Some problems in quantitative analysis with concentration-sensitive detectors in high-performance liquid chromatography

Means of validation of HPLC systems and components

“…The requirements from the chromatographic column are symmetrical peaks, (thermostating is recommended). By measuring retention times and peak areas it is possible to determine the long and short term flow accuracy of the pump and the repeatability of injection [1]. At least six consecutive injections are required.…”

“…Therefore, system suitability tests have been described, which permit the control of the correct Presented at the 21 st ISC held in Stuttgart, Germany, 15th-20 th September, 1996 functioning of the whole system. A convenient way for routine system validation is to use a chromatographic column and a mixture of aromatic hydrocarbons [1,2]. This tests complete systems, especially the repeatability of injection and the short term flow accuracy of the eluent delivery system.…”

Fundamental Relationships of Chromatography

“…Leaving aside the special case of fused peaks, discussed above, there is no simple answer to this question since the precision and accuracy of peak height and area measurements depend on several Chromatographie variables, including sample size, mobile phase composition, flow rate and column temperature [270,[282][283][284][285][286], From a purely theoretical viewpoint there is no doubt that when a mass sensitive detector is used quantitation should be based on peak area since precision of the measurement will be independent of flow rate, temperature stability and any other Chromatographie factors that can alter either the elution volume or peak shape. Leaving aside the special case of fused peaks, discussed above, there is no simple answer to this question since the precision and accuracy of peak height and area measurements depend on several Chromatographie variables, including sample size, mobile phase composition, flow rate and column temperature [270,[282][283][284][285][286], From a purely theoretical viewpoint there is no doubt that when a mass sensitive detector is used quantitation should be based on peak area since precision of the measurement will be independent of flow rate, temperature stability and any other Chromatographie factors that can alter either the elution volume or peak shape.…”

Deductive Prediction of Measurement Precision from Signal and Noise in Photomultiplier and Photodiode Detector of Liquid Chromatography

“…The precision is usually expressed in statistical terms as RSD or SD and is a universal criterion for evaluating the analytical fulfillment of a newly developed method as well as diverse traditional methods.1 In HPLC, however, relatively few publications can be found describing and discussing the statistical aspect and its cause and effect. [2][3][4][5][6][7][8][9][10][11][12][13][14][15] To the authors' knowledge, only three reviews have treated the HPLC precision from the theoretical viewpoint. 2,3,15 Quite recently, a theory to predict the actual RSD values in an analytical instrument has been proposed and the application to an HPLC apparatus equipped with a photomultiplier has been demonstrated.…”