RESUMEN
Polymer mediated capillary electrophoresis (CE) using poly(diallyldimethylammonium chloride) (PDDAC) in the electrolyte with end-column single reflection attenuated total internal reflectance (ATR) Fourier transform infrared (FT-IR) microspectroscopy is presented. The terminus of the capillary is placed approximately 1 microm from the internal reflectance element (IRE), at the focus of the ATR infrared microscope. Electrophoretic separations of benzenesulfonate, 1-naphthalenesulfonate and 1,5-naphthalenesulfonate in a NaCl and PDDAC electrolyte using either -11 or -15 kV are demonstrated with the CE-ATR FT-IR spectra providing identification of these compounds. Running electrolyte salt concentrations in the 0.85-1.7 M range are required due to the 50-80 mg mL(-1) sample concentrations. Increasing the NaCl salt concentration improves peak resolution but also increases analysis time. A PDDAC concentration as low as 0.0075% can facilitate the separation of the aromatic sulfonates while maintaining reasonable electroosmotic flow. Etching of the germanium IRE by the applied current, which can affect the intensity of the infrared beam, is corrected by fabrication of a plastic mounting post for the IRE to prevent current conduction through the IR instrument.
Asunto(s)
Compuestos Alílicos/análisis , Técnicas de Química Analítica/métodos , Polímeros/química , Compuestos de Amonio Cuaternario/análisis , Electroforesis Capilar/métodos , Microespectrofotometría/métodos , Espectroscopía Infrarroja por Transformada de Fourier/métodosRESUMEN
We have biopsied the papillae of patients who have cystine stones asking if this stone type is associated with specific tissue changes. We studied seven cystine stone formers (SF) treated with percutaneous nephrolithotomy using digital video imaging of renal papillae for mapping and obtained papillary biopsies. Biopsies were analyzed by routine light and electron microscopy, infrared spectroscopy, electron diffraction, and micro-CT. Many ducts of Bellini (BD) had an enlarged ostium, and all such were plugged with cystine crystals, and had injured or absent lining cells with a surrounding interstitium that was inflamed to fibrotic. Crystal plugs often projected into the urinary space. Many inner medullary collecting ducts (IMCD) were dilated with or without crystal plugging. Apatite crystals were identified in the lumens of loops of Henle and IMCD. Abundance of interstitial Randall's plaque was equivalent in amount to that of non-SF. In the cortex, glomerular obsolescence and interstitial fibrosis exceeded normal. Cystine crystallizes in BD with the probable result of cell injury, interstitial reaction, nephron obstruction, and with the potential of inducing cortical change and loss of IMCD tubular fluid pH regulation, resulting in apatite formation. The pattern of IMCD dilation, and loss of medullary structures is most compatible with such obstruction, either from BD lumen plugs or urinary tract obstruction from stones themselves.