Trial Outcomes & Findings for Eating Chocolate at the Right Time Benefits the Circadiam Sytem and Metabolic Efficiency. (NCT NCT03949803)

Results Overview

The investigators will measure by indirect calorimetry the resting energy expenditure after each condition (no chocolate, Morning Chocolate, Night Chocolate) Changes in Resting Energy Expenditure (Indirect Calorimetry) were determined by changes in resting energy expenditure occurred in the participants between baseline and after 2 weeks in each of the three different experimental conditions. Oxygen (O2) consumption (mL/min) and carbon dioxide (CO2) production (mL/min) were measured. The Respiratory Quotient (RQ) was calculated out of O2 consumption (mL/min) and CO2 production (mL/min). Energy expenditure values (kcal/day) were calculated according to the Weir equation: Metabolic rate (kcal per day) = 1440 (3.9 VO2 + 1.1 VCO2)\*

Recruitment status

COMPLETED

Study phase

NA

Target enrollment

19 participants

Primary outcome timeframe

Baseline and after 2 weeks in each condition

Results posted on

2025-04-15

Participant Flow

Participant milestones

Participant milestones
Measure	Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Third condition is eating no milk chocolate or other chocolate	Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime	Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate	Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Second condition is eating no milk chocolate or other chocolate Third condition is eating 100gr chocolate within one hour of habitual waketime.	Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate First condition is eating no milk chocolate or other chocolate Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime	Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate First condition is eating no milk chocolate or other chocolate Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Third condition is eating 100gr chocolate within one hour of habitual waketime.
First Intervention (2 Weeks) STARTED	3	3	5	2	3	3
First Intervention (2 Weeks) COMPLETED	3	3	5	2	3	3
First Intervention (2 Weeks) NOT COMPLETED	0	0	0	0	0	0
Washsout (1 Week) STARTED	3	3	5	2	3	3
Washsout (1 Week) COMPLETED	3	3	5	2	3	3
Washsout (1 Week) NOT COMPLETED	0	0	0	0	0	0
Second Intervention (2 Weeks) STARTED	3	3	5	2	3	3
Second Intervention (2 Weeks) COMPLETED	3	3	5	2	3	3
Second Intervention (2 Weeks) NOT COMPLETED	0	0	0	0	0	0
Third Intervention (2 Weeks) STARTED	3	3	5	2	3	3
Third Intervention (2 Weeks) COMPLETED	3	3	5	2	3	3
Third Intervention (2 Weeks) NOT COMPLETED	0	0	0	0	0	0

Reasons for withdrawal

Withdrawal data not reported

Baseline Characteristics

Eating Chocolate at the Right Time Benefits the Circadiam Sytem and Metabolic Efficiency.

Baseline characteristics by cohort

Baseline characteristics by cohort
Measure	Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Third condition is eating no milk chocolate or other chocolate.	Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime.	Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate n=5 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Second condition is eating 100gr chocolate within one hour of habitual waketime.	Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate n=2 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Second condition is eating no milk chocolate or other chocolate Third condition is eating 100gr chocolate within one hour of habitual waketime.	Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate Second condition is eating 100gr chocolate within one hour of habitual waketime.	Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime Third condition is eating 100gr chocolate within one hour of habitual waketime.	Total n=19 Participants Total of all reporting groups
Age, Customized Age	56.00 years STANDARD_DEVIATION 7.81 • n=5 Participants	54.00 years STANDARD_DEVIATION 4.58 • n=7 Participants	50.20 years STANDARD_DEVIATION 3.56 • n=5 Participants	51.50 years STANDARD_DEVIATION 9.19 • n=4 Participants	51.00 years STANDARD_DEVIATION 0.00 • n=21 Participants	52.67 years STANDARD_DEVIATION 0.58 • n=10 Participants	52.37 years STANDARD_DEVIATION 4.59 • n=115 Participants
Sex: Female, Male Female	3 Participants n=5 Participants	3 Participants n=7 Participants	5 Participants n=5 Participants	2 Participants n=4 Participants	3 Participants n=21 Participants	3 Participants n=10 Participants	19 Participants n=115 Participants
Sex: Female, Male Male	0 Participants n=5 Participants	0 Participants n=7 Participants	0 Participants n=5 Participants	0 Participants n=4 Participants	0 Participants n=21 Participants	0 Participants n=10 Participants	0 Participants n=115 Participants

