Chemical and Sensory Characterization of a Drink Based on Coffee and Agave Inulin and Its Effect on Appetite, Dietary Intake, and Biochemical Profile in Subjects With Obesity
Phase: NA
Conditions: Obesity
Sponsor: University of Guadalajara

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
University of Guadalajara 
Health Sciences University Center 
Institute for Translational Nutrigenetics and 
Nutrigenomics 
 
 
 
June 2024 – July 2025 
 
Effect of coffee and 
agave inulin-based 
beverage on appetite 
sensations, food 
intake, and ghrelin, 
GLP-1, and PYY 
concentrations in 
adults with obesity 

ABSTRACT 
Obesity is a disease that currently represents a great challenge from different areas, such 
as health, economy, society, and politics since it represents a problem with different negative 
consequences in each of these areas, not to mention that its prevalence worldwide is 
increasing. The main treatment for this problem is a lifestyle change, including diet and 
physical activity modifications. Specifically, nutritional treatment is usually based on 
balanced meal plans with caloric restriction to generate a negative energy balance. 
However, when a nutritional treatment is carried out, adherence to the treatment can be 
compromised by experiencing low satiety, or the patient's remaining hungry with the meal 
plan. 
  
In this sense, some studies show evidence of increased satiety and reduced energy intake 
when coffee is consumed in certain doses. In the case of inulin, a decrease in appetite, an 
increase in GLP-1 and PYY concentrations, and even a decrease in energy intake have 
been reported. These effects have been attributed to certain bioactive compounds present 
in coffee, such as caffeine and chlorogenic acids, and in the case of inulin, to its effect on 
the microbiota that produces certain short-chain fatty acids, which in turn stimulate the 
secretion of peptides related to satiety and, being a fiber, slows gastric emptying. However, 
with coffee and inulin, there is contradictory evidence; in many cases, no significant changes 
are observed. In other words, there are still many questions to be answered.   
Therefore, more scientific evidence is needed to contribute to nutritional recommendations 
for hunger control and satiety. Therefore, this study aims to evaluate the chemical and 
sensory characteristics of a coffee and agave inulin-based beverage and its effect on 
appetite, dietary intake, and biochemical profile in subjects with obesity. For this purpose, a 
randomized crossover clinical trial with an estimated n of 30 participants has been proposed 
to evaluate the effect of daily and two-week consumption of a coffee beverage with agave 
inulin in comparison with the daily and two-week consumption of a placebo beverage for two 
weeks (Sedes: Instituto de Nutrigenética y Nutrigenómica Traslacional (INNUGET) of the 
CUCS and the Bariatric and Metabolic Surgery Unit of the Civil Hospital "Juan I. Menchaca"). 
The measurement of appetite hormones (acetylated ghrelin, GLP-1, and PYY) will be 
measured by ELISA assays. Visual analog scales measure appetite sensations, and the 
biochemical profile is determined by dry chemistry. The study will be conducted from April 
2024 to March 2025. The results of this research will be disseminated at international 

congresses in a high-impact international journal, and its completion will allow several 
students to complete doctoral and bachelor's theses. 
 
BACKGROUND 
 
Globally, obesity has almost tripled since 1975, as more than 650 million adults had obesity 
in 2016, representing about 13% of the world's population (1). In Mexico, obesity is a health 
problem that continues to increase, having in 2018, 75.2% of adults aged 20 years and older 
with overweight and obesity (2). In Jalisco, in the same year, it was reported a prevalence 
of obesity of 39.2% in women and 29.5% in men (3). 
  
