HPLC/CHEMILUMINESCENCE OF OPIOID PEPTIDES

阿片肽的 HPLC/化学发光

• 批准号: 3210411
• 负责人: JOHN F STOBAUGH
• 金额: $ 9.43万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-04-01 至 1992-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3210411
• 关键词: enkephalins; fluorescent dye /probe; high performance liquid chromatography; laboratory rat; luminescence; neuropeptides

The opioid peptides are biologically important products resulting from the biochemical processing of three precursor proteins: preproopiomelanocortin, preproenkephalin A and preproenkephalin B. The primary objective of this research is the development of direct chemical assay methodology which has the required selectivity and sensitivity to unambiguously quantify opioid peptides, especially, Leu-enkephanlin and Met-enkephalin, in biologically relevant tissues. This overall objective is being accomplished via the unique combination of fluorogenic derivatization, multidimensional HPLC and chemiluminescence detection. Specifically, the primary amine groups of the peptides are reacted with the novel NDA reagent (naphthalene-2,3- dialdehyde plus cyanide) to form stable and intensely fluorescent N-substituted 1-cyanobenz f isoindole (CBI) derivatives. Next, multidimensional chromatography is employed to achieve the desired selectivity. The CBI derivatives are initially fractionated on an HPLC column which retains the analytes by one mechanism (eg., reversed phase) and then switched automatically to a second column with a completely different mechanism (eg., ion-pair). Finally, the resolved analytes are detected via chemiluminescence of the CBI moiety, generated by an aryloxalate/H2O2 post-column reaction detector. The following studies will be carried out to optimize the approach: 1) the rate and yield of the NDA reaction will be studied using several peptides including those with multiple reaction sites, 2) the luminescent properties of the CBI peptides wil be characterized paying particular attention to the products of multiple derivatization, 3) the systematic characterization of the retention of CBI peptides on HPLC columns with different retention mechanisms will be carried out, 4) the characterization of the post-column chemiluminescent process will include: reactor design, flow-cell design, use of novel aryloxalate esters and reaction conditions. The results obtained will permit the unification of these various components into a system that will permit the direct chemical determination of opioid peptides with excellent selectivity and sensitivity (limits of detection of 1-10 femtomole on-column). The resulting methodology will be applicable to the quantification of endogenous or exogenously administered opioid peptides that could be easily transferred to other laboratories engaged in this area of peptide research without the addition of highly specialized equipment.

阿片肽是重要的生物学产物

项目成果

Fluorogenic derivatization of peptides with naphthalene-2,3-dicarboxaldehyde/cyanide: optimization of yield and application in the determination of leucine-enkephalin spiked human plasma samples.

用萘-2,3-二甲醛/氰化物对肽进行荧光衍生化：产量优化及其在测定亮氨酸脑啡肽加标人血浆样品中的应用。

• DOI: 10.1016/0731-7085(90)80070-6
• 发表时间: 1990
• 期刊: Journal of pharmaceutical and biomedical analysis
• 影响因子: 3.4
• 作者: deMontigny,P;Riley,CM;Sternson,LA;Stobaugh,JF
• 通讯作者: Stobaugh,JF

Reversed-phase liquid chromatography of the opioid peptides. 3. Development of a microanalytical system for opioid peptides involving microbore liquid chromatography, post-column derivatization and laser-induced fluorescence detection.

阿片肽的反相液相色谱。

• DOI: 10.1016/0731-7085(91)80106-j
• 发表时间: 1992
• 期刊: Journal of pharmaceutical and biomedical analysis
• 影响因子: 3.4
• 作者: Dave,KJ;Stobaugh,JF;Rossi,TM;Riley,CM
• 通讯作者: Riley,CM

Reversed-phase liquid chromatography of the opioid peptides--2. Quantitative structure-retention relationships and isocratic retention prediction.

阿片肽的反相液相色谱--2。

• DOI: 10.1016/0731-7085(92)80010-k
• 发表时间: 1992
• 期刊: Journal of pharmaceutical and biomedical analysis
• 影响因子: 3.4
• 作者: Dave,K;Stobaugh,JF;Riley,CM
• 通讯作者: Riley,CM

Multidimensional liquid chromatography of opioid peptides: fluorogenic labelling, retention prediction and separation optimization.

阿片肽的多维液相色谱：荧光标记、保留预测和分离优化。

• DOI: 10.1016/0731-7085(90)80125-9
• 发表时间: 1990
• 期刊: Journal of pharmaceutical and biomedical analysis
• 影响因子: 3.4
• 作者: Nicholson,LM;Patel,HB;Kristjansson,F;CrowleyJr,SC;Dave,K;Stobaugh,JF;Riley,CM
• 通讯作者: Riley,CM

Reversed-phase ion-pair liquid chromatography of the angiotensins.

血管紧张素的反相离子对液相色谱。

• DOI: 10.1016/s0021-9673(01)89270-0
• 发表时间: 1991
• 期刊: Journal of chromatography
• 作者: Patel,HB;Stobaugh,JF;Riley,CM
• 通讯作者: Riley,CM