The Effect of Methylphenidate Treatment on Oxidative Stress Levels in Children Diagnosed With Attention Deficit Hyperactivity Disorder (ADHD)
Conditions: Attention Deficit Hyperactivity Disorder (ADHD)
Sponsor: Saglik Bilimleri Universitesi

Study Title 
The Effect of Methylphenidate Treatment on Oxidative Stress Levels in Children 
Diagnosed With Attention Deficit Hyperactivity Disorder (ADHD) 
  
Observational Study 
 
  
 
Content 
Study Protocol 
Statistical Analysis Plan (SAP) 
Informed Consent Form (ICF) 
  
Medicine Doctor Enes Faruk ALTUNKILIÇ 
 
 
 
18.03.2024 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

Research Protocol 
 
 
Location of the Study 
Health Sciences University Istanbul Bakırköy Prof. Dr. Mazhar Osman Mental Health and 
Neurological Diseases Training and Research Hospital. 
 
Purpose of the Study. 
The aim of the study is to investigate the effect of methylphenidate treatment on oxidative 
stress by measuring the levels of Total Oxidant Status (TOS), Total Antioxidant Status (TAS), 
Oxidative Stress Index (OSI = TOS/TAS), Malondialdehyde (MDA), Oxidized LDL (Ox-LDL), and 
Superoxide Dismutase (SOD) in serum samples from patients diagnosed with Attention 
Deficit Hyperactivity Disorder (ADHD) who have either started or are planned to start 
methylphenidate treatment. Measurements will be taken at baseline (before the start of 
treatment) and at the 3rd month after the treatment begins. 
 
Material and Methods of the Study 
The study population will consist of patients who are diagnosed with "Attention Deficit 
Hyperactivity Disorder (ADHD)" according to DSM-5-TR, who have applied to the child and 
adolescent psychiatry outpatient clinics of Bakırköy Prof. Dr. Mazhar Osman Mental Health and 
Neurological Diseases Training and Research Hospital and have been started or planned to start 
methylphenidate treatment by the physician they have been examined by. After an in-clinic 
announcement, these patients will be directed to the researcher, Enes Faruk Altunkılıç. After 
being informed both verbally and in writing through the informed consent process, those who 
agree to participate, sign the informed consent form, and meet the inclusion and exclusion criteria 
will be included in the study. It will also be explained to the patients and their families that their 
participation or non-participation in this study will not affect the treatment they will receive. 

After informed consent, a structured clinical interview for DSM-5-TR will be conducted using 
the "Mood Disorders and Schizophrenia Form for School-Aged Children - Now and Lifetime 
DSM-5 - Turkish Adaptation (ÇDŞG-ŞY-DSM-5-T)." To obtain sociodemographic and clinical 
data for the participants, the "Sociodemographic and Clinical Data Form" created by the 
researchers will be completed. For the ADHD diagnosed group, the "Conners Parent Rating Scale 
- Revised Short" and "Conners Teacher Rating Scale - Revised Short" will be applied to 
determine the severity of the disorder, symptoms, and predominant subtyping. After the diagnosis 
is made and evaluated according to the exclusion criteria, blood samples will be taken from the 
patient after a 10-12 hour fasting period, between 9-12 AM, in a yellow-capped tube, before the 
routine methylphenidate treatment is used. After waiting for 10-20 minutes at room temperature, 
the sample will be centrifuged at 3000 RPM for 20 minutes, and the serum will be collected in 
Eppendorf tubes and stored at -80°C until analysis. After 3 months of treatment, blood samples 
will be taken again in the same manner and stored. Once all the samples are collected, the serum 
samples will be analyzed for total antioxidant status (TAS), total oxidant status (TOS), 
malondialdehyde (MDA), superoxide dismutase (SOD), and oxidized LDL levels according to 
the human ELISA kit protocols at the Biochemistry Laboratory of Bakırköy Dr. Sadi Konuk 
Training and Research Hospital by biochemist Dr. Hacer Eroğlu İçli. After 3 months of treatment, 
the Conners Parent Rating Scale - Revised Short and Conners Teacher Rating Scale - Revised 
Short forms will be applied again. In addition to the markers, the Oxidative Stress Index (OSI = 
TOS/TAS) will be calculated and included in the evaluation before and 3 months after the 
treatment. 
 
Inclusion criteria for the study: 
• A diagnosis of "Attention Deficit Hyperactivity Disorder (ADHD)" according to DSM-5-TR 
and the initiation or planned initiation of methylphenidate treatment as part of routine care. 
• Age between 6-11 years. 
• Agreement to participate in the study after being informed about the study. 
Exclusion criteria for the study 

• Presence of any psychiatric disorder other than "Attention Deficit Hyperactivity Disorder (ADHD)." 
• Diagnosis of "Attention Deficit Hyperactivity Disorder (ADHD)" without the initiation or planned 
initiation of methylphenidate treatment. 
• Age below 6 years or above 11 years. 
• Presence of organic brain damage, mental retardation, autism spectrum disorder, neurological diseases, 
or physical illnesses affecting neurocognitive functions. 
• History of alcohol and/or psychoactive substance use. 
• Ongoing active infection, allergic diseases, or chronic illnesses. 
• Previous use of psychiatric medication. 
• Presence of chronic diseases. 
• Use of regular medications.  
 
