Fatty acids are essential for the structure and function of biological systems, and they are the primary source of energy. Avocado oil has high levels of omega-6 and omega-9 fatty acids, as well as natural antioxidants, which help lower total cholesterol, triacylglycerol, and low-density lipoprotein (LDL) cholesterol while maintaining healthy high-density lipoprotein (HDL) cholesterol levels. Avocado's fatty acid profile varies depending on cultivar, plant portion, ripening stage, geographical location, and sampling procedures. The purpose of this study was to examine the effect of cultivar and altitude on the fatty acid composition of avocado grown in Murang'a County, Kenya. Four different avocado cultivars were sampled from Murang'a County's three climate zones: Zone 1 (above 2,200m a.s.l.), Zone 2 (1,700m-2,100m a.s.l.), and Zone 3 (below 1,600m a.s.l). The oil was extracted from dry powdered pulp using Soxhlet extraction with petroleum ether 40-60. The fatty acid profile was evaluated using GC-MS. Oil output dropped as altitude decreased from Zone 1 to Zone 3. A total of 11 different fatty acids were identified, namely 9-hexadecenoic (palmitoleic), hexadecanoic (palmitic), 9-octadecenoic (oleic), stearic, 12-hydroxy oleic acid (ricinoleic), nonanedioic (azelaic), tridecanedioic (brassylic), eicosanoic (arachidic), undecylenic, heptadecanoic (margaric), and dodecanoic (lauric). All cultivars had four fatty acids: 9-octadecenoic, octadecanoic, 12-hydroxy oleic, and 9-hexadecenoic, with the most prevalent being 9-octadecenoic (oleic). Except for the Giant cultivar, the amount of oleic acid reduced as altitude fell. The fatty acid makeup was comparable to that of other avocados published before.