Journal of Dietary Supplements

ISSN: 1939-0211 (Print) 1939-022X (Online) Journal homepage: https://www.tandfonline.com/loi/ijds20

A systematic review of the effect of L-tryptophan

supplementation on mood and emotional
functioning

Asako M. Kikuchi, Aya Tanabe & Yoshihiro Iwahori

To cite this article: Asako M. Kikuchi, Aya Tanabe & Yoshihiro Iwahori (2020): A systematic review
of the effect of L-tryptophan supplementation on mood and emotional functioning, Journal of Dietary
Supplements, DOI: 10.1080/19390211.2020.1746725

To link to this article: https://doi.org/10.1080/19390211.2020.1746725

Published online: 10 Apr 2020.

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JOURNAL OF DIETARY SUPPLEMENTS
https://doi.org/10.1080/19390211.2020.1746725

REVIEW

A systematic review of the effect of L-tryptophan

supplementation on mood and emotional functioning
Asako M. Kikuchi, PhDa, Aya Tanabe, PhD, DDSa, and Yoshihiro Iwahori, PhDa,b
a
LLC Okutoeru, 4-18-21-314, Minamiaoyama, Minato-ku, Tokyo, Japan; bDepartment of Pharmaceutical
and Medical Business Sciences, Nihon Pharmaceutical University, Kitaadachi-gun, Saitama, Japan

ABSTRACT KEYWORDS
L-tryptophan (TRP), one of the essential amino acids in humans, is a L-tryptophan; serotonin;
precursor of serotonin, and hence its intake is closely related to the emotion; depression;
suppression of depressed and anxious moods. We did a systematic healthy adults
review of RCTs to examine the effects of tryptophan intake on the
mood of healthy adults by searching PubMed, the Cochrane Library,
and Ichu-shi according to PRISMA guidelines. As a result, 11 RCTs
met the criteria and were accepted. Four RCTs showed the effects of
tryptophan intake on negative feelings and happy feelings in healthy
individuals, with significant differences between the treatment and
the control groups. This suggests that TRP intake may be an effect-
ive approach to decrease anxiety and increase positive mood in
healthy individuals. On the other hand, the effectiveness of TRP for
aggressive feelings was not recognized. Reviewing these 11 RCTs, we
concluded that taking 0.14–3 g of TRP per day in addition to the
usual meal can be expected to improve the mood of healthy individ-
uals. In order to estimate the optimum amount of TRP intake more
accurately, further studies need to be conducted with more appropri-
ate settings of intake period, intake frequency, and intake method.

Introduction
In modern society, mental disorders due to stress, such as depression and anxiety, are
increasing (Hulsken et al. 2013), and ingredients that improve moods without side
effects are required (Mohajeri et al. 2015). The nascent field of nutritional psychiatry
offers some opportunities for clinical intervention for individuals who suffer from
depression and anxiety. L-tryptophan and probiotic supplementation are good examples
of nutritional interventions. Ng et al. found that probiotic supplementation produced
significant improvements in the moods of individuals with mild to moderate depressive
symptoms (Ng et al. 2018). Also, L-tryptophan (TRP), which is included in dietary pro-
teins, is expected to improve the mood of individuals with depressive feelings.
L-tryptophan, which is a precursor for serotonin, has been used as dietary
supplements in many countries for many years (Murphy et al. 2006). Serotonin (5-
hydroxytryptamine), a bioactive substance and one of the neurotransmitters, is involved
in emotion processing, memory, attention and information processing (Riedel et al.

CONTACT Asako M. Kikuchi info@okutoeru.com LLC Okutoeru, 4-18-21-314, Minamiaoyama, Minato-ku, Tokyo,
107-0062, Japan.
ß 2020 Taylor & Francis Group, LLC
2 A. M. KIKUCHI ET AL.