Obesity results from a combination of genetic, biological, sociocultural, and lifestyle factors 
(4). In that sense, two lifestyle habits that are very important in its development are high 
consumption of sugary beverages and low fiber intake. According to the National Health and 
Nutrition Survey (ENSANUT) 2018, 85.8% of the Mexican population over 20 years of age 
consumes sweetened beverages, and a large part of the Mexican population has a deficient 
fiber intake (2). On the other hand, appetite regulation also plays an important role in body 
weight control (5). Under normal conditions, hunger and satiety are well connected to allow 
a balance between energy consumed and energy expended; however, this does not occur 
in obesity (6). When there is a dysregulation in the hunger signals given by peptides such 
as ghrelin and in the satiety signals given by peptides such as PYY, GLP-1, and CCK, among 
others, or when there are alterations in the cognitive and hedonic regulation of hunger, 
excessive consumption of kilocalories is produced, which are stored in the form of fat, that 
is obesity is predisposed (7). 
  
The increase in body fat, mainly at the visceral level, leads to metabolic alterations such as 
insulin resistance and dyslipidemia, increasing the risk of type 2 diabetes mellitus and 
cardiovascular disease (8). Therefore, to combat obesity and its consequences, the first 
thing recommended is a lifestyle change, such as an adequate diet, regular exercise, and 
psychotherapeutic interventions (7, 8). However, at present, it is a challenge to have 
strategies that allow successful nutritional interventions in patients with obesity. In this 
context, it has been observed that coffee consumption in overweight and people with obesity 
has been positively associated with satiety (9), reduction of body weight and hunger, and 

even with a lower energy intake, partly due to mechanisms that directly or indirectly regulate 
the appetite mentioned above hormones (10,11). 
 
In addition, caffeine can amplify the appetite-suppressing effect if combined with other 
compounds, such as fibers (12) like inulin. It has been observed that in overweight and 
subjects with obesity who had consumed a beverage with 24 g of inulin diluted in 300 ml of 
water with glucose, there was a decrease in ghrelin levels 6 hours after consumption 
compared to the ingestion of water with glucose (13). Inulin has also been associated with 
reduced hunger and energy intake (14) and increases in GLP-1 and PYY concentrations 
(15). However, the joint effect of coffee and inulin has not been studied, and many separate 
studies are inconclusive. 
 
STATEMENT OF THE PROBLEM 
The accelerated increase in the prevalence of overweight and obesity in our country has 
generated concern and the need to generate strategies that provide solutions for its 
prevention and treatment (16-18). The cost of various medical complications for a person 
with obesity and prediabetes, if he/she does not change his/her lifestyle habits, can reach 
up to 1.9 million pesos in 30 years. However, if he or she modifies his or her lifestyle, in 30 
years, the total expense would decrease to only 92,860 pesos (19). 
  
In that sense, a lifestyle habit that favors the development of obesity is the consumption of 
sugar-sweetened beverages. According to the National Health and Nutrition Survey 
(ENSANUT) 2018, 85.8% of the Mexican population over 20 years of age consumes 
sweetened beverages, and 16.8% consume sweetened dairy beverages (17). Likewise, the 
obesogenic environment in which we live favors easy access to this type of beverage, and 
there are very few healthy beverage options within the food industry (20). On the other hand, 
the average fiber consumption in Mexico is 27 g per day in men and 22 g per day in women, 
while the daily fiber recommendation for adults is 30 g per day. 
 
High consumption of sugar-sweetened beverages combined with low fiber intake are risk 
factors for developing obesity and other chronic noncommunicable diseases (21). Obesity 
may arise from inappropriate food choices stemming from a strong urge to eat or weak 
hunger inhibition (22). In addition, there are hormones and molecules involved in appetite 
control, and when their functionality is altered (as occurs in obesity), they lead to overeating 

and the progression of obesity. Among these hormones are peptide YY (PYY), glucagon-
like peptide 1 (GLP-1), and ghrelin (23,24). 
  
Currently, it is a challenge to have strategies that allow successful nutritional interventions 
in patients with obesity. In this context, coffee consumption in overweight and obese people 
has been positively associated with satiety (25), reduction of body weight and hunger, and 
even with a lower energy intake, partly due to mechanisms that directly or indirectly regulate 
the appetite mentioned above hormones (26, 27). For its part, caffeine can amplify the 
appetite-suppressing effect if combined with other compounds, as is the case with fibers 
(28) such as inulin, which has been associated with a reduction in hunger and energy intake 
(29), as well as with increases in GLP-1 and PYY concentrations (30). However, the joint 
effect of coffee and inulin has not been studied, and many separate studies are inconclusive. 
  
