Introduction

Determination of detection and quantification limits of a given analytical procedure can be used
to express its reliability and accuracy in analytical measurements. These limits certainly become
more important in trace analysis and are closely related to the sensitivity and selectivity of the
analytical procedure of choice or the analytical instrument of interest. Health related
manufacturing sectors require highly sensitive and selective instruments because meaningful data
has to be collected at very low sample concentrations. For example, high performance liquid
chromatography (HPLC) is fairly a powerful and versatile technique that can be used to analyze
pharmaceutical products now that many individual components in a sample can be analyzed in a
single procedure while the above mentioned figures of merit are maintained at high levels. The
US Food and drug Administration (FDA) and the United States Pharmacopeia (USP) have in the
recent past increased analytical measurement standards in pharmaceutical organizations in order
to ensure that meaningful and reliable data about pharmaceutical products and drug development
processes are available. This paper is a review of the work that was done by Ribani M. et al.
where they evaluated the detection and quantification limits of a High performance liquid
chromatography method in the determination of impurities in Omeprazole. Here, analytical
parameters that are important in separation methods and especially in HPLC include selectivity,
linearity and range, precision, accuracy, detection limit, quantification limit and robustness. As it
will be shown later, among the two methods that were used, (that is signal-to-noise ratio and the
analytical curve techniques), it was realized that signal-to-noise ration technique is subjective
and prone to errors and deviations while analytical curve technique proved to be more
metrologically reliable [1,2 and 3].

In this paper, detection limit (DL) of an analyte is described as that concentration which gives an
instrumental signal that is significantly different from the “blank” or background signal while
quantification limit (QL) is described as the smallest value of the analyte that can be determined
quantitatively, with a certain limit of confidence [1].
Analytical Method:

Although various articles have been published regarding validation of analytical methods, Ribani
M. et al. applied the signal-to-ratio and standard deviation of the response techniques to validate
their analytical method. As far as the signal to noise ratio is concerned, a comparison was made
between the signals from sample components of low known concentrations that had been
prepared in the matrix of interest and a “white” or component free sample in the same matrix. On
the other side, in the standard deviation of the response approach, the detection and
quantification limits were calculated based on the residual standard deviation of the weighted
regression for LD and on the Eurachem approach for LQ [1,4, and 5].

In the experimental part, HPLC-grade acetonitrile, Merck ultrapure grade-1 water, ACS reagent
grade phosphoric acid, Meck and 35% p.a. ammonium hydroxide, Merck, were used to prepare
mobile phases which were filtered before use using a 0.22 μm membrane before use. The
standards used were

Summary of the analytical method and the application:

Summary of the analytical data and other figures of merit reported and your evaluation of its
completeness

Discussion of the effectiveness of the analytical method for the application that it has been used
for
My own evaluation of the effectiveness of the method

Research and application activities that were performed well

Research and application activities that could be improved upon

References

1. Marcelo Ribani, Carol H. Collins and Carla B.G. Bottoli. Journal of Chromatography A,
1156 (2007) 201-205

2. Zeinab Abdelaziz El-Sherif, Afaf Osman Mohamed, Mohamed Galal El-Bardicy and
Mohamed Fayez El-Tarras. J. Chem. Pharm. Bull 54 (6) 814 – 818 (2006)

3. Ghulam A. Shabir. Journal of Chromatography A, 987 (2003) 57-66

4. J. Vial, A. Jardy, Chromatographia 53 (2001) S-141

5. J.Vial, A. Jardy, Anal. Chem. 71 (1999) 2672