Exploring the Impact of Menstrual Cycle Phases on Agility Performance in Semi-Professional Female Soccer Players

Abstract

Background/Objectives: To analyze how the different phases of the menstrual cycle affect agility in female football players. Methods: A total of 11 female football players were selected from the third tier of the Spanish Football Federation (Third RFEF) and an agility test (t-test) was conducted to measure agility during the three phases of the menstrual cycle: the menstrual, late follicular, and mid-luteal phases. These phases were determined through self-reporting and the use of ovulation test strips for luteinizing hormone detection. Perceptual variables, such as sleep quality, stress, muscle pain, and fatigue, as well as the rating of perceived exertion, were measured. Results: There were no statistically significant differences in agility performance across menstrual cycle phases (F_(2,20) = 1.86; p = 0.18). However, performance in the mid-luteal phase was slightly better compared to other phases. Similarly, no significant differences were found in perceptual variables such as fatigue, sleep quality, stress, and muscle soreness (p > 0.05), although slightly better perceptual responses were observed in the late follicular phase. Conclusions: No significant differences were found when analyzing the influence of menstrual cycle phases on agility, although performance appeared slightly better in the mid-luteal phase. No significant differences were observed in the perceptual variables. Both objective and perceptual variables should be considered in future studies or training programs based on the menstrual cycle.

1. Introduction

Greater emphasis should be placed on studies involving women in the field of sports science, as representation is notably low, with participation rates below 35% [1,2]. It is essential to consider the complexity in women’s sports as well, given the unique characteristic of women experiencing a menstrual cycle (MC), with hormonal fluctuations each month [3]. Nevertheless, the number of women engaging in sports continues to rise, with football being one of the most prominent disciplines, both at training and elite levels. Consequently, the MC emerges as a critical factor to consider, not only for athletic performance but also when planning training [4].

The MC is a physiological process of the body with different hormonal variations. This cycle spans the time between two menstrual or bleeding phases, encompassing distinct stages, and is considered normal or eumenorrheic if it lasts between 21 and 35 days [5]. During this period, women undergo the different phases of the cycle: menstrual, follicular, ovulatory, and luteal. These phases are characterized by hormonal fluctuations [3] that influence the function of organs and tissues in the body, as well as energy levels, perceived exertion, mood, and various other factors that may impact athletic performance [6].

During the menstrual phase or the early follicular phase (EFP), the hormone levels of estrogen and progesterone are low (days 1–5 of the MC). In the late follicular phase (LFP), there is an estrogen peak (days 6–12 of the MC), which is often associated with an enhanced anabolic response to exercise. In the mid-luteal phase (MLP), characterized by a progesterone peak (days 20–24 of the MC), a possible catabolic effect is observed [7,8,9]. Therefore, these phases are more likely to exhibit variations or differences in performance [9,10].

Currently, scientific literature has not identified clear evidence regarding how MC phases influence athletic performance. For example, Colenso-Semple et al. [11] found no influence on acute strength performance, whereas Niering et al. [12] reported slight improvements towards the late follicular phase. In contrast, Taylor et al. [13] found no differences in endurance-trained athletes across phases. Importantly, most of these studies focus on strength or endurance, with agility performance remaining largely unexplored, despite being critical in football.

Performance in football is influenced by numerous physical, physiological, psychological, social, technical, and tactical variables [14], making its analysis through performance indicators inherently complex [15]. Agility is defined as the ability to change body position quickly and effectively [16]. This skill is essential in football, as it enhances players’ ability to move rapidly and efficiently across the field. Given that the MC is an additional variable to consider in women’s sports, it may influence athletes’ agility due to hormonal fluctuations [17], alterations in muscle elasticity and tone [18], or other physical and physiological changes occurring throughout the cycle. It is not clear how exactly the MC phases affect performance in football [19]. Understanding how the MC affects agility could serve as an effective tool for designing training plans aimed at improving agility, thereby enhancing performance, preventing injuries, improving directional changes, dribbling skills, and even reaction times in unexpected situations [20].

Due to the controversy, lack of knowledge, and limited research regarding how the phases of the MC may influence athletic performance [1,3,9,11,12,21,22], there is a recognized need to examine their impact on agility in female athletes.