Coffee is a popular beverage. Worldwide consumption is estimated at 168 million bags of 
60 kilograms (31). At the national level, Mexicans consume an average of two and a half 
cups daily. However, inulin is not a food consumed significantly by the Mexican population; 
fiber consumption generally is below the nutritional recommendations. That is why, if its 
effects are proven, a beverage as widely consumed as coffee in combination with agave 
inulin could help people with obesity regulate their appetite. 
 
JUSTIFICATION 
The prevalence of obesity in 1988 in Mexico was only 9.5%, compared to 36.1% in 2018 
(17), so it is necessary to start acting to improve the welfare of the Mexican population. In 
the search for evidence to complement a nutritional plan, the combined consumption of 
coffee and inulin has been proposed. Coffee consumption in our country increases by 2% 
each year and according to data from PROFECO (Procuraduría Federal del Consumidor), 
85% of Mexicans beverage one to three cups a day (32). Agave tequilana Weber, the blue 
variety, is the most widely cultivated of the 117 species of agaves native to Mexico. This 
variety is an important source of inulin (33). 
  
Based on previous research, these two products of the Mexican countryside have been 
shown, each in their own right, to provide health benefits. In the case of coffee, it has been 
shown to improve endurance and exercise capacity, it has antioxidant capacity, it has been 
associated with a lower risk of Parkinson's and Alzheimer's disease, and several studies 

report a hepatoprotective effect (34); regarding appetite control and weight loss, there is still 
great controversy within the scientific community about its effects, especially due to the lack 
of randomized clinical trials. On the other hand, inulin has been shown to improve intestinal 
transit, lower LDL cholesterol, and be an important prebiotic for the intestinal microbiota, 
although its relationship with appetite control is not clear (35). These effects on regulating 
appetite processes such as hunger and satiety have been studied in other countries, but not 
in Mexico, and much less so using inulin from agave tequilana Weber. 
 

OBJECTIVE 
To evaluate the chemical and sensory characteristics of coffee and agave inulin-based 
beverages and their effects on appetite, dietary intake, and biochemical profiles in subjects 
with obesity. 
 
b. Specific  
1. To compare the concentrations of appetite hormones when consuming a coffee and agave 
inulin-based beverage vs a control beverage. 
2. To compare the subjective sensations of appetite when consuming a coffee and agave 
inulin-based beverage vs a control beverage.  
3. To compare the dietary intake when consuming a coffee and agave inulin-based beverage 
vs a control beverage. 
4. To compare the biochemical profile when consuming a coffee and agave inulin-based 
beverage vs a control beverage. 
 
HYPOTHESIS 
A beverage based on coffee and agave inulin modifies appetite, dietary intake, and 
biochemical profile in subjects with obesity. 
 
METHODOLOGICAL DESIGN 
a. Type of study 
Double-blind randomized crossover clinical trial. 
 
b. Research location 
Instituto de Nutrigenética y Nutrigenómica Traslacional (INNUGET), Centro Universitario 
de Ciencias de la Salud, Universidad de Guadalajara. 
 
Unidad de Cirugía Bariátrica y Metabólica del Nuevo Hospital Civil de Guadalajara <Dr. 
Juan I. Menchaca=. 
 
c. Study period 
From June 2024 to July 2025 
 
d. Inclusion and non-inclusion criteria 

Inclusion criteria 
Men and women 
25 to 55 years of age 
BMI ≥ 30 and <35 kg/m2 
Habitual coffee consumption (1 - 3 cups of 240 ml daily). 
Willingness to stop consuming food and/or supplements with coffee, caffeine, and/or inulin 
during the study except for the intervention. 
Availability of time in the mornings 
Residents of the Guadalajara Metropolitan Area 
Signed informed consent form 
 
