Tests for Urea Quantification:

1.Blood Urea Nitrogen (BUN) Test

The BUN test is a measure of the amount
of nitrogen in the blood that comes from
urea.

2 methods to determine BUN-:

i) Diacetyl Monoxime method

ii) Berthelot Enzymatic method

 Diacetyl Monoxime method
Principle Fearon reaction

The pink-red coloured

product is quantified using a
Spectrophotometer at 540nm.

(𝒂𝒃𝒔𝒐𝒓𝒃𝒂𝒏𝒄𝒆 𝒐f 𝒔𝒂𝒎𝒑𝒍𝒆 )/(𝒂𝒃𝒔𝒐𝒓𝒃𝒂𝒏𝒄𝒆 𝒐𝒇

𝒔𝒕𝒂𝒏𝒅𝒂𝒓𝒅)×𝐜𝐨𝐧𝐜.𝐨𝐟 𝐬𝐭𝐚𝐧𝐝𝐚𝐫𝐝=𝐮𝐫𝐞𝐚
𝐜𝐨𝐧𝐜.𝐢𝐧 𝐬𝐚𝐦𝐩𝐥𝐞

Normal Value - 2.86 to 7.14 mmol/L (or 8 to 20 mg/dL)

 Berthelot Enzymatic method
Principle Urea Hydrolysis

The final blue coloured product

(indophenol) is quantified using a
spectrophotometer at 540nm.

(𝒂𝒃𝒔𝒐𝒓𝒃𝒂𝒏𝒄𝒆 𝒐f 𝒔𝒂𝒎𝒑𝒍𝒆 )/(𝒂𝒃𝒔𝒐𝒓𝒃𝒂𝒏𝒄𝒆 𝒐𝒇

𝒔𝒕𝒂𝒏𝒅𝒂𝒓𝒅)×𝐜𝐨𝐧𝐜.𝐨𝐟 𝐬𝐭𝐚𝐧𝐝𝐚𝐫𝐝=𝐮𝐫𝐞𝐚
𝐜𝐨𝐧𝐜.𝐢𝐧 𝐬𝐚𝐦𝐩𝐥𝐞
 Spectrophotometry
Principle lambert Beer law

A=klc
concentration
Absorbance
Path length

 Use to quantify compound

present in a mixture as
every compound has
specific wavelength for
max. absorbance (λmax)
 Impaired
renal
function

Excessive
protein
intake
Congestive
heart failure Increased
AZOTEMI
A
value of
Shock
Hemorrhage
into the
BUN
gastrointestin
al tract

Dehydration
 Impaired
nutrient
absorption
Syndrome of
inappropriate
anti-diuretic Liver
secretion
(SIADH)
failure
Decreased
DECREASE
D VALUE
value of
Pregnancy Malnutrition
BUN
Over Anabolic
hydration steroid use
2.Urea Clearance Test
Volume of blood or plasma which are completely cleared of urea by the kidney
per minute.

= urea clearance

U = concentration of urea in urine (in mg/dL)

V = Volume of urine in ml/min
P = Concentration of urea in blood (in mg/dL)

Normal Value – 12 to 20 grams/24 hours

Thank you