This study is original in its approach as it examines agility performance—a fundamental skill in football—across different MC phases in semi-professional female football players. Additionally, by combining objective agility testing with perceptual wellness variables, this research offers a more comprehensive understanding of performance fluctuations, potentially guiding individualized training and injury prevention strategies in female athletes.

Therefore, the objective of this study was to analyze how the different phases of the MC affect agility in female football players and to analyze the influence of the MC phases on perceptual variables.

2. Results

2.1. t-Test and MC Phases

No significant differences were found between menstrual cycle phases in the agility test (F_(2,20) = 1.86; p = 0.18). However, performance in the mid-luteal phase (11.34 s) was slightly better compared to the menstrual (11.58 s) and follicular (11.47 s) phases, with the largest differences observed in the first part of the t-test (

Table 1. t-test results based on MC Phases.

Variables	Menstrual Phase		Late Follicular Phase		Mid-Luteal Phase		ANOVA
	Mean	SD	Mean	SD	Mean	SD	F(2,20)	P	η2p
Total time	11.58	0.53	11.47	0.6	11.34	0.42	1.86	0.18	0.16
Sprint out	2.67	0.15	2.57	0.25	2.58	0.18	1.16	0.33	0.1
Shuffle right	1.6	0.13	1.54	0.12	1.51	0.13	3.27	0.06	0.25
Shuffle left	2.87	0.18	2.92	0.17	2.85	0.16	3.24	0.06	0.24
Shuffle right (2)	1.59	0.1	1.63	0.1	1.6	0.1	0.74	0.49	0.07
Sprint back	2.84	0.11	2.8	0.15	2.79	0.1	0.72	0.5	0.07

Note: SD: standard deviation; Total time (s); Sprint out (s); Shuffle right (s); Shuffle left (s); Shuffle right (2) (s); Sprint back (s); ANOVA: analysis of variance; F: Snedecor’s F; P: p-value; η²p = eta cuadrado parcial.

).

2.2. Perceptual Variables and MC Phases

No significant differences were found between any of the perceptual variables or RPE and the phases of the MC (p > 0.05). Better results were obtained in the late follicular phase for all perceptual variables related to the WQ (

Table 2. Comparative analysis of perceptual variables.

Variables	Menstrual Phase		Late Follicular Phase		Mid-Luteal Phase		ANOVA
	Mean	SD	Mean	SD	Mean	SD	F(2,20)	P	η2p
Fatigue	3.18	0.98	2.36	0.81	3.09	1.58	2.43	0.11	0.19
Sleep quality	3.27	1.01	2.91	1.38	3.18	0.87	0.37	0.69	0.04
Stress	2.91	1.04	2.36	1.12	2.82	1.17	1.22	0.32	0.11
Muscle soreness	2.82	0.75	2.45	0.93	3.09	1.3	1.71	0.21	0.15
WQ	12.2	1.72	10.1	3.05	12.2	4.14	2.52	0.11	0.2
RPE	5.09	1.51	5.36	1.36	5.27	1.27	0.48	0.63	0.04

Note: SD: standard deviation; Fatigue (scale of 1 to 7); Sleep quality (scale of 1 to 7); Stress (scale of 1 to 7); Muscle soreness (scale of 1 to 7); RPE (Rating of Perceived Exertion, scale 1 to 10); WQ (Wellness Questionnaire, sum of perceptual variables: fatigue, sleep, stress, and muscle pain); ANOVA: analysis of variance; F: Snedecor’s F; P: p-value; η²p = eta cuadrado parcial.

).

3. Discussion

The results obtained show that participants performed better in the agility test (t-test) during the MLP compared to the EFP (0.24 s) and the LFP (0.13 s), although no significant differences (p = 0.18) were observed. The perceptual variables showed better results in the LFP, with greater differences in fatigue, stress, and the total WQ score.

