1. Introduction
Choline has established itself as an important nutrient essential for proper functioning of many organs in the human body [
1]. As part of it being a critical component of the neuronal phospholipid bilayer, choline is connected to signal transduction and methylation of DNA, histones, and neurons. Choline is a precursor of acetylcholine (ACh), a neurotransmitter involved in memory, attention, and skeletal muscle contraction [
1]. As such, ACh is a key component of communication throughout the nervous system, thus creating its theorized connection to cognition while ACh also sits at the coalface between the nervous and muscular systems at the neuromuscular junction. Previous work has indicated that supplementation with alpha-glycerylphosphorylcholine (A-GPC), a choline-containing phospholipid, can increase choline levels, and in doing so may augment cognition, physical performance, and endogenous hormone production [
2,
3,
4].
Due to A-GPC’s ability to increase choline levels [
5] and other evidence that demonstrates disruption in cholinergic transmission leading to learning and memory deficits in times of reduced ACh availability [
6], the interest in A-GPC supplementation to support cognition has been evident for several years. Currently, research findings are mixed regarding A-GPC’s potential to impact cognition [
7,
8]. When investigated in clinically compromised or healthy aged populations, A-GPC studies have indicated that A-GPC supplementation enhances memory and cognitive function and is an effective treatment for neurodegenerative disorders such as Alzheimer’s disease and dementia [
1,
9,
10,
11]. A-GPC has also been proposed to offer treatment support for traumatic brain and other cerebrovascular accidents [
12]. Research in healthy populations, however, is scant and, to date, the majority of these studies have failed to highlight cognitive benefits in healthy individuals [
13,
14]. One of the initial studies exploring A-GPC for its ability to impact cognition used 32 healthy young volunteers who were administered an intramuscular injection of 1000 mg A-GPC or placebo for ten days before having cognition evaluated [
15]. The authors reported A-GPC administration was able to antagonize memory and attention impairment induced by scopolamine. Unfortunately, the method of administration (intramuscular injection) in this study challenges its translation to larger segments of the population. A 2021 study by Tamura et al. [
4] administered daily 400 mg doses of A-GPC or a placebo for 14 days to healthy volunteers who completed a scale to evaluate emotional states. After the treatment was administered, motivation levels were significantly improved while anxiety levels were not impacted. Two other studies in healthy individuals examined the potential for A-GPC to impact cognition. The first study was completed by Parker et al. [
14] who used a randomized, double-blind, placebo-controlled, crossover design where they had 20 healthy men and women complete visual analog scales, a serial subtraction test, and a battery of physical performance tests after ingesting either 200 mg of A-GPC, 200 mg of caffeine, 400 mg of A-GPC, or a placebo. No statistically significant changes were observed in the serial subtraction test although the 200 mg dose of A-GPC scores were 18.1% and 10.5% faster than the caffeine and placebo groups, respectively. Additionally, Bunn and colleagues [
13] acutely supplemented 21 college-aged males in a randomized, crossover manner to receive either a placebo or a combination of 500 mg A-GPC, 250 mg of uridine-5’-monophosphate, and 1,500 mg DHA). During each condition, participants had their cognition assessed using the ImPACT protocol along with vertical jump and maximal bench press repetitions. Results from this study also indicated that a single day of supplementation exerted no impact on their outcomes.
Other studies have also explored the potential for A-GPC to positively influence various indicators of muscular performance [
3,
5,
14,
16]. The results from these studies are currently mixed, which may be due to the length and amount of supplementation provided in each study. Three peer-reviewed studies are available that support the ability of A-GPC to augment exercise performance. Bellar et al. [
3] supplemented 13 young, healthy males with either 600 mg A-GPC or placebo for six days and reported significant improvements in lower body force production. Another study involving young, healthy males required the participants to supplement with either a placebo, 250 mg A-GPC, or 500 mg A-GPC in a randomized, double-blind fashion for seven consecutive days [
5]. Serum choline was increased along with significantly greater maximum velocity and maximum power values during a countermovement jump. Finally, Harrington et al. [
17] reported that muscular power was improved in 30 male trained cyclists after seven days of supplementing with 300 mg of A-GPC, but the A-GPC was provided as part of a combination that also included BCAAs and citrulline, so the independent impact of A-GPC was not able to be clearly discerned from this study.
