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Purpose: This study examined young women's actual timing of use of a microbicide surrogate gel (vaginal moisturiser (VM)) compared with assigned timing conditions., Methods: Participants used a VM with coitus during 4-week cycles over a 3-year period in random timing sequences: 1h before coitus, 10 min before coitus and 10 min after coitus. Daily diaries collected information related to coital behaviours, VM use and timing, and participants' and partners' VM assessments. Descriptive and mixed-effects model analyses were conducted., Results: At least three VM timing conditions were completed by 109 women aged 18-22 years old. Of 17?772 diary days collected, coitus was reported on 2128 (1252 with VM use; 59%). Median times between VM application and coitus were: 60 min before coitus (mean=68.2; s.d.=76.9) for the 1-h pre-coital group, 13.5 min before coitus (mean=44.9; s.d.=117.1) for the 10-min pre-coital group and 5 min before coitus (mean=24.5; s.d.=205.1) for the 10-min post-coital group. Women reported that the VM was very easy to use (68%), it was somewhat messy (61%), they were very wet during sex (81%), sex was very good (80%) and their partners liked using the VM (38%)., Conclusions: Overall, the VM was rated positively. There was substantial deviation in application time across timing conditions, with significantly greater variability in the post-coital group. These findings contribute to understanding of how VMs are accepted and used, with implications for HIV prevention with microbicides requiring specific application timing.