PRIMARY outcome

Timeframe: Baseline and after 2 weeks in each condition

The investigators will measure by indirect calorimetry the resting energy expenditure after each condition (no chocolate, Morning Chocolate, Night Chocolate) Changes in Resting Energy Expenditure (Indirect Calorimetry) were determined by changes in resting energy expenditure occurred in the participants between baseline and after 2 weeks in each of the three different experimental conditions. Oxygen (O2) consumption (mL/min) and carbon dioxide (CO2) production (mL/min) were measured. The Respiratory Quotient (RQ) was calculated out of O2 consumption (mL/min) and CO2 production (mL/min). Energy expenditure values (kcal/day) were calculated according to the Weir equation: Metabolic rate (kcal per day) = 1440 (3.9 VO2 + 1.1 VCO2)\*

Outcome measures

Outcome measures
Measure	Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating no milk chocolate or other chocolate.	Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime.	Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate n=5 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate	Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate n=2 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr chocolate within one hour of habitual waketime.	Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime	Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating 100gr chocolate within one hour of habitual waketime.
Resting Energy Expenditure (Indirect Calorimetry) Resting energy expenditure at Baseline	1249.33 kcal/day Standard Deviation 147.56	1377.67 kcal/day Standard Deviation 158.80	1360.40 kcal/day Standard Deviation 137.06	1468.50 kcal/day Standard Deviation 272.23	1427.33 kcal/day Standard Deviation 121.71	1259.67 kcal/day Standard Deviation 83.29
Resting Energy Expenditure (Indirect Calorimetry) Resting energy expenditure Morning Condition	1354.33 kcal/day Standard Deviation 227.25	1344.00 kcal/day Standard Deviation 115.47	1413.80 kcal/day Standard Deviation 151.61	1389.00 kcal/day Standard Deviation 255.97	1289.67 kcal/day Standard Deviation 153.05	1354.00 kcal/day Standard Deviation 60.23
Resting Energy Expenditure (Indirect Calorimetry) Resting energy expenditure Evening Condition	1292.33 kcal/day Standard Deviation 126.02	1459.67 kcal/day Standard Deviation 30.14	1323.60 kcal/day Standard Deviation 174.69	1497.50 kcal/day Standard Deviation 208.60	1281.00 kcal/day Standard Deviation 187.20	1351.67 kcal/day Standard Deviation 60.14
Resting Energy Expenditure (Indirect Calorimetry) Resting energy expenditure Control Condition	1342.00 kcal/day Standard Deviation 45.64	1376.67 kcal/day Standard Deviation 95.36	1439.20 kcal/day Standard Deviation 175.72	1464.50 kcal/day Standard Deviation 27.58	1420.67 kcal/day Standard Deviation 69.87	1397.00 kcal/day Standard Deviation 105.79

PRIMARY outcome

Timeframe: Baseline and after 2 weeks in each condition

The investigators will measure total body weight after each condition (no chocolate, Morning Chocolate, Night Chocolate)

Outcome measures

Outcome measures
Measure	Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating no milk chocolate or other chocolate.	Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime.	Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate n=5 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate	Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate n=2 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr chocolate within one hour of habitual waketime.	Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime	Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating 100gr chocolate within one hour of habitual waketime.
Total Body Weight Weight at Night condition	63.22 kilograms (kg) Standard Deviation 2.28	62.00 kilograms (kg) Standard Deviation 8.66	67.68 kilograms (kg) Standard Deviation 10.91	74.25 kilograms (kg) Standard Deviation 22.98	65 kilograms (kg) Standard Deviation 6.24	62.67 kilograms (kg) Standard Deviation 15.04
Total Body Weight Weight at Baseline	64.32 kilograms (kg) Standard Deviation 3.43	61.97 kilograms (kg) Standard Deviation 8.09	67.87 kilograms (kg) Standard Deviation 10.37	74.08 kilograms (kg) Standard Deviation 24.50	64.42 kilograms (kg) Standard Deviation 7.13	61.95 kilograms (kg) Standard Deviation 14.90
Total Body Weight Weight at Morning condition	63.02 kilograms (kg) Standard Deviation 1.76	62.07 kilograms (kg) Standard Deviation 8.42	67.73 kilograms (kg) Standard Deviation 10.68	73.35 kilograms (kg) Standard Deviation 22.98	64.58 kilograms (kg) Standard Deviation 7.28	63.2 kilograms (kg) Standard Deviation 15.24
Total Body Weight Weight at Control condition	63.33 kilograms (kg) Standard Deviation 2.08	61.00 kilograms (kg) Standard Deviation 7.81	68.00 kilograms (kg) Standard Deviation 10.49	74.00 kilograms (kg) Standard Deviation 24.04	64.33 kilograms (kg) Standard Deviation 7.77	60.67 kilograms (kg) Standard Deviation 14.22