Currently, there are few studies that have analyzed how the phases of the menstrual cycle influence agility performance. Agility is essential as it combines accelerations and decelerations as quickly as possible, reflecting the developed force ratio capacity [23]. According to hormonal impact, strength capacity is greatly influenced by estrogen and progesterone; estrogen has a positive, neuroexcitatory effect and progesterone inhibits cortical excitability [24]. In contrast, Ref. [9], after analyzing 78 studies, found only a trivial effect of worse performance in the menstrual phase compared to the others. Similarly, Carmichael et al. [10] did not find differences in anaerobic performance. These findings are consistent with the results of the present study, which also did not show significant differences in agility performance. However, the observed improvements in our study were slightly greater in the MLP, which may have practical relevance despite the lack of statistical significance.

More specifically, in studies analyzing agility tests, Juillard et al. [25] observed that in the Illinois agility test performed with football players, the results were very similar across the different phases of the MC. This was also observed by Oğul et al. [26] in physically active women, although with a slight tendency toward the MLP. This could be due to the fact that stiffness does not seem to vary significantly during the MC [27]. On the other hand, Sawai et al. [28] found that agility performance worsened during the EFP, possibly due to fluid retention during this phase. Our results are consistent with the current evidence, showing no significant differences. However, there was a slight improvement in the MLP, which may have practical relevance. Further research is needed to better understand the effects of the menstrual cycle phases on agility performance.

Sports performance should not only be measured objectively. The subjective and perceptive aspects should also be taken into account, as these perceptual variables are an important aspect in studies related to women and the MC [10]. In general, women experience more negative symptoms related to the MC during the EFP, with over 50% of athletes reporting such symptoms [29], followed by the late luteal phase, which can affect up to 83% of athletes, leading to more fatigue or poorer physical performance perceptions [30]. In our findings, an improvement in all perceptual variables was observed during the late follicular phase, possibly due to the increased production of estrogen, which indirectly leads to an increase in serotonin [31], potentially improving athletes’ mood and reducing fatigue.

Sleep quality can be affected by the phases of the MC, mainly showing poorer quality during the premenstrual and menstrual phases [32], with possible improvement during the transition from the follicular to the luteal phase [33]. In this case, the perceived sleep quality of our athletes was worse during the menstrual phase, as was also observed by Osmani et al. [34], possibly due to menstrual symptoms. The RPE does not show differences based on the phases of the menstrual cycle, which was corroborated by several studies reviewed by Ekenros et al. [35], who also found no differences in this variable. According to the results obtained, perceptual variables based on the menstrual cycle should be taken into account to properly periodize training. This aligns with the comments made by Julian et al. [36], but further studies should continue to focus on objective performance while considering perceptual variables in order to eventually apply this knowledge in practical contexts with appropriate individualization for each athlete.

• Limitations

Despite the findings obtained, some limitations should be considered. First, hormone concentration measurements were not performed through blood analysis; however, LH ovulation kits and self-recording of menstrual cycle phases via a mobile application were used to determine the phases of the menstrual cycle as objectively as possible, as recommended by McNulty et al. [37]. Second, the number of participants was limited (n = 11), but the sample had “moderate statistical power”. Third, although the participants were familiar with the agility test and performing direction changes, there may have been a learning effect across the different test attempts.

4. Materials and Methods

• Experimental Approach to the Problem

A cross-sectional cohort experimental study with repeated measures was conducted to analyze the effect of the menstrual cycle (MC) phases on agility, perceptual variables from the Hopper scale, and the subjective perception of effort (RPE). The MC phases were determined using two tools: a calendar and a luteinizing hormone (LH) urine kit [38]. Participant assessments were conducted during the three phases of the menstrual cycle: the menstrual phase or early follicular phase, the pre-ovulatory phase or late follicular phase, and the post-ovulatory phase or mid-luteal phase.

