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Temporal changes in blood oxidative stress biomarkers across the menstrual cycle and with oral contraceptive use in active women

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To examine the temporal changes in blood oxidative stress biomarkers in recreationally-trained women that were naturally-cycling (WomenNC) or using oral contraceptives (WomenOC) across one month.


Blood samples were acquired at three timepoints of the menstrual cycle (1: early-follicular, 2: late-follicular and 3: mid-luteal) and oral contraceptive packet (1: InactiveOC, 2: Mid-activeOC and 3: Late-activeOC) for determination of estradiol, progesterone, oxidative stress, C-reactive protein (CRP) and other cardiometabolic biomarkers in plasma and serum.


There was a Group by Time effect on estradiol (p < 0.001, partial η2 = 0.64) and progesterone (p < 0.001, partial η2 = 0.77). Malondialdehyde, lipid hydroperoxides and CRP concentrations were higher in WomenOC during Late-activeOC compared to InactiveOC (+ 96%, + 23% and + 104%, respectively, p < 0.05). However, there were no changes in these biomarkers across the menstrual cycle in WomenNC (p > 0.05). At all timepoints (i.e., 1, 2 and 3), WomenOC had elevated lipid hydroperoxides (+ 28, + 48% and + 50%) and CRP (+ 71%, + 117% and + 130%) compared to WomenNC (p < 0.05, partial η2 > 0.25). There was no Group by Time effect on non-enzymatic antioxidants or glutathione peroxidase; however, glutathione peroxidase was lower in WomenOC, i.e., main effect of group (p < 0.05, partial η2 > 0.20).


These findings demonstrate that WomenOC not only have higher oxidative stress and CRP than WomenNC, but also a transient increase across one month of habitual oral contraceptive use. Since changes in oxidative stress and CRP often relate to training stress and recovery, these outcomes may have implications to workload monitoring practices in female athletes.

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Alanine aminotransferase


Analysis of variance


Aspartate aminotransferase


C-reactive protein


Coefficient of variation


Free oxygen radical defence


Free oxygen radical test


Gamma-glutamyl transferase


Glutathione peroxidase


No synthetic hormone ingestion (placebo pills)


Between 14 and 19 days of exogenous sex steroid ingestion


Lactate dehydrogenase


Luteinizing hormone


Multivariate analysis of variance




Between 5 and 7 days of exogenous sex steroid ingestion


Oral contraception


Reactive oxygen and nitrogen species


Standard deviation


Total antioxidant capacity


Total iron binding capacity


Unsaturated iron binding capacity

\(\dot{V}\)O2 :

Oxygen uptake


Peak oxygen uptake


Women with a natural, regular menstrual cycle


Women using combined, monophasic oral contraceptives


  • Baltgalvis KA, Greising SM, Warren GL, Lowe DA (2010) Estrogen regulates estrogen receptors and antioxidant gene expression in mouse skeletal muscle. PLoS ONE 5:e10164

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Becatti M et al (2017) Redox status alterations during the competitive season in élite soccer players: focus on peripheral leukocyte-derived ROS. Intern Emerg Med 12:777–788

    Article  PubMed  Google Scholar 

  • Bell HK, Bloomer RJ (2010) Impact of serum estradiol on postprandial lipemia, oxidative stress, and inflammation across a single menstrual cycle. Gend Med 7:166–178

    Article  PubMed  Google Scholar 

  • Bellanti F, Matteo M, Rollo T, De Rosario F, Greco P, Vendemiale G, Serviddio G (2013) Sex hormones modulate circulating antioxidant enzymes: impact of estrogen therapy. Redox Biol 1:340–346

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Blum CA et al (2005) Low-grade inflammation and estimates of insulin resistance during the menstrual cycle in lean and overweight women. J Clin Endocrinol Metab 90:3230–3235

    Article  CAS  PubMed  Google Scholar 

  • Borg GA (1982) Psychophysical bases of perceived exertion. Med Sci Sports Exerc 14:377–381

    Article  CAS  PubMed  Google Scholar 

  • Braakhuis AJ, Hopkins WG, Lowe TE, Rush EC (2011) Development and validation of a food-frequency questionnaire to assess short-term antioxidant intake in athletes. Int J Sport Nutr Exerc Metab 21:105–112

