Rare excitatory amino acid from flowers of zonal geranium responsible for paralyzing the Japanese beetle.

The Japanese beetle (JB), Popillia japonica, exhibits rapid paralysis after consuming flower petals of zonal geranium, Pelargonium x hortorum. Activity-guided fractionations were conducted with polar flower petal extracts from P. x hortorum cv. Nittany Lion Red, which led to the isolation of a paralysis-inducing compound. High-resolution-MS and NMR ((1)H, (13)C, COSY, heteronuclear sequential quantum correlation, heteronuclear multiple bond correlation) analysis identified the paralytic compound as quisqualic acid (C(5)H(7)N(3)O(5)), a known but rare agonist of excitatory amino acid receptors. Optical rotation measurements and chiral HPLC analysis determined an L-configuration. Geranium-derived and synthetic L-quisqualic acid demonstrated the same positive paralytic dose-response. Isolation of a neurotoxic, excitatory amino acid from zonal geranium establishes the phytochemical basis for induced paralysis of the JB, which had remained uncharacterized since the phenomenon was first described in 1920.

Measurement of glutamate and aspartate in Planaria.

INTRODUCTION: The major excitatory neurotransmitters in the mammalian central nervous system are glutamate and aspartate. We developed a rapid and efficient method for the extraction and measurement of these amino acids in Planaria--a valuable model for mammalian processes because of their simple, centralized nervous system and similar neurotransmitter systems. METHOD: The method utilized buffer extraction (perchloric acid containing 0.025% of L-cystine and Na2EDTA), simple derivatization, high-pressure liquid chromatography (HPLC), and fluorescence detection. RESULTS: The mean+/-S.E.M. amounts of glutamate and aspartate were 322.6+/-43.6 and 188.6+/-27.6 pmol/mg-planarian, respectively. DISCUSSION: The method provides the ability to investigate changes in glutamate and aspartate in response to drug administration or withdrawal.

A facile and sensitive chemiluminescence detection of amino acids in biological samples after capillary electrophoretic separation.

It was found that native amino acids enhanced the chemiluminescence (CL) reaction between luminol and BrO(-) in an alkaline aqueous solution. This has led to the development of a facile and highly sensitive CL detection scheme for the determination of amino acids in biological samples after capillary electrophoretic (CE) separation. The CE-CL conditions were optimized. An electrophoretic buffer of 2.5 x 10(-2) M sodium borate (pH 9.4) containing 1 x 10(-4) M luminol was used. The oxidizer solution of 8 x 10(-4) M NaBrO in 0.1 M sodium carbonate buffer solution (pH 12.5) was introduced post-column. Under the optimal conditions, the detection limits were 1.0 x 10(-7) M for glutamic acid (Glu) and 1.3 x 10(-7) M (S/N = 3) for aspartic acid (Asp). The relative standard deviations (RSDs) of peak area and migration time were in the ranges of 3.8-4.3% and 1.4-1.6%, respectively. The present method was applied to the determination of excitatory amino acids (i.e., Asp and Glu) in rat brain tissue and monkey plasma. The levels of these major excitatory amino acids in monkey plasma were quantified for the first time and found to be 1.17 +/- 0.17 x 10(-5) M (mean +/- SD, n = 6) for Glu and 1.64 +/- 0.19 x 10(-6) M for Asp, which were comparable with the levels in human plasma.

Isolation and characterization of four novel bioactive peptides from a polychaete annelid, Perinereis vancaurica.

Four bioactive peptides were purified from an extract of the polycheate annelid Perinereis vancaurica using an HPLC system for fractionation of the extract and the esophagus of the worm as bioassay system. The sequences of the peptides were determined to be H-Trp-Val-Val-Gly-Asp-Val-Gln-OH, H-Ala-Thr-Trp-Leu-Asp-Thr-OH, H-Trp-Met-Val-Gly-Asp-Val-Gln-OH and H-Phe-Tyr-Glu-Gly-Asp-Val-Pro-Tyr-OH. These peptides showed an excitatory activity on the esophagus of P. vancaurica. The excitatory effect of the second and fourth peptides was marked. They may be neuropeptides involved in regulation of the esophagus.