کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9748852 | 1493794 | 2005 | 9 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Rapid analysis of porphyrins at low ng/l and μg/l levels in human urine by a gradient liquid chromatography method using octadecylsilica monolithic columns
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کلمات کلیدی
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Rapid analysis of porphyrins at low ng/l and μg/l levels in human urine by a gradient liquid chromatography method using octadecylsilica monolithic columns Rapid analysis of porphyrins at low ng/l and μg/l levels in human urine by a gradient liquid chromatography method using octadecylsilica monolithic columns](/preview/png/9748852.png)
چکیده انگلیسی
Rapid gradient RP-HPLC method with fluorimetric detection for trace analysis of diagnostically significant porphyrins in human urine was developed for clinical and diagnostic purposes. Results show that optimized high-pressure gradient elution and monolithic column Chromolith SpeedRod RP18e enabled separation of seven urine porphyrins including baseline separation of I and III positional isomers of uro- and coproporphyrins within 3.2 min. Problems associated with high metal cation complexing ability of the analytes and common stainless steel based instrumentation were substantially reduced by use of 0.1 mol/l ammonium citrate buffer (pH 5.47) and methanol as a mobile phase components. Good reproducibilities of retention times (within ±0.36% RSD) and peak areas (from ±0.6 to ±2.5% RSD) at 5-20 μg/l level of the analytes were achieved. Determined LOQ (10 Ã S/N) values of diagnostically important porphyrins using fluorimetric detection (ex.405 nm/em.620 nm) were 82 pmol/l (65 ng/l, 1.30 pg/injection) for uroporphyrin I, 44 pmol/l (33 ng/l, 0.66 pg/injection) for uroporphyrin III, 50 pmol/l (40 ng/l, 0.80 pg/injection) for coproporphyrin I and 47 pmol/l (39 ng/l, 0.78 pg/injection) for coproporphyrin III. Attained LOQ concentration level is approximately 20-120 times lower than concentration of porphyrins in a urine of healthy person. Calculated LOD's (3 Ã S/N) were at a low ng/l levels, what enabled quantification of carry-over effect to be from 2.0% to 0.2% in each of three consecutive blank runs and from 2.5% to 7% in total after injection of mixed standard of porphyrins with 5-20 μg/l concentrations. Recovery of porphyrins at low μg/l concentration levels was from 93% to 97.5%. Devised method increases productivity of clinical laboratory from 2 to 10 times in dependence of duration of currently used method.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1084, Issues 1â2, 19 August 2005, Pages 24-32
Journal: Journal of Chromatography A - Volume 1084, Issues 1â2, 19 August 2005, Pages 24-32
نویسندگان
Peter Božek, Milan Hutta, Barbora Hrivnáková,