    Article  CAS  PubMed  Google Scholar 

  • Bruinvels G, Burden R, Brown N, Richards T, Pedlar C (2016) The prevalence and impact of heavy menstrual bleeding (menorrhagia) in elite and non-elite athletes. PLoS ONE 11:e0149881

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Capobianco G et al (2010) Plasma levels of C-reactive protein, leptin and glycosaminoglycans during spontaneous menstrual cycle: differences between ovulatory and anovulatory cycles. Arch Gynecol Obstet 282:207–213

    Article  CAS  PubMed  Google Scholar 

  • Cauci S, Buligan C, Marangone M, Francescato MP (2016) Oxidative stress in female athletes using combined oral contraceptives. Sports Med Open 2:40

    Article  PubMed  PubMed Central  Google Scholar 

  • Cauci S, Francescato MP, Curcio F (2017) Combined oral contraceptives increase high-sensitivity C-reactive protein but not haptoglobin in female athletes. Sports Med 47:175–185

    Article  PubMed  Google Scholar 

  • Chung SC, Goldfarb AH, Jamurtas AZ, Hegde SS, Lee J (1999) Effect of exercise during the follicular and luteal phases on indices of oxidative stress in healthy women. Med Sci Sports Exerc 31:409–413

    Article  CAS  PubMed  Google Scholar 

  • Clancy KB, Baerwald AR, Pierson RA (2013) Systemic inflammation is associated with ovarian follicular dynamics during the human menstrual cycle. PLoS ONE 8(5):e64807.

  • Cook NR, Buring JE, Ridker PM (2006) The effect of including C-reactive protein in cardiovascular risk prediction models for women. Ann Intern Med 145:21–29

    Article  CAS  PubMed  Google Scholar 

  • Cornelli U, Belcaro G, Cesarone MR, Finco A (2013) Analysis of oxidative stress during the menstrual cycle. Reprod Biol Endocrinol 11:74

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Critchley H, Kelly RW, Brenner RM, Baird DT (2001) The endocrinology of menstruation: a role for the immune system. Clin Endocrinol 55:701–710

    Article  CAS  Google Scholar 

  • d Kerdelhué B, Brown S, Lenoir V, Queenan JT Jr, Jones GS, Scholler R, Jones HW Jr (2002) Timing of initiation of the preovulatory luteinizing hormone surge and its relationship with the circadian cortisol rhythm in the human. Neuroendocrinology 75:158–163

    Article  PubMed  Google Scholar 

  • De Groote D, d’Hauterive SP, Pintiaux A, Balteau B, Gerday C, Claesen J, Foidart J-M (2009) Effects of oral contraception with ethinylestradiol and drospirenone on oxidative stress in women 18–35 years old. Contraception 80:187–193

    Article  PubMed  CAS  Google Scholar 

  • Decroix L, De Pauw K, Foster C, Meeusen R (2016) Guidelines to classify female subject groups in sport-science research. Int J Sports Physiol Perform 11:204–213

    Article  PubMed  Google Scholar 

  • Dekany M, Nemeskeri V, Györe I, Harbula I, Malomsoki J, Pucsok J (2006) Antioxidant status of interval-trained athletes in various sports. Int J Sports Med 27:112–116

    Article  CAS  PubMed  Google Scholar 

  • Fallah S, Sani FV, Firoozrai M (2009) Effect of contraceptive pill on the selenium and zinc status of healthy subjects. Contraception 80:40–43

    Article  CAS  PubMed  Google Scholar 

  • Fernández-Lázaro D, Fernandez-Lazaro CI, Mielgo-Ayuso J, Navascués LJ, CórdovaMartínez A, Seco-Calvo J (2020) The role of selenium mineral tace element in exercise: antioxidant defense system, muscle performance, hormone response, and athletic performance. A systematic review. Nutrients 12:1790

    Article  PubMed Central  CAS  Google Scholar 

  • Finaud J, Scislowski V, Lac G, Durand D, Vidalin H, Robert A, Filaire E (2006) Antioxidant status and oxidative stress in professional rugby players: evolution throughout a season. Int J Sports Med 27:87–93

