Development and validation of RP-HPLC method for simultaneous estimation of hydrochlorothiazide and thiocolchicoside in bulk and in a synthetic mixture


  • Nagunath S
  • Alekya
  • Priyanka
  • Srivani
  • Pallavi
  • Rajeshekar
  • vennela


Hydrochloro thiazide, Thiocolchicoside, HPLC, Validation.


A simple, rapid and precise reverse phase liquid chromatographic (RP-HPLC) method was developed and subsequently
validated for simultaneous estimation of Hydrochloro thiazide (HCT) and Thiocolchicoside (TCS) in bulk drug and in a
synthetic mixture. The analysis was carried out using Zodiac ODS C18 (4.6 x 250mm, 5µm, Make: Zodiac Life Sciences),
pre-packed column. The separation was carried out using a mobile phase containing a buffer of pH 4.0, Acetonitrile (80:20
v/v), was pumped at a flow rate of 1.2 mL/min with UV-detection at 267 nm. Both the drugs were well resolved on the
stationary phase and the retention times were around 5.170 minute for Hydrochloro thiazide and 7.477 minute for
Thiocolchicoside. The method was validated and shown to be linear for Hydrochloro thiazide and Thiocolchicoside. The
correlation coefficients for Hydrochloro thiazide and Thiocolchicoside are 0.997 and 0.999 respectively. The relative
standard deviations for five replicate measurements in two sets of each drug in the tablets is always less than 2% and mean
% error of active recovery not more than ±1.5%. The method was validated for precision and accuracy. The proposed
method was successfully applied to the synthetic mixture containing the above-mentioned drug combination without any
interference by the excipients.