Enhancing Biochemistry Education Through Movement-Based Learning: Investigating Oxygen Delivery Using Personal Health Data

Abstract

Students in undergraduate biochemistry courses often struggle to connect molecular mechanisms with dynamic physiological processes. To bridge these ideas, a novel laboratory module titled Cardio Chemistry: The Human Element was implemented to investigate oxygen delivery through the use of real-time personal health data. Using wearable fitness trackers and metabolic monitoring, students collected individual data on oxygen saturation, oxygen uptake, carbon dioxide production, and respiratory exchange ratio during a guided aerobic fitness assessment. Designed as a mixed-methods, within-subject case study, the intervention was evaluated through pre/postlab assessments, exam scores, reflective responses, and surveys. Students who completed the activity demonstrated significant gains in conceptual understanding, with postlab Bohr effect comprehension reaching 100% alongside significantly higher final exam scores compared to nonparticipants (p < 0.05). Self-reported familiarity with physiological metrics increased across all variables (p < 0.001), and reflections indicated enhanced engagement and relevance. These findings suggest that integrating physiological data into biochemistry instruction improves mechanistic understanding, fosters interdisciplinary thinking, and supports knowledge transfer to clinical and real-world contexts.

Introduction

The effective teaching of complex biochemical principles to undergraduate students in introductory courses presents ongoing pedagogical challenges. Found across the chemistry curricula, these courses often serve a broad population of students with diverse academic backgrounds and career goals. Biochemistry is one of the five foundation courses in the ACS Guidelines for Bachelor’s Degree Programs and a common requisite for those preparing for careers in medicine, dentistry, or veterinary science. − To prepare this diverse range of students, Introductory Biochemistry often surveys biomolecules and biochemical principles, and is frequently paired with a laboratory course designed to reinforce content and introduce foundational experimental techniques. Despite supports, students frequently have difficulty connecting molecular-level content to dynamic, whole-body biochemical processes. This disconnect underscores the need for instructional approaches that contextualize biochemical concepts in relevant, applied settings.

Biochemical and physiological concepts may remain abstract for even the best-prepared students unless reinforced through direct, experiential learning. Traditional instruction often relies on static models or textbook diagrams, offering limited opportunities for real-time, meaningful exploration and application. One concept that exemplifies this pedagogical disconnect is the Bohr effect, an essential yet conceptually challenging topic linking protein structure to physiological adaptation. The Bohr effect describes how increasing carbon dioxide concentration or decreasing pH reduces hemoglobin’s affinity for oxygen, promoting oxygen release in metabolically active tissues. Experimentally, this relationship is observed as a rightward shift in the oxygen–hemoglobin dissociation curve, which can be quantified by measuring oxygen saturation or partial pressure of oxygen under varying physiological conditions. In biochemical curriculum, the Bohr effect builds upon acid–base chemistry to explain how changes in physiological parameters alter the oxygen-binding affinity of hemoglobin, the oxygen transport protein in blood, to facilitate oxygen delivery in metabolically active tissues. Changes to the structure of hemoglobin are essential for its ability to pick up oxygen in the lungs and deliver oxygen to tissues in need, such as muscles during exercise, inflammation, or during acidosis. As the Bohr effect integrates concepts including pH, molecular structure, and protein action, the effect on hemoglobin serves as a foundational case study for understanding how chemical principles manifest in biological systems. − In addition, hemoglobin provides a clear example of how local biochemical environments regulate protein function via ligand interactions and allosteric modulation, a recurring theme in biochemistry. Despite its physiological relevance, students often perceive the Bohr effect as a static curve rather than a dynamic regulatory mechanism, failing to understand clinical or physiological implications. As a result, students struggle to apply the concept when analyzing real-world scenarios such as respiratory distress, exercise tolerance, or acid–base imbalance, all of which rely on the principles of the Bohr effect.