SECONDARY outcome

Timeframe: Baseline and after 2 weeks in each condition

The investigators will measure the fragmentation parameter derived from wrist temperature after each condition (no chocolate, Morning Chocolate, Night Chocolate). Intradaily variability (IV) characterizes the rhythm fragmentation. Changes in daily rhythm of wrist temperature fragmentation were measured by changes in the fragmentation parameter derived from wrist temperature along the two weeks of intervention in each of the three conditions. The participants wore a wristwatch during the last 7 days of each condition on the non-dominant wrist to record body temperature and activity. Wrist temperature parameters were measured to represent the wrist body temperature pattern, which provides an objectively measure of sleep and siesta (together with activity rhythm). A low fragmentation involves a low intraday rhythmicity of the circadian rhythm. Its values oscillated between 0, when the wave was perfectly sinusoidal, and 2, when the wave was as Gaussian noise.

Outcome measures

Outcome measures
Measure	Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating no milk chocolate or other chocolate.	Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime.	Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate n=5 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate	Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate n=2 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr chocolate within one hour of habitual waketime.	Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime	Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating 100gr chocolate within one hour of habitual waketime.
Fragmentation of Wrist Temperature (WT) Daily Rhythm Fragmentation of WT rhythm at Control condition	0.11 arbitrary units Standard Deviation 0.03	0.24 arbitrary units Standard Deviation 0.14	0.12 arbitrary units Standard Deviation 0.04	0.15 arbitrary units Standard Deviation 0.04	0.15 arbitrary units Standard Deviation 0.03	0.13 arbitrary units Standard Deviation 0.04
Fragmentation of Wrist Temperature (WT) Daily Rhythm Fragmentation of WT rhythm at Baseline	0.23 arbitrary units Standard Deviation 0.11	0.15 arbitrary units Standard Deviation 0.05	0.25 arbitrary units Standard Deviation 0.13	0.18 arbitrary units Standard Deviation 0.08	0.13 arbitrary units Standard Deviation 0.05	0.18 arbitrary units Standard Deviation 0.07
Fragmentation of Wrist Temperature (WT) Daily Rhythm Fragmentation of WT rhythm at Morning condition	0.13 arbitrary units Standard Deviation 0.03	0.18 arbitrary units Standard Deviation 0.04	0.15 arbitrary units Standard Deviation 0.06	0.31 arbitrary units Standard Deviation 0.06	0.18 arbitrary units Standard Deviation 0.08	0.16 arbitrary units Standard Deviation 0.09
Fragmentation of Wrist Temperature (WT) Daily Rhythm Fragmentation of WT rhythm at Night condition	0.15 arbitrary units Standard Deviation 0.07	0.24 arbitrary units Standard Deviation 0.09	0.21 arbitrary units Standard Deviation 0.10	0.25 arbitrary units Standard Deviation 0.14	0.17 arbitrary units Standard Deviation 0.04	0.20 arbitrary units Standard Deviation 0.14

SECONDARY outcome

Timeframe: Baseline and after 2 weeks in each condition

The investigators will measure the regularity parameter derived from wrist temperature after each condition (no chocolate, Morning Chocolate, Night Chocolate). RegularIty is measured as interdaily stability (IS), that means rhythm stability over different days; it varied between 0 for Gaussian noise to 1 for perfect stability, where the rhythm repeated itself exactly day after day. Changes in daily rhythm of wrist temperature regularity were measured by changes in the regularity parameter derived from wrist temperature along the two weeks of intervention in each of the three conditions. A regular rhythm means recurring at uniform intervals. The participants wore a wristwatch during the last 7 days of each condition on the non-dominant wrist to record body temperature and activity. Wrist temperature parameters were measured to represent the wrist body temperature pattern, which provides an objectively measure of sleep and siesta (together with activity rhythm).