    Article  CAS  PubMed  Google Scholar 

  • Finco A, Belcaro G, Cesarone MR (2011) Assessment of the activity of an oral contraceptive on the levels of oxidative stress and changes in oxidative stress after co-treatment with two different types of physiological modulators with antioxidant action. Contraception 84:418–422

    Article  CAS  PubMed  Google Scholar 

  • Gaskins AJ et al (2012) Endogenous reproductive hormones and C-reactive protein across the menstrual cycle: the BioCycle Study. Am J Epidemiol 175:423–431

    Article  PubMed  PubMed Central  Google Scholar 

  • Goldzieher JW, Stanczyk FZ (2008) Oral contraceptives and individual variability of circulating levels of ethinyl estradiol and progestins. Contraception 78:4–9

    Article  CAS  PubMed  Google Scholar 

  • Hartigan JA, Wong MA (1979) Algorithm AS 136: A k-means clustering algorithm. Appl Stat 28:100–108

    Article  Google Scholar 

  • Iida T et al (2015) Comparison of urinary levels of 8-hydroxy-2’-deoxyguanosine between young females with and without depressive symptoms during different menstrual phases. Acta Med Okayama 69:45–50

    PubMed  Google Scholar 

  • Janse De Jonge X, Thompson B, Han A (2019) Methodological recommendations for menstrual cycle research in sports and exercise. Med Sci Sports Exerc 51:2610–2617

    Article  PubMed  Google Scholar 

  • Jilma B et al (1997) Menstrual cycle-associated changes in blood levels of interleukin-6, α1 acid glycoprotein, and C-reactive protein. J Lab Clin Med 130:69–75

    Article  CAS  PubMed  Google Scholar 

  • Kassambara A (2017) Practical guide to cluster analysis in R: unsupervised machine learning, vol 1. Sthda

  • Kassambara A, Mundt F (2017) Factoextra: extract and visualize the results of multivariate data analyses. R Package Version 107(1):337–354

    Google Scholar 

  • Kidane D, Chae WJ, Czochor J, Eckert KA, Glazer PM, Bothwell AL, Sweasy JB (2014) Interplay between DNA repair and inflammation, and the link to cancer. Crit Rev Biochem Mol Biol 49:116–139

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Knez W, Jenkins D, Coombes J (2014) The effect of an increased training volume on oxidative stress. Int J Sports Med 35:8–13

    CAS  PubMed  Google Scholar 

  • Kowalska K, Milnerowicz H (2016) Pro/antioxidant status in young healthy women using oral contraceptives. Environ Toxicol Pharmacol 43:1–6

    Article  CAS  PubMed  Google Scholar 

  • Lacut K et al (2003) Differential effects of oral and transdermal postmenopausal estrogen replacement therapies on C-reactive protein. Thromb Haemost 90:124–131

    Article  CAS  PubMed  Google Scholar 

  • Landau RL, Poulos JT (1971) The metabolic influence of progestins. In: Advances in metabolic disorders, vol 5. Elsevier, pp 119–147

  • Landgren B, Unden A, Diczfalusy E (1980) Hormonal profile of the cycle in 68 normally menstruating women. Acta Endocrinol (copenh) 94:89–98

    CAS  Google Scholar 

  • Larsen B et al (2020a) Inflammation and oral contraceptive use in female athletes before the Rio olympic games. Front Physiol.

  • Larsen B, Morris K, Quinn K, Osborne M, Minahan C (2020b) Practice does not make perfect: a brief view of athletes’ knowledge on the menstrual cycle and oral contraceptives. J Sci Med Sport 23:690–694

    Article  PubMed  Google Scholar 

  • Lê S, Josse J, Husson F (2008) FactoMineR: an R package for multivariate analysis. J Stat Softw 25:1–18

    Article  Google Scholar 

  • Le Moal E et al (2016) Redox status of professional soccer players is influenced by training load throughout a season. Int J Sports Med 37:680–686

