HPLC method development and validation for in vitro and in vivo quantification of vancomycin in rabbit plasma

Faryal Jahan; Shahiq-uz Zaman; Muhammad Ali Syed; Rabia Arshad; Oswa Amjad; Ijaz Ali; Rabia Gul; Gul Shahnaz; Muhammad Naeem Aamir; Sana Hanif

Faryal Jahan Department of Pharmaceutics, Riphah Institute of Pharmaceutical Sciences, Riphah International University Islamabad, Pakistan;Shifa College of Pharmaceutical Sciences Shifa Tameer-e-Millat University Islamabad, Pakistan

Shahiq-uz Zaman Department of Pharmaceutics, Riphah Institute of Pharmaceutical Sciences, Riphah International University Islamabad,,Pakistan

Muhammad Ali Syed The University of Lahore,Lahore,Pakistan

Rabia Arshad The University of Lahore,Lahore,Pakistan

Oswa Amjad The University of Lahore,Lahore,Pakistan

Ijaz Ali The University of Lahore,Lahore,Pakistan

Rabia Gul Shifa College of Pharmaceutical Sciences Shifa Tameer-e-Millat University Islamabad,,Pakistan

Gul Shahnaz Department of Pharmacy, Quaid-i -Azam University,Islamabad,Pakistan

Muhammad Naeem Aamir Department of Pharmaceutics, Faculty of Pharmacy, The Islamia University of Bahawalpur,Bahawalpur,Pakistan

Sana Hanif College of Pharmacy, University of Sargodha,Sargodha,Pakistan

Vancomycin (VAN) is an effective antibiotic due to its broad-spectrum bactericidal action. High performance liquid chromatography (HPLC), a powerful analytical technique is used for the in vitro/ in vivo quantification of VAN. The current study was aimed to detect the VAN from in vitro as well as the plasma after the extraction from blood of rabbits. The method was developed and validated according to International Council on Harmonization (ICH) Q2 R1 guidelines. Results showed that the peak of VAN was recorded at 2.96 and 2.57 min, respectively in vitro and serum. The coefficient of VAN turned out to be >0.9994 each for in vitro and in vivo samples. VAN was found linear in the range of 6.2-25000ng/mL. The values of accuracy and precision in terms of coefficient of variation (CV) were less than 2%, indicating the validity of the method. The values for LOD and LOQ were estimated to be 1.5 and 4.5ng/mL, correspondingly, which were lower than the values calculated from in vitro media. Furthermore, the score of the greenness found out to be 0.81, depicting good score using AGREE tool. It was concluded that the developed method was found accurate, precise, robust, rugged, linear, detectable and quantifiable at prepared analytical concentrations and could be used for in vitro and in vivo VAN determination.