IPC-USP 7th Annual Scientific Meeting

February 6 - 7, 2008
Hyderabad International Convention Center
Hyderabad, India

Revised USP System Suitability Parameters

Horacio N. Pappa, Ph.D.

Senior Scientist
Anthony J. DeStefano, Ph.D.
Vice President, General Chapters
System Suitability – General Principles

‹System Suitability Test are an integral part of

gas and liquid chromatographic methods.

‹They are used to verify that the

chromatographic system is adequate for the
intended analysis.

‹The Concept: Equipment, electronics,

analytical operations and samples constitute an
integral system that can be evaluated as such.
System Suitability – General Principles, cont.

‹ Injections of the appropriate preparation(s)

required to demonstrate adequate system
suitability are made to demonstrate the
system is suitable throughout the run.

‹ Whenever there is a significant change in

equipment or in a critical reagent, suitability
testing should be performed before the
injection of samples.
System Suitability – General Principles

‹ No sample analysis is acceptable unless the

requirements of system suitability have been
met.

‹ Sample analyses obtained while the system

fails requirements are unacceptable.
System Suitability – General Concepts

‹ Specification of parameters in a monograph does

not preclude the use of other suitable operating
conditions to meet System Suitability
requirements.

‹ Adjustments permitted only if

 Standards are available for each system
suitability component.
 Standards show quality of the
chromatography has improved.

‹ Not to compensate for column failure or system

malfunction.
Suitability Testing – Factors to Consider

‹ Repeatability
Capacity factor (k’) N ⎛ α − 1 ⎞⎛ 1 + kb' ⎞
Rs =
‹
⎜ ⎟⎜ ⎟
‹ Resolution 4 ⎝ α ⎠⎝ kb' ⎠
‹ Tailing factor at W0.05 Efficiency Selectivity Retentivity
‹ Theoretical plates (N)
Repeatability

USP:
‹ Data from five replicate injections of the analyte are used to
calculate the relative standard deviation if the requirement is
2.0% or less.
‹ Data from six replicate injections are used if the relative
standard deviation requirement is more than 2.0%

EP:

Number of individual injections

3 4 5 6
Upper limit Maximum permitted relative standard deviation

2.0 0.41 0.59 0.73 0.85

2.5 0.52 0.74 0.92 1.06
3.0 0.62 0.89 1.10 1.27
Capacity Factor

tr − tm
k'=
tm

8
Resolution

2(t 2 − t1 )
R=
W2 + W1

>1.5 for the critical pair

R = 1.0 R = 1.5 R = 2.0

FDA (CDER), Reviewer Guidance: Validation of
Chromatographic Methods, Nov. 1994
Tailing Factor

( a + b)
USP/EP Tailing Factor: T=
2a

a b 5 % of Peak Height

10
Relative Retention Times

<621> Chromatography (USP 30 second supp.):

Relative retention times may be provided in

monographs for informational purposes
only, to aid in peak identification. There are
no acceptance criteria applied to relative
retention times.
System Suitability – Allowed variations -
HPLC

Ratio of Components in Mobile Phase

‹ Adjustments to the composition of mobile phase in

gradient elution are not recommended.

‹ The amount of minor component can be adjusted by

±30% relative.

‹ However, the change in any component cannot exceed

±10% absolute (i.e., in relation to the total mobile
phase).

‹ Adjustment can be made to one minor component in a

ternary mixture.
System Suitability – Allowed Variations –
HPLC, cont.

‹ pH of Mobile Phase: The pH of the aqueous

buffer can be adjusted to within ±0.2 units of
the value or range specified.

‹ Concentration of Salts in Buffer: Can be

adjusted ±10% provided permitted pH variation
is met.

‹ Column Temperature: Temperature can be

adjusted by as much as ±10°. Thermostatting
is recommended.
System Suitability – Allowed Variations

‹ Particle size (HPLC): Can be reduced by

as much as 50%.
‹ Column length (GC, HPLC): As much as
±70%.
‹ Injection volume (GC/HPLC): can be
reduced as far as is consistent with accepted
precision and detection limits.
System Suitability – Allowed Variations -
GC

‹ Column inner diameter: Up to ±50%

‹ Film thickness (Capillary GC): -50% to
100%
‹ Oven Temperature: As much as ±10%
‹ Oven Temperature Program:
Temperature as above, ramps and holds, up
to ±20%.
‹ Flow rate: As much as ±50%.
HPLC System Suitability – Proposal in PF
34(1)

‹ Column Inner Diameter (HPLC): can be

adjusted provided that the linear velocity is
kept constant.
‹ When column dimensions have been
modified, the flow rate can be adjusted
using the following formula:

l2d22
F2 = F1 2
l1d1

‹ Deadline for comments April 15, 2008

Need to Focus on Method Performance

<621> Chromatography – Official USP 30/Supp. 2

The user should verify the suitability of the

method under the new conditions by
assessing the relevant analytical
performance characteristics potentially
affected by the change.

‹ Additional validation data may be necessary.

Conclusions

‹ System Suitability is critical to demonstrating

method acceptability.

‹ USP is aware that small changes to

chromatographic parameters occur due to
changes in ambient laboratory conditions and
deliberately in an effort to optimize the
chromatography.

‹ The current USP chapter allows for reasonable

changes in these parameters, if acceptable
method performance can be demonstrated.
Theoretical Plates

2
t
N= f r
2
w 1
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
f 0.1 t
-3 -2 -1

0 0 1 2 3

-0.1
w =2.355σ
w =4 σ

Neue, U. HPLC Columns

Theoretical Plates

Method Peak width Factor

60.3% of peak
Inflection point 4.000
height
50% of peak
Half-height 5.545
height
13.4 of peak
4 Sigma method 16.00
height
4.4% of peak
5 sigma method 24.00
height
Intersection of
Tangent method tangents with 16.00
baseline