The disconnect is further compounded by low engagement and limited personal relevance in traditional curricula, negatively impacting students’ intrinsic motivation and learning outcomes in STEM fields. Recent trends in science education emphasize the integration of active learning, inquiry-based experiments, and use of personal data to enhance student understanding and engagement. , In the context of biochemistry education, the use of wearable or real-time physiological sensors has created new opportunities to incorporate approaches from physiology courses and bridge theoretical learning with authentic biological feedback – visualizing biochemical regulation in their own bodies. Prior studies have demonstrated that student-centered activities involving personal health data can significantly improve comprehension of cardiovascular fitness and systems biology but are primarily focused on applied physiology or exercise science rather than foundational biochemical principles. ,

To create a more tangible and immersive learning experience, a guided-inquiry laboratory module titled Cardio Chemistry: The Human Element was designed, executed, and evaluated alongside undergraduate researchers. The intervention was designed to actively engage students generating their own health data while investigating regulation of oxygen delivery in real time. Students observed and analyzed their own physiological responses, including oxygen saturation (SpO₂), heart rate (HR), oxygen uptake (VO₂), carbon dioxide output (VCO₂), and respiratory exchange ratio (RER) using noninvasive sensors during participation in light to moderate exercise. In exploring these elements, students were able to connect physiological data to biochemical processes to enhance comprehension, engagement, and retention.

The study was embedded in BCH 301L: Introduction to Biochemistry Lab at Canisius University, a Jesuit primarily undergraduate institution (PUI) in Buffalo, New York. This course lies within the Department of Chemistry and Biochemistry, enrolling upper-level undergraduates across multiple majors. Small class sizes provided an environment particularly well-suited to adapting a new, interdisciplinary learning approach. As seen in Figure , the activity introduces variables atypical to biochemistry, drawing on the closely related discipline of physiology. Both classes are common to STEM curricula: in the preceding ten years, 30.5% (47 of 154) of undergraduate students that completed biochemistry had credit for one or more physiology courses during their time at the university. By tracking data during controlled physical exertion and recovery, students are provided a personalized lens through which to explore hemoglobin function and oxygen delivery. By newly integrating real-time personal health data into the biochemistry lab, the intervention reinforces core biochemical principles while increasing student engagement and promoting health literacy. For students pursuing healthcare careers, the Bohr effect underlies essential clinical phenomena, from why oxygen therapy works in hypoxia to how blood gas readings are interpreted in respiratory illness. By mastering this concept through direct experience, students gain not only biochemical understanding but also diagnostic intuition. The cross-disciplinary approach introduces a rarely emphasized yet pedagogically rich connection between biochemistry and human physiology, empowering students to explore protein–ligand interactions within the context of their own metabolic responses.

1.

Conceptual framework for integrating personal physiological data into biochemistry instruction. The diagram outlines the interdisciplinary approach of the activity, using real-time physiological measurements to support student understanding of oxygen transport through experiential learning.

Grounded in experiential learning and constructivist theory, the activity encourages deeper conceptual understanding through direct data interaction, hypothesis testing, and reflection. The integration of wearable sensors and health metrics engages students as both scientists and subjects, increasing cognitive and emotional investment in the learning process. Using pre- and postlaboratory assessments alongside a comprehensive student survey, the educational impact of the lab on students’ conceptual understanding, engagement, and perceived relevance of biochemistry to their own lives was assessed. Postlaboratory reflections and exam scores relative to student peers who did not participate in the activity provide further insight into the efficacy of integrating the activity into the laboratory curriculum. The included laboratory activity offers an easily adaptable, evidence-based model for integrating physiological data into biochemistry instruction to advance interdisciplinary understanding and translational thinking in the undergraduate science curriculum. This study explores whether incorporating a physiology-based experiment into an introductory biochemistry laboratory can measurably enhance student comprehension of oxygen transport mechanisms and foster transferable skills in mechanistic reasoning, clinical relevance, and health literacy.

