High-performance liquid chromatographic analysis of arginine-containing peptides in biological fluids by means of a selective post-column reaction with fluorescence detection.
High-performance liquid chromatographic analysis of arginine-containing peptides in biological fluids by means of a selective post-column reaction with fluorescence detection.
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通过选择性柱后反应和荧光检测，对生物液体中的含精氨酸肽进行高效液相色谱分析。
DOI：
10.1016/s0021-9673(01)94015-4
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
V. Boppana
中科院分区：
文献类型：
--
作者：
G. Rhodes;V. Boppana
描述了用于生物基质中含精氨酸肽的荧光检测的高灵敏度和特异的高性能液相色谱方法。 HPLC 分离后，精氨酸部分的胍基与茚三酮在碱性条件下发生柱后反应，生成荧光肽产物，可以高灵敏度测量。仔细优化柱后反应条件，并使用内径较小的 HPLC 色谱柱，使柱上检测限低至 50 fmol。应用于测定人血浆中合成的含精氨酸的加压素拮抗剂，产生了在 0.5-100 pmol/ml 范围内呈线性的定量响应。该测定方法足够灵敏、准确和精确，可用于这些合成肽的药代动力学研究。该方法在天然存在的含精氨酸肽的检测中也具有普遍适用性。
Highly sensitive and specific high-performance liquid chromatograhic methodology for the fluorescence detection of arginine-containing peptides in biological matrices is described. Following HPLC separation, a post-column reaction of the guanidino group of the arginine moiety with ninhydrin under basic conditions is utilized to generate a fluorescent peptide product which can be measured at high sensitivity. Careful optimization of the post-column reaction conditions, and the use of HPLC columns of reduced internal diameter, resulted in on-column detection limits as low as 50 fmol. Application to the determination of synthetic arginine-containing vasopressin antagonists in human plasma resulted in a quantitative response which is linear over the range 0.5–100 pmol/ml. The assay method is sufficiently sensitive, accurate, and precise for use in pharmacokinetic studies of these synthetic peptides. The methodology also has general applicability in the detection of naturally occurring arginine-containing peptides.