Research Questionnaire  
1. Informed Patient Consent Form: It is prepared to verify the patient's consent and to 
inform the participant. 
 
2. Sociodemographic and Clinical Data Form: This is a form prepared by the researchers to 
record the sociodemographic and clinical data of the patient volunteers for the purpose of 
the study. It includes questions about participants' sociodemographic characteristics such 
as age, gender, and education level, as well as questions regarding family history, any 
chronic illness or psychiatric disorder history, and inquiries about smoking and 
alcohol/substance use. 
 
3. Mood Disorders and Schizophrenia Form for School-Aged Children - Current and 
Lifetime Version DSM-5 - Turkish Adaptation (ÇDŞG-ŞY-DSM-5-T): The first version of this 
form was created by Chambers et al. in 1985 according to DSM-3. It was updated based on 
DSM-5 diagnostic criteria by Kaufman et al. In the first section, the sociodemographic 
information of the child and family, complaints, developmental and general health status, 
and functionality are questioned. The second section contains screening questions covering 
over 200 specific symptoms from both the past and the last two months. The third section 
consists of evaluation and observation results by the clinician to verify DSM-5 diagnoses. 

Ünal and colleagues conducted the Turkish translation and validity-reliability studies in 
2019. 
 
4. Conners Parent Rating Scale-Revised Short Form (CPRS-RS): The Conners Parent Rating 
Scale-Revised Short Form (CPRS-RS) was developed by Conners et al. in 1997 to measure 
the severity of ADHD symptoms and behavioral problems, and it is administered by parents. 
In 2013, Kaner and colleagues conducted the Turkish validity and reliability study. The scale 
consists of 27 items, including 3 subscales (Oppositional (KG), Cognitive Problems-
Inattention (BP-D), Hyperactivity (H)) and 1 additional scale (ADHD index to assess children 
at risk for ADHD) answered by the parent. 
 
5. Conners Teacher Rating Scale-Revised Short Form (CTRS-RS): The Conners Teacher 
Rating Scale-Revised Short Form (CTRS-RS) was developed by Conners et al. in 1997 to 
measure the severity of ADHD symptoms and behavioral problems, and it is administered by 
the teacher. In 2013, Kaner and colleagues conducted the Turkish validity and reliability 
study. The scale consists of 28 items, including 3 subscales (Oppositional (KG), Cognitive 
Problems-Inattention (BP-D), Hyperactivity (H)) and 1 additional scale (ADHD index to 
assess children at risk for ADHD) answered by the teacher. 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
 
 
 
 
 
 
 
 
Statistical Analysis 
The statistics of this study were analyzed using the JAMOVI (The jamovi project (2024). jamovi 
(Version 2.5) [Computer Software]. Retrieved from https://www.jamovi.org) package program 
and SPSS 26.0 (IBM Corp. Released 2019. IBM SPSS Statistics for Windows, Version 26.0. 
Armonk, NY: IBM Corp) package program. 
First of all, descriptive statistics (mean, standard deviation, median, lowest-highest values, 
percentage) of the variables in the study were determined and these statistics were given as tables 
or bar graphs.  
The normality of the variables was checked with the 'Shapiro-Wilk' test. In paired samples 
analyses, if the differences of the variables before and after the normality test were observed to fit 
the normal distribution, 'paired sample t-test' was performed; if it was observed that it did not fit 
the normal distribution, 'Wilcoxon Signed Ranks' test was performed. In SOD measurement, 2 
samples did not react and did not change color and these values could not be read. For this 
reason, statistics were made over 37 sample numbers for SOD. 
In order to determine the relationship between clinical change before and after treatment and 
changes in biomarkers, correlations were examined between the changes in the total score and 
subscale scores (before and after) and the changes in the biomarkers (before and after) of CADÖ-
YK and CÖDÖ-YK. 
In correlation analyses, the 'Pearson Correlation' test was performed for normally distributed 
variables, and the 'Spearman Correlation' test was used for variables that did not fit the normal 
distribution. 
 
 
 

