HPLC laboratory

The sixth item is a series of three HPLC laboratory experiments. The first familiarizes the student with getting a system up and running and calibrating a column with standards. The second experiment shows how to clean a column and pacify a system. The last is a first, quick look at methods development. 

Preventive maintenance helps the chromatographer to reduce instrument downtime, allowing for more efficiency and cost effectiveness in the HPLC laboratory. 

Size-exclusion chromatography is an important separation method. It provides an easy way of solving many types of problem and as such should form an integral part of any HPLC laboratory set-up. There are four main areas of application ... 

An HPLC laboratory which has to deal with a range of different problems must be stocked with the following stationary phases ... 

Today, Fast LC is the easiest way to increase HPLC laboratory productivity by 3-5 fold without sacrificing resolution, sensitivity, or assay reliability.19 Figures 3.18-3.20 demonstrate the versatility and dramatic speed achievable with Fast LC. Chapter 4 discusses the effect of extracolumn band-broadening... 

Figure 9.1. A schematic diagram listing the various tools, systems, and procedures used to ensure HPLC data accuracy and regulatory compliance under a cGMP environment. The three circles represent the cGMP organization governing the HPLC laboratory and its daily operation.

This chapter discusses regulatory issues in HPLC laboratories with a focus on procedures and requirements for system qualification, calibration, method validation, and system suitability testing. Examples in a cGMP pharmaceutical environment are used to illustrate the various tools and systems used to ensure the degree of HPLC data accuracy necessary to achieve the delicate balance of regulatory compliance and laboratory productivity. 

While automated tools and software are available for system qualification, method validation, and system suitability testing, achieving regulatory compliance for an HPLC laboratory remains a complex, costly, and time-consuming process. Many companies are adopting a risk-based approach to balance productivity and compliance. For instance, many pharmaceutical research... 

The principles of green chemistry by adopting the most efficient and environmentally friendly processes should be practiced whenever possible in the HPLC laboratory. One obvious approach is reduction of solvent consumption by using solvent recycling for isocratic analysis and narrowbore LC columns. Another area is to find ways to reduce sample size and the number of sample preparation steps without sacrificing method performance.2 A case study to illustrate this principle for a tablet assay is shown in Table 11.1. Another example is an environmental analysis of soil/sediment sample3 is shown in Table 7.7. 

HPLC Laboratory Data Ground Soybeans Whole Soybeans ... 

Table 6.16. HPLC laboratory Reference (ppm) Values for the Total of the Three Forms of Three Types of Isoflavones and Total Isoflavones. ...

In the following, the most important HPLC activities are described in more detail, just in case you do not have a description at hand. Otherwise, make the following assumption if you have to follow an existing method (SOP, System Operation Procedure), stick to this method Your creativity and experimental skills are an invaluable asset in the HPLC laboratory, but please, at the right time. 

Specialization, a phenomenon of our times, also happens in analytical chemistry. The widening gap between routine and research analysis strongly influences the daily work and the environment of an HPLC laboratory. 

Based on typical workflows and documents of an HPLC laboratory, fundamental procedures are presented, aiming at efficient laboratory documentation that meets the regulatory requirements e.g., data integrity. 

Table 8.1 Analytic process in an HPLC laboratory, according to the SIPOC-model. 

A typical HPLC laboratory workflow is exemplified in Table 8.3, specifying which meta-information is usually communicated and must also be filed with the actual measurement data in order to ensure traceability. 

Objectives of Documentation 311 Table 8.3 Meta-data model of data traceability in an HPLC laboratory. 

In this book, we cover different topics in the field of modern HPLC. The purpose is to demonstrate current developments and dwell on techniques which recently found their way to the HPLC-laboratory or will do in near future. 

During the daily routine in an HPLC laboratory, problems often arise that may result from a lack of robustness of the RP-HPLC methods used. 

It is common for the test mixture used by a manufacturer to check the performance and quality of an HPLC column to be totally umelated to the assay for which the column was purchased because they are made up of quite simple compounds, which are readily available in the laboratory. When setting up a test procedure for monitoring column performance over its lifetime, the same procedure as the manufacturer should be used if possible. It is useful to note that a different result for the efficiency of a column will occur for each different test method, and it is therefore important for each HPLC laboratory to standarize on one dy. Suggested test mixtures, along with their respective test conditions, are outlined in Table 12. Commonly, two or three components are used, one with a very short retention time [typically with a k of about 0.2, where k is the column capacity factor (see Figure 19) to assess band broadening caused by... 

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