Citrulline malate supplementation does not improve German Volume Training performance or reduce muscle soreness in moderately trained males and females

We measured the effects of an acute 8 g dose of CM on anaerobic performance/recovery using a GVT protocol performed with an isokinetic chair in 15 subjects. The acute dose of CM was administered one hour before resistance training and did not significantly affect the number of repetitions performed. This finding is contrary to similar studies, which found significant effects from CM of the same dose [18, 19, 22]. In the present investigation not only did the CM not attenuate the marker of muscle soreness, but was associated with greater soreness over the 72 h following exercise. Moreover, CM supplementation had no effect on blood lactate concentrations following exercise. More recently, a trial using a lower dose (6 g versus 8 g in the present investigation) did not find any significant difference in any dependent variables (including number of repetitions, soreness, creatinine kinase levels and rating of perceived exertion [25]. The present investigation also included chemical analysis of the supplement used, which concluded that the ratio of citrulline to malate was not equivalent to the ratio stated by the manufacturer. Analysis suggests that the supplement contained just over half the manufacturers stated dose of citrulline. Subsequent analysis of four additional commercially available CM products, with a purported ratio of 2:1 (citrulline to malate), shows that only one of these products had a value close to the ratio stated by the manufacturers. This is the first study, to our knowledge, to report the citrulline to malate ratio of a number of these supplements.

Although no difference was found for the main outcome measure of repetitions, this design utilised an acute dose of CM and our data supports the findings of recent investigations of resistance training performance [2527]. In each instance, a 6 to 8 g acute dose CM had no effect on the number of repetitions achieved when performing either upper or lower-body resistance exercises compared to a placebo. By way of comparison, our results are in contrast to other trials, which investigated the effects of 8 g of acutely administered CM on the number of repetitions achieved during both upper and lower body exercise challenges [17, 18, 22]. It is however worth pointing out that there are differences in the study design, subject populations and mode of delivery between the present investigation and previous trials. The present study and utilised a single-leg leg extension exercise, and although the subjects had some experience with weight training the lack of familiarisation with the isokinetic chair protocol may have influenced performance. We did accommodate for this however with our counterbalanced design to negate any potential training effect. It is plausible that CM may be more effective in highly trained resistance trained populations. Differences between fixed range of motion single joint exercises and multi joint exercises employed in the different modalities may have also resulted in the differences between this study and previous trials findings. Although most athletes will likely utilise free weight exercises in their training, the use of an isokinetic chair should be considered a strength for its ability to deliver a consistent resistance and standardisation across subjects.

Citrulline malate supplementation also had no effect on the loss of force production and similar results were noted by Martinez-Sanchez et al. [29], who found CM was ineffective at preserving mean average force, peak average force, or the decline in force production following 8 sets of 8 half squats at 80% RM. The addition of pomegranate juice containing ellagitannins however did improve efficacy of the supplement [29]. Similarly, a recent trial of a pre-workout supplement which compared 3 supplements; one containing BCAA’s, creatine, caffeine and a 6 g dose of CM; one containing the same (but without caffeine); and a placebo, failed to find any significant improvement in loss of force during resistance exercise from either supplement [30]. In contrast, CM was effective in retaining muscle power measured by counter measure jump in runners following a marathon Martinez-Sanchez et al. [29] but not at retaining peak power following maximal cycle sprints in well-trained cyclists, [20] these results suggest the effectiveness of CM to retain power may be dependent exercise modality.

The GVT protocol utilised was relatively novel and although it isn’t generally employed by athletic populations such programmes are common amongst recreational weight trainers likely to utilise and benefit from CM. The protocol effectively reduces the number of repetitions achieved across set and was effective at increasing lactate concentrations in both the placebo and CM group. Citrulline malate however had no effect on lactate buffering when compared to a placebo which might explain the lack of difference in repetition performance between the placebo and CM group. We did not measure citrulline, arginine or NOS, following supplementation. However, previous investigations have demonstrated there is a dose response relationship between ingestion of CM and plasma citrulline, ornithine, arginine and glutamine 1 h after consumption [10, 20, 29]. L-arginine is the key substrate in NO synthesis and it has been suggested that CM may increase NO synthesis indirectly: and subsequently muscle waste-product clearance e.g. lactate [25]. Previous trials have produced mixed results in response to CM on lactate acid. It seems exercise modality may have a bearing on CMs ability to influence blood lactate concentrations. Martinez-Sanchez et al. [29] for example found a reduction in lactate and an increase in lactate dehydrogenase, responsible for converting lactate to pyruvate following marathon running using a Watermelon CM drink containing 3.45 g of citrulline. While, Kiyici et al. [5] reported a reduction in lactate concentrations in handball players immediately following exercise after supplementing with 4 g of CM 4 times per week for 4 weeks. In contrast Cunniffe et al. [20] found no effect of 12 g of CM on blood pH or lactate following 10 sets of 15 s anaerobic sprints on a cycle ergonometer. Likewise, a 6 g per day, 15 supplementation protocol of CM had no effect on muscle pH in patients suffering from fatigue following aerobic finger flexion exercise [2]. Furthermore, in trials similar to this one, Wax et al. [18], Wax et al. [19] and de Silva et al. [25] all found no effect of 6 to 8 g of CM on lactate concentrations following resistance training exercise. While Martinez-Sanchez et al. [29] reported no difference in lactate dehydrogenase concentrations compared to placebo following half-squat exercises. Interestingly however Martinez-Sanchez et al. [29] did report a different in plasma urea concentrations when citrulline was consumed in combination with pomegranate extract high in ellagitannins suggesting CM play some role in ammonia metabolism, proteolysis and recovery.