Non-inclusion criteria 
Hypersensitivity or intolerance to coffee or inulin 
Allergy or intolerance to any of the test foods 
Current or within the last 6 months use of weight-loss or appetite-altering medications 
appetite altering medications 
Diagnosis of diabetes, hypertension or any heart disease, thyroid disease, infectious 
disease (COVID-19, flu, etc.), digestive or inflammatory bowel disease or dysgeusia 
Smoking 
Women who are pregnant, want to become pregnant or are breast-feeding 
Peri-menopausal or menopausal women 
Who is on any dietary regimen for weight loss 
Currently or within the last 12 months, participated in another study 
Consumption of prebiotics or probiotics in the last three months 
 
e. Exclusion criteria 
Incomplete data and/or measurements 
Lack of expected compliance (drink at least 5 of the 7 days of each week is considered 
compliance). 
Individuals who decide to leave the study 
 
f. Sample size calculation 
The information reported in the article by Rahat et al., which had a double-blind randomized 
crossover design, was used to calculate the sample size (13). In this study, a significant 

increase in the active PYY response was found when consuming 24 g of inulin mixed with 
75 g of glucose dissolved in water (increase in the area under the curve from 0 to 4 h 
(iAUC04) = 5.69 pmol×h/L ± 9.8 pmol×h/L), compared with consumption of only 75 g of 
glucose dissolved in water ((iAUC04) = 1.06 pmol×h/L ± 10 pmol×h/L) with a p=0.003.   
The program G*Power v.3.1.9.7 (36) was used for the sample size calculation, and an n=24 
was obtained using the formula for dependent mean difference and considering a loss of 
25% (which is common in this type of study); 30 subjects would be required for this project. 
 
g. Dependent and independent variables 
 
Independent variable 
Consumption of coffee and agave inulin-based beverage 
 
Dependent variables 
Appetite hormones 
Subjective appetite sensations 
Dietary intake 
Biochemical profile 
 
Confounding variables 
Sex 
Age 
Habitual coffee consumption 
Self-perceived stress 
Score in the "Food restriction" section of the three-factor eating questionnaire (TFEQ) 
 
h. Ethical considerations 
This protocol adheres to the ethical principles of the General Health Law on Health Research 
regulations. According to Article 17 of this law, it is classified as "with higher than minimal 
risk" since it uses a method of assignment to therapeutic schemes. Likewise, each 
participant will be given an informed consent form based on the requirements of Article 22 
(37). 
 

On the other hand, the Good Clinical Practice (GCP) standards, issued by the International 
Conference on Harmonization (ICH), will be considered, which detail guidelines for the 
design, conduct, performance, monitoring, auditing, recording, analysis, and reporting of 
clinical trials, to ensure that the data and results obtained are accurate and credible and that 
the rights, integrity, and confidentiality of the study subjects are protected (38). 
  
In addition to the above, the international ethical guidelines for health-related research on 
human subjects elaborated by the Council for International Organizations of Medical 
Sciences (39) will be considered, in particular, guideline 5 (choice of control mechanism in 
clinical trials), guidelines 9 and 10 (on informed consent), guideline 11 (collection, storage 
and use of biological materials and related data), guideline 12 (collection, storage and use 
of data), guideline 20 (in disease outbreak situations (for the context of COVID-19)) and 
guideline 22 (use of online data and digital tools). 
 
i. Biosafety Considerations 
Instituto de Nutrigenética y Nutrigenómica Traslacional 
This institute has a laboratory safety regulation that requires the use of protective clothing 
such as lab coats, mouth masks, gloves according to the technique to be performed, 
appropriate footwear, and safety glasses. In addition, all students working on this project will 
receive a laboratory safety course in which the previously mentioned information is 
presented, training in good laboratory practices, and their practice will be constantly 
supervised and advised by trained personnel from the institute. INNUGET has a specific 
area for storing biohazardous and infectious waste, a specific area for storing reagents, a 
shower, separate areas for different types of techniques, emergency telephones, evacuation 
route signs, and a route for transporting biohazardous and infectious waste. 
  
