Validation of UV-HPLC method for simultaneous quantification of organic acids in disinfectants for haemodialysis machines

About author
¹Mahmood Ahmed*, ¹Muhammad Abdul Qadir, ¹Shabnam Shahzad, ¹Rabia Waseem, ²Muhammad Saqlain Tahir
¹Institute of Chemistry, University of Punjab, Lahore-Pakistan
²Highnoon Laboratories Limited, Lahore- Pakistan

Abstract
A UV-HPLC method was validated for quantification of citric, lactic and malic acidsin disinfectants. The chromatographic separation was performed at 40°C by using mobile phase comprised a mixture (90:10) of potassium phosphate monobasic (50mM) and acetonitrile and pH was adjusted to 2.8 with ortho phosphoric acid, on C₁₈-5µm and 100⁰A pore size reverse phase column (250mm×4.6mm) at λ=210 nm using UV-Vis detector with 1.25ml/min flow rate. ICH guidelines were adopted to perform the validation studies that the analytical method’s characteristics are reliable throughout the time during analysis. Validation parameters showed efficiency and adequate linearity with correlation coefficient (r²) 0.9997, 0.9998 and 0.9997for 20-80 μg/ml citric acid, 2-8μg/ml lactic acid and malic acid respectively. The repeatability and intermediate precision (%RSD<2) indicated the method reproducibility. The limits of detection (LOD) 1.93μg/ml, 0.11μg/ml,0.21μg/ml and the limit of quantification (LOQ) were 6.46μg/ml, 0.34μg/ml, and 0.68μg/ml for citric acid, lactic acid and malic acid respectively.% Recovery ± SD for citric, lactic and malic acids was98.22±0.24 – 100.3±0.03, 98.26±0.03 – 101.26±0.03 and 97.42±0.03 – 100.5±0.07 respectively for three spikes levels.
Key words: Organic acids, HPLC, Disinfectants, Haemodialysis