2003). Previous studies have shown that increased serotonin in the brain improves
mood, reduces emotional disturbances in healthy subjects, and a decrease in brain sero-
tonin causes depression and anxiety (Maes and Meltzer 1995; Markus et al. 1999; Markus
2003; van Praag 2004). Serotonin synthesis in the brain has long been recognized as diet-
ary therapy, which has been attributed to the increased use of TRP in the brain (Young
et al. 1985; Young 2013). To date, many TRP-depleted studies have been conducted to
mimic depressed moods. Continuing a low TRP diet can lead to a decrease in blood TRP
concentration (>70%) within 4 to 6 h. There is also a report that TRP induces an
unpleasant mood (Silber and Schmitt 2010). This reduction in TRP usage in the brain
tends to resemble the cognitive bias seen in depressed people in terms of memory and
attention to positive/negative information (Mathews and MacLeod 2005).
The precursor of serotonin, TRP, is one of the essential amino acids that cannot
be synthesized in the body and must be obtained from the diet, so there has been a
long-standing interest in the effects of TRP-containing diet (Mohajeri et al. 2015). In
addition, the rate-limiting step for serotonin synthesis involves the conversion of TRP
to 5-hydroxytryptophan by the enzyme tryptophan hydroxylase, which is not fully satu-
rated by the substrate under normal conditions. Thus, by increasing brain TRP levels,
serotonin synthesis can be enhanced in the brain (Mohajeri et al. 2015).
TRP-rich foods or supplements have been studied for their effects on mood and anx-
iety symptoms. There is a secondary analysis to determine the association of daily TRP
intakes and with biochemical markers of health- and safety-related outcomes, self-
reported depression, and sleep-related variables (Lieberman et al. 2016). It has been
reported that an inverse correlation between tryptophan intake and self-reported depres-
sion (p < 0.01), and a positive correlation with self-reported sleep duration (p ¼ 0.02)
(Lieberman et al. 2016). On the other hand, it is also reported that there is an inverse
U-shaped relationship between the brain TRP levels and the state of mood/recognition.
Therefore, it is feared that excessive intake of TRP would have an adverse effect
(Hulsken et al. 2013).
In a 2010 review paper, TRP intake was found to improve the mood of subjects who
are vulnerable to stress, such as those who have experienced depression (Silber and
Schmitt 2010). It has also been reported that TRP improves the sleep of adults with sleep
disorders and delays in reaction time due to its sedative effects (Silber and Schmitt 2010).
Although there are some secondary analyzes and reviews on mood improvement by
TRP intake, there is still no systematic review including qualitative evaluation. In add-
ition, it is necessary to obtain knowledge not only on the effectiveness but also on the
optimum dose because adverse effects due to overdose have been reported. The purpose
of this SR is to prevent the depression of healthy people through food and supplements.
The effectiveness and optimal TRP intake are investigated by conducting a systematic
review of the effects of TRP intake on anxiety and mood in healthy adult populations.

Methods
This systematic review was reported according to the PRISMA reporting guidelines for
meta-analysis and systematic reviews (Knobloch et al. 2011). The PRISMA checklist was
provided in Additional file 1.
 JOURNAL OF DIETARY SUPPLEMENTS 3

Data sources and searches

We searched PubMed, the Cochrane Library, and the Japanese medical magazines
named “Ichu-shi”, all of which are highly reliable and rich publications/citations
(Ichushi-Web). The search has been conducted in October 2018. The search terms for
PubMed and The Cochrane Library were “tryptophan,” “healthy,” and “adult.” In
PubMed, Article types are further limited to Randomized Controlled Trial (the search
term was “Search (Tryptophan) AND ((healthy) AND adult) Filters: Randomized
Controlled Trial”). The search term for “Ichu-shi” was “tryptophan,” and the type of
paper (PT) ¼ original paper, research design (RD) ¼ randomized or quasi-randomized
controlled trial, and subject (CK) ¼ human. The search was performed by two inde-
pendent researchers using the same search terms and filters. All duplicate articles were
identified and deleted. Reviewers individually rated all articles that might be relevant. In
the case of different evaluations, two people consulted, and if the evaluations were still
different, another reviewer made the final decision.

Research question (RQ)

“Does oral tryptophan intake affect improving mood depression? If so, what is the
optimal dose?”

PICOS
P (Participant): Healthy adult men and women, excluding patients with depression and
other related diseases
i (Intervention): Intervention of tryptophan ingestion, excluding cases involving
5-hydroxytryptophan intervention, other interventions, and tryptophan depletion
C (Comparison): A control group is a group without tryptophan intervention or a
group with less tryptophan intervention
O (Outcome): Depressed feelings, positive/negative moods, recognition to positive/
negative expressions, excluding items related to sleep, workability, and fatigue
S (Study design): Randomized controlled trial, quasi-randomized controlled trial,
excluding reviews, observational studies, case reports, and in vitro/vivo human studies

Inclusion criteria
The study was selected based on RQ and PICOS. All papers were reviewed by two
researchers and the data were extracted independently in a standard predefined way.
Reviewers assessed all the articles individually. If there was a difference in evaluation
regarding the adoption of the paper, two people discussed, and if the opinion was still
different, another reviewer judged it. Duplicated papers were deleted from papers
that were hit by database search. We reviewed the publication titles and summaries of
the remaining papers, excluding those that were clearly irrelevant, reviewed the full text
of the remaining papers, examined eligibility, and selected studies that met the
review criteria.
4 A. M. KIKUCHI ET AL.

Methodological evaluation of RCTs using the modified Jadad scale

The Jadad scale was used to assess the methodological quality of the included clinical
trials. This scale was developed to assess the quality of articles in systematic reviews by
avoiding selection bias and examining the effects of blinding in randomized controlled
trials (Jadad et al. 1996). Table 1 shows the modified Jadad scale (Dimitriou et al.
2017), some of which were changed from the original scale so that it is more suitable
for the RCT’s evaluation examining the effects of TRP on mood and emotional func-
tion. Scores in the range of 0–8 are possible, and higher scores mean higher quality.
Studies with a score of less than four are considered low quality, while studies with 4–8
points are considered high quality. Two reviewers independently evaluated all 32 items
and there was no disagreement regarding the score.

