Using Ion Chromatography and Subsequent UV/VIS Detection to Determine Nitrite and Nitrate in Tobacco

The appearance of tobacco-specific nitrosamines is facilitated by the nitrite present in tobacco. Since most of these nitrosamines are carcinogenic, it is essential to determine the presence of nitrite content in tobacco.

This application note describes the application of ion chromatographic separation and subsequent UV/VIS detection after sequential suppression to determine the presence of nitrite and nitrate contents in acetic acid extracts of tobacco.


Tobacco extract (5% acetic acid)

Sample preparation

Dilution 1:10 in ultrapure water.


. .
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Metrosep A Supp 4/5 - Guard/4.0 6.1006.500


Eluent 3.6 mmol/L sodium carbonate
Regenerant 100 mmol/L sulfuric acid
Rinsing STREAM


Sequential suppression followed by UV/VIS detection.


. .
Flow rate 0.7 mL/minutes
Injection volume 20 μL
Pmax 15 MPa
Recording time 25 minutes
Column temperature 45 °C
Wavelength 220 nm


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944 Professional UV/VIS Detector Vario 2.944.0010
858 Professional Sample Processor 2.858.0020
MSM Rotor A 6.2832.000
Adapter sleve for Suppressor Vario 6.2842.020


Anions Concentration [mg/kg] RSD [%, n = 3]
1 Acetate - -
2 Nitrite 0.23 8.9
3 Nitrate 337 1.0

About Metrohm

At Metrohm is one of the world’s most trusted manufacturers of high-precision instruments for chemical analysis. Metrohm was founded in 1943 by engineer Bertold Suhner in Herisau, Switzerland. Today, Metrohm is represented in 120 countries by subsidiaries and exclusive distributors. The global Metrohm Group also includes the Dutch companies Metrohm Applikon and Metrohm Autolab, manufacturers of online analyzers and instruments for electrochemical research, respectively. Recently, the Metrohm Group was joined by Metrohm Raman, a leading manufacturer of handheld Raman spectrometers.

Metrohm is the global market leader in analytical instruments for titration. Instruments for ion chromatography, voltammetry, conductivity, and stability measurement make the Metrohm portfolio for ion analysis complete. Instruments for Near-infrared and Raman spectroscopy are another, strongly growing segment of the Metrohm portfolio.

Metrohm is a problem solver, both in the laboratory and within the industrial process. To this end, the company offers their customers complete solutions, including dedicated analytical instrumentation as well as comprehensive application know-how. More than 30% of the company’s employees at the Metrohm international headquarters in Herisau work in R&D.

Metrohm has been owned 100% by the non-profit Metrohm Foundation since 1982. The Metrohm Foundation, which does not exert any influence on the company’s business operations, sponsors gifted students in the natural sciences, supports charitable and philanthropic purposes and, above all, ensures the independence of the company.

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Last updated: Apr 2, 2019 at 9:26 AM

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