The Mediterranean Diet Based on Local Foods for Obese Patients: A Study of Anthropometric Parameters, Inflammatory Markers, and Gut Microbiota
Phase: NA
Conditions: Obese Subjects
Sponsor: Indonesia University

Indonesia University 
RESEACRH PROTOCOL 
The Mediterranean Diet Based on Local Foods for Obese 
Patients: A Study of Anthropometric Parameters, 
Inflammatory Markers, and Gut Microbiota 
Protocol version:  3.0 
Current protocol date: 15 December 2025 
Principal Investigator: 
Dr. Diyah Eka Andayani, M.Gizi, SpGK(K) 
at the Department of Nutrition, Faculty of Medicine, 
University of Indonesia, Jakarta, Indonesia 
Email: diyahekaandayani@gmail.com 
Telephone: number 081310637429 

2 
Table of Contents 
Protocol Summary ................................................................................................................. 3 
1.
Background and rationale ............................................................................................... 6 
2.
Hypothesis ..................................................................................................................... 7 
3.
Study Objectives ............................................................................................................. 7 
4.
Study Design ................................................................................................................... 8 
5.
Participant Population .................................................................................................... 9 
6.
Treatment of Participant............................................................................................... 10 
7.
Study Procedures .......................................................................................................... 11 
8.
Clinical and Toxicity Management Guideline ................................................................. 12 
9.
Adverse Event Recoding and Reporting ......................................................................... 12 
10. Biological samples ........................................................................................................ 13 
11. Analysis Plan ................................................................................................................ 14 
12. Data Collection, Source Documents, Record Retention, and Confidentiality of Data ...... 15 
13. Ethics Committee/Regulatory Approval and Informed Consent ..................................... 15 
14. Financing and Insurance ............................................................................................... 16 
15. Quality Control and Quality Assurane ........................................................................... 16 
LIST OF REFENCES ................................................................................................................ 17 
Apendix 1 Participant Information sheet and consent………….……………………………………….……….21 

3 
Protocol Summary 
Title 
The Mediterranean Diet Based on Local Foods for Obese Patients: A Study of Anthropometric 
Parameters, Inflammatory Markers, and Gut Microbiota 
Protocol registration number 
25-09-1322
Principal Investigator 
dr. Diyah Eka Andayani, M.Gizi, Sp.GK (K) 
Background and Rationale 
Obesity is a global problem with incidents increasing every year and estimated to reach 21% 
by 2025. Based on data from the 2023 Indonesian Health Survey, the prevalence of obesity in 
Indonesia is 23.4%. This prevalence is highest in the Jakarta, at 31.8%. In addition, a report on 
health examinations at the Indonesia University found that 16.2% (50/321) of employees were 
obese. The modern diet-includes fast food, sweet, and fatty foods-is associated with metabolic 
diseases such as hypertension, glucose intolerance, and obesity. Weight loss in obesity can be 
achieved through behavioral modification, such as improving dietary patterns and increasing 
physical activity. Weight loss in the early stages can help predict the success of long-term 
weight loss management.  
The Mediterranean diet is one diet that can be used to reduce the risk of cardiovascular 
disease, but some studies show that the Mediterranean diet has a positive impact on weight 
loss in obese patients. The Mediterranean diet includes a plant-based foods, with olive oil as 
the main source of additional fat, as well as high intake of fish and seafood, moderate 
consumption of eggs, poultry, and dairy products (cheese and yogurt), and low consumption 
of red meat. In addition, the Mediterranean diet also includes activity management. Compared 
to other diets, the Mediterranean diet shows a decrease in BMI and weight, HbA1c levels, 
fasting blood sugar, fasting insulin, inflammatory markers, and can improve gut microbiota 
dysbiosis.  
The implementation of the Mediterranean diet often faces challenges related to cultural 
factors and the availability of local food ingredients. Considering the potential of the 
Mediterranean diet in modulating gut microbiota and metabolic status, as well as the 
challenges of its adaptation in Indonesia, this study aims to examine the effectiveness of a 
Mediterranean diet based on local Indonesian foods on changes in gut microbiota, body 
composition, and metabolic biomarkers in obese.  
Study Objectives 
To examine the efficacy of a Mediterranean diet adapted to local Indonesian foods on changes 
in gut microbiota, body composition, and metabolic biomarkers in obese individuals. 