Quality assessment using the Cochrane Collaboration risk of bias tool

The methodological quality of the included studies is assessed according to the “risk of
bias table” of the Cochrane Collaboration (Cochrane Handbook for Systematic … ),
including seven items. The assessment degree of each item is divided into three levels:
low risk, unclear risk, and high risk. According to the seven items, the specific condi-
tions included in the literature are assessed by two reviewers independently. In the case
of different assessment, the two people consulted, and if the assessment was still differ-
ent, another reviewer made the final decision.

Table 1. Modified Jadad scale with eight items for randomized controlled trials.
Question/Response SCORE
Was the study described as randomized?
Yes þ1
No 0
Was the method of randomization appropriate?
Yes þ1
No 1
Not described 0
Was the study described as blinded?
Yes (Single blinded) þ0.5
Yes (Double blinded) þ1
No 0
Was the method of blinding appropriate?
Yes þ1
No 1
Not described 0
Was there a description of withdrawals and dropouts?
Yes þ1
No 0
Was there a clear description of inclusion/exclusion criteria?
Yes þ1
No 0
Was the method used to assess adverse effects described?
Yes þ1
No 0
Is the method of statistical analysis described?
Yes þ1
No 0
 JOURNAL OF DIETARY SUPPLEMENTS 5

Results
Search and screening results
Total of 556 articles matched the search, 213 of which were deleted due to duplication.
The algorithm that was used to select papers is shown in Figure 1. As a result of evalu-
ating titles or summaries of the remaining 343 papers, 328 articles (205 in PubMed, 297
in Cochrane, 8 in “Ichu-shi”) were rated as irrelevant. As a result of obtaining and
examining the remaining 15 articles, four articles were excluded (reasons for exclusion
are shown in Figure 1). Eventually, 11 RCTs conducted between 1999 and 2015 met the
review criteria and were included in this systematic review (Mohajeri et al. 2015;
Murphy et al. 2006; Gibson et al. 2014; Merens et al. 2005).

Methodological quality and Jadad score

The eleven RCTs were rated according to the modified Jadad scale (Table 2), 10 of
which were rated high quality (Jadad score 4–6) and the remaining one was rated low

Figure 1. Flowchart of exclusion process and final selection of RCTs included in the systematic review.
 6

Table 2. Modified Jadad score for the methodological evaluation of the randomized controlled trials.
Description of
Appropriate Description of Method of method of
A. M. KIKUCHI ET AL.

method Appropriate Description of inclusion/ adverse data

Author (year of Description of of Description method of withdrawals exclusion effects statistical Jadad
publication) randomization randomization of blinding blinding and dropouts criteria assessment analysis score
Mohajeri et al (2015) Yes ND Yes (double blinded) ND Yes Yes Yes Yes 5
Gibson et al (2014) Yes Yes Yes (double blinded) ND Yes Yes Yes Yes 6
Firk and No ND Yes (double blinded) Yes No Yes No Yes 4
Markus (2009)
Murphy et al (2009) Yes ND Yes (double blinded) ND No Yes No Yes 4
Markus et al (2008) Yes ND Yes (double blinded) Yes No Yes No Yes 5
Aan Het Rot No ND Yes (double blinded) ND Yes Yes No Yes 3
et al (2006)
Murphy et al (2006) Yes ND Yes (double blinded) ND Yes Yes Yes Yes 5
Attenburrow Yes (stratified ND Yes (double blinded) ND No Yes No Yes 4
et al (2003) randomization)
Wingrove et al (1999) Yes ND Yes (double blinded) Yes Yes Yes No No 4
Lindseth et al (2015) Yes ND Yes (double blinded) Yes - (No dropout) Yes No Yes 5
Merens et al (2005) Yes (described only ND Yes (double blinded) Yes, partially Yes Yes No Yes 4.5
in abstract)
 JOURNAL OF DIETARY SUPPLEMENTS 7

quality (Jadad score 3). Nine studies had randomization descriptions (one of them was
stratified randomization), only one of which was described on the method of random-
ization. There were only five studies described the blinding methods (one of which only
described the blinding method of the subjects). Withdrawal and dropout rates were
reported in 6 studies, whereas there was no mention of dropout in 4 studies, and there
was no dropout in the other study. The side effects assessment method was mentioned
in 3 studies, although subject inclusion criteria were described in all studies. There was
one study without a description of the statistical methods used.