Results and Discussion

Pedagogical Motivation and Framework

The motivation for this study arose from the desire to make complex biochemical concepts more accessible and personally relevant to undergraduate students. The Bohr effect is particularly well suited to an active, data-driven lab as its outcomes are readily observable in a noninvasive fitness test. Unlike many biochemical mechanisms, the Bohr effect produces measurable changes in physiological parameters, including oxygen saturation and carbon dioxide output, within minutes. This allows students to directly connect molecular concepts to their own bodily responses, reinforcing learning through lived experience. Thus, by leveraging wearable fitness trackers (WFTs) and real-time personal health data, this work sought to provide students with an immersive and tangible experience of actively changing biological parameters to witness the Bohr effect in action. The activity aimed to promote deeper conceptual understanding through first-person data interaction, hypothesis testing, and reflection. The activity structure also mirrors best practices in experiential and guided-inquiry learning, which have been shown to improve STEM student performance and motivation. ,

Table presents a summary of participating student demographics, including major and academic standing. The cohort represented a diversity of life science disciplines and prehealth interests. All procedures were reviewed and approved by the Canisius University Institutional Review Board (IRB). Students participating in the study signed an informed consent form which outlined the purpose, procedures, potential risks, and benefits of participation, including the voluntary nature of involvement and the availability of a nonexercise alternative to fulfill course requirements. The full consent form is available in Supporting Information. Though the present analysis includes a single-semester cohort, findings consistently demonstrated learning gains and positive engagement, providing a strong foundation for longitudinal validation in subsequent semesters.

1. Activity Participant Demographics (n = 14) .

variable	count
enrollment
lecture + lab	11
lecture only	3
academic major
biochemistry	7
chemistry	4
biology	1
animal behavior, ecology, conservation	2
academic standing
junior	10
senior	4

^aData includes enrollment status, academic major, and academic standing for students participating in the activity.

As part of the activity, students were first asked to complete a prelab quiz using only their existing knowledge. After completing the prelab assessment, students participated in a submaximal aerobic fitness test to monitor their cardiorespiratory responses to exercise. The fitness test was administered by exercising on a treadmill while connected to a heart rate monitor, metabolic cart, pulse oximeter, and optional WFT. During the test, real-time data were collected on VO₂, VCO₂, RER, METs, HR, and SpO₂ at 20 s intervals. During exercise, elevated CO₂ production leads to increased formation of carbonic acid (H₂CO₃) via the reversible hydration reaction catalyzed by carbonic anhydrase, which dissociates into H⁺ and HCO₃ ^–, thereby lowering blood pH. Although direct blood pH was not measured in this lab for ethical and logistical reasons, students inferred acid–base changes from increases in VCO₂ and RER, which together reflect systemic acidification. The fitness test continued until the student reached volitional fatigue or physiological end points indicating maximal effort. Average test time was 11.04 min, allowing a standard fitness test to be performed every 30 min when accounting for setup and cleanup between students. Following the activity students analyzed their data, completed a postlab quiz and written reflection questions, and submitted a survey evaluating their understanding and experience. Full versions of all assessment instruments are available in the Supporting Information. As students exercised, the hands-on intervention and subsequent evaluation allowed students to directly observe an increase in their respired carbon dioxide levels. Students were then asked to relate this observation to predicted changes in acidity and the Bohr effect, stating how changes in CO₂ production and pH influence oxygen delivery. With their data students could observe a shift in gas levels in real time, reinforcing a previously static concept in a dynamic, personalized context, as demonstrated by a marked improvement in assessment scores.

Assessment Outcomes: Knowledge Gain and Conceptual Mastery

The Bohr effect is a central concept in biochemistry describing how pH and CO₂ modulate oxygen affinity through conformational changes in hemoglobin. These interactions drive the shape of the oxygen-hemoglobin dissociation curve and have been mathematically modeled to predict shifts in oxygen delivery across a wide range of environmental and metabolic conditions. , On the postlab quiz, binary questions assessing the effect of pH and pCO₂ on hemoglobin affinity improved from 63.6% (7/11 students prelab) to 100% (11/11 postlab) correct responses. While an improvement was observed, the high scores on these questions indicate that prior lecture coverage of this topic may have been sufficient for comprehension.

