- Frequency: Once in 6 months.
- Operate the instrument as per the respective standard operating procedure of HPLC.
- Prepare the mobile phase consisting of 70% methanol. : 30 % of water.
- Prepare a mixture of 0.1 % Benzene and 0.1% Toluene in methanol.
- Set flow rate at 1.0 ml/min.
- Allow
the system to be saturated with the mobile phase for at least 15 minutes before
injecting the test
sample.
- Record the area and retention time of both Benzene and Toluene in methanol.
- Calculate the system suitability parameters with the help of software such as resolution, tailing factor and theoretical plate.
- Now calculate the tailing factor, resolution and theoretical plate manually by following formula:
Theoretical
Plate : N = 16 (t/w)2
Where t = R.T of the component
w = width of the component
Resolution : 2(t2 –
t1)/ w1 + w2
Where t1 = R.T of Ist components
t2 = R.T of IInd components
w1 =
Width of Ist components
w2 = Width of IInd component
Tailing : T = w/2f
Where w =
width of peak
f = width of half peak at 5 % height
The
performance of software of the instrument is satisfactory if all the
parameters of the system suitability test are within the acceptance criteria.
Verification of system suitability parameters
1. Resolution :
As per software =
_________________
2 (t2 – t1)
By manually = R
= ------------------
w1 + w2
= ----------------- ( +/- 10 %)
2.
Tailing :
As per software = ________________
By manually = T
= w/2f
= ------------------------( +/- 10%)
3)
Theoretical Plate :
As per software =______________
By manually = N
= 16 (t/w)2
= ------------------------ ( +/- 10%)
Remarks: Comply/Does not comply
Acceptance criteria: Difference should be within
+/- 10.0 %
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