HPLC Calibration Process Parameters in Terms of System Suitability Test

Mini Review

Austin Chromatogr. 2014;1(2): 4.

# HPLC Calibration Process Parameters in Terms of System Suitability Test

Anirbandeep Bose*

Acharya and BM Reddy College of Pharmacy, India

*Corresponding author: Anirbandeep Bose, Acharya and Bm Reddy college of Pharmacy, India

Received: September 01, 2014; Accepted: September 24, 2014; Published: October 07, 2014

## Abstract

During the routine analysis of drug and analytes System Suitability Test (SST) is one of the most important and integral parts of HPLC method development and calibration. System Suitability Test (SST) is generally performed to evaluate the suitability and effectiveness of the entire chromatographic system not only prior to use but also during the time of analysis. Due to several reasons the performance and the capacity of the entire chromatographic system may abruptly or mildly change during their regular uses. This modification of the system can in turn affect the reliability of the entire HPLC analytical results. For this reasons different parameters which is related to the operation of the whole chromatographic system can be monitored and investigated to find out the integrity and reliability of the whole HPLC systems. These parameters are used to determine characteristic chromatographic parameters, such as the number of effective theoretical plates, resolution, asymmetry, detection limit and selectivity. After the checking of the parameters, the system is then only can be declared suitable if the responses are within permitted limits. The objective of our present review article is to emphasize the importance of the System Suitability Test (SST) for development and validation of the HPLC analysis. In addition, we also discuss about the measurement and calculation of different SST parameters in very simpler way.

Keywords: HPLC; LC-MS; Pharmacokinetics; Bioanalytical method development

## Introduction

Chromatography, specifically liquid chromatography in the form of HPLC is used enormously in pharmaceutical drug analysis. In pharmaceutical analysis generally 4 steps is important for the successful validation of HPLC analysis. The first step and second step can be defined as Initial System Qualification followed by Periodic Calibration. The third step is Method Validation, where performance of the entire analytical procedure including sample preparation has been verified and justified. The final and last step is the determination of System Suitability Testing (SST), which significantly contribute to determine the suitability of chromatographic system on a day-to-day basis. In this review article, we emphasize on this final validation step and the main focus of our entire review article is to discuss how to perform SST and define the exact limits according to the latest regulatory guidelines [1-3].

## What is SST?

SST is mainly applied to determine the column efficiency, resolution, and repeatability of a particular chromatographic system to verify its ability for a defined analysis. According to the United States Pharmacopeia and the International Conference on Harmonization (ICH), SST is an essential feature of all HPLC analytical procedures. Although USP and ICH are not the mandatory part of regulatory agencies, but their guidelines are the integral parts of the industry because of its acceptance by the FDA. SST is based on the concept that the equipment, electronics, analytical operations, and samples to be analyzed constitute an integral system that can be evaluated as a whole. The chromatographic systems used for most pharmaceutical analyses such as assays of the active ingredients, impurity determinations, and dissolution testing (measuring the dissolution rate for a particular form of dosage) must pass a set of predefined acceptance criteria (SST limits) before sample analysis can commence [1-5].

## Why SST is necessary?

USP and ICH guidelines have specially emphasize on the use of SST to satisfy the latest regulatory requirements. One of the most important facts about the SST is that it should be performed not only before the experiment but also at different time interval throughout the all HPLC assays. The reason behind the SST analysis at different time interval is based on the concept that it should not be assumed that the system will behave properly throughout experiment. In addition to this, SST studies corresponding to compound of our interest is not enough to check system suitability because the system’s separation capacity is not investigated thoroughly. But, System Suitability Samples (SSSs) or resolution test mixtures of components of interest and expected impurities is required. For this reasons SSTs are analysed before and during testing.

The most important SST parameters which are investigated for different HPLC analysis is Resolution (R), repeatability (RSD— relative standard deviations—of peak response and retention time), column efficiency (N), and Tailing factor (T). These parameters are possessing para amount interest as they indicate system specificity, precision, and column stability. Other parameters include capacity factor (k) and Signal-to-noise ratio (S/N) for impurity peaks. All most all HPLC data systems are based on the software dta system which can calculate the measurement and report of these SST parameters. most “official” analytical methods has fixed the acceptance criteria or SST limits which may vary with different tests and are typically less stringent for biologics and trace impurities [1-7].

## Setting limits

SST limits should emphasize the minimum criterion rather than optimal values. Analytical methods simply adopted the general limits from the CDER guidance document [2]. Different guidelines related to SST limits are shown (Table 1)