4 
Study Design 
This study will be conducted using a randomized, single-blind, clinical trial design. The 
intervention group consists of obese patients who receive dietary education and a 
Mediterranean diet adapted to local Indonesian foods. The control group is composed of obese 
patients who will receive low calorie diet and dietary education.  
Participant Population 
The inclusion criteria 
1. Women who were obese, as defined by a BMI of at least 27.
2. Aged 18-50 years
3. Subjects with a non-shift work schedule
4. Willing to follow the research procedures and sign the informed consent form.
The exclusion criteria 
1. Subjects with a history of type 2 diabetes, kidney disease, liver disease, or thyroid
disorders, as indicated by medical history or medical records.
2. Contraindications for the MF-BIA examination include the use of pacemakers or implants
and a history of amputation.
3. Pregnant or within 40 days of childbirth
4. Adults with edema
5. Taking medications with hyperglycemic effects, such as beta-blockers, thiazides,
corticosteroids, calcineurin inhibitors, anti-dyslipidemic medications (e.g., statins, fibrates,
niacin, and bile acid sequestrants), or antibiotics.
6. History of food allergies
Treatment of participants 
The research subjects received diet around 1500 calories according to their randomized 
groups, consisting of breakfast, lunch, dinner, and snacks.   
-
Arm 1: The intervention group provide the composition was 50% carbohydrates, 30-
34% fat (SFA ≤8%, MUFA 15-20%, PUFA 5-10%), and 17-20% protein.
-
Arm 2: The control group receive 50-54% carbohydrates, 25-30% fat (SFA <12, MUFA
8-12%, PUFA 5-8%), 19-20% protein.  The researchers developed the diet composition
supplied to the catering service. Then, the catering service translated these
compositions into meals.
Study Procedur 
Screening, Randomisation and Follow-up 
At screening: informed consent; Demographics (year of birth, race and gender at birth); 
allergies and medical history. 
At washout period: participants were not permitted to consume foods or beverages containing 
probiotics or prebiotics (such as yogurt), weight-loss medications, or fiber supplements. 

5 
 
Follow up visit: Participants received meals according to their assigned group allocation. They 
were required to consume the meals within 15–20 minutes and reported to researcher every 
meal time. 
At week 0 and week 4 examination: anthropometric measures (BMI, WHR), body composition 
parameters (fat mass, fat-free mass, visceral fat), IL-6 levels, I-FABP levels, the TyG index, and 
gut microbiota composition. 
 
Statistical consideration 
- 
Univariate Analysis: Ratio scales (age, BMI) and dependent variables (anthropometry, FFM, 
FM, visceral fat, TyG index, IL-6 levels, gut microbiota diversity, I-FABP levels) are presented 
with a normal distribution in the form of mean±SD, but if the data distribution is not 
normal, it is presented in the form of median (minimum–maximum values). Data normality 
was assessed using the coefficient of variation (CoV) and the Kruskal-Wallis test. 
- 
Bivariate Analysis: If dependent variables were normally distributed, an unpaired t-test was 
performed. However, if the dependent variable data are not normally distributed (changes 
in all variables studied between the intervention group and the control group), the Mann-
Whitney test will be performed. Analysis of the difference in the mean of the dependent 
variable data between baseline and week 4 of the study: if the data distribution is normal, 
a t-test will be performed, but if the data distribution is not normal, a Wilcoxon test will be 
performed.  
- 
Multivariate Analysis: Linear regression test to control for confounding variables. Bivariate 
tests are required (if p <0.25) and statistical tests are considered significant if the p-value 
is <0.05. Linear mixed model analysis is used to analyze the relationship between insulin 
resistance variables (TyG index) and changes in I-FABP or microbiota composition. 
Statistical tests are considered significant if the p-value is <0.001. 
 
 
 

6 
 
1. Background and rationale 
Obesity is a global problem, with cases increasing each year and projected to reach 21% by 
2025. Based on the 2023 Indonesian Health Survey, obesity prevalence in Indonesia is 23.4%, 
with the highest prevalence (31.8%) recorded in the Jakarta. Additionally, Indonesia 
University health examination report found that 16.2% (50 out of 321) of employees were 
obese. A study reported that obese female employees at Cipto Mangunkusumo Hospital 
(RSCM) had a mean BMI of 30.67 ± 4.91. Dietary patterns high in fast food, sweets and fatty 
foods are associated with increased risk factors for metabolic diseases, including 
hypertension, glucose intolerance, and obesity. Obesity can be addressed through behavioral 
modifications, including dietary improvement and increased physical activity. Early weight 
loss may serve as a predictor of long-term weight loss success. 
Adherence to the Mediterranean diet has been associated with a reduced risk of 
cardiovascular disease and has demonstrated beneficial effects on weight loss among 
individuals with obesity. The Mediterranean diet is characterized by a high intake of plant-
based foods, the use of olive oil as the main source of dietary fat, moderate consumption of 
fish, seafood, eggs, poultry, and dairy products, and limited intake of red meat. Furthermore, 
it integrates lifestyle behaviors such as regular physical activity. Compared with other dietary 
patterns, the Mediterranean diet is associated with reductions in BMI, body weight, HbA1c 
levels, fasting blood glucose, fasting insulin, and inflammatory markers, while also improving 
gut microbiota dysbiosis.  
Gut microbiota is an important factor in obesity, as the diversity of its composition and 
metabolic activity is closely associated with nutritional intake, dietary patterns, and obesity. 
Gut microbiota dysbiosis is one of the underlying contributors to obesity-related 
susceptibility to chronic inflammation. Another condition is adipose tissue stimulates the 
release of proinflammatory adipokines—such as interleukin-6 (IL-6), tumor necrosis factor-α 
(TNF-α), monocyte chemoattractant protein-1 (MCP-1), and resistin—which contribute to 
chronic inflammation. 