Assessment of overall risk of bias

Based on the Cochrane review manager, we assessed bias risk for each outcome (Figure 2).
As a result, none of the outcomes (facial expression recognition/POMS/BDHI/PANAS) had
a high bias risk. The description of the blinding method was not described in all the papers
in the study on facial expression recognition, and the bias risk was unknown. None of the
outcomes have adequately documented assignment concealment, and the risk of conceal-
ment assignment was unknown. For other biases (incomplete outcomes, selective reporting,
other bias risks), the bias risk was often low.
On the other hand, in the review manager of Cochrane, a study by Mohajeri et al. in
2015 and Gibson et al. in 2014, whose bias risk was low, included authors of companies
who supply supplements including tryptophan, and there was a risk of bias due to con-
flicts of interest.

Heterogeneity of the comparators

As expected, there was no homogeneity regarding the group of subjects compared to the
11 RCTs. Eleven trials were conducted in the United Kingdom (6), the Netherlands (3),
the United States (1), and Canada (1), including a total of 413 participants, 401 of which
were completed each study. Two studies included only adult participants and seven stud-
ies included 18-19 years-old participants, which was very minor. There were no partici-
pants whose ages were less than 18. In the Firk and Markus study in 2009 participants
were university students, with a mean age of 21.7 ± 1.9 years for the high cognitive reactiv-
ity group and 21.1 ± 2.1 years for the low cognitive reactivity group. Participants in the
study of Aan Het Rot et al. described only the average age, which was 30.1 ± 9.1 years for
women, and 34.1 ± 10.1 years for men. Nine studies included both male and female partic-
ipants, whereas two studies included only female participants.
In 7 studies, all subjects were healthy subjects, while the remaining four studies
included participants who were prone to depression or diagnosed as having temper or
aggression. Interventions of three studies were tryptophan alone, six were tryptophan-
rich proteins derived from eggs and dairy products, and the others were meals contain-
ing more tryptophan. In addition, five studies examined the short-term effects (0.5 to
4.5 h after tryptophan intake), while the other six studies examined the long-term effects
(continuous intake daily for 4 to 28 days).
8 A. M. KIKUCHI ET AL.

(a) Facial expression recognition (C) BDHI

blinding of data analyst

blinding of data analyst

allocation concealment

allocation concealment
blinding of participants

blinding of participants
incomplete outcome

incomplete outcome
selective reporting

selective reporting
random sequence

random sequence
stopped early for

stopped early for

and personnel

and personnel
benefit, etc

benefit, etc
generation

generation
data

data
Mohajeri MH
Murphy SE et
et al (2015-
al (2006-UK)
UK)
Gibson EL et Wingrove J et
al (2014-UK) al (1999-UK)
Murphy SE et
al (2006-UK)
Attenburrow
MJ et al (2003-
UK)

(b) POMS (D) PANAS

blinding of data analyst

blinding of data analyst

allocation concealment

allocation concealment
blinding of participants

blinding of participants
incomplete outcome

incomplete outcome
selective reporting

selective reporting
random sequence

random sequence
stopped early for

stopped early for

and personnel

and personnel
benefit, etc

benefit, etc
generation

generation
data

data
Firk C and Firk C and
Markus CR Markus CR
(2009- (2009-
Netherlands) Netherlands)
Markus CR et
Murphy SE et
al (2008-
al (2009-UK)
Netherlands)
Merens W et
Murphy SE et
al (2005-
al (2006-UK)
Netherlands)

risk of bias
low
unclear
high

Figure 2. Cochrane risk of bias of the studies included in this systematic review.

The effects of tryptophan intake

The effects of tryptophan intake were observed both in the short and long term. To facilitate the
interpretation of RCT results, the included studies were grouped into four categories (negative
emotion, happiness, aggression, and risk management) by the outcome. Classification criteria
were effect indicators. The results of all included studies are shown in Table 3.

The evaluation of the effect on negative emotions

Four RCTs evaluated the negative emotional effects of tryptophan intake in healthy
individuals. In a study published in 2003 (Attenburrow et al. 2003), women took 1.8 g
Table 3. Brief summary of the studies included in the systematic review.
Parients
total
number (N) Inervention Comparator
Author Intervention Age number number Adverse
(Year-Country) category Intervention Control Duration Trp intake Sex of patients (N) of patients (N) Measures Outcomes Effects
Short duration Gibson et al TRP-rich protein Beverage containing Beverages 1 hour 140 mg, 280 mg N ¼ 60 LumiVida TM 2g placebo N ¼ 19 Happiness 2 g intake None
(2014-UK) 2 g or 4 g of egg containing 3.11 g 45–65 y N ¼ 20 suppresses the
white protein casein Healthy female LumiVida TM 4g Feeling tired increase in
hydrolyzate rich hydrolyzate N ¼ 21 fatigue and
in TRP Once/day happiness
associated with
Once/day testing

