMA Yoshiharu Tohoku University Hospital, Pharmaceatical Sciences, Assistant, 医学部附属病院・薬剤部, 助手 (30233290)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
HISHINUMA Takanori Tohoku University Hospital, Phamaceatical Sciences, Assistant Professor, 医学部附属病院・薬剤部, 助教授 (20199003)
MIZUGAKI Michinao Tohoku University Hospital, Phamaceatical Sciences, Professor, 医学部附属病院・薬剤部, 教授 (60004595)
|
|---|
| Project Period (FY) |
1994 – 1995
|
| Keywords | 20HETE / 19HETE / GC / ECNICIMS / PFB / BDMS / glucuronide conjugate |
|---|
| Research Abstract |
19-HEHE and 20-HETE is a major metabolite of arachidonic acid from cytochrome P-450 omega-hydroxylase in neutrophils and renal microsomes. 20-HETE is a potent vasoconstrictor and promotes sodium excretion. The in vitro biosynthesis of 20-HETE from exogenous AA by renal microsomes was increased during the development of hypertension in the SHR rat. This suggested a possible role for 20-HETE in blood pressure control. In order to assess whether 19-HETE and 20-HETE are formed in vivo in humans, we have developed a highly sensitive gas chromatography/electron capture negative ionization (EC) mass spectrometry assay, based on stable isotope dilution, for is quantification in human urine. Deuterated HETEs were used as an internal standard. The assay involves solid phase extraction of urine with subsequent TLC and HPLC purification. 19-HETE and 20-HETE were analyzed as the PFB ester, tert-butyldimethylsilyl ether derivative. EC mass spectra of derivatized 20-HETE and 19-HETE gave base peak ions at m/z433. In this studies we have observed that 20-HETE is excreted primarily as a glucuronide conjugate but 19-HETE is not excreted primarily as a glucuronide conjugate. We established whether glucuronidation had occurred on the hydroxyl moieties by CID-FABMS/MS.
|