In the narrative section, students were asked to describe at least one factor other than pH and pCO₂ that can affect hemoglobin’s affinity for oxygen in the human body. While scores saw a moderate increase following the activity from 72.6 ± 18.4 to 82.6 ± 26.8%, the most common response remained the effect of 2,3-bisphosphoglycerate and the corresponding reduction in hemoglobin’s oxygen affinity. Introducing other variables that can affect affinity, such as temperature, carbon monoxide, or the presence of fetal hemoglobin presents an opportunity for future activity improvement. When asked to “explain the Bohr effect and how it relates to oxygen transport in the human body, including hemoglobin’s propensity to bind or release oxygen,” student scores increased from 67.2 ± 26.9% prelab to 82.5 ± 26.6% postlab. While these findings were not statistically significant (p > 0.05), qualitative improvements were noted in explanations of hemoglobin-oxygen affinity as it relates to pH and carbon dioxide concentration.

To evaluate whether participation in the physiology-based laboratory experience had an impact on content retention and application, examination responses in lecture were compared between students who completed the activity in lab, and those who were enrolled only in the paired lecture and therefore did not complete the activity. Students who participated in the lab intervention scored significantly higher on Bohr effect-related questions, with an average score of 76.4 ± 22.9% (n = 11) compared to 36.7 ± 32.1% among nonparticipants (n = 3) (p < 0.05). Significant differences in exam scores were not observed on questions of similar rigor on topics unrelated to oxygen delivery. While the small sample size limits statistical inference, the nearly 40-point difference in exam scores suggests active engagement with personal physiological data enhances conceptual understanding and long-term retention. These findings align with prior work indicating that contextual and experiential learning can improve assessment outcomes in STEM education. Taken together, results support the pedagogical value of integrating physiological data into biochemical instruction, particularly to bridge molecular processes to whole-body physiological function.

Application to Clinical and Personal Contexts

The Bohr effect serves as a model system for broader biochemical reasoning, as it illustrates how local conditions modulate macromolecular function. This principle recurs throughout biochemistry in contexts such as enzyme regulation, drug binding, and signal transduction, making mastery of the Bohr effect a channel to deeper conceptual understanding. Written postlab reflection questions were designed to prompt students to analyze their own physiological data, compare observed trends to their original hypothesis, and reflect on real-world changes in O₂ delivery to reinforce this conceptual understanding through applied inquiry. Responses demonstrated a strong conceptual grasp of the Bohr effect and changes in oxygen affinity. All 11 students who participated in the activity correctly described how increased carbon dioxide and decreased pH during exercise facilitate oxygen release. Several students explicitly referenced conformational changes in hemoglobin, with many noting the transition between the relaxed and tense states. This level of mechanistic detail, absent in most prelab responses, supports the observed learning gains in biochemical reasoning. One student summarized: “Before this experiment, I lacked a full understanding of how exercise impacted levels of oxygen and carbon dioxide in the body and my hypotheses were mostly speculative. After completing the experiment and analyzing my data, I believe I now have a much clearer understanding about the effects of exercise and hyperventilation on physiological variables like oxygen consumption, carbon dioxide production, and more.”

At the conclusion of the fitness test, students were provided multiple measures of metabolic activity and aerobic fitness, including RER/VO_2,/VCO₂ as a function of time (Figure ), VCO₂ vs VO₂, and HR vs VO₂. Students were thus able to integrate their own real-time physiological data into their understanding of oxygen transport and their body’s response to exercise. Most participants noted that both VO₂ and VCO₂ increased during exercise, and several noted that VCO₂ increased at a greater rate, an observation that reflects elevated metabolic activity and respiratory compensation. When reflecting on their original hypotheses, another student wrote: “I was surprised with my VCO ₂ max being higher than my VO ₂ max, but understand in retrospect.” Observations were frequently tied back to the Bohr effect, as students explained how increased CO₂ production contributes to local acidosis and promotes oxygen unloading in active tissues. Conversely, responses to SpO₂ trends revealed a nuanced learning moment: while many students expected a decline in oxygen saturation during intense activity, the data often showed minimal change. A third student specifically reflected, “I was surprised that my SpO ₂ did not significantly decrease during exercise.” Interpreting oxygen-binding data can be challenging, however rather than dismissing the discrepancy students reflected on possible explanations, including efficient ventilatory response and cardiovascular compensation. Overall, students found the activity easy to complete, with a score of 4.27 ± 0.79 out of 5.

