Background: Primary care physicians (PCP) play an important role in early detection of Familial Hypercholesterolaemia (FH). However, knowledge, awareness and practice (KAP) regarding FH among Malaysian PCP are not well established, and there was no validated tool to assess their FH KAP. Therefore, this study aimed to adapt an FH KAP questionnaire and to determine its validity and reliability among Malaysian PCP. Methods: This cross-sectional validation study involved PCP with ≥1-year working experience in the Malaysian primary care settings. In Phase 1, the original 19-item FH KAP questionnaire was content validated and adapted by 7 experts. The questionnaire was then converted into an online survey instrument and was face validated by 10 PCP. In Phase 2, the adapted questionnaire was distributed via e-mail to 1500 PCP for self-administration. Data were collected on their KAP, demography, qualification and work experience. The construct validity was tested using known-groups validation method. PCP with postgraduate qualification (PCP-PG-Qual) were hypothesized to have better FH KAP compared with PCP without postgraduate qualification (PCP-noPG-Qual). The internal consistency reliability was tested using Kuder Richardson formula-20 (KR-20) and test-retest reliability was tested on 26 PCP using kappa statistics. Results: During content validation and adaptation, 10 items remained unchanged, 8 items were modified, 1 item was moved to demography and 7 items were added. The adapted questionnaire contained 25 items (11 knowledge, 5 awareness and 9 practice items). A total of 130 out of 1500 PCP (response rate: 8.7%) completed the questionnaire. The mean percentage knowledge score was significantly higher in PCP-PG-Qual compared to PCP-noPG-Qual (53.5, SD±13.9 vs. 35.9, SD±11.79), t(128)=6.90, p<0.001. The median percentage awareness score was significantly higher in PCP-PG-Qual compared to PCP-noPG-Qual (15.4, IqR±23.08 vs. 7.7, IqR±15.38), p=0.030. The mean percentage practice score was significantly higher in PCP-PG-Qual compared to PCP-noPG-Qual (69.2, SD±17.62 vs. 54.4, SD±19.28), t(128)=3.79, p<0.001. The KR-20 was 0.79 (moderate reliability) and average Kappa was 0.796 (substantial agreement). Conclusion: The robust validation methods and findings suggest that the adapted 25-item questionnaire is a valid and reliable tool to measure FH KAP among Malaysian PCP. This would aid to improve FH care in the community.