Simultaneous Detection of Cholyl Adenylate and Coenzyme A Thioester Utilizing Liquid Chromatography/Electrospray Ionization Mass Spectrometry.
-
- MANO Nariyasu
- Graduate School of Pharmaceutical Sciences, Tohoku University
-
- UCHIDA Masashi
- Graduate School of Pharmaceutical Sciences, Tohoku University
-
- OKUYAMA Hiroko
- Graduate School of Pharmaceutical Sciences, Tohoku University
-
- SASAKI Ikuko
- Graduate School of Pharmaceutical Sciences, Tohoku University
-
- IKEGAWA Shigeo
- Faculty of Pharmaceutical Sciences, Kinki University
-
- GOTO Junichi
- Graduate School of Pharmaceutical Sciences, Tohoku University
この論文をさがす
抄録
It has been proposed that acyl adenylate is first formed during activation of the carboxy group into the acyl CoA thioester, an intermediate in the formation of amino acid conjugates. Acyl CoA synthetases may be responsible for this acyl adenylate formation. Recently, we hypothesized the preferential formation of cholic acid adenylate, a major bile acid, preceding production of the corresponding CoA thioester in incubations with rat liver microsomal fractions. To verify this biosynthetic mechanism, monitoring of the incubation mixture of acyl adenylate together with both substrate and acyl CoA thioester is needed. We have developed a detection method for the simultaneous detection of these cholic acid derivatives utilizing liquid chromatography/electrospray ionization mass spectrometry. The CoA thioester of cholic acid forms a chelation complex with the divalent cations remaining on the silica gel packed into the analytical column. Both the addition of a chelating agent, such as EDTA, to the mobile phase and an adjustment of the mobile phase pH to a weak alkaline effectively removed such chelate formation, producing a sharp CoA thioester peak. For a simultaneous mass spectrometric analysis of cholic acid, the corresponding adenylate and CoA thioester, the combined use of a 300 Å particle diameter ODS column and 20 mM ammonium acetate buffer (pH 9.0)/2-propanol/acetonitrile as the mobile phase have been proved to be preferable. To avoid any degradation of the chemically unstable adenylate produced in the incubation, we employed a direct injection of the sample onto a preconcentration column. The obtained results indicated a high sensitivity of this method.
収録刊行物
-
- Analytical Sciences
-
Analytical Sciences 17 (9), 1037-1042, 2001
社団法人 日本分析化学会
- Tweet
キーワード
詳細情報 詳細情報について
-
- CRID
- 1390282679234074752
-
- NII論文ID
- 10007177608
-
- NII書誌ID
- AA10500785
-
- COI
- 1:CAS:528:DC%2BD3MXntFWqtbs%3D
-
- ISSN
- 13482246
- 09106340
-
- NDL書誌ID
- 5911560
-
- PubMed
- 11708055
-
- 本文言語コード
- en
-
- データソース種別
-
- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
-
- 抄録ライセンスフラグ
- 使用不可