Detection of dexamethasone in the cornea and lens by NMR spectroscopy.

BACKGROUND: To study the penetration and metabolism of dexamethasone phosphate in the cornea and lens from rabbit following topical administration. METHODS: After topical administration of 0.1, 1 and 10% dexamethasone phosphate solutions, respectively, the cornea and lens were removed and extracted with either perchloric acid (PCA) or with chloroform/methanol to study the lipophilic and hydrophilic metabolites of the drug. The extracts were analyzed with 1H and 19F NMR spectroscopy. RESULTS: Using 19F NMR spectroscopy, both dexamethasone and dexamethasone phosphate were detected simultaneously in the extract of the cornea and lens obtained from the eyes denuded of the epithelium before the treatment. Using 10 and 1% drug solutions, the signals from dexamethasone were also detectable in the 1H NMR spectra among many endogenous metabolites. CONCLUSION: 1H and 19F NMR spectroscopy was shown to be a useful method in the study of penetration and metabolism of dexamethasone in the eye, particularly because both the lipophilic and hydrophilic metabolites of the drug could be detected simultaneously.

Dexamethasone and dexamethasone phosphate detected by 1H and 19F NMR spectroscopy in the aqueous humour.

To apply nuclear magnetic resonance (NMR) spectroscopy to study the penetration of dexamethasone phosphate into the aqueous humour from rabbit following topical administration. After topical administration of 0.1%, 1.0% and 10% dexamethasone phosphate solutions, respectively, samples of aqueous humour were aspirated, freeze-dried, redissolved in deuterium oxide and analyzed by high resolution 1H and 19F NMR spectroscopy. In order to study the lipophilic and hydrophilic metabolites of the drug, samples obtained after application of 1% dexamethasone phosphate were extracted with methanol/chloroform, and then extracted with perchloric acid. In all samples obtained from eyes denuded of the corneal epithelium prior to administration of dexamethasone, signals corresponding to the chemical shifts of the drug were identified in 19F NMR spectra. In the experiments performed with 1% dexamethasone phosphate, both dexamethasone and dexamethasone phosphate were detected in the aqueous humour. Using 10% dexamethasone phosphate solutions, signals from the drug were detected in 1H NMR spectra simultaneously with signals from about twenty other substances present in the aqueous humour. NMR spectroscopy appears to be a valuable method for studying dexamethasone metabolism and penetration into ocular tissues. It provides simultaneous detection of both the drug metabolites and other substances in the sample and might offer a complementary approach to other analytical methods.

Detection of different metabolites in the rabbit lens by high resolution 1H NMR spectroscopy.

PURPOSE: To investigate the metabolic profile of the rabbit lens using high resolution 1H NMR spectroscopy including two-dimensional shift correlated (COSY) technique. METHODS: Perchloric acid extracts of the rabbit lens were analysed with a Bruker AM-500 spectrometer and the metabolites were assigned in the spectra. Some of these were also quantified. RESULTS: More than 20 metabolites were detected in the perchloric acid extract of a single lens, including amino acids, nucleotides and other related compounds. Of particular importance is the ability to detect and identify glutathione, myoinositol, scyllo-inositol and taurine. CONCLUSIONS: The study demonstrated the potential of 1H NMR spectroscopy for monitoring the metabolic profile of the lens in normal and pathologic conditions.

Metabolic analysis of the rabbit cornea by proton nuclear magnetic resonance spectroscopy.

Perchloric acid extracts of the rabbit cornea were analyzed by high-resolution 1H NMR spectroscopy, including the two-dimensional shift-correlated (COSY) technique. Numerous metabolites including leucine/isoleucine, 3-hydroxybutyrate, valine, lactate, alanine, lysine, acetate, glutamate, creatine/phosphocreatine, taurine, glycine, inositol, glucose, tyrosine, phenylalanine, histidine, adenosine and formate have been simultaneously detected and identified in the NMR spectra. Concentrations of some of these were also determined. Of particular importance are the detection and identification of hypotaurine in the corneal tissue. Hypotaurine is a precursor of the synthesis of taurine and may act as an antioxidant. Moreover, the presence of choline, a precursor for acetylcholine, was established. This novel metabolite profile of the cornea obtained by high-resolution 1H NMR spectroscopy may be of value in the study of pathologic and toxic disorders.