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CompletedNCT06486623Updated Jul 3, 2024

Effect of Supplementation With Anthocyanins on Iron Metabolism in Female Swimmers

A Phase 1/2 interventional study of Black chokeberry extract in capsules with 18% standardization of the anthocyanins content and Chokeberry fiber in capsules in Iron Deficiencies, Inflammation and Oxidative Stress, sponsored by Poznan University of Physical Education. Completed at 1 site in Poland. Open to female participants aged 12 Years to 19 Years. Per ClinicalTrials.gov, last updated 2024-07-03.

Sponsored by Poznan University of Physical Education · Phase 1/2, Interventional, and Basic science

Phase
Phase 1/2
Study type
Interventional
Enrollment
22
Allocation
Randomized
Ages
12 Years to 19 Years
Sex
Female
01

Study summary

The study included 8-weeks supplementation of black chokeberry extract with 18% standardization of the anthocyanins content (dose 2×200 mg per day), or a placebo product that was made from chokeberry fiber.

Purpose:

The aim of the study is effectiveness check of black chokeberry intake as a potential factor which, due to its ability to reduce inflammation and chelate iron ions, will have an impact on minimizing undesirable changes in the red blood cell system.

Hypothesis:

Main hypothesis: Supplementation of black chokeberry (Aronia melanocarpa) has a positive effect on iron economy of female swimmers.

Auxiliary hypotheses:

  1. Supplementation of black chokeberry enhances the antioxidant status and has a positive effect on the prooxidant-antioxidant balance.
  2. Supplementation of black chokeberry reduce inflammation induced by intensive exercises, by reducing inflammatory markers.

Study plan:

Before first term of the study:

Control of Menstrual Cycle Phases since March 2023 (installing the application on the phone - Menstrual Period Tracker calendar).

First term of the study:

  1. Menstrual cycle control survey.
  2. Conduct a body composition analysis.
  3. Collection of blood samples before the exercise test for biochemical determinations.
  4. Exercise test in the swimming pool - crawl style intensive test (800 m + 200 m + 50 m).
  5. Collection of blood immediately after exercise and 3 hours later.
  6. Supplementation for nearly 56 days, depending on the length of the menstrual cycle.

Second term of the study:

Repeat the measurements from the 1st date of test.

Participants: The group of selective and homogeneous female swimmers, who train swimming intensively and attend the Sports Championship School Complex.

Read the detailed description

This study included 8-weeks supplementation of black chokeberry extract with 18% standardization of the anthocyanins content (dose 2×200 mg per day), or a placebo product that was made from chokeberry fiber. The first date of research completed the group of twenty-four and homogeneous female swimmers, who train swimming intensively and attend the Sports Championship School Complex. The second date of research completed the group of twenty-two selective and homogeneous female swimmers, who train swimming intensively and attend the Sports Championship School Complex.

Purpose:

The aim of the study is effectiveness check of black chokeberry intake as a potential factor which, due to its ability to reduce inflammation and chelate iron ions, will have an impact on minimizing undesirable changes in the red blood cell system.

Hypothesis:

Main hypothesis: Supplementation of black chokeberry (Aronia melanocarpa) has a positive effect on iron economy of female swimmers.

Auxiliary hypotheses:

  1. Supplementation of black chokeberry enhances the antioxidant status and has a positive effect on the prooxidant-antioxidant balance.
  2. Supplementation of black chokeberry reduce inflammation induced by intensive exercises, by reducing inflammatory markers.

The research is going to be carried out as a medicinal experiment with tissue interference (approval of Research Ethics Committee: 538/22 of 06/23/2022, with changes: 221/23 of 03/09/2023). All swimmers qualified to study based on inclusion and exclusion criteria and were informed of the extract course of the study and gave written consent to participate.

EXPERIMENTAL PROCEDURE, WHICH WAS APPLIED (STUDY PLAN):

A. Before first term of the study:

Control of Menstrual Cycle Phases since March 2023 (installing the application on the phone - Menstrual Period Tracker calendar) and three surveys in three terms.

B. First term of the study:

  1. Menstrual cycle control by application Menstrual Tracker Period Calendar and surveys, before swimming test.
  2. Body composition analysis (TANITA MC-780 S MA).
  3. Biochemical determinations - blood samples will be taken before and after exercise test as well as after 3-hour restitution, in order to determine:

    • morphological parameters,
    • value of indicators:

      1. iron status problems (iron, hepcidin, erythroferrone, total iron-binding capacity, unsaturated iron-binding capacity, ferritin, transferrin,
      2. oxidative stress (total antioxidant capacity),
      3. immunological indicators (interleukins: IL-6, IL-2, IL-10, C-reactive protein),
      4. hormonal balance (progesterone, estradiol, luteinizing hormone, follicle-stimulating hormone),
      5. other: lactic acid, myoglobin, hypoxanthine.