Laboratory techniques to be performed at INNUGET include handling blood, serum, plasma, 
and genomic DNA extraction (the latter for future use). Blood is collected in vacutainer tubes, 
and serum, plasma, and genomic DNA are packaged in 1.5 ml Eppendorf tubes at 80% of 
their capacity and stored at -80°C. Vacutainer tubes containing traces of liquid blood, as well 
as Eppendorf tubes containing serum, plasma, or genomic DNA that will not be stored, are 
disposed of in hermetically sealed red containers that comply with the characteristics 
specified in NOM 087-SEMARNAT-SSA1-2002 (40), temporarily in the area designated for 
this purpose within INNUGET. 

Sharps will be deposited in a rigid red polypropylene container with a lid. Disposable 
materials used to transfer, contain, inoculate, and mix biological waste will be deposited in 
red plastic bags. Finally, disposable gloves will be disposed of in the regular trash if not 
soaked with blood. All this is done according to the specifications of the aforementioned 
official standard (40). Once the red containers and bags are filled to 80% of their capacity, 
they are taken to the appropriate containers at the CUCS  where a company collects them 
for incineration. 
  
Likewise, the handling and disposal of chemical reagents with CRETI characteristics will 
follow the stipulations of NOM-052-SEMARNAT-2005 (41) and their corresponding safety 
data sheets. It is worth mentioning that before each investigator and student uses the 
chemical reagents, he/she should carefully read the manufacturers' instructions and make 
a flow chart of the entire procedure for their use, recorded in the laboratory logs. 
 
Personal protective equipment should be worn in any contingency, and precautionary signs 
should be followed. The institution's civil protection personnel will be called and asked for 
help if a spill occurs. They are trained to apply the cleaning and evacuation protocol (if 
required) based on the nature of the residue or spill. The contact information for this unit is 
as follows: Unidad Interna de Protección Civil CUCS, Edificio E, Tel. 3310585200. Ext. 
33964, e-mail proteccioncivil@cucs.udg.mx. 
 
Unidad de Cirugía Bariátrica y Metabólica (Hospital Civil de Guadalajara <Dr. 
Juan I. Menchaca= 
No chemical reagents will be used at this site. Blood samples will be taken from participants 
in vacutainer tubes, which will be transferred to INNUGET in a cooler for biological samples 
for handling and disposal as mentioned in the section of the institute. The sharps used for 
sample collection will be deposited in a rigid red polypropylene container with a lid, in the 
area where the sample was collected, in accordance with the specifications of NOM-087-
SEMARNAT-SSA1-2002 (40).  
The protocol has also been submitted to the Ethics Committee of the Civil Hospital "Juan I. 
Menchaca" (0602-23 HCJIM-2023). 
 
METHODOLOGY 
Study group 

All participants will be assigned to two interventions separately, according to the random 
assignment sequence corresponding to the crossover design. 
The interventions consist of consuming a beverage (intervention or control) 30 minutes 
before breakfast daily for 2 weeks, with an intermediate washout period of 2 weeks between 
each treatment, as shown in Figure 1. Participants will be assigned randomly to the 
sequence AB (intervention beverage/control) or BA (control/intervention beverage) by 
generating a table of random numbers in the SPSS software. 
 
 
Figure 1.Scheme of the crossover study design 
 
Intervention procedure 
Subjects who attend the Bariatric and Metabolic Surgery Unit of the Nuevo Hospital Civil de 
Guadalajara <Dr. Juan I. Menchaca= and who meet the selection criteria. Subjects attend six 
sessions, one each week. The times and procedures can be seen in the following figure: 
 
Control
Intervention
Wash-out
Phase 1
Phase 2
Sequence 
AB
Sequence 
BA
Randomization
Participants
2 weeks
2 weeks
2 weeks
Control
Intervention

 
Figure 2. Intervention study procedure 
Appetite assessment 
 
Appetite hormones 
The hormones and peptides ghrelin are measured in their acetylated form (Biovendor brand 
kit, catalog #: RA194062400R), GLP-1 (Biovendor brand kit, catalog #: YK160), and PYY 
(Biovendor brand kit, catalog #: YK080 ), by an ELISA assay and using a Thermo Scientific 
brand Multiskan Sky spectrophotometer. 
 