2.

Representative student data showing RER, VO_2, (L/min), and VCO₂ (L/min) over time during exercise. The graph highlights changes in metabolic parameters during the submaximal fitness test, illustrating how VO₂ and VCO₂ increase with exertion and demonstrating real-time physiological shifts relevant to the Bohr effect.

Metric understanding was reinforced by paired-sample t tests, which revealed a significant increase in familiarity scores (Table ). Average ratings for VO₂ rose from 1.64 to 4.09 (p < 0.001); VCO₂ from 1.45 to 4.18 (p < 0.001); RER from 1.54 to 3.64 (p < 0.001); and SpO₂ from 2.00 to 4.36 (p < 0.001). These significant increases in self-reported familiarity reflect a meaningful shift in student confidence and comprehension toward concepts not traditionally in the biochemistry curriculum. When considered with the improvement in student scores, these results indicate that integrating physiological variables into the biochemical curriculum enhances students’ ability to interpret the physiological basis of oxygen unloading in active tissues through the lens of biochemical regulation. This capability to process interdisciplinary content is an essential outcome for those pursuing careers in healthcare and biomedical research.

2. Changes in Self-Reported Familiarity with Physiological Metrics before and after the Lab Activity (n = 11),

metric (1–5 Likert Scale)	pre-lab	post-lab
VO2	1.64 ± 1.03	4.09 ± 0.83
VCO2	1.45 ± 0.69	4.18 ± 0.75
RER	1.54 ± 0.82	3.64 ± 0.92
SpO2	2.00 ± 1.18	4.36 ± 0.67

^aMean Likert-scale ratings indicate significant increases in student familiarity with VO₂, VCO₂, RER, and SpO₂ following participation in the activity.

^bValues reported as mean ± standard deviation.

Physiological mechanisms become especially critical in clinical settings involving hypoxia, respiratory illness, or high-altitude exposure, where efficient oxygen unloading is a compensatory necessity. , In their reflections, students extended their learning to applied settings, with personal relevance aiding knowledge transfer and retention. Several cited specific conditions highlighting the significance of O₂ delivery, while others drew connections to athletic performance and nutritional influence on RER and substrate metabolism. This transfer of knowledge beyond the immediate lab context is a key indicator of meaningful learning. As one student noted: “Respiratory illness, high-altitude exposure, or physical training may impact the Bohr effect by restricting the amount of oxygen entering the lungs and therefore increasing the demand for oxygen in tissues that are likely not receiving enough oxygen.” Another emphasized the personal impact of the experience: “This activity was very helpful in understanding the role of the Bohr effect. I was able to see the Bohr effect occurring in real life. Being able to view the results from the experiment helped prove the Bohr effect to be true. It was interesting to see the many different values that were tested and I never thought I would be able to be a part of an experiment like this.” These reflections suggest successful conceptual transfer from a biochemical process to physiological and pathological conditions, a hallmark of durable learning.

Survey-Based Remarks

Responses to a postactivity student survey (Table ) further supported an increase in student comprehension. All students reported a clearer postactivity understanding of the Bohr effect when asked to rate their understanding of the Bohr effect on a scale of 1–5, with understanding scores improving from 3.00 ± 0.77 to 4.54 ± 0.69 (p < 0.001). Nine out of 11 students (81.8%) found that using personal health data added value to their understanding of the Bohr effect, with ten students (90.9%) stating the lab activity improved their understanding of the relationship between oxygen, carbon dioxide, and pH in blood. All students agreed that using personal health data in the lab activity helped them to connect theoretical knowledge with real-world experiences. Overall, students reported being more likely to recall the concepts of the Bohr effect in future coursework, with a score of 4.45 ± 0.52. When asked how likely they would be to recommend this type of activity for future laboratories, students rated the activity at 4.45 ± 0.69, making it one of the most favorably rated activities in the course. To contextualize these findings, typical end-of-semester evaluations for biochemistry laboratory activities at Canisius University average 3.18 ± 1.21 (on a 5-point Likert scale) for recommendation, suggesting that incorporating personal physiological data meaningfully enhanced engagement when compared to traditional lab exercises.