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Implementing the Mediterranean diet can be challenging due to cultural factors and the 
availability of local ingredients. Mediterranean diet can modulate gut microbiota and 
metabolic status and the challenges of adapting it in Indonesia, this study aims to evaluate 
the effectiveness of a Mediterranean diet based on local Indonesian foods in altering gut 
microbiota, body composition, and metabolic biomarkers in obese. 
2. Hypothesis 
The provision of a Mediterranean diet modification based on local food ingredients can result 
in greater weight loss, greater reductions in inflammatory markers, improve gut microbiota 
diversity compared to a balanced low-calorie diet in obese patients. 
3. Study Objectives 
General Objective 
The implementation of a Mediterranean diet modified using local foods has the potential to 
improve anthropometric outcomes, inflammatory markers, and gut microbiota profiles in 
obese individuals, thereby providing a reference for dietary selection in obesity treatment. 
Specific Objectives 
- 
To determine the characteristics of the research subjects based on age, underlying 
disease, nutritional status, body composition, intake patterns, and physical activity. 
- 
To determine the energy, protein, fat, and carbohydrate intake of research subjects 
before and after diet administration. 
- 
To determine the effect of a Mediterranean diet based on local foods on the 
anthropometry of obese patients. 
- 
To determine the anthropometric data (weight, height, and waist circumference), 
nutritional status, and body composition of research subjects before and after diet 
administration. 
- 
To determine the differences in the mean anthropometry (weight, height, and waist 
circumference), nutritional status, and body composition of the research subjects in each 
group.  

8 
 
- 
To determine the effect of a modified Mediterranean diet based on local food 
ingredients on inflammatory marker values in obese patients. 
- 
To determine changes in IL-6, I-FABP, and TyG index levels in research subjects before 
and after diet administration. 
- 
To determine the differences in the mean levels of IL-6, I-FABP, and TyG index of the 
research subjects in each group. 
- 
To determine the effect of a Mediterranean-based diet modification using local food 
ingredients on improving gut microbiota diversity and metabolomic profile in obese 
patients. 
- 
To determine the gut microbiota diversity and metabolomic profile of the study subjects 
before and after the diet intervention. 
4. Study Design 
This study will be conducted using a single-blind randomized clinical trial to evaluate the 
effects of a locally adapted Mediterranean diet intervention on anthropometric measures 
(BMI, WHR), body composition parameters (fat mass, fat-free mass, visceral fat), IL-6 levels, 
I-FABP levels, the TyG index, and gut microbiota composition. The study will be conducted at 
the Nutrition Clinic of Cipto Mangunkusumo Hospital (RSCM) and the Faculty of Medicine, 
Indonesia University (FKUI), Jakarta. Analyses of gut microbiota diversity and metabolomics 
will be performed at the Department of Chemistry, FKUI, and at the Indonesian Medical 
Education and Research Institute (IMERI), FKUI Occupational and Bioinformatics Cluster. The 
study is scheduled to take place between 2025 and 2026, following approval from the Ethics 
Committee FKUI–RSCM. 
 