    Blood samples were collected from the elbow vein into two 4-ml tubes (to obtain serum) and centrifuged at 3,000 rpn and into 2,7-ml tubes to determine morphology. Morphology was determined on the same day the samples were collected, while serum was frozen and stored at -80 Celsius degrees until assays were performed. In addition, before and immediately after the exercise test, capillary blood was drawn from the earlobe to determine lactate levels.

    Lactate level were determined immediately after capillary blood sampling using a commercial available kit (Dr Lange, Germany). The concentration of lactate was determined in mmol/l. Other measurements were determined using ELISA kits (DRG Medtek).

  4. Intensive exercise test in the swimming pool, before supplementation - crawl style (800 m + 200 m + 50 m) with 15-minute active rest between test sections.
  5. Supplementation of black chokeberry - nearly 56-days supplementation of black chokeberry extract with 18% standardization of the anthocyanins content (connection with menstrual cycle phase).The swimmers were randomly divided into two groups: supplementation and placebo. The placebo contained chokeberry fiber. Whole products were manufactured by MLB BIOTRADE, Poznan. The swimmers took 2 capsules per day (before breakfast and dinner) and they sipped each capsule with glass of water.

C. Second term of the study:

  1. Menstrual cycle control by application Menstrual Tracker Period Calendar and surveys, before swimming test.
  2. Body composition analysis (TANITA MC-780 S MA).
  3. Biochemical determinations - blood samples will be taken before and after exercise test as well as after 3-hour restitution, in order to determine:

    • morphological parameters (erythrocytes, leukocytes, thrombocytes),
    • value of indicators responsible for:

      1. iron status problems (iron, hepcidin, erythroferrone, total iron-binding capacity, unsaturated iron-binding capacity, ferritin, transferrin),
      2. oxidative stress (total antioxidant capacity),
      3. immunological indicators (interleukins: IL-6, IL-2, IL-10, C-reactive protein (CRP)),
      4. hormonal balance (progesterone, estradiol, luteinizing hormone (LH), follicle-stimulating hormone (FSH)),
      5. other: lactic acid, myoglobin, hypoxanthine.

    Blood samples were collected from the elbow vein into two 4-ml tubes (to obtain serum) and centrifuged at 3,000 rpn and into 2,7-ml tubes to determine morphology. Morphology wa determined on the same day the samples were collected, while serum was frozen and stored at -80 Celsius degrees until assays were performed. In addition, before and immediately after the exercise test, capillary blood was drawn from the earlobe to determine lactate levels.

    Lactate level were determined immediately after capillary blood sampling using a commercial available kit (Dr Lange, Germany). The concentration of lactate was determined in mmol/l. Other measurements were determined using ELISA kits (DRG Medtek).

  4. Intensive exercise test in the swimming pool, after supplementation - crawl style (800 m + 200 m + 50 m) with 15-minute active rest between test sections.

Data collection, analysis in laboratory, statistical analysis and preparation articles to publication.

02

Conditions studied

  • Iron Deficiencies
  • Inflammation
  • Oxidative Stress
  • Female Athlete Triad
03

Who can participate

Ages eligible
12 Years to 19 Years
Sexes eligible
Female
Accepts healthy volunteers
No

Inclusion criteria

  • Current medical examinations issued by a sports medicine doctor
  • Training experience a minimum of three years
  • Student of a sports championship school team
  • Consent of the athlete/legal guardian to participate in the research
  • Completion of the exercise test in the swimming pool - crawl style intensive test (800 m + 200 m + 50 m) on two test deadline

Exclusion criteria

Exclusion Criteria:

  • Taking: oral contraceptive pills, supplements containing anthocyanin compounds during the study or a one month earlier
  • Health problems
  • Antibiotic therapy
04

Study design

Phase
Phase 1 / Phase 2
Primary purpose
Basic science
Allocation
Randomized
Intervention model
Single group
Masking
Double (Participant, Outcomes assessor)
Enrollment
22 participants (actual)

Study arms

  • Experimental
    1. Supplemented group

    The supplemented group (n=12) consumed capsules with highly concentrated 18% chokeberry extract (one capsule contained 200 mg). Each capsule also contained: chokeberry fiber, E460b magnesium salts of stearic acid (anti-caking agent), E551 silicon dioxide (anti-caking agent), hydrocypropyl methylcellulose (capsule shell) and E171 titanium dioxide (shell colour). The producer of all capsules is MLD Biotrade in Poznan. The swimmers took 2 capsules a day (before breakfast and dinner) for nearly 56 days (depending on length menstrual cycle phase). The swimmers sipped each capsule with glass of water.

    Drug: Black chokeberry extract in capsules with 18% standardization of the anthocyanins content

  • Placebo comparator
    2. Placebo group

    The control group (n=10) consumed capsules that were made from chokeberry fiber. The producer of all capsules is MLD Biotrade in Poznan. The swimmers took 2 capsules a day (before breakfast and dinner) for nearly 56 days (depending on length menstrual cycle phase). The swimmers sipped each capsule with glass of water.