3. Summary of Post-Lab Survey Responses Evaluating Student Experience, Engagement, and Learning Outcomes (n = 11),

category	survey Item	response summary
general experience	overall enjoyment	3.45 ± 1.21
	mental engagement	yes: 7; somewhat: 2; no: 2
	ease of lab	4.27 ± 0.79
	personal value	yes: 9; not sure: 2
understanding the Bohr effect	pre-lab understanding	3.00 ± 0.77
	post-lab understanding	4.55 ± 0.69
	understanding of O2/CO2/pH	yes: 10; no: 1
	personal data for visualization	yes: 9; not sure: 2
application & relevance	connect theory & real-world	yes: 11
	likely to recall	4.45 ± 0.52
learning outcomes	more confidence in Bohr effect	yes: 8; somewhat: 3
	concept clarity improved	yes: 9; somewhat: 2
lab environment & tools	user-friendliness	yes: 6; somewhat: 4; no: 1
	clear instructions	yes: 11
	sufficient time	yes: 7; somewhat: 3; no: 1
reflection	recommend activity	4.45 ± 0.69

^aItems include Likert-scale and binary items assessing students’ enjoyment, conceptual understanding, relevance of personal data, and perceived effectiveness of the lab in reinforcing biochemical concepts.

^bValues reported as mean ± standard deviation. All numeric responses based on 1–5 scale.

Students again returned positive reviews of the activity in narrative responses, particularly with its use of personal health data and the emotional impact. One student wrote “This was very valuable to my education on my personal health. I’ve never done something like this and being able to see my own data made me care about it more. I took it as a sign that I can make changes to be a healthier individual.” Another reflected on the application and impact: “Seeing how ‘easy’ it is to see the Bohr effect in action, even after just a 10 min run. This experiment was simple but gave me health data I can personally use in the future as I now know more ways to test aerobic fitness.” A small number of students did note unclear expectations, with another student writing “The activity felt more like gym class than biochem lab. Wouldn’t have minded a variety such as bike/rowing.” Responses suggest overall strong engagement while highlighting future opportunities to broaden accessibility and framing for different learner preferences, with the underlying benefit of promoting student physical fitness and health awareness.

These findings align with previous educational research emphasizing the effectiveness of active and inquiry-based learning strategies. The personalized nature of the activity enhanced student motivation, as supported by previous work on the role of personal relevance in science learning. Furthermore, this work builds on studies that have demonstrated the positive effects of student-generated physiological data in physiology laboratories. , By extending the approach to biochemistry and anchoring it in a mechanistic concept such as the Bohr effect, this study advances the integration of physiology and biochemistry in undergraduate education.

Limitations and Future Considerations

In 2019, nearly one-quarter of college-aged students reported using a WFT. As wearable technology becomes increasingly accessible, this model offers a scalable and impactful way to personalize biochemistry education. In the absence of health trackers common to physiology and kinesiology courses, a modified, low-cost version of the activity may be recreated at any fitness facility with a simple heart rate monitor and pulse oximeter. The educational use of personal data, however, does not come without concerns. Wearable technology in the lab offers students real-time health insights but may inadvertently introduce bias or misconceptions when self-interpreted without context. Future laboratories should consider student training in data literacy and the variability of physiological baseline.

Despite promising results, the study is not without limitations. The current study reflects data from a single semester (n = 14), and the small sample size restricts generalizability. Future iterations will expand to include data from multiple cohorts to enhance the statistical robustness of the findings. Additionally, as students self-identified when they had reached fatigue, varying levels of participation in the physical component of the lab may have influenced individual outcomes. Nevertheless, the consistency of the reported learning gains and student satisfaction supports the feasibility and value of such an intervention. While small in scope, the results provide a proof-of-concept that justifies broader adoption and further study. First centered on the Bohr effect, the approach offers a framework for integrating physiology and metabolism with core biochemical content. Using self-generated physiological data to explore biochemical principles is broadly adaptable across the life sciences. Similar strategies could be applied to topics such as metabolic regulation, nerve impulses, or enzyme kinetics, enabling students to observe the dynamic interplay between molecular mechanisms and systemic physiology. This approach is particularly well suited for prehealth and interdisciplinary science programs at PUIs, where connecting molecular content to human function enhances both engagement and retention.