 
 
 
 

9 
 
5. Participant Population 
Study population 
a.    Target Population 
The target population of this study includes all obese patients living in Jakarta, Depok, and 
surrounding areas. 
b.    Accessible Population 
The accessible population of this study consists of obese patients who are employees of FKUI 
and RSCM and are treated at the Nutrition Clinic. 
c.    Research Subjects  
 
The research subjects are part of the accessible population selected based on the established 
inclusion and exclusion criteria. 
a.   The inclusion criteria  
-  Women who were obese, as defined by a BMI of at least 27. 
-  Aged 18-50 years  
-  Subjects with a non-shift work schedule 
-  Willing to follow the research procedures and sign the informed consent form. 
b.   The exclusion criteria 
-  Subjects with a history of type 2 diabetes, kidney disease, liver disease, or thyroid 
disorders, as indicated by medical history or medical records. 
-  Contraindications for the MF-BIA examination include the use of pacemakers or 
implants and a history of amputation. 
-  Pregnant or within 40 days of childbirth 
-  Adults with edema 
-  Taking medications with hyperglycemic effects, such as beta-blockers, thiazides, 
corticosteroids, calcineurin inhibitors, anti-dyslipidemic medications (e.g., statins, 
fibrates, niacin, and bile acid sequestrants), or antibiotics. 
-  History of food allergies 
 

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c.  The DO criteria for this study are: 
-  If severe side effects occur 
-  The subject decides to withdraw from the study 
-  The subject does not comply with the established research protocol. 
 
6. Treatment of Participant 
Participants received a low-calorie diet of approximately 1.500 kcal/day according to their 
randomized group assignments, consisting of breakfast, lunch, dinner, and snacks. In the 
intervention group, the macronutrient distribution was 50% carbohydrates, 30–34% fat (SFA 
≤ 8%, MUFA 15–20%, PUFA 5–10%), and 17–20% protein. In the control group, the diet 
provided 50–54% carbohydrates, 25–30% fat (SFA < 12%, MUFA 8–12%, PUFA 5–8%), and 19–
20% protein. 
Table 1. Diet formulation and composition in the intervention group and control group 
Composition 
Intervention Group (%) 
Control group (%) 
Protein  
17-20 
19-20 
Carbohydrates   
50 
50-54 
Fat 
SFA 
MUFA 
PUFA 
30-34 
≤8 
15-20 
5-10 
25-30 
<12 
8-12 
5-8 
 
The diets for both groups will be prepared by a certified diet catering service under the 
supervision of a clinical nutritionist. This catering service meets established certification 
standards and routinely prepares meals for individuals with special nutritional needs, 
including obesity. The dietary composition will be designed by the researchers and 
subsequently translated into meal plans by the catering service for both study groups. 
Details of the diet will be known only to the catering service and the research team to 
maintain blinding. Meals for both groups will be provided by the catering service from 

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Monday to Saturday, while on Sundays, dietary intake for both groups will be supervised 
directly by the researchers. In addition, participants will receive education on hunger 
management and will be permitted to consume additional foods that do not alter the core 
composition of the Mediterranean diet, such as antioxidant-rich foods, vegetables, and 
fruits. 
Both groups will receive dietary education and physical activity guidance tailored to their 
weight loss needs. The dietary education includes instructions that participants are not 
permitted to consume foods, beverages, or supplements outside the provided catering 
menu without the researchers’ knowledge. The prescribed physical activities consist of 
aerobic exercises, such as brisk walking, swimming, cycling, jogging, and light aerobic 
workouts, performed 3–5 times per week, with each session lasting 30–45 minutes. 
7. Study Procedures 
Initial screening period. 
Data collection commenced after approval was obtained from the Ethics Committee FKUI–
RSCM and research permission was granted by the RSCM Research Department. Prospective 
participants were interviewed to collect baseline characteristics using a screening 
questionnaire. Clinical assessments conducted during the screening phase included: 
informed consent; Demographics (year of birth, race and gender at birth); allergies and 
medical history. 
Randomisation and wash out 
To proceed to the randomization and washout phases, all participants were required to meet 
the eligibility criteria based on screening evaluation results. Clinical assessments conducted 
during the randomization and washout phases included anthropometric measurements, 
collection of 10 mL blood samples, and collection of 250 mg fecal samples. Analyses of gut 
microbiota diversity and metabolomics were performed at the Department of Chemistry 
FKUI, and at the Indonesian Medical Education and Research Institute (IMERI), FKUI 
Occupational and Bioinformatics Cluster.  
A 7-day washout period was implemented. During this period, participants were not 
permitted to consume foods or beverages containing probiotics or prebiotics (such as 

12 
 
yogurt), weight-loss medications, or fiber supplements. Participants were instructed to 
perform light to moderate aerobic physical activity. Dietary intake during the washout 
period was assessed using food recall. 
 