    Article  PubMed  CAS  Google Scholar 

  • Lewis NA et al (2020) Increased oxidative stress in injured and ill elite international olympic rowers. Int J Sports Physiol Perform 15:625–631

    Article  PubMed  Google Scholar 

  • Margaritelis NV, Paschalis V, Theodorou AA, Kyparos A, Nikolaidis MG (2020) Redox basis of exercise physiology. Redox Biol 35:101499

  • Margonis K et al (2007) Oxidative stress biomarkers responses to physical overtraining: implications for diagnosis. Free Radic Biol Med 43:901–910

    Article  CAS  PubMed  Google Scholar 

  • Martin D, Sale C, Cooper SB, Elliott-Sale KJ (2018) Period prevalence and perceived side effects of hormonal contraceptive use and the menstrual cycle in elite athletes. Int J Sports Physiol Perform 13:926–932

    Article  PubMed  Google Scholar 

  • Massafra C, Gioia D, De Felice C, Picciolini E, De Leo V, Bonifazi M, Bernabei A (2000) Effects of estrogens and androgens on erythrocyte antioxidant superoxide dismutase, catalase and glutathione peroxidase activities during the menstrual cycle. J Endocrinol 167:447–452

    Article  CAS  PubMed  Google Scholar 

  • Massart A, Portier H, Rosado F, Toumi H, Filaire E (2012) Lipid peroxidation in judoists using oral contraceptives. Int J Sports Med 33:781–788

    Article  CAS  PubMed  Google Scholar 

  • Mcintosh JE, Matthews CD, Crocker JM, Broom TJ, Cox LW (1980) Predicting the luteinizing hormone surge: relationship between the duration of the follicular and luteal phases and the length of the human menstrual cycle. Fertil Steril 34:125–130

    Article  CAS  PubMed  Google Scholar 

  • McMillian M et al (2004) A gene expression signature for oxidant stress/reactive metabolites in rat liver. Biochem Pharmacol 68:2249–2261

    Article  CAS  PubMed  Google Scholar 

  • Miller PB, Soules MR (1996) The usefulness of a urinary LH kit for ovulation prediction during menstrual cycles of normal women. Obstet Gynecol 87:13–17

    Article  CAS  PubMed  Google Scholar 

  • Oostenbrug G, Mensink R, Hardeman M, De Vries T, Brouns F, Hornstra G (1997) Exercise performance, red blood cell deformability, and lipid peroxidation: effects of fish oil and vitamin E. J Appl Physiol 83:746–752

    Article  CAS  PubMed  Google Scholar 

  • Palan PR, Magneson AT, Castillo M, Dunne J, Mikhail MS (2006) Effects of menstrual cycle and oral contraceptive use on serum levels of lipid-soluble antioxidants. Am J Obstet Gynecol 194:e35–e38

    Article  CAS  PubMed  Google Scholar 

  • Pedlar CR, Newell J, Lewis NA (2019) Blood biomarker profiling and monitoring for high-performance physiology and nutrition: current perspectives, limitations and recommendations. Sports Med.

  • Pepys MB, Hirschfield GM (2003) C-reactive protein: a critical update. J Clin Invest 111:1805–1812

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Powers SK, Duarte J, Kavazis AN, Talbert EE (2010) Reactive oxygen species are signalling molecules for skeletal muscle adaptation. Exp Physiol 95:1–9

    Article  CAS  PubMed  Google Scholar 

  • Prokai L, Prokai-Tatrai K, Perjési P, Simpkins JW (2005) Mechanistic insights into the direct antioxidant effects of estrogens. Drug Dev Res 66:118–125

    Article  CAS  Google Scholar 

  • Quinn KM, Billaut F, Bulmer AC, Minahan CL (2018) Cerebral oxygenation declines but does not impair peak oxygen uptake during incremental cycling in women using oral contraceptives. Eur J Appl Physiol 118:2417–2427

    Article  CAS  PubMed  Google Scholar 

  • Quinn KM, Cox AJ, Roberts LA, Briskey D, Minahan C (2020) Reliability of a point-of-care device to determine oxidative stress in whole blood before and after acute exercise: a practical approach for the applied sports sciences. J Sports Sci.

  • Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C (1999) Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med 26:1231–1237

    Article  CAS  PubMed  Google Scholar 

  • Richardson JT (2011) Eta squared and partial eta squared as measures of effect size in educational research. Educ Res Rev 6:135–147

    Article  Google Scholar 

  • Ridker PM, Hennekens CH, Buring JE, Rifai N (2000) C-reactive protein and other markers of inflammation in the prediction of cardiovascular disease in women. N Engl J Med 342:836–843

    Article  CAS  PubMed  Google Scholar 

  • Schaumberg MA, Jenkins DG, de Jonge XAJ, Emmerton LM, Skinner TL (2016) Three-step method for menstrual and oral contraceptive cycle verification. J Sci Med Sport. 20(11):965–969.

  • Schisterman EF, Mumford SL, Sjaarda LA (2014) Failure to consider the menstrual cycle phase may cause misinterpretation of clinical and research findings of cardiometabolic biomarkers in premenopausal women. Epidemiol Rev 36:71–82

    Article  PubMed  Google Scholar 

  • Straub RH (2007) The complex role of estrogens in inflammation. Endocr Rev 28:521–574

    Article  CAS  PubMed  Google Scholar 

  • Tanskanen M, Atalay M, Uusitalo A (2010) Altered oxidative stress in overtrained athletes. J Sports Sci 28:309–317

    Article  PubMed  Google Scholar 

  • Team RC (2013) R: A language and environment for statistical computing R foundation for statistical computing, Vienna

  • Tsikas D (2017) Assessment of lipid peroxidation by measuring malondialdehyde (MDA) and relatives in biological samples: analytical and biological challenges. Anal Biochem 524:13–30

    Article  CAS  PubMed  Google Scholar 

  • Vincent HK, Powers SK, Stewart DJ, Demirel HA, Shanely RA, Naito H (2000) Short-term exercise training improves diaphragm antioxidant capacity and endurance. Eur J Appl Physiol 81:67–74

    Article  CAS  PubMed  Google Scholar 

  • Wessling-Resnick M (2010) Iron homeostasis and the inflammatory response. Annu Rev Nutr 30:105–122

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Westhoff CL, Torgal AH, Mayeda ER, Pike MC, Stanczyk FZ (2010) Pharmacokinetics of a combined oral contraceptive in obese and normal-weight women. Contraception 81:474–480

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wunder D, Yared M, Bersinger NA, Widmer D, Kretschmer R, Birkhauser M (2006) Serum leptin and C-reactive protein levels in the physiological spontaneous menstrual cycle in reproductive age women. Eur J Endocrinol 155:137–142

    Article  CAS  PubMed  Google Scholar 

  • Wyss M, Kaddurah-Daouk R (2000) Creatine and creatinine metabolism. Physiol Rev 80:1107–1213

    Article  CAS  PubMed  Google Scholar 

  • Zal F, Mostafavi-Pour Z, Amini F, Heidari A (2012) Effect of vitamin E and C supplements on lipid peroxidation and GSH-dependent antioxidant enzyme status in the blood of women consuming oral contraceptives. Contraception 86:62–66

    Article  CAS  PubMed  Google Scholar 

  • Zhang H et al (2007) Pharmacokinetic drug interactions involving 17α-ethinylestradiol. Clin Pharmacokinet 46:133–157

    Article  CAS  PubMed  Google Scholar 

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This study was supported by the Sport Performance Innovation and Knowledge Excellence unit, Queensland Academy of Sport.

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Conception or design of the work: KQ. Acquisition of data: KQ, AC, and LR. Biochemistry analysis: KQ, AC, LR, EP, DM, JF, and AP. Statistical analysis and interpretation of the data: KQ, AC, and CM. All authors approved the final version of the manuscript and agree with the order of presentation of the authors.

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Correspondence to Karlee M. Quinn.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

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Communicated by Michalis G Nikolaidis .

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Quinn, K.M., Cox, A.J., Roberts, L. et al. Temporal changes in blood oxidative stress biomarkers across the menstrual cycle and with oral contraceptive use in active women. Eur J Appl Physiol 121, 2607–2620 (2021).

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