Three reports are available that outline findings after a single dose was delivered. In an abstract presented as part of a conference proceedings, Ziegenfuss et al. [
16] used a randomized, double-blind, placebo-controlled, crossover approach with seven healthy men to examine the impact of consuming a single 600 mg dose of A-GPC 90 minutes prior to completing a bout of lower-body resistance exercise and reported that peak force production (p < 0.02) during a series of bench press throws was greater when A-GPC was provided while no changes in peak velocity or peak power were observed. Alternatively, no changes in vertical jump performance were reported in an abstract presented as part of a conference proceedings by Parker et al. [
14] who supplemented 20 healthy men and women with either 200 mg of A-GPC, 200 mg of caffeine, 400 mg of A-GPC, or a placebo. Finally, Bunn and colleagues [
13] acutely supplemented 21 college-aged males in a randomized, crossover manner to receive either a placebo or a combination of 500 mg A-GPC, 250 mg of uridine-5’-monophosphate, and 1,500 mg DHA) and reported that supplementation did not impact vertical jump performance or the maximal number of bench press repetitions completed.
A final area of interest has been in the ability of A-GPC to heighten endogenous production of growth hormone secondary to activation of the hypothalamic-pituitary axis. Kawamura et al. [
2] supplemented eight healthy males with either 1000 mg A-GPC or a placebo in a randomized, double-blind, crossover fashion and reported that growth hormone levels were significantly increased 60 minutes after taking A-GPC. Another study was completed by Maldonado et al. [
18] as part of a graduate thesis and has not been published in a peer-reviewed journal that required overweight adults to supplement with 1200 mg of A-GPC each day for eight weeks. No changes in growth hormone were identified in this study. Finally, Ziegenfuss et al. [
16] reported a significantly greater area under the curve for growth hormone in response to a heavy bout of lower-body resistance exercise when a 600 mg dose of A-GPC was provided when compared to placebo. With the popularity of A-GPC being used in an acute fashion as part of various nutritional formulations ingested prior to workouts and competitions, more research is needed to examine the ability of A-GPC to impact cognition and exercise performance. Therefore, the purpose of this study was to evaluate the impact of two different doses of A-GPC on cognitive and physical performance in healthy resistance-trained individuals.
4. Discussion
Choline availability can have widespread physiological impact by way of impacting acetylcholine status across the body and facilitating other intracellular communication. A-GPC supplementation has been documented in studies to positively influence choline levels [
5,
35] and exert influence over mood [
4], cognition [
1,
9,
12], and physical performance [
3,
5]. The primary findings from the current study highlight the statistically significant improvement in Stroop total score when participants supplemented with both HD and LD when compared to PL (
Figure 3). Additionally, outcomes revealed that HD spent significantly less time (they completed the test faster) on the test when compared to PL (
Figure 4). Forced post-hocs also revealed that HD exhibited faster responses times during the Flanker test when compared to PL (
Table 5). Finally, although faster, HD exhibited decreased accuracy when compared to LD, but accuracy was similar when compared to PL (
Table 5). These results are meaningful because they represent the first time that A-GPC supplementation has been documented to increase parameters of cognitive performance in a young, healthy cohort. Previous investigations using this population were unable to identify such changes [
13,
14] while studies involving older and clinically compromised populations [
1] have reported on the ability of A-GPC to positively impact cognitive performance. The reasons for why these outcomes deviate from previous studies involving healthy populations are not entirely known. The current study and the two other investigations using single doses of oral ingestion both used robust randomized, double-blind, crossover, placebo-controlled study designs and made acute assessments of physical and cognitive performance. The Stroop color and matching test is a clinically validated and popular assessment of selective attention, cognitive flexibility, processing speed, and inhibition while the other investigations used the ImPACT [
13] and serial subtraction [
14] tests. The higher A-GPC dosage (HD: 630 mg) during the present study was higher than the dosages provided during both the Parker (200 mg and 400 mg) and Bunn (500 mg) investigations [
13,
14], which may have impacted the outcomes, but the lower dose (LD: 315 mg) also exerted some positive influence, which somewhat rebuts this as an explanation for our outcomes. It is worth mentioning that the A-GPC delivered as part of the Bunn investigation was combined with two other ingredients (UMP and DHA), which further challenges one’s ability to compare the results of these investigations. Collectively, each of these studies used young, healthy cohorts (20 – 30 years) and completed measurements within a similar time frame of administration (30 – 90 minutes after ingestion). Another difference may be the manufacturing method utilized for the A-GPC provided in the current investigation versus the other investigations. The A-GPC used in the present study (GeniusPure, NNB Nutrition) is commercially produced at a higher purity rate in a manner that is free of soy and other common allergens. The extent to which commercial production methods may have impacted the bioavailability or absorption kinetics is not currently known.