Increased
response to
positive
language and
reduced bias to
negative facial
expressions
Firk and Markus TRP-rich protein Beverages Casein beverage 2 hours 0.8g/Day N ¼ 38 18 subjects with a 18 subjects with a Mood Significant difference –
(2009-Netherlands) containing containing Average age of low low CR score assessment in POMS
0.8 g tryptophan 0.4 g tryptophan depressed group: CR score (LEIDSscore <24) item vitality
21.7 (LEIDSscore and 20 subjects Cortisol
Average age of <24) and 20 with a high measurement
group with no subjects CR score (LEIDS
depression: 21.1 with a high CR score > 47)
score (LEIDS
score > 47)
Markus et al TRP-rich protein
15 g a-lactalbumin Casein protein 20 g 30, 60, 90, 0.8 g/Day N ¼ 18 N ¼ 18 N ¼ 18 Plasma amino Slight improvement –
(2008-Netherlands) whey protein containing 120, 180, 18–30 y acid in POMS
containing 0.8 g tryptophan 0.4 g 210 minutes 9 males and measurement
tryptophan and neutral 9 females
Or, 4 g mix of amino acid 10 g Mood
tryptophan 0.8 g assessment
and
synthetic
peptide
Attenburrow et al 80%Tryptophane 1.8 g nutritionally – 4 hours – N ¼ 24 N ¼ 12 placebo N ¼ 12 Venous sampling There is no –
(2003-UK) sourced TRP , Average age: significant
80% tryptophan 36.2 Facial expression difference in
hydrolyzed by Female recognition test facial expressions
JOURNAL OF DIETARY SUPPLEMENTS

milk and purified other than fear

by HPLC and happiness
(continued)
9
 10

Table 3. Continued.
Parients
total
number (N) Inervention Comparator
Author Intervention Age number number Adverse
(Year-Country) category Intervention Control Duration Trp intake Sex of patients (N) of patients (N) Measures Outcomes Effects
Wingrove et al TRP-rich protein Drink containing Tryptophan- 4.5 hours N.D. N ¼ 28 N ¼ 14 placebo N ¼ 13 Serum prolactin Reduces aggression –
(1999-UK) 10.3 g free drink 18-44 y measurement and hostility
of tryptophan Healthy male
and female
Long duration Lindseth et al Meal Meal containing Meal containing 4days Calculate the target N ¼ 25 N ¼ 25 N ¼ 25 Salivary cortisol Reduced anxiety –
A. M. KIKUCHI ET AL.

(2015-USA) tryptophan of tryptophan of amount from 18 y old or older Zung’s Self-

10 mg/kg body 5 mg/kg body the average Healthy male Rating Anxiety
weight/day weight/day weight of the and female Scale
subject Zung’s Self-
Rating
Depression Scale
Positive Affect
Negative Affect
Schedule test
Murphy et al Tryptophane Tryptophan 1 g 1 g placebo 14 days 3 g/Day N ¼ 30 N ¼ 15 placebo N ¼ 15 Risky Choice Test Tendency to make –
(2009-UK) 18–40 y decisions that
3 times a day 3 times a day Healthy 14 underestimate
for 14 days for 14 days males risk of loss in
and 16 females risky decisions
No significant
impact on
anxiety that took
significantly
longer to make
a decision
Murphy et al Tryptophan Tryptophan 1g 1 g placebo 14 days 3 g/Day N ¼ 38 N ¼ 19 placebo N ¼ 19 Emotion The woman reacts None
(2006-UK) 18–40 y processing to a happy
3 times/day 3 times/day for Healthy male evaluation expression
14 days 14 days and female Decreased
reaction to
disgusting facial
expressions
There is not
much tendency
for men
Aan Het Rot et al (2006 Tryptophan Tryptophan 1g 1 g placebo 15 days 3 g/Day N ¼ 39 N ¼ 39 N ¼ 39 Positive and The effect of –
-Canada) Average age of negative affect reducing conflict
3 times/day for 3 times/day for 20 healthy adjectives on a and increasing
15 days 15 days males: 34.1 scale from 0 to 6 synchrony,
(continued)
Table 3. Continued.
Parients
total
number (N) Inervention Comparator
Author Intervention Age number number Adverse
(Year-Country) category Intervention Control Duration Trp intake Sex of patients (N) of patients (N) Measures Outcomes Effects
Average age of Mark on a 9 1 especially
19 healthy 9 affect grid reducing the
females: 30.1 Check off 12 dominant
behaviors attitude in men
11 1 11 There was no
interpersonal significant
grid difference when
tryptophan was
taken for the
first time in the
crossover, and
the joy value
was significantly
increased in the
second
intake group
TM
Mohajeri et al TRP-rich protein Drink containing Drink containing 19 days 140 mg/Day N ¼ 59 LumiVida N ¼ 29 placebo N ¼ 30 Active mood Increased happiness None
(2015-UK) 0.5 g 0.5 g of casein 45–65 y Reaction to when taken
supplement hydrolyzate Healthy female words 60–90 minutes
containing Evaluation of before bedtime
approximately 2 times/day facial expressions Delayed reaction
70 mg Sleep quality to negative
tryptophan words
Reduces anger
2 times/day and disgust
response
Increased
response to
happiness
and surprise
Merens et al (2005 TRP-rich protein Oral administration Oral administration 4 weeks – N ¼ 43 N ¼ 23 N ¼ 23 Stress test Mood did –
-Netherlands) of of casein male and female N ¼ 20 N ¼ 20 not improve
a-lactalbumin with a history of
major depression
and healthy
male and female
JOURNAL OF DIETARY SUPPLEMENTS
11
12 A. M. KIKUCHI ET AL.

