Listening to specific music frequencies is associated with distinct biological changes in students experiencing exam-related stress. Recent research published in brain and behavior The 528 Hertz frequency tends to promote proteins associated with brain health, while the 432 Hertz frequency suggests that it triggers a cellular stress response. Despite these biological changes, short listening sessions do not immediately change cognitive performance on attentional tasks.
University students often face intense pressure and anxiety during exam periods. This stress affects both your body and your mental focus. To cope with this pressure, many students turn to simple ways to relax, such as listening to music. Research shows that music can influence the nervous system, relieve stress, and support overall brain health.
“The reality is that university students experience a significant increase in stress during exam periods, so we were interested in whether a simple, low-cost intervention of listening to music could be tracked using non-invasive salivary biomarkers,” said Ummü Gülçen Bozok, assistant professor of physiology at Ankara Medipol University in Turkey.
Bozok and her colleagues wanted to investigate whether certain sound frequencies had distinct biological effects. Sound is measured in Hertz. Hertz refers to the number of sound waves per second. Standard music on the radio is usually tuned to 440 hertz. Alternative tunings such as 432 Hertz and 528 Hertz are popular for relaxation tracks and meditation playlists.
“We decided to compare two musical tunings that are commonly discussed as experimental auditory stimuli, primarily to see if the different conditions are associated with measurable differences in stress- and plasticity-related markers,” Bozok said.
To measure these effects, the authors focused on three specific proteins found in human saliva. The first is brain-derived neurotrophic factor. This protein acts like fertilizer for the brain. It helps brain cells grow, survive and form new connections, supporting learning and memory.
The second protein is the cyclic AMP response element binding protein. This molecule helps turn on the genes needed to form long-term memory and absorb new information. Both of these proteins are associated with brain plasticity, the brain’s ability to adapt, heal, and change over time.
The third protein is glucose-regulated protein 78. This particular molecule acts as an indicator of cellular stress, especially within the cell’s protein production factories. When cells are overworked or damaged, levels of this protein increase to repair misfolded proteins and protect the cells from death.
The researchers designed a study to observe how short-term exposure to different musical tunings affected these three specific proteins. They wanted to assess the biological changes occurring beneath the surface during stressful school years. They also aimed to see if the biological changes would lead to immediate improvements in concentration and attention right before a test.
The study involved 162 healthy college students between the ages of 18 and 25. The authors recruited these volunteers from universities in Turkey and conducted the experiment during official exams to capture their natural stress. Participants were randomly divided into three equal groups of 54 students.
The first group served as a control condition and sat quietly without music. The second group listened to instrumental music tuned to a frequency of 528 hertz. The third group listened to instrumental music tuned to a frequency of 432 hertz.
For the experimental group, participants listened to their assigned music on headphones for exactly 20 minutes. These sessions took place approximately one hour before students were scheduled to take their university exams. All participants were seated in the same environmental conditions so that no external visual or auditory factors influenced the results.
Immediately after the 20-minute session, researchers collected saliva samples from each participant. They used a passive drooling technique to collect the liquid. This method allowed us to preserve the samples and measure the accurate concentrations of the three target proteins.
“Saliva is attractive because it is easy to collect and well tolerated,” Bozok told SciPost. Using saliva, you can easily and painlessly check biological markers without having to draw blood.
After providing a saliva sample, each student completed a psychological test called the Stroop Color Word Test. This psychological task assesses selective attention, mental flexibility, and the ability to process conflicting information. During testing, the clinician presented participants with color words printed in non-matching ink colors.
For example, a student might see the word “blue” written in red ink. Participants first read the plain text words out loud as quickly as possible. They were then asked to look at the unmatched words and say the name of the ink color, rather than read the words themselves. The researchers recorded the time it took to complete each part of the test to assess students’ cognitive processing speed.
Biological analysis revealed clear differences between the three groups of students. Students who listened to 528 Hz music maintained high levels of brain-derived neurotrophic factor, comparable to the control group. They also showed the highest levels of memory-building proteins compared to all other participants.
Furthermore, the 528 Hz group showed lower levels of cellular stress markers than the control and 432 Hz groups. This particular pattern suggests that the 528 Hertz frequency promotes biological conditions associated with brain growth. It also appears to reduce the overall stress burden on cells.