Follow Up Visits 
Participants received meals according to their assigned group allocation. They were required 
to consume the meals within 15–20 minutes at the agreed mealtime. Participants were 
instructed to complete a food intake record and report their consumption to the researchers 
using intake forms and photographs of the food packaging. Weekly meetings were held via 
Zoom to evaluate dietary compliance, and participants were monitored daily through a 
WhatsApp group, with compliance verified using photographic evidence. Blood samples and 
fecal specimens were collected at baseline (week 0) and at week 4 of the study. 
At week 0 and week 4 examination: anthropometric measures (BMI, WHR), body 
composition parameters (fat mass, fat-free mass, visceral fat), IL-6 levels, I-FABP levels, the 
TyG index, and gut microbiota composition. 
 
8. Clinical and Toxicity Management Guideline 
The Mediterranean diet has been widely consumed and recommended for individuals at risk 
of cardiovascular disease and obesity, with no reported adverse effects to date, provided 
that individuals are not allergic to the ingredients used. Potential risks associated with blood 
sample collection include pain, bruising, swelling, or infection at the venipuncture site. These 
risks will be minimized by using small-gauge needles, ensuring that blood is drawn by trained 
personnel, and implementing appropriate infection prevention measures prior to sample 
collection. 
 
9. Adverse Event Recoding and Reporting 
The researchers collaborated with food providers that met Hazard Analysis and Critical 
Control Points (HACCP) standards and operated under the supervision of a clinical 
nutritionist. In the event that any adverse conditions arose as a result of the provided food, 

13 
 
participants were instructed to immediately contact the researchers, who would facilitate 
access to appropriate medical care. 
 
Timely and complete reporting of all adverse events (AEs) helps to identify any adverse 
medical occurrences, thereby enabling:  
-  Protection of the safety of study participants 
-  A better understanding of the overall safety profile of the study intervention 
-  Recognising food-related allergies or toxicities 
-  Appropriate modification of study protocols 
-  Improvements in study design or procedures 
-  Adherence to regulatory requirements 
 
10. Biological samples 
Blood and Feces Sample Collection Procedure 
When blood is drawn, the subject must sit for at least 5 minutes before sampling. Blood 
sampling is performed on the anterior cubital vein, sterilizing the area with an alcohol swab. 
The tourniquet should not be applied for more than 2 minutes. 10 ml of blood is drawn and 
placed in a red vacutainer. Collected blood must be labeled with the patient's identity, date 
of collection (format DD/MMM/YYYY), and time of collection (24-hour format). Samples 
must arrive at the laboratory within 3 hours of blood collection (the time required for 
centrifugation of storage samples and biochemical and lipid testing within 4 hours of blood 
collection). All samples must be stored and transported from the local site at room 
temperature (20°C-25°C) to the responsible laboratory. 
Fecal samples were collected by subjects at home in the morning using sterile containers 
provided. Instructions for collection and storage were provided in writing. After collection, 
samples were immediately stored in a coolbox with dry ice, then frozen at -80°C in the 
laboratory to prevent microbial DNA degradation. Collected feces must be labeled with the 
patient's identity, date of collection (DD/MMM/YYYY format), and time of collection (24-
hour format). Collected stool samples must be immediately stored at a low temperature, 

14 
 
usually -80°C, to prevent metabolite degradation. In addition, stool can be stored at 4°C in 
an anaerobic atmosphere for less than 24 hours, and can be stored at -20°C for several days 
(anaerobic atmosphere or ambient environment). 
Processing of Samples 
The TyG index was calculated based on fasting blood glucose and triglyceride levels obtained 
from venous blood serum samples. Fasting glucose and triglyceride concentrations were 
measured using the glycerol phosphate oxidase–phenol aminophenazone (GPO–PAP) 
enzymatic method with a clinical spectrophotometer, in accordance with standard 
laboratory protocols. 
 
Interleukin-6 (IL-6) levels were measured using the sandwich enzyme-linked immunosorbent 
assay (ELISA) method with a Human IL-6 ELISA Kit. Absorbance was read at a wavelength of 
450 nm using a microplate reader. 
Intestinal fatty acid–binding protein (I-FABP) levels were determined using a Human I-FABP 
ELISA Kit based on the sandwich ELISA method, following the manufacturer’s protocol. 
Metabolomic profile analysis was performed using the Shimadzu GCMS-QP2020 NX system. 
Gut microbiota diversity was assessed using the QIAGEN QIAamp PowerFecal Pro DNA Kit. 
All procedures were conducted according to the manufacturer’s instructions. DNA 
concentration and purity were evaluated using a NanoDrop spectrophotometer and agarose 
gel electrophoresis. 
 
