A UV-visible spectrophotometer has a minimum detectable absorbance of 0.02. The minimum concentration of a protein sample that can be measured reliably in this instrument with a cuvette of 1 cm path length is _______ μM. The molar extinction coefficient of the protein is 10,000 L mol⁻¹ cm⁻¹.
GATE 2019 BT Q.45: solution and answer
Answer and solution
ANSWER
The minimum concentration that can be measured reliably is 2 μM.
Official answer: 2Source: Official answer key, GATE 2019 BT (IIT Madras)
Beer-Lambert law: , where is absorbance, is the molar extinction coefficient, is concentration, and is the path length.
Given: minimum detectable absorbance , , .
Rearranging for concentration:
Substituting the values:
Converting to (since ):
So the minimum concentration that can be measured reliably is .
KEY IDEA
The Beer-Lambert law (A = εcl) directly relates absorbance to concentration through the molar extinction coefficient and path length; a spectrophotometer's detection limit in absorbance units translates into a minimum measurable concentration via this same relation.
WATCH OUT FOR
Forgetting to convert the resulting mol/L answer into μM (a factor of 10⁶), or mixing up which quantity is being solved for (rearranging incorrectly to isolate ε or l instead of c).
From the GATE 2019 BT paper. Every question of this paper · Download the full paper · All 8 Microscopy & spectroscopy questions, with notes
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