    Drug: Chokeberry fiber in capsules

Interventions

  • DrugBlack chokeberry extract in capsules with 18% standardization of the anthocyanins content

    Supplement group: supplement of diet - black chokeberry extract (capsules) with 18% standardization of the anthocyanins content (one capsule contained 200 mg). Each capsule also contained: chokeberry fiber, E460b magnesium salts of stearic acid (anti-caking agent), E551 silicon dioxide (anti-caking agent), hydrocypropyl methylcellulose (capsule shell) and E171 titanium dioxide (shell colour). The producer of all capsules is MLD Biotrade in Poznan.

    Also known as: Aronia melanocarpa

  • DrugChokeberry fiber in capsules

    Placebo group: placebo product - chokeberry fiber (placebo capsules, which were made from chokeberry fiber). The producer of all capsules is MLD Biotrade in Poznan.

    Also known as: Placebo product

05

What researchers measure

Primary outcomes

  1. Physiological parameters of blood morphology I

    erythrocytes: RBC \[10\^6/mL\], HGB \[g/dL\], HCT \[%\], MCV \[mm\^3\], MCH \[pg\], MCHC \[g/dL\], RDW \[%\],

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  2. Physiological parameters of blood morphology II

    leukocytes: WBC \[10\^3/mL\], LYM \[10\^3/mL\], MON \[10\^3/mL\], GRA \[10\^3/mL\], LYM \[%\], MON \[%\], GRA \[%\],

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  3. Physiological parameters of blood morphology III

    thrombocytes: PLT \[10\^3/mL\], MPV \[mm\^3\], PCT \[%\], PDW \[%\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  4. Physiological parameters of iron metabolism indicator I

    iron \[µg/dl\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  5. Physiological parameters of iron metabolism indicator II

    hepcidin \[ng/ml\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  6. Physiological parameters of iron metabolism indicator III

    transferrin \[mg/ml\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  7. Physiological parameters of iron metabolism indicator IV

    total iron-binding capacity (TIBC) \[µg/dl\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  8. Physiological parameters of iron metabolism indicator V

    ferritin \[ng/ml\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  9. Physiological parameters of iron metabolism indicator VI

    erythroferrone \[ng/mL\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  10. Physiological parameters of antioxidant-prooxidant indicator

    total antioxidant capacity \[mmol/l\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  11. Physiological parameters of immunological indicator I

    interleukin 6 \[ng/L\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  12. Physiological parameters of immunological indicator II

    interleukin 2 \[ng/L\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  13. Physiological parameters of immunological indicator III

    interleukin 10 \[ng/L\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  14. Physiological parameters of immunological indicator IV

    C-reactive protein \[mg/dL\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  15. Physiological parameter of hormonal indicator I

    progesterone \[ng/ml\]

    Time frame: At each exercise test (up to 15 minute before exercise)

  16. Physiological parameter of hormonal indicator II

    estradiol \[pg/ml\]

    Time frame: At each exercise test (up to 15 minute before exercise)

  17. Physiological parameter of hormonal indicator III

    luteinizing hormone (LH) \[mIU/ml\]

    Time frame: At each exercise test (up to 15 minute before exercise)

  18. Physiological parameter of hormonal indicator IV

    follicle-stimulating hormone (FSH) \[mIU/ml\]

    Time frame: At each exercise test (up to 15 minute before exercise)

  19. Other physiological parameter I

    myoglobin \[ng/ml\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  20. Other physiological parameter II

    hypoxanthine \[mmol/l\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

  21. Other physiological parameter (with capillary blood - earlobe)

    lactic acid \[mmol/l\]

    Time frame: At each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

Secondary outcomes

  1. Physiological parameters of iron metabolism indicator VII

    unsaturated iron-binding capacity (UIBC)

    Time frame: Calculated with blood samples at each exercise test (up to 15 minute before exercise, up to 1 minute post exercise, 3 hours after)

06

Study locations

1 site
  • University of Physical Education in Poznan
    Poznań, Poland
07

References and documents

Individual participant data

Plan to share: No

No publications or documents are linked to this record.

08

Registry details

Key details

Study ID
NCT06486623
Lead sponsor
Poznan University of Physical Education
Responsible party
Sponsor
First posted
Jul 3, 2024
Start date
Mar 1, 2023
Primary completion
Dec 28, 2023
Completion
Dec 28, 2023
Last update
Jul 3, 2024

Study contacts

Marta Mydłowska, PhD student
principal investigator · Poznań University of Physical Education, Faculty of sport sciences in Gorzów Wielkopolski

Oversight

Data monitoring committee
Yes
FDA-regulated drug
No
FDA-regulated device
No
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