11. Analysis Plan 
Univariate Analysis: Ratio scales (age, BMI) and dependent variables (anthropometry, FFM, 
FM, visceral fat, TyG index, IL-6 levels, gut microbiota diversity, I-FABP levels) are presented 
with a normal distribution in the form of mean±SD, but if the data distribution is not normal, 
it is presented in the form of median (minimum–maximum values). Data normality was 
assessed using the coefficient of variation (CoV) and the Kruskal-Wallis test for normality. 
Bivariate Analysis: If dependent variables were normally distributed, an unpaired t-test was 
performed. However, if the dependent variable data are not normally distributed (changes 

15 
 
in all variables studied between the intervention group and the control group), the Mann-
Whitney test will be performed. Analysis of the difference in the mean of the dependent 
variable data between baseline and week 4 of the study: if the data distribution is normal, a 
t-test will be performed, but if the data distribution is not normal, a Wilcoxon test will be 
performed.  
Multivariate Analysis: Linear regression test to control for confounding variables. Bivariate 
tests are required (if p <0.25) and statistical tests are considered significant if the p-value is 
<0.05. Linear mixed model analysis is used to analyze the relationship between insulin 
resistance variables (TyG index) and changes in I-FABP or microbiota composition. Statistical 
tests are considered significant if the p-value is <0.001. 
 
12. Data Collection, Source Documents, Record Retention, and Confidentiality of Data 
The Principal Investigator or designee is responsible for preparing and maintaining adequate 
and accurate data pertinent to the investigation on each individual in the study. All data 
collected in this study will be kept confidential. Data processing and storage will only record 
identities in coded form. Furthermore, the presentation of research results at scientific 
meetings/conferences and publications in scientific journals will not include the names of 
research subjects. Soft files will be stored in password-protected folders, while hard files will 
be stored in locked research cabinets. Only the research team will have access to research 
documents. 
 
13. Ethics Committee/Regulatory Approval and Informed Consent 
This study will be conducted in compliance with the ethical principles outlined in the 
National Statement on Ethical Conduct in Research Involving Humans and the Declaration of 
Helsinki, and in accordance with Good Clinical Practice (GCP) guidelines and all applicable 
regulatory requirements. The study will be reviewed by the relevant local ethics committees, 
in accordance with current local guidelines.  
 

16 
 
Informed consent and procedures 
The principal investigator will prepare the informed consent which must: include all 
elements required by ICH-GCP; all applicable regulatory requirements; and adhere to the 
ethical principles that have their origin in the Declaration of Helsinki.  
The Principal Investigator is responsible for ensuring that participants, or their legally 
authorized representatives (LARs), receive comprehensive and clearly communicated 
information regarding the study objectives, potential risks, and all other relevant aspects of the 
clinical trial. 
 
14. Financing and Insurance 
Research funding is fully covered by the Researcher.   
 
15. Quality Control and Quality Assurane 
For implementing and maintaining quality control and quality assurance systems with 
written standard operating procedures to ensure the study is conducted and data are 
generated, documented and reported in compliance with the protocol, Good Clinical 
Practice standards and all applicable local laws and regulations relating to the conduct of a 
clinical study. 
 
 

17 
 
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the gut microbiome and metabolome independently of energy intake. Gut. 2020;69(7):1258–68.  
19.  
Cunningham AL, Stephens JW, Harris DA. A review on gut microbiota: a central factor in the 
pathophysiology of obesity. Lipids Health Dis. 2021;20(1):1–13.  
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Sikora M, Stec A, Chrabaszcz M, Waskiel-Burnat A, Zaremba M, Olszewska M, et al. Intestinal fatty 
acid binding protein, a biomarker of intestinal barrier, is associated with severity of psoriasis. J Clin 

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El-Mikkawy DME, EL-Sadek MA, EL-Badawy MA, Samaha D. Circulating level of interleukin-6 in 
relation to body mass indices and lipid profile in Egyptian adults with overweight and obesity. Egypt 
Rheumatol Rehabil. 2020;47(1). 
 
 
 
 
 

20 
 
Abbervation List 
BMI 
: Body Mass Index 
DNA 
: Deoxyribonucleic Acid  
ELISA  : Enzyme-Linked Immunosorbent Assay 
FFM 
: Fat-Free Mass 
FM 
: Fat Mass 
I-FABP  : Intestinal Fatty Acid Binding Protein 
IL-6 
: Interleukin-6 
TyG 
: Triglyceride-Glucose Index 
MCP-1 : Monocyte Chemoattractant Protein-1  
MF-BIA: Multi-frequency Bioelectrical Impedance Analysis 
MUFA : Monounsaturated Fatty Acids 
PUFA : Polyunsaturated Fatty Acid 
RSCM : Cipto Mangunkusumo Hospital 
SFA 
: Saturated Fatty Acid 
TNF-α : Tumor Necrosis Factor Alpha 
WHO : World Health Organization  
WHR : Waist-to-Hip Ratio  
 
