Determination of nitrite and nitrate in cell culture medium by reversed-phase high-performance liquid chromatography with simultaneous UV-VIS and fluorescence detection
Author
Source
Issue
Vol. 3, Issue 1 (31 Mar. 2008), pp.69-75, 7 p.
Publisher
Yarmouk University Deanship of Research and Graduate Studies
Publication Date
2008-03-31
Country of Publication
Jordan
No. of Pages
7
Main Subjects
Abstract EN
A reliable method for determination of nitrite and nitrate in cell culture medium has been developed.
The method is based on pre-column derivetization of nitrite with 2, 3- diaminonaphthalene (DAN) under acidic medium to yield 2, 3-naphthotriazole (NAT).
For determination of nitrate, it was converted to nitrite by nitrate reeducates before the derivetization step.
NAT was separated on a 5 μm C18 column (250 x 4.6 mm, I.D) guarded by a 10 μm C18 column (20 X 4.6 mm, I.D), and eluted with 15 mM phosphate buffer (pH 7.5) containing 35 % acetonitrile (flow rate 1.0 Ml / min).
Average retention time of NAT was 12.4 min.
Absorbance of NAT was monitored at 369 nm and its fluorescence was monitored at 375 nm for excitation and 415 nm for emission.
Calibration curves of nitrite or nitrate were linear in the range 0.03-2.0 μM for fluorescence detection and 0.5-50 μM for UV-VIS detection.
Correlation coefficients were better than 0.994 and mean recoveries were 96 ± 7 %.
American Psychological Association (APA)
Tahbub, Yahya R.. 2008. Determination of nitrite and nitrate in cell culture medium by reversed-phase high-performance liquid chromatography with simultaneous UV-VIS and fluorescence detection. Jordan Journal of Chemistry،Vol. 3, no. 1, pp.69-75.
https://search.emarefa.net/detail/BIM-275393
Modern Language Association (MLA)
Tahbub, Yahya R.. Determination of nitrite and nitrate in cell culture medium by reversed-phase high-performance liquid chromatography with simultaneous UV-VIS and fluorescence detection. Jordan Journal of Chemistry Vol. 3, no. 1 (Mar. 2008), pp.69-75.
https://search.emarefa.net/detail/BIM-275393
American Medical Association (AMA)
Tahbub, Yahya R.. Determination of nitrite and nitrate in cell culture medium by reversed-phase high-performance liquid chromatography with simultaneous UV-VIS and fluorescence detection. Jordan Journal of Chemistry. 2008. Vol. 3, no. 1, pp.69-75.
https://search.emarefa.net/detail/BIM-275393
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references : p. 75
Record ID
BIM-275393