 
INFORMED CONSENT FORM 
Bakırköy Prof. Dr. Mazhar Osman Mental Health and Neurological Diseases Training and 
Research Hospital 
You have been invited to participate in a clinical study. After reading the information below, if 
you wish to participate, please sign this form. If you choose not to participate, there will be no 
interruption in your treatment. Your signature on this form indicates that you have been informed 
about the study and that you have freely made your decision. If you do not understand any of the 
terms in the form, please ask the doctors involved in the study for clarification. The title of the 
study is “The Effect of Methylphenidate Treatment on Oxidative Stress Levels in Children with 
Attention Deficit Hyperactivity Disorder (ADHD).” 
Your clinical diagnosis has already been established, and the treatment plan prescribed by your 
physician has been determined. Therefore, your participation or non-participation in this study 
will not affect the treatment you will receive. This study aims to investigate the potential role of 
methylphenidate on oxidative stress. Ethical approval for the research has been obtained. Dr. 
Enes Faruk ALTUNKILIÇ will assist you with any questions or problems you may have (Phone: 
05077362191). 
For the research, a psychiatric interview will be conducted, and certain scales will be applied. 
There will be a follow-up for 3 months. The follow-up will occur at most once a month, 
depending on your clinical condition. Before starting methylphenidate treatment, venous blood 
will be drawn from the inner side of your elbow between 9 AM and 12 PM after a 10-12 hour 
fasting period. This will be done again 3 months after the treatment begins. A total of two blood 
samples will be collected. Your participation in the research will take a maximum of 30 minutes, 
which is the duration required during your routine tests. You will not have any additional 
responsibilities. This research does not offer any direct benefit to your treatment at this time, and 
it is not expected to alter the course of your treatment. The results obtained from this research are 
expected to contribute to the scientific community and potentially benefit the treatment of others 
in the future. There will be no costs for the procedures involved, and no costs will be charged to 
you or any institution under your insurance. No payment will be made to you for participating in 
this research. Your personal and medical information will remain confidential. Even if the 
research is published, your identity will not be shared. However, the research observers, record 
keepers, ethics committees, and official authorities may have access to your medical information 
when necessary. You may also access your own medical information upon request. The total 
number of volunteers for this research will be 39 individuals. 
You may withdraw from the study at any time without penalty or loss of any rights. If you fail to 
comply with the treatment protocol, interrupt the research program, or experience any adverse 
effects related or unrelated to the research, the responsible researchers may remove you from the 

study without your consent. The results of the study will be used for scientific purposes. In case 
of withdrawal or removal from the study, your medical data may be used for scientific purposes if 
necessary. 
Participant's Statement 
I have read the above information and fully understand the scope and purpose of the study I am 
being asked to participate in. I have been informed that the research results will be used for 
educational and scientific purposes, and that my personal information will remain confidential. 
Written and verbal explanations about the study have been provided by the researcher mentioned 
below, and I have asked questions and received satisfactory answers. The potential risks and 
benefits of the study have been explained to me. I understand that I may leave this study at any 
time without giving a reason and without facing any negative consequences. Under these 
conditions, I voluntarily agree to participate in this study without any pressure or coercion. 
For minors or those under guardianship or custody, a parent or legal guardian must sign: 
Name-Surname: 
Phone: 
Date and Signature: 
Witness of the researcher's explanation: 
Name-Surname: 
Phone: 
Date and Signature: 
Researcher's information: 
Name-Surname: Enes Faruk ALTUNKILIÇ 
Position: Assistant Doctor 
Phone: 05077362191 
Date and Signature: 
Note: This form is prepared in two copies. One copy is given to the volunteer with their 
signature, and the other is kept by the researcher. 
 
 
 
 
 
 

 
PARTICIPANT'S STATEMENT: 
I have been informed by Assistant Doctor Enes Faruk ALTUNKILIÇ, that a medical research 
study will be conducted at the Children and Adolescent Psychiatry Clinic of Bakırköy Prof. 
Dr. Mazhar Osman Psychiatry and Neurology Training and Research Hospital of the Health 
Sciences University. The above information regarding this study was provided to me. After 
receiving this information, I have been invited to participate as a "participant" in this 
research study. 
If I participate in this research, I believe that confidentiality will be maintained regarding my 
personal information during the study, and I am assured that my personal information will 
be carefully protected in any educational or scientific use of the study results. 
I understand that I can withdraw from the study at any time without providing a reason. 
However, I am aware that it is appropriate to inform the researchers in advance of my 
decision to withdraw. I also understand that I may be removed from the study by the 
researchers for my medical well-being. I have no financial responsibility for the costs 
associated with the study, and no payment will be made to me. In the event of any health 
issue arising from the study, either directly or indirectly, I am assured that all necessary 
medical intervention will be provided, and I will not incur any financial burden related to 
these interventions. 
If I encounter a health issue during the study, I know that I can contact Assistant Doctor 
Enes Faruk ALTUNKILIÇ at the Children and Adolescent Psychiatry Clinic of Bakırköy Prof. 
Dr. Mazhar Osman Psychiatry and Neurology Training and Research Hospital, or reach him 
at (0212) 4091515-3510. I understand that I am not obligated to participate in this study and 
that I may choose not to participate. I have not encountered any coercion to participate. If I 
refuse to participate, I am aware that this will not affect my medical care or relationship 
with my doctor. 
I have read all the explanations in the Informed Consent Form. A written and verbal 
explanation about the study, its purpose, and the details mentioned above has been 
provided to me by the physician named below. I understand all the explanations provided to 
me. After taking adequate time to think, I have decided to participate as a "participant" in 
this research project. I am voluntarily participating in this study, and I am aware that I can 
withdraw at any time, with or without reason. I agree to participate in the study without any 
pressure or coercion. 
 

Participant’s Name/Signature 
Date 
Researcher's Name/Signature 
Date 
Witness Name/Signature 
Date