Outcome measures

Outcome measures
Measure	Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating no milk chocolate or other chocolate.	Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime.	Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate n=5 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate	Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate n=2 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr chocolate within one hour of habitual waketime.	Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime	Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating 100gr chocolate within one hour of habitual waketime.
Regularity of Wrist Temperature (WT) Daily Rhythm Regularity of WT daily rhythm at Baseline	0.44 arbitrary units Standard Deviation 0.12	0.45 arbitrary units Standard Deviation 0.06	0.49 arbitrary units Standard Deviation 0.16	0.32 arbitrary units Standard Deviation 0.04	0.56 arbitrary units Standard Deviation 0.05	0.35 arbitrary units Standard Deviation 0.17
Regularity of Wrist Temperature (WT) Daily Rhythm Regularity of WT daily rhythm at Morning condition	0.49 arbitrary units Standard Deviation 0.14	0.43 arbitrary units Standard Deviation 0.13	0.41 arbitrary units Standard Deviation 0.07	0.30 arbitrary units Standard Deviation 0.18	0.30 arbitrary units Standard Deviation 0.02	0.44 arbitrary units Standard Deviation 0.16
Regularity of Wrist Temperature (WT) Daily Rhythm Regularity of WT daily rhythm at Night condition	0.41 arbitrary units Standard Deviation 0.22	0.45 arbitrary units Standard Deviation 0.12	0.41 arbitrary units Standard Deviation 0.06	0.40 arbitrary units Standard Deviation 0.07	0.39 arbitrary units Standard Deviation 0.20	0.32 arbitrary units Standard Deviation 0.08
Regularity of Wrist Temperature (WT) Daily Rhythm Regularity of WT daily rhythm at Control condition	0.43 arbitrary units Standard Deviation 0.27	0.34 arbitrary units Standard Deviation 0.15	0.35 arbitrary units Standard Deviation 0.05	0.24 arbitrary units Standard Deviation 0.03	0.42 arbitrary units Standard Deviation 0.22	0.29 arbitrary units Standard Deviation 0.26

SECONDARY outcome

Timeframe: Baseline and after 2 weeks in each condition

The investigators will measure the amplitude parameter derived from wrist temperature after each condition (no chocolate, Morning Chocolate, Night Chocolate). In the cosinor analysis to characterize the WT rhythm, we calculate amplitude (difference between the maximum \[or minimum\] value of the cosine function and mesor) Changes in daily rhythm of wrist temperature amplitude were measured by changes in the amplitude parameter derived from wrist temperature along the two weeks of intervention in each of the three conditions. The participants wore a wristwatch during the last 7 days of each condition on the non-dominant wrist to record body temperature and activity. Wrist temperature parameters were measured to represent the wrist body temperature pattern, which provides an objectively measure of sleep and siesta (together with activity rhythm). A healthy circadian rhythm is considered as the one with high amplitude.

Outcome measures

Outcome measures
Measure	Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating no milk chocolate or other chocolate.	Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime.	Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate n=5 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate	Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate n=2 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr chocolate within one hour of habitual waketime.	Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime	Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating 100gr chocolate within one hour of habitual waketime.
Amplitude of Wrist Temperature (WT) Daily Rhythm Amplitude of WT daily rhythm at Baseline	0.95 ºC Standard Deviation 0.59	1.42 ºC Standard Deviation 0.57	0.92 ºC Standard Deviation 0.29	0.89 ºC Standard Deviation 0.55	1.67 ºC Standard Deviation 0.73	0.94 ºC Standard Deviation 0.90
Amplitude of Wrist Temperature (WT) Daily Rhythm Amplitude of WT daily rhythm at Morning condition	1.00 ºC Standard Deviation 0.97	1.35 ºC Standard Deviation 0.47	1.45 ºC Standard Deviation 0.38	0.71 ºC Standard Deviation 0.68	0.73 ºC Standard Deviation 0.22	0.95 ºC Standard Deviation 0.67
Amplitude of Wrist Temperature (WT) Daily Rhythm Amplitude of WT daily rhythm at Night condition	1.50 ºC Standard Deviation 0.83	1.05 ºC Standard Deviation 0.31	1.16 ºC Standard Deviation 0.33	0.54 ºC Standard Deviation 0.46	1.12 ºC Standard Deviation 0.26	0.86 ºC Standard Deviation 0.51
Amplitude of Wrist Temperature (WT) Daily Rhythm Amplitude of WT daily rhythm at Control condition	1.83 ºC Standard Deviation 1.05	0.85 ºC Standard Deviation 0.56	1.51 ºC Standard Deviation 0.47	0.77 ºC Standard Deviation 0.34	0.95 ºC Standard Deviation 0.63	1.06 ºC Standard Deviation 0.97

