INTRODUCTION:
Nitrate is found only in small
amounts in fresh domestic wastewater but in the effluent of nitrifying
biological treatment plants nitrate may be found in concentration up to 30mg
nitrate as nitrogen/L. It is essential nutrient for many photosynthetic
autotrophs and in some cases has been identified as the growth-limiting
nutrient. Generally 10mg nitrate nitrogen/L of drinking water is recommended to
prevent disorder called methemoglobinemia in infants. Determination of nitrate
nitrogen is difficult because of the relatively complex procedure. Although UV
technique that measures the absorbance of nitrate at 220nm is suitable for
measuring nitrate in wastewater.
PRINCIPLE:
This technique is useful for the
screening of sample that has low organic content that is uncontaminated natural
water and potable water supplies. The nitrate calibration curve follows Beer's
Law up to 11 mgN/L. Measurement of UV absorbance at 220nm enables rapid
determination of nitrate. Because dissolved organic matter may also absorb at
220nm and nitrate does not absorb at 275nm, a second measurement made at 275nm
may be used to correct nitrate value.
REQUIREMENTS:
·
Stock nitrate solution: Dissolve 0.7218gm KN03
in water and dilute to 1000m1 (1ml = 100mcg nitrate nitrogen). Preserve this
with 2ml chloroform/L.
·
Standard
nitrate solution: Dilute 100 ml stock solution to 1000 ml distilled water (1
ml= 10mcg nitrate nitrogen').
PROCEDURE:
·
Take
different aliquots of standard solution from 0.4, 0.8,………, 2.0 ml and one
blank.
·
Take
three different aliquots (0.4, 1.2, 2.0ml) of unknown sample.
·
Make
up the final volume to 5ml with distilled water.
·
Take
the O.D. of all the tubes at 220nm (if organic matter is present take the O.D.
at 275 nm also)
·
Prepare
a standard graph and plot the absorbance of unknown to find the concentration
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