Subjective sensations of appetite 
The subjective sensations of appetite consist of 9 questions that are asked in the fasting 
state and after drinking the beverage. Specifically, 30 minutes before drinking the beverage, 
immediately after consuming it, 30 and 60 minutes after consumption. These scales assess 
hunger, fullness, satiety, desire to eat, prospective consumption, and the desire for sweet, 
salty, fatty, and savory foods. Visual analogue scales are made up of lines, usually 100 mm 
long, with the term <None= or <Not at all= at one end and <Yes, very much= at the other end. 
or "As much as I have never felt." The subject marks a point between these two extremes, 
and the quantification is done by measuring the distance from the left end of the line to the 
mark to which a score is given (49). 
 
Sensory specific satiety (SSE) 

SSE will be evaluated by visual analog scales of liking and wanting of 100 mm each. The 
subjective taste or liking of certain sensory characteristics of the food before and after 
consumption is asked; it is complemented by the evaluation of desire, which is a motivational 
aspect of appetite or the inclination to consume a specific food (15). 
 
Specific sensory desire (SSD) 
Fasting, post-beverage, and post-food test participants will be instructed to rate their desire 
to eat something sweet, salty, sour, bitter, fatty, and spicy using visual analog scales. 
 
Food testing 
It consists of serving 9 samples of foods with certain sensory properties: 
1. Sweet: peach in syrup (13 g) 
2. Sour: green apple (8 g) with lemon drops (4 drops) 
3. Salty: salted biscuit (4 g) 
4. Bitter: cocoa nibs (3 pieces) 
5. Spicy: chilled peanuts (3 g) 
6. Fatty: avocado (11 g) 
7. Ultra-processed sweet: sandwich-type chocolate cookie with flavored filling 
vanilla (10g) 
8. Ultra-processed salty: cheese-flavored corn chips (4 g) 
9. Test beverage: it will be the intervention beverage or the control beverage, depending on 
the phase in which each participant is (20 ml) 
 
Participants will evaluate these foods using scales for sensory-specific satiety and sensory-
specific craving. Finally, they will be asked to choose the 3 test foods they would eat the 
most and the one they would eat the least at that time (15,16). 
 
Dietary intake 
 
Dietary record 
In session 1, participants will receive instructions to complete the dietary record form to 
evaluate their diets. They are asked to record all food and beverages consumed 3 days: a 
weekday, a weekend day, and the day before the session. This is done in each of the six 

sessions. The Nutritionist ProTM Diet Analysis software (Axxya Systems, Stafford, TX, USA) 
will be used for analysis. 
 
Biochemical profile 
 
Blood sample collection 
Of the 6 sessions that the participants attend, two blood samples will be taken at the 
beginning and end of each phase, that is, in 4 sessions. 
The first is in the morning, after 8 to 12 hours of fasting by venipuncture, and later 30 minutes 
after ingestion of the beverage. Two red and one purple vacutainer tubes are obtained, each 
with 5 ml of blood, centrifuged at 3500 rpm for 15 minutes at 4ºC to separate the serum and 
plasma. 
The serum will be used for glucose and lipid profile determinations, the plasma will be used 
for determining appetite hormones, 
 
Glucose and lipid profile determinations 
The determinations are carried out on the VITROS® 350 automated dry chemistry analyzer 
equipment. 10 μl of biological sample (serum) is used, which is applied to a slide that 
contains the reagents that allow the detection of the analyte. These measurements are 
carried out while fasting and later 30 minutes after ingestion of the beverage. 
In addition, capillary glucose measurements will be made with a glucometer (Accu-Check® 
Roche) while fasting and 30 minutes after ingestion of the beverage. The measurement is 
done through a puncture with a new, sterile lancet, which obtains a drop of blood that is 
applied to the reactive area of a strip (Accu-Check® Roche) to be read on the glucometer. 
 