SECONDARY outcome

Timeframe: Baseline and after 2 weeks in each condition

Microbiota diversity changes were measured using Inverse Simpson Index, accounting for both species richness (number of species) and evenness (distribution of species). A higher Inverse Simpson Index indicates greater diversity because it reduces the impact of dominant species and gives more weight to rarer species. Unlike the regular Simpson Index (which decreases as diversity increases), the inverse form is more intuitive: higher values mean a more diverse microbiota. This index is widely used in microbiome studies to compare microbial diversity across different samples, such as in health versus disease states.

Outcome measures

Outcome measures
Measure	Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating no milk chocolate or other chocolate.	Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate n=2 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime.	Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate n=1 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate	Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate n=1 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr chocolate within one hour of habitual waketime.	Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate n=1 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime	Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate n=2 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating 100gr chocolate within one hour of habitual waketime.
Changes in Microbiota Diversity (Inverse Simpson Index) Inverse Simpson Index at Baseline	39.72 Index Standard Deviation 7.35	32.21 Index Standard Deviation 11.26	25.37 Index Standard Deviation 0	30.08 Index Standard Deviation 0	52.15 Index Standard Deviation 0	33.64 Index Standard Deviation 22.46
Changes in Microbiota Diversity (Inverse Simpson Index) Inverse Simpson Index Control condition	30.31 Index Standard Deviation 11.49	34.44 Index Standard Deviation 3.81	37.38 Index Standard Deviation 0	32.65 Index Standard Deviation 0	41.94 Index Standard Deviation 0	42.67 Index Standard Deviation 1.39
Changes in Microbiota Diversity (Inverse Simpson Index) Inverse Simpson Index Morning condition	30.53 Index Standard Deviation 19.08	29.60 Index Standard Deviation 12.83	30.12 Index Standard Deviation 0	18.70 Index Standard Deviation 0	31.87 Index Standard Deviation 0	34.49 Index Standard Deviation 34.08
Changes in Microbiota Diversity (Inverse Simpson Index) Evening condition	28.70 Index Standard Deviation 13.26	38.91 Index Standard Deviation 3.75	37.73 Index Standard Deviation 0	35.58 Index Standard Deviation 0	28.25 Index Standard Deviation 0	27.14 Index Standard Deviation 19.15

SECONDARY outcome

Timeframe: After 2 weeks in each condition

Short-chain fatty acids (SCFAs) in fecal samples were analyzed using gas-liquid chromatography (GLC) with a GC-FID system. Samples were mixed with NaOH, lyophilized, and homogenized with formic acid, methanol, and an internal standard (2-ethyl butyric acid). After ultrasonic treatment and centrifugation, the supernatant was filtered and injected into an Agilent 7890A GC system equipped with a Nukol capillary column. Helium was used as the carrier gas, and the FID temperature was set at 220°C. SCFAs were identified by comparing retention times with authentic standards, and quantification was performed using calibration curves with a high correlation (R² = 0.99). Total average values of SCFA (acetate, propionate, other minorities) in each condition were calculated.

Outcome measures

Outcome measures
Measure	Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate n=2 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating no milk chocolate or other chocolate.	Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime.	Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate n=1 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate	Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate n=1 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr chocolate within one hour of habitual waketime.	Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate n=1 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime	Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate n=1 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating 100gr chocolate within one hour of habitual waketime.
Concentration of Total Short-chain Fatty Acids (SCFAs) Total SCFA Control condition	419.93 mmol/mg feces Standard Deviation 160.81	293.15 mmol/mg feces Standard Deviation 107.81	588.49 mmol/mg feces Standard Deviation 0	196.93 mmol/mg feces Standard Deviation 0	659.92 mmol/mg feces Standard Deviation 0	596.45 mmol/mg feces Standard Deviation 0
Concentration of Total Short-chain Fatty Acids (SCFAs) Total SCFA Morning condition	533.10 mmol/mg feces Standard Deviation 171.38	450.68 mmol/mg feces Standard Deviation 209.02	482.40 mmol/mg feces Standard Deviation 0	876.56 mmol/mg feces Standard Deviation 0	608.80 mmol/mg feces Standard Deviation 0	426.75 mmol/mg feces Standard Deviation 0
Concentration of Total Short-chain Fatty Acids (SCFAs) Total SCFA Evening condition	377.78 mmol/mg feces Standard Deviation 152.54	661.70 mmol/mg feces Standard Deviation 217.39	552.32 mmol/mg feces Standard Deviation 0	754.51 mmol/mg feces Standard Deviation 0	890.21 mmol/mg feces Standard Deviation 0	580.86 mmol/mg feces Standard Deviation 0