• Subjects

A total of 11 female football players were selected from the third tier of the Spanish Football Federation (3rd RFEF), corresponding to Tier 3 level [39]. Their characteristics (mean ± standard deviation) were: age = 18.7 ± 3.3 years; height = 1.62 ± 0.07 m; weight = 58.6 ± 6.7 kg. The inclusion criteria were: (i) having a regular MC (21–35 days) in the last 6 months; (ii) at least 2 years of football experience; (iii) not using contraceptives; (iv) not suffering from any disease or injury. The procedure and instruments to be used were explained to the participants prior to the start of the protocol. Informed consent was signed by each participant, which addressed the protection of personal data, ethical criteria for human experimentation, and ensured the highest confidentiality and scientific rigor as detailed in the 1975 Declaration of Helsinki. Furthermore, this study was approved by the Ethics Committee (CEI 03-2024). The sample size was selected based on convenience, and a post hoc power analysis was conducted using G*Power software (version 3.1.9.7), with α = 0.05, power (1 − β) = 0.8, and an effect size of 0.25. Using the statistical test ANOVA: repeated measures within factors, the analysis yielded a moderate power of 0.37.

• Protocol

The participants were scheduled 3 times during the study, corresponding to their training field and during the 3 phases of the MC to be analyzed: (i) menstrual phase, between days 1–5; (ii) late follicular phase, between days 6–12; and (iii) mid-luteal phase, between days 20–24.

Determination of the MC phases: The MC of the participants was monitored using a mobile application (Clue^® Period Tracker, BioWink GmbH, Berlin, Germany) and an LH ovulation kit (Ovulation LH Test Strip, Cuckool, Nantong, China). The tests were conducted during the menstrual phase (2.82 ± 1.08 days), late follicular phase (10.2 ± 1.17 days), and mid-luteal phase (21.8 ± 1.33 days). Once a positive LH result was confirmed, the test was repeated a few days later, approximately between days 20–24 of the menstrual cycle. The average day on which the players tested positive with the ovulation kit was day 14.7 ± 1.27 days.

Agility Test (t-test): This test was chosen to assess the agility of each player, and is a valid and reliable test for this purpose [40,41]. Two valid attempts were made, with a 3-min rest between them to ensure complete recovery [41,42]. The players started the test with both feet behind the sensor (BlazePod^® Reaction Training System, BlazePod Ltd., Tel Aviv, Israel), ran forward 9.14 m, and touched the next sensor. Then, the players moved 4.57 m to the right to touch another sensor with their right hand, followed by a lateral movement to the left for 9.14 m to touch the sensor with their left hand. They then moved again 4.57 m to the right to touch the second-to-last sensor. Finally, the players ran backward to reach the final sensor. Tests in which players crossed their feet, missed a BlazePod sensor, or did not face forward were considered invalid [43].

Perceptual Variables: During the intervention days, the wellness questionnaire (WQ) test was administered before the agility test. It was also used the day after the test to assess participants’ sleep quality, fatigue, muscle soreness, and stress variables, with values ranging from 1 to 7, where 1 represented very good and 7 represented very bad [44]. The range of perceived effort (RPE) (scale 1–10) [45] was also recorded after each t-test to determine participants’ subjective effort perception. All participants were familiarized and trained on the proper use of these tools.

• Statistical analysis

A descriptive analysis was conducted, with data presented as mean ± standard deviation (SD). All data were recorded in an Excel database, and the statistical analyses were performed using Jamovi software (2.5.5). A repeated measures ANOVA was conducted to compare execution time, t-test sector performance, and perceptual variables across the three phases of the CM. Normality was assessed using the Shapiro–Wilk test (p < 0.05).

5. Conclusions

No significant differences were found regarding the influence of the menstrual cycle phases on agility performance. However, performance was slightly better in the mid-luteal phase, although this difference was not statistically significant. Similarly, perceptual variables showed slight improvements in the late follicular phase, without reaching statistical significance. These findings suggest that such variations may be worth considering when designing individualized training periodization in real-world athletic contexts. Both objective and perceptual variables should be taken into account in studies or training programs based on the menstrual cycle.

• Future practical applications

To build on the findings of the present study, further randomized controlled trials with larger sample sizes are necessary to increase the statistical power and improve the generalizability of the results. Also, it is highly recommended to incorporate more objective hormonal monitoring techniques, such as blood assays or salivary hormone analysis, to accurately determine the menstrual cycle phases or hormone concentrations of athletes. This approach would allow a better understanding of the physiological fluctuations and their potential impact on performance variables. Furthermore, given the importance of agility in many sports disciplines, future research should place a stronger emphasis on the detailed and specific analysis of agility performance during different menstrual phases.