Effect of 528 HZ Music On The Endocrine System and
Effect of 528 HZ Music On The Endocrine System and
Effect of 528 HZ Music On The Endocrine System and
http://www.scirp.org/journal/health
ISSN Online: 1949-5005
ISSN Print: 1949-4998
Department of Hospital Administration, Graduate School of Medicine, Juntendo University, Tokyo, Japan
1
Center for Advanced Kampo Medicine and Clinical Research, Graduate School of Medicine, Juntendo University, Tokyo, Japan
2
Keywords
Music, Stress, Salivary Assay, Autonomic Nervous System
1. Introduction
It is well known that listening to music reduces stress. Research has investigated
creases early after stress is relieved [7]. Furthermore, others have examined the
effect on both cortisol and chromogranin A of listening to music [8] [9]. Specif-
ically, they report that salivary cortisol and chromogranin A decrease signifi-
cantly before and after listening to music [8]. In contrast, other researchers ob-
serve that, although salivary chromogranin A decreases significantly, there is no
significant difference in cortisol levels [9].
In addition to the above indicators, we also focused on oxytocin. The effect of
listening to music on oxytocin has recently attracted attention, with a previous
study indicating that salivary oxytocin increases when participants listen to
slow-tempo music compared to fast-tempo music [10].
As an additional objective index, we measured the activity of the autonomic
nervous system. Finally, we also administered the Japanese short version of the
Profile of Mood State, 2nd edition (POMS 2) to participants as a subjective index
of stress. Both of these measures have also been used in many previous studies
[11] [12]. We consider these measures to support the data provided by the
above-mentioned salivary biomarkers of stress marker.
The 528 Hz music that we used in this study was soothing piano music. Some
researchers report that, depending on the mental state of the listener, music pre-
ferences vary [13]. However, they also find that piano music is appreciated re-
gardless of the listener’s circumstances. Thus, we examined the stress-mitigating
effect of 528 Hz music from various perspectives. This study offers a new piece
of evidence for use in music therapy.
2. Methods
2.1. Participants
Participants were nine healthy adults, one man and eight women, aged 26 - 37
years (mean age: 31.1; SD = 1.35). We recruited participants as volunteers re-
gardless their previous musical education. Everyone participated with full un-
derstanding of the study and gave their informed consent. They were asked to
abstain from caffeine and smoking for one hour before starting the experiment.
They did not apprise of which music they were listening to. This study was ap-
proved by the Ethics Committee of Shiba Palace Clinic, Tokyo, Japan (Protocol #
2018040001).
2.2. Procedure
Figure 1 shows an outline of the experimental procedure. The study was carried
out in a conference room, where the room temperature was 25.0˚C - 25.5˚C and
humidity was 67% - 76%. In consideration of circadian variation in cortisol and
chromogranin A [14] [15], we started the experiment at 2 PM for each condi-
tion. Participants listened to the music from a seated position beside a speaker.
The music was soothing piano music, and the only difference between the two
musical conditions was in frequency, which was either 528 Hz or 440 Hz. Each
participant took part in each of the conditions on separate days.
2.3. Measures
2.3.1. Salivary Assay
We collected salivary samples at five time points (immediately before listening to
music, immediately after listening to music, 15 minutes later, 30 minutes later,
and 45 minutes later) using the Saliva Collection Aid (Salimetrics LLC, USA).
We measured cortisol, chromogranin A, and oxytocin as salivary biomarkers of
stress. These were measured using the Salivary Cortisol Enzyme-Linked Immu-
nosorbent Assay (ELISA) Kit (Salimetrics LLC, USA), Human Chromogranin A
ELISA Kit (Yanaihara Institute Inc., Japan), and Oxytocin ELISA Kit (Arbor
Assays, USA), respectively.
used in a variety of fields such as clinical, workplace, and school. For example, it
has been applied to the course of treatment of mental disorders, mental changes
of people with physical disorders, screening in the workplace, and exercise or
relaxation effect.
The POMS 2 evaluates seven mood scales of anger-hostility (AH), confu-
sion-bewilderment (CB), depression-dejection (DD), fatigue-inertia (FI), ten-
sion-anxiety (TA), vigor-activity (VA), friendliness (F) at the same time. TMD is
a general indicator such as mood disorders, psychological distress, and subjec-
tive well-being. The TMD score is calculated by the sum of the elementary scores
of AH, CB, DD, FI, TA, and VA. (Since VA is weighted of negative, it is sub-
tracted from the sum of the other five). Therefore, the score is a comprehensive
evaluation of expression of negative mood.