 
 
 
 
 
 
 
 
 
 

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PARTICIPANT INFORMATION AND CONSENT FORM 
 
I am dr. Diyah Eka Andayani, M.Gizi, SpGK(K), from the Department of Nutrition, Faculty of 
Medicine, Indonesia University–Cipto Mangunkusumo Hospital. Under the supervision of Dr. dr. 
Diana Sunardi, M.Gizi, SpGK(K), I will conduct a study entitled The Mediterranean Diet Based on 
Local Foods for Obese Patients: A Study of Anthropometric Parameters, Inflammatory Markers, 
and Gut Microbiota. 
I will first provide you with detailed information about this study and invite you to participate. 
Individuals who agree to participate will be asked to sign an informed consent form in the 
presence of a witness. You may withdraw from the study at any time without penalty. You also 
have the right to receive updated information from the research team regarding the intervention 
being tested, if applicable. Choosing not to participate will not affect the care or services you 
receive at this hospital, nor will it affect your relationship with the researchers. 
If you do not understand any statement in this form, you may ask me about it.    
1. Research objectives 
The objective of this study is to determine whether a modified Mediterranean diet based on local 
foods can improve body weight, waist circumference, body composition (including body fat 
mass), inflammatory markers, and gut microbiota composition in obese patients, thereby 
providing a reference for dietary selection in obese management. 
The Mediterranean diet is a diet rich in fiber sources such as vegetables and fruits with olive oil 
as the main source of additional fat, as well as high intake of fish and seafood. Moderate 
consumption of eggs, poultry, and dairy products (cheese and yogurt), and low consumption of 
red meat. In addition, the Mediterranean diet also includes physical activity such as regular 
exercise. 
 
2.  Participation in the study 
This study will last approximately 28 days and will be preceded by a 7-day pre-study period. 
During the assessment phase, measurements of body weight, height, waist circumference, hip 
circumference, and body composition will be taken, which will require approximately 20 minutes. 
Blood sampling will take approximately 5–10 minutes. You will also be asked to collect stool 
samples according to procedures that will be explained to you. 
Following these assessments, participants will receive meals prepared by the research team from 
Monday to Saturday, consisting of three main meals and two snacks per day. The Sunday meal 

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menu will be supervised by the researcher. On the 29th day, the baseline examinations will be 
repeated. 
The diet will be arranged and provided by a catering service from Monday through Saturday, 
while on Sundays the meal menu will be supervised by the researchers. Participants are expected 
to consume only the provided meals and follow the dietary recommendations, as well as engage 
in physical activity as advised. By agreeing to participate in this study, you will be asked to adhere 
to the study schedule and confirm your ability to comply with all study requirements. 
 
3.  Reasons you have been invited to participate in this study 
a. Women who are obese based on a nutritional status of ≥ 27 
b. Aged 18-50 years 
c. Willing to follow the research procedures and sign the informed consent form. 
d. No history of type 2 diabetes mellitus, kidney, liver, or thyroid disorders based on medical 
history or medical records 
e. No contraindications for body composition measurement: use of pacemakers, implants, 
history of amputation 
f. Not pregnant or in the postpartum period (up to 40 days after giving birth) 
g. Not experiencing edema 
h. Not taking medications that increase blood sugar (e.g., beta blockers, thiazides, 
corticosteroids, and calcineurin inhibitors) or lower blood lipids (e.g., statins, fibrates, 
niacin, bile acid sequestrants) 
 
4.  Research procedures 
You will undergo the following procedures: 
a. Interview and identification of dietary history using a food record sheet completed by a 
nutritionist covering food consumed during the previous 24 hours.  This interview will last 
30 minutes 
b. A physical examination will be conducted by a trained measurement officer to assess body 
weight, height, and body composition, including fat mass, muscle mass, and body fluids. 
The examination will take approximately 15 minutes. 
c. Blood sampling of 2 tablespoons. 
Blood sampling will be performed by a trained and experienced officer. Prior to blood 
collection, the participant will be asked to sit for at least 5 minutes. Blood will be drawn 
from the antecubital vein (located in the crease of the arm at the elbow) after the area 
has been cleaned with alcohol. A tourniquet will be applied to the upper arm to facilitate 
venous access. A total of 10 mL of blood (approximately 2 tablespoons) will be collected 
and placed into appropriate blood collection tubes. 