of nutritionally sourced TRP. When the effect on emotion was evaluated using vein
sampling and facial expression recognition test 4 h after ingestion, there was a signifi-
cant difference in the recognition of feared facial expressions between the intake group
and the control group. Murphy et al. evaluated the effects of taking 1 g tryptophan three
times a day for 14 days (Murphy et al. 2006). In the evaluation of emotional processing
between the two groups, only females showed a significant difference in disgusted facial
expressions. In a study of women who took a drink containing 2 g or 4 g of TRP-rich
egg white protein hydrolyzate once a day, in the group that took 2 g, there was a signifi-
cant difference in the recognition of facial expressions of fear, sadness, and disgust with
stress (Gibson et al. 2014). In a study by Mohajeri et al. (2015), the effect of taking a
drink containing 0.5 g of a supplement containing approximately 70 mg of tryptophan
twice a day for 19 days was evaluated. In this study, there was a significant difference in
the recognition of angry facial expressions between the test group and the con-
trol group.

The evaluation of the effect on happiness

Four RCTs evaluated the effect of tryptophan intake on happiness in healthy individu-
als. In a study published in 2003 (Attenburrow et al. 2003), women took 1.8 g of nutri-
tionally sourced TRP. When the effect on emotion was evaluated using vein sampling
and facial expression recognition test 4 h after ingestion, there was a significant differ-
ence in recognition of happy facial expressions between the intake group and the con-
trol group. Murphy et al. evaluated the effects of taking 1 g tryptophan three times a
day for 14 days (Murphy et al. 2006). In the evaluation of emotional processing between
the two groups, only women recognized a significant difference in their perception of
happiness. In a study of women who took a beverage containing 2 g or 4 g of TRP-rich
egg white protein hydrolyzate once a day (Gibson et al. 2014), the reduction in well-
being caused by stress was significantly suppressed in the 2 g group. The reaction to
positive words has increased. In a study by Mohajeri et al. (2015), the effect of ingesting
a drink containing 0.5 g of a supplement containing approximately 70 mg of tryptophan
twice a day for 19 days was evaluated. In this study, there was a significant difference in
the recognition of happiness between the test group and the control group.

The evaluation of the effect on risk management

One RCT evaluated the impact of tryptophan intake on risk management in healthy
individuals. In this study published in 2009 (Murphy et al. 2009), 1 g tryptophan was
taken three times a day for 14 days, and the risk management was significantly different
between the intake and control groups. In “Gains-only gamble,” subjects were asked to
select either A or B: (A) 50% chance of getting 60 points or nothing, (B) always get 30
points. In “Loses-only gamble,” subjects were asked to select either A or B: (A) 50%
chance of losing 60 points or nothing, (B) always lose 30 points. As a result, the trypto-
phan intake group chose to gain a solid gain in Gains-only gamble, and they chose to
avoid a loss even if it took the risk for the bigger loss in Loses-only gamble. Also, the
time for making the decision was significantly longer in the tryptophan intake group.
 JOURNAL OF DIETARY SUPPLEMENTS 13

The evaluation of the effect on aggression

In trials evaluating the impact of tryptophan intake on aggression, there was no signifi-
cant difference between the interventions (Murphy et al. 2006; Wingrove et al. 1999).