The POMS 2 consists of 35 items questions and a respondent answer each of
question with the evaluation scale of POMS 2 (0 = not at all, 1 = slightly, 2 =
modestly, 3 = considerably, 4 = very). Although this evaluation scale is constant,
since the number of items differs depending on the scale, the degree indicated by
each scale score is not constant. In short, even though the absolute value of the
score is the same, the meaning of the score differs between scales. Since it is im-
possible to properly compare the scales at this score, the score is converted to a
standardized score, that is, a T score. The T score is a normalized standard of
assessment (average value is 50 and the standard deviation is 10) so that the
same value has equivalent meaning. The measurer interprets the score by con-
verting the graded evaluation scale into the T score.
Regarding the TMD score and negative mood state (AH, CB, DD, FI, and
TA), the higher the T score, the stronger the emotion concerning negative emo-
tion or mood disorder. Regarding the positive emotional state (VA, F), the high-
er the T score, the more positive emotion means [16].
3. Results
3.1. Salivary Assay
3.1.1. Cortisol
In consideration of the response time lag of cortisol [17] [18], we took the mean
level of salivary cortisol measured immediately after music exposure as
representing the level immediately before music exposure, and so on; therefore,
data were only available until 30 minutes after music exposure, rather than 45
(Figure 2). After exposure to 528 Hz music, mean levels of salivary cortisol de-
creased with the passage of time and were significantly reduced 30 minutes later
(0.43 ± 0.04 → 0.25 ± 0.02, p < 0.011). In contrast, after listening to 440 Hz mu-
sic, mean levels of cortisol slightly decreased after 30 minutes, but there was no
significant difference between the time points.
3.1.2. Chromogranin A
There was no significant difference in chromogranin A levels following exposure
to either 528 Hz music or 440 Hz music (Figure 3). However, in the 528 Hz mu-
sic condition, mean levels of chromogranin A showed a tendency to fall until 30
minutes after music exposure. In contrast, in the 440 Hz music condition, mean
levels of chromogranin A showed a tendency to increase until 30 minutes later.
3.1.3. Oxytocin
Figure 4 illustrates the effects of music on oxytocin. Mean levels of oxytocin in-
creased significantly immediately after listening to 528 Hz music (37.57 ± 1.53 →
73.58 ± 5.04, p < 0.038). After listening to 440 Hz music, mean levels of oxytocin
also increased, but the difference was not significant.
3.3. POMS 2
After exposure to 528 Hz music, all negative mood scores decreased. In particu-
lar, tension-anxiety (48.44 ± 2.28 → 43.67 ± 2.45, p < 0.0091) and Total Mood
Disturbance (45.56 ± 1.99 → 42.00 ± 2.00, p < 0.0487) decreased significantly.
Additionally, all positive mood scores tended to increase. In contrast, after ex-
posure to 440 Hz music, almost all scores showed a similar tendency to change
in the same direction as observed in the 528 Hz music condition, but there was
no significant difference for any score. Furthermore, anger-hostility scores (in-
dicating negative mood) slightly increased, and friendliness scores (indicating
positive mood) decreased (Table 1).
4. Discussion
The purpose of this study was to examine the effect of 528 Hz music on the en-
docrine system and the autonomic nervous system using objective and subjective
measures. Based on salivary biomarkers, electrocardiogram, and a mood state
questionnaire, we found that stress levels were reduced following five minutes’
exposure to 528 Hz music, whereas this was not the case for 440 Hz music.
We found that mean levels of cortisol decreased and mean levels of oxytocin
increased after listening to 528 Hz music. In general, cortisol increases via the
Table 1. Effects of music on mood as assessed by subscales of the Profile of Mood State,
Second Edition.
Total Mood Disturbance 45.6 (1.99) 42.0 (2.00)* 43.6 (1.99) 41.9 (2.48)
*p < 0.05; **p < 0.01 in pre-exposure vs post-exposure comparison using the Wilcoxon signed-rank test.
Figure 2. Effects of music on mean levels of salivary cortisol. Error bars represent
standard errors. Time points are as indicated in Figure 1 (Mean ± SE) *p < 0.05.
Figure 5. Effects of music on the autonomic nervous system, as measured by the ratio of
low frequency to high frequency (LF/HF) and coefficient of variation of R-R intervals
(CVRR). Time points are as indicated in Figure 1 (Mean ± SE) *p < 0.05.
5. Conclusion
We found that music of different frequencies had different effects on the endo-
crine system, especially oxytocin and cortisol. Our findings suggest that 528 Hz
music reduces stress even if participants listen to the music for only a short time.
Acknowledgements
We thank the participants for volunteering and the Institute of Man and Science
Inc. for analyzing the electrocardiogram data.
Conflicts of Interest
The authors declare no conflicts of interest regarding the publication of this pa-
per.
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