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d. Stool sample collection: 1 teaspoon 
Stool samples will be collected by the participant at home in the morning using a sterile 
container provided by the study team. Participants are advised to urinate before stool 
collection. The stool should first be collected in a large plastic container provided, after 
which approximately 250 mg (about 1 teaspoon) of stool should be transferred using a 
spoon into the sterile container. The sample should then be submitted to the study staff 
within 2 hours of collection. 
All of these tests will be conducted at the beginning and end of the research day. 
 
5.  Risks, side effects, and management 
The Mediterranean diet has been widely consumed and promoted among individuals at risk of 
cardiovascular disease and obesity. To date, no adverse effects have been reported, provided 
that individuals are not allergic to the ingredients used. Potential risks associated with blood 
collection include pain, bruising, swelling, or infection at the puncture site. These risks can be 
minimized through the use of small-gauge needles, procedures performed by trained personnel, 
and adherence to infection prevention measures prior to blood collection. 
 
6.  Benefits 
The benefits to you include measurements of body weight, waist circumference, and body 
composition (such as body fat mass), laboratory tests for inflammatory markers and intestinal 
bacterial composition, as well as provision of balanced meals. All tests are provided free of 
charge, and all related costs are covered by the researchers. 
 
7.  Compensation 
You will receive transportation compensation of Rp 50,000 for each visit to the research site and 
compensation of Rp 100,000 for time spent participating in the study. These compensations will 
be provided at the beginning and at the end of the research period. If you experience any injury 
or illness as a result of participating in this research, please contact the study physician 
immediately, who will assist you in obtaining the necessary medical care. 
 
8.  Funding  
Research funding is fully covered by the Researcher.  
 
 
 

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9.  Confidentiality 
All data collected in this study will be kept confidential. Data processing and storage will not 
include your name and will use coded identifiers only. The presentation of research results at 
scientific meetings or conferences and publication in scientific journals will not include any 
information that could identify you.  
 
10. Obligations of research subjects 
As a research participant, you are required to follow the study rules and instructions outlined 
above. If anything is unclear, you may ask the research team for further clarification. During the 
study period, you are not permitted to take any supplements or herbal medicines other than 
those provided by the researchers. 
 
11. Right to refuse and withdraw 
As a research participant, you are required to follow the study rules and instructions outlined 
above. Even if you agree to participate, you have the right to withdraw from the study at any 
time. This decision will not affect your relationship with the investigator or the standard of care 
provided at this hospital. At the end of this explanation, you will be given time to consider the 
decision you wish to make. 
 
12. Post-trial access 
At the end of the study, all participants will receive follow-up monitoring of body weight, waist 
circumference, and body composition (including body fat mass), as well as one month of dietary 
education, provided that the study results demonstrate benefits for obese patients. 
 
13. Additional Information 
You will be given the opportunity to ask any questions you may have regarding this study. If at 
any time you experience side effects or require further explanation, you may contact dr. Diyah 
Eka Andayani, M.Gizi, SpGK(K), at 0813-1063-7429, Department of Nutrition, Faculty of Medicine, 
University of Indonesia / Dr. Cipto Mangunkusumo Hospital, Jakarta. 
 
 
 
 
 

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CONSENT FORM 
All of these explanations have been conveyed to me and all of my questions have been 
answered by Dr. Diyah Eka Andayani, M.Gizi, SpGK(K). I understand that if I need further 
clarification, I can ask dr. Diyah Eka Andayani, M.Gizi, SpGK(K). 
 
 
 
Researcher Information: 
Principal Investigator: dr. Diyah Eka Andayani, M.Gizi, SpGK(K) 
Address: Permata Permai Housing Complex Block B 03, Pancoran Mas, Depok 
Email: diyahekaandayani@gmail.com 
Ethic Committee FKUI-RSCM    : Jalan Salemba 6, Central Jakarta, 10430 
              
 
               Phone: 021 3157008 
       
 
 
               Email: ec_fkui@yahoo.com 
Consent 
 
I have read all the explanations about this 
study. I have been given the opportunity to ask 
questions and all my questions have been 
answered clearly. I am willing to participate in 
this research study voluntarily. 
 
 
____________________________ 
Name of subject 
 
 
____________________________ 
Sign  
 
Date_________________________ 
day/month/year 
I confirm that participants were given the 
opportunity to ask questions about this 
study, and all questions were answered 
correctly. I confirm that consent was 
given voluntarily. 
 
 
____________________________ 
Name of researcher/approval requester  
 
 
____________________________ 
Sign 
 
Date_________________________ 
day/month/year