The application of clinical pharmacokinetics is the responsibility of all pharmacists providing pharmaceutical care. An appropriately applied clinical pharmacokinetic is expected to result in improved patient outcomes: decreased mortality, reduced length of treatment, reduced length of hospital stays and cost-savings. Data on the extent of pharmacokinetic application in Libyan hospitals remain scarce but available subjective evidence suggests that services related to clinical pharmacokinetics are mostly provided and performed by professionals other than clinical pharmacists. To explore the training background and perceptions of pharmacists on the pharmacokinetic course contents they received during their undergraduate pharmacy programs. Also, to determine the attitudes and barriers experienced by the pharmacists when applying pharmacokinetic principles in their current practice. This is a cross-sectional, descriptive study that was conducted between July 2018 and November 2019 using a self-administered survey. The study targeted hospital pharmacists practicing in different hospitals in Libya. A total of 104 pharmacists completed the questionnaire and submitted it back. The majority of participants learned pharmacokinetic courses as a mandatory course during undergraduate courses 81.0% with 37.0% selecting that course was taught as a separate course or courses. Around 80.0% of the participants agreed or strongly agreed that pharmacokinetic courses received in undergraduate studies are useful in pharmacy practice with over 75.0% of the participants agreeing or strongly agreeing on the relevance of those courses to their current clinical practice. About 40.0% of the participants described their current skills in allowing optimal patient care as being better. Different barriers were highlighted by the participants to allow sufficient clinical pharmacokinetic practice including lack of sufficient information, and lack of awareness of pharmacists’ role and skills in applying clinical pharmacokinetic by other pharmacists and by other health care providers. Thus, this study shows that most practicing pharmacists showed a positive attitude to current pharmacokinetic practice and to the impact of their undergraduate studies on their successful practice. However, they have clearly addressed the room for improvement.