SECONDARY outcome

Timeframe: Baseline and after 2 weeks in each condition

The investigators will measure changes in total energy intake measured as kilocalories between baseline and after 2 weeks in each condition (no chocolate, Morning Chocolate, Night Chocolate) At baseline and during the 2 weeks of each condition, time-stamped photographs were captured with a cell phone app. Total energy intake during the 14 days of each condition was analyzed with the nutritional evaluation software (Grunumur 2.0 8).

Outcome measures

Outcome measures
Measure	Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating no milk chocolate or other chocolate.	Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime.	Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate n=5 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate	Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate n=1 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr chocolate within one hour of habitual waketime.	Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime	Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating 100gr chocolate within one hour of habitual waketime.
Energy Intake Energy intake at Baseline	1177.20 kcal Standard Deviation 402.25	1987.20 kcal Standard Deviation 404.29	1558.15 kcal Standard Deviation 271.12	2044.61 kcal Standard Deviation 0	1843.39 kcal Standard Deviation 208.39	2031.93 kcal Standard Deviation 271.14
Energy Intake Energy intake at Morning condition	1596.74 kcal Standard Deviation 340.18	2214.31 kcal Standard Deviation 122.01	2374.69 kcal Standard Deviation 248.37	2034.83 kcal Standard Deviation 0	1933.39 kcal Standard Deviation 109.42	2248.11 kcal Standard Deviation 220.18
Energy Intake Energy intake at Night condition	1929.59 kcal Standard Deviation 295.25	2225.40 kcal Standard Deviation 26.95	2237.69 kcal Standard Deviation 570.40	2487.92 kcal Standard Deviation 0	2248.46 kcal Standard Deviation 64.52	2448.73 kcal Standard Deviation 589.64
Energy Intake Energy intake at Control condition	1797.58 kcal Standard Deviation 431.61	1843.50 kcal Standard Deviation 405.89	2173.95 kcal Standard Deviation 722.95	1882.36 kcal Standard Deviation 0	1548.47 kcal Standard Deviation 97.43	1766.66 kcal Standard Deviation 358.91

SECONDARY outcome

Timeframe: Baseline and after 2 weeks in each condition

The investigators will measure macronutrient composition measured as and grams per day between baseline and after 2 weeks in each condition (no chocolate, Morning Chocolate, Night Chocolate). At baseline and during the 2 weeks of each condition, time-stamped photographs were captured with a cell phone app. Macronutrient composition during the 14 days of each condition was analyzed with the nutritional evaluation software (Grunumur 2.0 8).