Discussion
To our knowledge, this study is the first systematic review that includes a qualitative
assessment of mood improvement by TRP. This systematic review includes 11 RCTs
that examined the effects of TRP intake on positive and negative mood in healthy adult
populations. We have targeted healthy adult men and women in this review. This is
because people who suffer from mild depressive symptoms, whose symptoms are not
severe enough to go to the hospital, should improve their depressive symptoms with
food intake rather than medicine. There were four studies each that showed significant
differences in the effect of negative and positive emotions between the tryptophan
intake group and the control group. This suggests that TRP intake may be an effective
approach to decrease negative mood and increase positive mood in healthy individuals.
In 4 RCTs, TRP intake significantly increased positive emotions such as happiness.
One of them reported pre-sleep well-being when taking low-dose supplements including
TRP in 60 to 90 min before bed (Mohajeri et al. 2015). Gibson et al. (2014) reported
that low doses of TRP-rich protein hydrolyzate had a beneficial effect on mental func-
tion. Specifically, it could promote a healthy mood and show resistance to bad mood
and depressing episodes in healthy subjects. Attenburrow et al. (2003) also reported the
effectiveness of TRP on emotion processing. A study by Murphy et al. found a signifi-
cant difference in the perception of happiness only in women (Murphy et al. 2006).
Thus, the intake of TRP could increase the availability of TRP for brain serotonin syn-
thesis and may bring neuropsychologically favorable changes in the brain. Qualities of
these four studies were high according to the Jadad scale.
Four studies showed the effect of TRP intake on resistance to negative emotions.
Since each study showed a significant difference between the TRP intake group and the
control group, the use of TRP as a coping with negative emotions can be expected. In a
study published in 2003 (Attenburrow et al. 2003), there was a significant difference in
perceptions of fear after TRP consumption. In a study by Murphy et al. (2006), only
females showed a significant difference in disgust. In a study ingesting TRP-rich egg
white protein hydrolyzate, there was a significant difference in the perception of fearful,
sad, and disgust facial expressions (Gibson et al. 2014). In a study by Mohajeri et al.
(2015), there was a significant difference in the recognition of angry facial expressions
between the TRP intake group and the control group. Qualities of these studies were
high according to the Jadad scale.
There was a significant change in risk management with TRP intake (Murphy et al.
2009). All the choices have the same expected points, but the TRP intake group tended
to avoid certainties for gain and avoid risks for loss. The TRP intake group significantly
took longer to make decisions. Tha quality of this study was high according to the
Jadad scale.
Aggression was not significantly affected by TRP intake (Wingrove et al. 1999;
Murphy et al. 2006).
14 A. M. KIKUCHI ET AL.

In studies with significant differences in mood effectiveness, the daily intake of tryp-
tophan was 1.8 g/day (Attenburrow et al. 2003), 3 g/day (Murphy et al. 2006), about 2 g/
day (Gibson et al. 2014), and 0.14 g/day (Mohajeri et al. 2015). In the study of the
0.14 g/day TRP intervention, the effective dose seemed less than 0.14 g/day because of
the placebo control (casein) also include TRP. This dose of 0.14 g/day is extremely small
compared with other studies and it seems like a statistical outlier. Considering that this
study is a risk-biased study of conflicts of interest, it should be considered carefully
whether to incorporate the results of this study. Considering the knowledge gained
from these studies and their credibility, it is recommended to take 1–3 g of tryptophan
daily in addition to the usual meal. These studies included ingestion periods from the
short-term to the long-term. To some extent, tryptophan availability to the brain can be
enhanced by ingestion of carbohydrates (Fernstrom and Wurtman 1972) and trypto-
phan (Fadda 2000) and reduced by ingestion of branched amino acids (Choi et al.
2013). Carbohydrate ingestion does not change the levels of circulating tryptophan, but
it does decrease concentrations of competing amino acids (CAAs) through activation of
insulin, which increases the relative availability of tryptophan for transport.
When taking protein regularly, the average daily amount of protein for adults is
around 50 g, and the intake of tryptophan is around 10 g (Ministry of Health, Labour
and Welfare). It should also be noted that since the subjects under the test are also tak-
ing tryptophan from the normal diet, the intake of tryptophan is in addition to the nor-
mal diet.
In some studies, the control group also consumed diets and supplements containing
tryptophan such as casein. This lack of control of the amount of tryptophan taken may
have affected the outcome. The following experiments are considered to be the most
appropriate: the test group: only tryptophan, and the control group: placebo. Therefore,
the experiments of Wingrove (1999), Murphy (2009), Aan Het Rot (2006), and Murphy
(2006) are considered to be appropriate.
Furthermore, branched amino acids (Val, Leu, Ile) have been shown to have an effect
of reducing the concentration of tryptophan in the brain (Choi et al. 2013). The control
group of the Markus et al. (2008-Netherlands) study consumed neutral amino acids
(including branched amino acids), which may act to reduce the concentration of trypto-
phan in the brain. Although the direction of regulation of tryptophan concentration in
the brain is the same, attention should be paid to the fact that the results came from
the mixture of the two different interventions.

Study limitations
The result may have been affected by several limitations. First of all, the number of tri-
als included and the number of subjects in each study was small, and there was no
homogeneity regarding the group of subjects compared in the 11 RCTs. This may be
due to a variety of reasons, including strict selection criteria, willingness to participate,
and RCT study design. In addition, the risk of bias related to the concealment of assign-
ments was unclear in 10 studies, which is a general trend in any clinical study.
According to the Modified Jadad Score used for the evaluation, out of the 11 studies
reviewed, 10 articles had a score of four or higher and were of high quality. The
remaining one had a score of three and was of slightly lower quality. Although blinding
 JOURNAL OF DIETARY SUPPLEMENTS 15

