Cysteinylglycine in urine determined by high-performance liquid chromatography

Determination of endogenous thiols and thiol drugs in urine by HPLC with ultraviolet detection

“…Several methods have been reported for the determination of urinary thiols, including radioenzymic analysis [49], flow-injection analysis with biamperometric detection [50], isotachophoresis [44,45], CE [38], GC-MS [43], capillary-column liquid chromatography with amperometric detection [46], and, first of all, liquid chromatography with tandem mass spectrometry [29,51,48], fluorescent [28,[30][31][32][33]52,47] and UV-vis [34][35][36][37]39] detection. But in this review only liquid chromatography methods with ultraviolet detection will be reviewed.…”

“…For this purpose, a reductive cleavage of the disulfides is made before the derivatization and instrumental final analysis steps. In urine samples, for reduction dithiotreitol [28,29], sodium borohydride (NaBH 4 ) [30][31][32][33][34], tri-n-butylphosphine [35,36] or tris-(2-carboxyethyl)phosphine (TCEP) [37][38][39] are used.…”

New fluorogenic reagent having halogenobenzofurazan structure for thiols: 4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole

“…Under the selected condition, the detection limits (signal to noise ratio of 3) for ABD-cysteine, ABD-glutathione, ABD-2V-acetylcysteine, ABD-homocysteine, and ABD-cysteamine were 0.6, 0.4, 1.9, 0.5, and 0.5 pmol, respectively. DISCUSSION Thiols are easily oxidized by dissolved oxygen in alkaline medium (23); mild conditions (neutral pH, low temperature, short reaction time) are therefore desirable for the derivatization. Although NBD-X (X = Cl or F, 4-halogeno-7-nito- ro-2,l,3-benzoxadiazole), which was developed as the fluorogenic reagent for amine (14-17), also reacts with thiol at low pH (pH 4-7), the S -* N migration of the NBD group (18) renders it unsuitable.…”

Sulphur amino‐acid degradation in subjects with hereditary glutathione synthetase deficiency (5‐oxoprolinuria)

“…Cysteinylglycine in urine was determined by a highsensitive HPLC method [14]. The method includes reduction of urine disulphides with thiopropyl-Sepharose, clean-up of the thiol compound, derivatization with a fluorogenic reagent and finally HPLC determination.…”