On the day of the experimental trial, participants were Ro 61-8048 nmr asked to ingest 568 ml of water to maintain euhydration, and arrive in a fasted condition. On the morning of each trial, participants presented at an indoor sprint track to perform a standardized warm up (10-min), which consisted of jogging, cruising, sprinting, dynamic

stretching and the RSA protocol. This RSA was used as part of the warm-up and not as a measurement test. Temperature and relative humidity were recorded (Testo, Hampshire, UK) at the start and at the end of each experimental trial to check for changes in environmental conditions. Following the warm-up period, participants initiated the testing phase of the trial by performing the RSA test, followed by a 2-min recovery. Participants then completed the

LIST [16]. The LIST was comprised of 15-min sections of intermittent shuttle running over a 20-m distance. Each section of the LIST consisted of 11 cycles of a set running protocol. One cycle was comprised of three 20-m CX-5461 purchase walks (mean speed: 1.54 m · s-1), one 20 metre sprint, ~ 3 sec of rest, three 20 metre cruises (mean speed: 3.33 m · s-1) and three 20 metre jogs (mean speed: 2.86 m · s-1). Following each section, there was a 3-min AZ 628 nmr recovery period. Appropriate speeds for the walk, cruise and jog shuttles of the LIST were dictated by audible signals from a pre-recorded disc. On completion of the 3-min recovery of the second and fourth section of the LIST, participants completed the RSA test, followed by 2-min recovery period (Figure 1). Throughout the experimental protocol, every attempt was made to ensure that the participants were not distracted. No interaction or encouragement occurred between the investigator and the participants, except for mouth rinse administration. Carbohydrate solutions The CHO solution was a 6.4% maltodextrin solution, containing 64 g of maltodextrin Carnitine palmitoyltransferase II (HighFive, Bardon, England) per 1000 ml

of water. Maltodextrin was used because it is a non-sweet and colourless [5]. The PLA solution was water. To make solutions indistinguishable both treatments contained a non-calorific artificial sweetener consisting of sucralose (FlavDrops, MyProtein, Norwich, England). Each rinse solution was provided as a 25-ml bolus in a pre-weighed plastic cup. Participants were instructed to swirl all of the solution in their mouth for ~ 5 sec, before expectorating the solution back into the cup. Participants rinsed a solution 30 sec prior to each section of the LIST and each RSA test. Participants were also instructed to rinse a solution during the first 20 metre shuttle of the second, fourth, sixth, eighth and tenth cycles of each LIST section. In total, this equated to 27 rinses and 675 ml of solution being rinsed and expectorated during each trial (Figure 1). On completion of the study, participants were asked whether they could distinguish which solution contained CHO.