Participants exposed to 432 Hz music had different biological responses. Their saliva samples contained significantly lower levels of brain growth proteins compared to other groups. These students also had the lowest levels of proteins that build memory.
The 432 hertz group had significantly higher levels of cellular stress markers, instead promoting brain growth markers. Their stress protein levels were higher than both the control and 528 Hertz groups. The authors suggest that this increase may indicate a cellular defense response.
The body may be trying to manage environmental stress by increasing production of proteins that repair damaged cells. It acts as an adaptive mechanism to maintain cellular balance during demanding academic events. This suggests that 432 Hz music may trigger biological defense processes.
Despite these different biological responses, mental ability tests showed no significant differences between the three groups. Students in the control group, 528 Hertz group, and 432 Hertz group all took approximately the same amount of time to complete the reading and color naming tasks.
“The most interesting aspect for us was the dissociation. We saw differences at the level of salivary biomarkers, but no corresponding differences in the cognitive tasks,” Bozock explained. “This served as a reminder that acute biochemical signals do not automatically imply behavioral or cognitive effects, and as a result, we were cautious about how we interpreted our findings.”
Biological changes occurring at the cellular level do not immediately lead to faster processing in the brain. Brain plasticity and memory formation often require sustained or repetitive stimulation to produce significant behavioral changes. “The honest conclusion is modest. In our sample, different music conditions were associated with somewhat different patterns of salivary biomarkers, but these differences were not translated into measurable differences in cognitive (Stroop test) performance,” Bozock said.
Readers may think that listening to 432 Hertz music is inherently harmful due to the elevated stress markers observed in this study. This increase in stress proteins does not necessarily mean that the cells are damaged. It may simply reflect an adaptive process in which the body prepares cells to defend and repair themselves from external pressures.
Similarly, the benefits of 528 Hertz adjustment should not be overstated. “We caution readers not to conclude that certain frequencies ‘boost the brain,’ reduce stress, or cure anything. Our data do not support such claims,” Bozock warned. “We have intentionally avoided framing any particular tuning as a ‘healing frequency,’ and we encourage our readers to maintain the same healthy skepticism that we have tried to apply to ourselves.”
“If there is a practical message, it’s the familiar one that music is a reasonable and accessible way to relax during stressful times. The specific biology we looked at requires more research before anyone can draw firm conclusions,” Bozock added.
This study is characterized by several limitations that provide context for the findings. “This was a small, short-term pilot study in a narrow group (healthy college students aged 18 to 25) with only one 20-minute exposure, so the results are not generalizable to other populations or long-term use,” Bozok told PsyPost.
Another limitation includes the use of saliva for measuring brain-related proteins. “Importantly, the levels of these proteins in saliva should not be read as a direct window into the brain. Any association between what we measure in saliva and what is happening in the central nervous system is hypothetical and not something we have demonstrated,” Bozock cautioned.
“The markers we examined are not yet established salivary measurements, so absolute values must be interpreted with caution and confirmed with validated methods,” Bozok noted. The scientists also did not take into account the students’ personal musical tastes. “We also did not control for participants’ personal musical tastes or backgrounds. For all these reasons, we explicitly framed the work as hypothesis-generating.”
Future studies should follow participants over time to see if long-term listening habits lead to significant changes in mental performance. “We want to see whether any of these signals persist under more demanding conditions, including larger and more diverse samples, longer or repeated exposures, validated assays, and ideally complementary measures such as blood markers or neuroimaging,” Bozock said.
By studying diverse age groups and using advanced tools, researchers can build a more comprehensive understanding of how specific sound frequencies shape human biology. “The aim is not to confirm these preliminary associations, but to test whether they are real and meaningful,” Bozok concluded, noting that he hoped the study would be seen as a small preliminary step rather than a definitive finding.
The study, “Acute music frequency exposure modulates salivary stress and neurotrophic markers in young adults: a randomized controlled trial,” was authored by Ümmü Gülşen Bozok, Gülbahar Böyük Özcan, Bülent Bayraktar, and Doğukan Özen.