subjects is an important issue in RCT design, the method of blinding has been
described in only about half of the studies.
The most important factor limiting the strength of the basis for conclusions was
the wide variability of intervention methods. The reviewed RCTs included various
kinds of intakes, intake periods, and intake frequencies. There was one study in which
the age range was unknown, although subject demographics did not differ significantly
from the other studies. Also, Murphy et al. reported that the effects of TRP on emo-
tional processing were only seen in the female volunteers (Murphy et al. 2006). They
discussed that it is consistent with the fact that the mood-lowering effect of acute
TRP depletion (ATD) is greater in women, compared to men, and it is because ATD
has a greater biochemical effect in women than in men, as demonstrated using radio-
labelled alpha-methyl-L-tryptophan with positron emission tomography (Nishizawa
et al. 1997; Murphy et al. 2006). All these factors complicate the interpretation of
the results.
The possibility of publication bias cannot be denied. Because there were not enough
studies to determine the publication bias for each outcome, analysis of publication bias
using funnel plots has not been conducted.
Meta-analysis was not possible due to the small number of papers per outcome.

Implications for clinical practice and research

Since the 1970s, it has been discovered that TRP affects the rate of formation of the
neurotransmitter serotonin at nerve terminals and is also the precursor of melatonin.
Therefore, the idea to reduce mental stress and improve sleep by taking TRP has been
studied. Jenkins et al. have used the tryptophan depletion model to investigate the idea
that low serotonin synthesis is associated with depressed mood (Jenkins et al. 2016).
Moreover, influence of gut microbiota on behavior is becoming increasingly evident, as
is the extension to tryptophan and serotonin, producing a possibility that alterations in
the gut may be important in the pathophysiology of human central nervous system dis-
orders (Jenkins et al. 2016). There are various reports on the relationship between tryp-
tophan and behavior, but there is still no consensus on the use of TRP as a supplement
for behavior. In order to confirm the effectiveness of TRP intake on anxiety and mood,
strictly designed large-scale RCTs are required. Furthermore, the protocol for TRP
intake, intake period, intake frequency, and intake method should be established
through research.
Our findings are useful in considering the effects of TRP intake on anxiety and
mood. Over-the-counter (OTC) medications/supplements place patients at risk for drug
interactions. In particular, the combination of tryptophan or 5-HTP with monoamine
oxidase inhibitors (MAOIs) can produce serious interactions, including serotonin syn-
drome/neurotoxicity. There have been a number of neurobiological studies on the
effects of TRP on animals and the human body. More detailed studies on TRP pharma-
cokinetics, particularly the mechanism of brain translocation, and the conversion and
metabolism of TRP to serotonin and methionine may help to make effective use
of TRP.
16 A. M. KIKUCHI ET AL.

Conclusions
In modern society where mental disorders due to stress are increasing, ingredients that
improve mood are attracting much attention. This systematic review found that TRP
intake is useful on moods and feelings and that the several reports showed TRP
represses negative feelings and enhances positive feelings. This shows that TRP can be
expected to deal with emotions. In the studies that showed a significant effect on mood,
daily TRP intake ranged from 0.14 to 3 g. In the future, in order to accurately estimate
the amount of TRP required to obtain the preferable effect, further studies need to be
conducted with appropriate settings of the intake period, intake frequency, and
intake method.

Author contributions
AMK and YI designed the study. AMK and AT wrote the manuscript and conducted
the statistical analysis. Both authors contributed to and approved the final manuscript.

Declaration of interest
The authors declare no conflicts of interest associated with this manuscript.

About the authors

Dr. Asako M. Kikuchi was born in Japan in 1982. In 2005-2012, she was in the
Graduate School of Pharmaceutical Science at the University of Tokyo. In 2012 to 2015,
she was a project researcher at the University of Tokyo. In 2015, she received her Ph.D.
(Pharmaceutical Sciences, Pharmacy) from the University of Tokyo. Now she works as
an academic officer at LLC Okutoeru from 2018. Her research interests are pharmaceut-
ical sciences, nutrition, and mental health.
Dr. Aya Tanabe was born in Japan in 1980. In 2005, she became a dentist after passing
the National Examination for Dentists, and in 2008, she received her Ph.D. and Doctor
of Dental Surgery from Tokyo Dental College in Chiba. She was an assistant professor
at Tokyo Dental College from 2008 to 2011, and an assistant to director at Ministry of
Health, Labour and Welfare from 2011 to 2017. Now she works as an academic officer
at LLC Okutoeru from 2018. Her research interests are functional foods and den-
tal medicine.
Dr. Yoshihiro Iwahori was born in Japan in 1972. In 2001, he became a pharmacist
after passing the National Examination for Pharmacist, and he received his Ph.D. in
Pharmacy from the Meiji Pharmaceutical University. He has worked as a researcher at
Keio University School of Medicine from 2002 to 2003, and as a senior partner at LLC
Okutoeru in 2006. Now he has worked as a visiting professor at Nihon Pharmaceutical
University from 2018. His research interests are traditional medicine, herbal medicine,
natural medicine and functional foods.
 JOURNAL OF DIETARY SUPPLEMENTS 17

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