The eccentric component of exercise is thought to be a prominent factor in causing DOMS [31], despite employing a concentric only protocol, the GVT was effective in increasing muscle soreness following the intervention. The CM did not attenuate the muscle soreness and actually appeared to increase soreness. This lack of effect on muscle soreness is in agreement with other investigations [25], and in contrast to other investigations [17, 29]. Muscle soreness was also significantly reduced following administration with citrulline infused watermelon drinks following both a half marathon [29] and repeated sprints on a cycloergonometer (32). The fact that soreness was reduced following both aerobic and anaerobic exercise trials may make a case for CM being more effective in reducing soreness across modalities. The lack of soreness seen in this study is consistent with the fact we also found no difference in repetition performance, force loss, or lactate between the placebo and CM group. Presumably greater muscle soreness would likely have been observed had more repetitions been achieved from either group, although CM is proposed to offset this soreness by increased ammonium and lactate clearance. We however did not measure ammonium concentrations but did note a failure in CM to resist fatigue.

Chemical analysis of the CM product used in the study suggested that the citrulline:malate composition ratio was lower than that claimed by the manufacturer. The present investigation therefore used a lower dose of citrulline than we had intended administering (around 4.2 g instead of 5.3 g, based on a 2:1 ratio), this is still higher than the dose contained in many pre-exercise supplements and comparable doses administered in other studies [18, 19]. The lack of significant findings may therefore reflect a lower dosing of CM by comparison to other investigations. Interestingly citrulline has been infused into watermelon in other trials [29, 32] and appears to be more readily absorbed from a matrix of watermelon compared to a functional drink. Many manufacturers state the use of 2:1 formulas and it is unclear how much citrulline was administered in other investigations as the CM ratios and chemical analysis is often unreported. Further investigations should therefore explore the composition of the product being tested, as this may influence measurements and outcomes. Citrulline has been purported for its performance enhancing effect, while malate has been cited to improve performance via being an intermediate in the citric acid cycle. It seems more likely that citrulline is the active ingredient as malic acid is utilised to allow the supplement to form a stable salt for storage and neutralise the basicity of the supplement. This stability issue may be overcome by infusion into a watermelon matrix. Performance-enhancing effects have been found from using citrulline in isolation [14] and researchers may wish to concentrate on using pure citrulline, to avoid under dosed products.

Finally, given a number of trials suggesting a positive effect of CM on anaerobic performance (for e.g.’s see [19, 22]) further trials should be carried out utilising both acute and loading doses of CM for both isokinetic and multi joint protocols, using experienced resistance trained populations. There were a number of limitations of the present investigation include no measure of CK or ammonia to accompany the data gathered on lactate and muscle soreness. The subjects who participated in the study were not a homogenous group, and subjects varied in their training experience and included both men and women not controlling for the impact of menstrual cycling; however all participants had at least six months regular resistance training experience and are likely customers for the supplement in question. We also did not perform the experiment fasted, but subjects were instructed to consume the same breakfast on both occasions, however we did note no difference in 24 h dietary intake between laboratory visits and point out that exercisers are unlikely to perform in the fasted state. We also did not control or audit dietary supplements other than asking subjects to not stop or begin supplement regimes throughout the trial the rationale for this is that stopping or starting a new supplement could either positively or negatively affect performance. Furthermore, the isokinetic chair induces an eccentric and concentric force which is not the same, for example, as a standard leg extension exercise performed in a commercial fitness facility; field studies with commonly used equipment and exercises are needed to help answer questions over the efficacy of CM. Additionally particular attention should be paid to the chemical analysis of supplement content carried out in the present paper, which raises concerns over manufacturer’s claims. Athletes and coaches should remain sceptical as to the efficacy of CM.