Maternal food neophobia, a significant influencing factor in modeling maternal feeding practices, also plays a vital role in regulating preschoolers’ food neophobia. This cross-sectional study was conducted to culturally adapt and validate the Food Neophobia Scale for Pakistani mothers (FNS-PK), assess the prevalence of maternal food neophobia, and explore the correlation between maternal food neophobia, maternal controlling feeding practices and preschoolers’ food neophobia. The 10-item food neophobia scale was translated into the Urdu language following WHO guidelines, and various measures of validity (construct validity and discriminant validity) and reliability (internal consistency, test–retest consistency, and composite reliability) were used to test the scale. Afterward, a cross-sectional study was performed to assess the prevalence and correlation between maternal food neophobia, feeding practices and preschoolers’ food neophobia. A total of 260 preschool children and their mothers were recruited for the cultural adaptation of the FNS-PK. The original 10-item FNS was modified to an 8-item FNS-PK, which exhibited good scale validity (χ2/df = 1.113; TLI = 0.998; RMSEA = 0.021; AVE > 0.05) and reliability (Cronbach’s α = 0.857). In a cross-sectional study involving 347 mother–child pairs, 15.9 % of the mothers and 8.0 % of the preschoolers exhibited food neophobia. Furthermore, there was a significant negative correlation between maternal food neophobia and controlling feeding practices, while there was a significant positive correlation between maternal and preschoolers’ food neophobia. The adapted 8-item FNS-PK is a reliable tool for assessing food neophobia in Pakistan, and the significant correlation between maternal and preschoolers’ food neophobia warrants further research.