-
A protein is to be purified using ion- exchange column chromatography. The relationship between HETP (Height Equivalent to Theoretical Plate) and the linear liquid velocity of mobile phase is given by :
H = A + Bu + C u
where H is HETP (m) and u is linear liquid velocity of mobile phase (m.s–1). The values of A, B and C are 3 × 10–8 m2.s–1, 3 s and 6 × 10–5 m, respectively. The number of theoretical plates based on minimum HETP for a column of 66 cm length will be ___________.
-
- 2000
- 500
- 1000
- All of the above
Correct Option: C
(i)
| H = | + Bu + C | |
| u |
The minimum HETP occurs when
u = √A / B = 10–4 m / sec
A = 3 × 10–8 m2 / sec
B = 3 sec
C = 6 × 10–5 m
(ii) ∴ H = 6.6 × 10–4
Now, = 6.6 × 10–2 cm
(iii)
| H = | , N = No. of theoretical plates | |
| N |
L = Length of the column
| ∴ H = | ||
| N |
| = | = 1000 | |
| 6.6 × 10-2 cm |