Body composition and blood pressure 
Height measurement is done with a Seca stadiometer which has a precision of 0.1 cm. The 
subject remains standing with feet together (heels together and toes slightly apart). The 
heels, glutes, upper back, and occipital are in contact with the stadiometer. The head is 
placed in the Frankfort plane. 
 
Waist and hip circumference 
Waist circumference measurement is performed with a Lufkin brand anthropometric tape, 
which has an accuracy of 1 mm. The subject keeps his arms crossed on his chest, heels 

together with the toes slightly apart, and eyes in the Frankfort plane. The measurement is 
made at the narrowest level, between the edge of the lower costal rib (tenth rib) and the iliac 
crest. The subject breathes normally, and the measurement is taken at the end of an 
exhalation. For waist measurement, the circumference is taken at the maximum posterior 
level of the gluteal bulge. 
 
Electrical bioimpedance 
Measurements of fat mass, fat percentage, fat-free mass, and bone mass are obtained 
through the InBody 370 equipment, under the same conditions described for measuring 
body weight. 
 
Blood pressure 
Blood pressure measurement is performed in the morning, after 10 minutes of rest, with a 
LifeSource digital sphygmomanometer. The patient must be relaxed, sitting, not stretched 
out, with the back well supported on the back of the chair. His legs should be touching the 
floor, not crossed, and his hand should be relaxed, without squeezing, and in a resting 
position. The measurement is carried out on the reference or dominant arm supported more 
or less at the height of the heart. The subject's clothing does not cover the arm or fit tightly 
to it. 
 
Questionnaires 
Medical history 
A clinical history will be applied in the first session to obtain information on pathological 
history, clinical data, nutritional aspects, sociodemographic aspects, and lifestyle. 
 
Physical Activity Questionnaire (IPAQ) short version 
This instrument is a physical activity evaluation format; it examines different dimensions of 
the patient's physical activity. Weekly physical activity is measured by recording METs per 
minute per week. Subjects are classified into 3 categories of physical activity: high, 
moderate, and low (50). This questionnaire will be applied in the second and third sessions. 
 
The Three-Factor Eating Questionnaire 

This questionnaire consists of 51 items, is applied in the first session, and evaluates three 
dimensions of eating behavior: food restriction, disinhibition, and susceptibility to hunger 
(51). This questionnaire is included in the clinical history. 
 
Obtaining a stool sample 
At the beginning and the end of each intervention, participants will be asked to provide a 
stool sample (according to NOM-087-SEMARNAT-SSA1-2002, they are not considered an 
RBPI). They will be given a kit (sterile collection cup with buffer, collection strip, gloves, and 
stirrer) and written instructions for the collection procedure. Once subjects submit the 
sample, aliquots are made into Eppendorf tubes and stored at -80°C for later use. 
 
Statistical analysis 
Quantitative variables will be expressed as mean ± standard deviation or median and 
interquartile range for descriptive statistics, and qualitative variables as frequency or 
percentage. The normality of the variables is evaluated with the Shapiro-Wilk test. The 
general linear mixed model will be used to compare the two treatments. The data will be 
emptied into an Excel spreadsheet to be later imported into the SPSS v.21 program for 
analysis, considering a value of p<0.05 as statistically significant. The graphs will be made 
with Gradhpad software version 9.3.1. 
 
VIABILITY 
a. Financing 
This project is funded by <Fondo para Proyectos de Impulso a la Investigación, 2022 III=. 
 
b. Infrastructure 
There is access to the 2 study sedes and collaboration with the researchers responsible for 
the laboratories and hospital. These spaces already have the necessary equipment to carry 
out this project. 
 
c. Human resources training 
This research project will train two PhD students in Translational Nutrition Sciences and two 
undergraduate students. Furthermore, given its scope, it will allow the acceptance of 
professional internships and social service students. 
 

 
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