The present study also examined the potential impact of A-GPC’s ability to augment physical performance using a study approach that closely mimicked the study design previously utilized by Ziegenfuss et al. [
16]. Using this approach, peak force produced during the vertical jumps was different between groups with post-hoc comparisons indicating that LD was greater than HD (p = 0.0.27, d = 0.54) and tended to be greater than PL (p = 0.085, d = 0.41) (
Table 7). A trend was observed for vertical jump peak power using oneway ANOVA (p = 0.085) and forced LSD post-hocs with LD exhibiting a tendency to be greater than PL (p = 0.068, d = 0.43) and HD (p = 0.071, d = 0.43). Changes in upper-body performance were similar between all three groups using oneway ANOVA (
Table 7) but forced post-hocs indicated a statistically significant difference was present between HD and PL (p = 0.043, d = 0.49). These results align somewhat with the previous studies examining physical performance changes in a young, healthy cohort [
3,
5,
16]. Briefly, Bellar et al. [
3] reported statistically significant improvements in peak force production while Marcus et al. [
5] reported greater peak power production when A-GPC was provided. In addition, and in alignment with the present study, Ziegenfuss et al. [
16] measured a series of upper-body and lower-body outcomes and reported significant increases in upper-body force versus placebo and a statistical trend for A-GPC to be greater than PL. Both the Bellar and Marcus studies dosed orally for six and seven days, respectively, while Ziegenfuss delivered single oral doses as what was completed in the present study. Thus, it seems plausible that if a longer supplementation protocol was followed in the present study that force and power production may have been further augmented to yield even stronger outcomes in the present study related to performance. Future research should examine this as a possibility.
The present study also sought to examine changes in perception of affect (mood, motivation, etc.) and growth hormone changes in response to an intense, acute bout of lower-body resistance exercise. Using visual analog scales, perceptions of mood, alertness, concentration, and motivation towards physical and mental exercise were assessed and no changes were observed between any groups. The working hypothesis for these measurements suggested that the known increases in choline which occur with A-GPC supplementation may have improved these outcomes, particularly when surrounded by a challenging acute bout of exercise. Moreover, Tamura et al. [
4] previously showed that 14 days of 400 mg of A-GPC given orally can positively impact mood, but no changes in these variables were observed in the present study. Thus it seems prudent to suggest that future studies should examine if longer dosing protocols would demonstrate changes in these outcomes. Circulating growth hormone levels were also evaluated after A-GPC supplementation as it has been shown in previous studies to increase growth hormone levels [
2,
16,
36]. In contrast to the results provided by Ziegenfuss et al. [
16], no differences in growth hormone concentrations were observed in the present study. While our observed changes in growth hormone align with previous investigations that reported on growth hormone changes after acute bouts of resistance exercise [
37,
38], the lack of changes secondary to A-GPC delivery was somewhat unexpected considering the similar dosages used and the study design replication that was employed to the Ziegenfuss study [
16]. A ceiling effect surrounding growth hormone response is evident in this literature, which may have impacted any potential for A-GPC to further augment this response. Whether or not differences existed within the A-GPC remains to be seen and future studies using prolonged supplementation regimens of higher doses should be completed to continue to explore the ability of A-GPC to function as a growth hormone secretagogue
The current paper has a few key strengths to highlight starting with the randomized, double-blind, placebo-controlled, crossover study design used to examine the measured outcomes. In addition, the current study also provides insight into any dose-response outcomes which may occur with A-GPC supplementation. Other key strengths lie with the completion of both cognitive and physical performance outcomes after one dose of supplementation. A few limitations should also be highlighted for the reviewer to consider when evaluating the findings. We only recruited males into this study to best align our outcomes with the current literature and as result more research involving females should be completed to establish how well these results hold true for females. A full discovery of pairwise comparisons using LSD approaches was completed for primary and secondary outcomes to clearly understand all potential differences. It is acknowledged and should be considered by the reader that using more rigorous correction for pairwise comparisons may have impacted our final outcomes. Another key limitation was the single dose delivered as part of this study design. While results after a single dose have obvious value, being able to understand how the findings from our current study design are impacted by longer supplementation regimens would also be of interest, particularly if longer doses are also considered.