Outcome measures

Outcome measures
Measure	Morning Chocolate, Then Evening/Night Chocolate and Control Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating no milk chocolate or other chocolate.	Morning Chocolate, Then Control Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating 100gr chocolate within one hour of habitual waketime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime.	Evening/Night Chocolate, Then Morning Chocolate and Control Chocolate n=5 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating no milk chocolate or other chocolate	Evening/Night Chocolate, Then Control Chocolate and Morning Chocolate n=1 Participants First condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Second condition is eating no milk chocolate or other chocolate. Third condition is eating 100gr chocolate within one hour of habitual waketime.	Control Chocolate, Then Morning Chocolate and Evening/Night Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr chocolate within one hour of habitual waketime. Third condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime	Control Chocolate, Then Evening/Night Chocolate and Morning Chocolate n=3 Participants First condition is eating no milk chocolate or other chocolate. Second condition is eating 100gr of milk chocolate within 1 hours of habitual bedtime. Third condition is eating 100gr chocolate within one hour of habitual waketime.
Macronutrient Composition Protein at Baseline	45.56 grams/day Standard Deviation 10.26	76.59 grams/day Standard Deviation 10.76	58.37 grams/day Standard Deviation 17.75	75.46 grams/day Standard Deviation 0	75.28 grams/day Standard Deviation 6.49	71.53 grams/day Standard Deviation 11.08
Macronutrient Composition Carbohydrates at Night condition	174.53 grams/day Standard Deviation 21.70	237.02 grams/day Standard Deviation 10.58	232.31 grams/day Standard Deviation 50.15	233.36 grams/day Standard Deviation 0	207.00 grams/day Standard Deviation 27.04	233.52 grams/day Standard Deviation 76.51
Macronutrient Composition Protein at Night condition	75.34 grams/day Standard Deviation 10.11	76.99 grams/day Standard Deviation 5.70	76.04 grams/day Standard Deviation 20.96	71.53 grams/day Standard Deviation 0	76.30 grams/day Standard Deviation 9.20	87.47 grams/day Standard Deviation 21.96
Macronutrient Composition Fat at Baseline	58.78 grams/day Standard Deviation 13.50	101.32 grams/day Standard Deviation 28.72	66.44 grams/day Standard Deviation 19.62	103.12 grams/day Standard Deviation 0	92.87 grams/day Standard Deviation 10.77	116.83 grams/day Standard Deviation 15.56
Macronutrient Composition Carbohydrates at Baseline	111.39 grams/day Standard Deviation 45.57	198.72 grams/day Standard Deviation 30.66	182.84 grams/day Standard Deviation 45.09	202.74 grams/day Standard Deviation 0	164.86 grams/day Standard Deviation 40.63	171.72 grams/day Standard Deviation 39.89
Macronutrient Composition Carbohydrates at Morning condition	156.78 grams/day Standard Deviation 21.09	256.53 grams/day Standard Deviation 36.88	239.23 grams/day Standard Deviation 39.60	214.30 grams/day Standard Deviation 0	174.29 grams/day Standard Deviation 31.04	217.07 grams/day Standard Deviation 29.02
Macronutrient Composition Fat at Morning condition	76.97 grams/day Standard Deviation 19.28	102.66 grams/day Standard Deviation 14.92	122.65 grams/day Standard Deviation 14.30	99.44 grams/day Standard Deviation 0	103.31 grams/day Standard Deviation 19.58	119.00 grams/day Standard Deviation 9.83
Macronutrient Composition Protein at Morning condition	67.17 grams/day Standard Deviation 15.70	74.10 grams/day Standard Deviation 6.43	80.39 grams/day Standard Deviation 10.18	66.61 grams/day Standard Deviation 0	70.88 grams/day Standard Deviation 2.34	83.28 grams/day Standard Deviation 7.52
Macronutrient Composition Fat at Night condition	102.36 grams/day Standard Deviation 21.85	109.84 grams/day Standard Deviation 2.46	112.92 grams/day Standard Deviation 34.86	139.51 grams/day Standard Deviation 0	115.74 grams/day Standard Deviation 7.39	131.45 grams/day Standard Deviation 25.01
Macronutrient Composition Carbohydrates at Control condition	141.53 grams/day Standard Deviation 30.34	212.31 grams/day Standard Deviation 91.16	218.36 grams/day Standard Deviation 76.08	179.15 grams/day Standard Deviation 0	137.00 grams/day Standard Deviation 43.42	166.02 grams/day Standard Deviation 28.94
Macronutrient Composition Fat at Control condition	97.20 grams/day Standard Deviation 32.61	80.99 grams/day Standard Deviation 8.32	110.66 grams/day Standard Deviation 39.32	97.12 grams/day Standard Deviation 0	76.80 grams/day Standard Deviation 12.32	90.09 grams/day Standard Deviation 25.22
Macronutrient Composition Protein at Control condition	75.15 grams/day Standard Deviation 19.68	68.27 grams/day Standard Deviation 16.37	78.30 grams/day Standard Deviation 22.18	70.04 grams/day Standard Deviation 0	65.44 grams/day Standard Deviation 10.44	70.04 grams/day Standard Deviation 10.08

Deaths: 0 deaths

Serious adverse events

Adverse event data not reported

Other adverse events

Adverse event data not reported

Additional Information

Prof. Marta Garaulet Aza

Universidad de Murcia

Phone: +34868889478

Email: [email protected]

Results disclosure agreements

Principal investigator is a sponsor employee
Publication restrictions are in place