Conclusions

The integration of real-time physiological data collection into a biochemistry laboratory course measurably enhanced student comprehension of oxygen transport mechanisms and biochemical regulation. The intervention provided a dynamic and personalized learning experience, linking molecular-level concepts surrounding oxygen delivery to students’ own physiological responses during exercise. Students reported stronger engagement, demonstrated improved assessment performance, and expressed greater confidence in applying biochemical principles to clinical and fitness-related contexts. Beyond measurable learning gains, this movement-based laboratory represents a replicable framework for embedding physiological context into biochemistry education, aligning with broader efforts to humanize molecular science through experiential learning. By situating biochemical mechanisms in an experiential framework, this study supports the use of inquiry-based, interdisciplinary methods to reinforce complex content. These findings offer an adaptable, evidence-based model for advancing biochemistry education and can serve as a blueprint for curricular innovation in undergraduate STEM courses.

Methods

Study Design and Participants

The study was conducted at Canisius University, a Jesuit, midsized (Fall 2023 enrollment of 1,798) PUI in Buffalo, New York, United States of America. Eleven students enrolled in both the Introduction to Biochemistry lecture and laboratory courses voluntarily completed the study, with exam data compared to an additional three students enrolled in only the lecture course. The intervention was designed as a mixed-methods, within-subject case study to explore student engagement and conceptual understanding.

Pre-Laboratory Preparation

Informed consent was obtained from each participant using a signed prelaboratory consent form approved by the Canisius University IRB (see Supporting Information for full consent form). Participation was voluntary and students could opt out without penalty by completing an alternative written assignment; however, all laboratory students elected to take part in the study. Prior to the laboratory activity, students completed an online prelab quiz using the campus learning management system, Desire2Learn (D2L). The assessment evaluated baseline understanding of O₂ delivery, the Bohr effect, oxygen-hemoglobin binding, and related physiological variables. Students were instructed to complete the activity independently and without the use of external resources.

Laboratory Activity Overview

The instructional module, Cardio Chemistry: The Human Element was designed to link the biochemical mechanism of the Bohr effect to real-time physiological changes during exercise. To begin, students reviewed the laboratory document, which included a brief review of hemoglobin allostery and oxygen transport. Students then had the option to engage in a controlled submaximal fitness assessment using a treadmill. During the activity, physiological data were collected using a Dr. Talbot’s pulse oximeter, a Scosche Rhythm 2.0 heart rate monitor, and an iWorx TA-ROAM metabolic cart with a PEAK gas analyzer using LabScribe software. Students also had the option to use personal WFTs to supplement data collection and storage, though the accuracy of WFTs between brands remains uncertain.

Parameters recorded included SpO₂ (%), HR (beats per minute), VO₂ (L/min), VCO₂ (L/min), RER, and METs. Direct pH measurement was not performed; however, changes in CO₂ output and RER were used as physiologically relevant proxies for acid–base shifts associated with the Bohr effect. The treadmill protocol adhered to existing submaximal aerobic fitness test guidelines and was terminated upon volitional fatigue or once students reached 85% of age-predicted maximum HR, or RER ≥ 1.15. Data were continuously recorded and plotted at twenty-second intervals. All equipment was calibrated prior to use, and all procedures followed institutional safety guidelines. The complete lab protocol is provided in the Supporting Information.

Assessment Instruments

Following the activity, students completed a parallel postlaboratory quiz administered through D2L, along with a survey that evaluated engagement, clarity, and perceived relevance. The survey included Likert-scale items, binary (option 1/option 2) questions, and open-ended questions (see Supporting Information). Likert-scale responses were analyzed using mean and standard deviation. Open-ended responses were scored using a faculty-designed rubric focused on accuracy of biochemical reasoning, use of appropriate terminology, and conceptual application to physiological data. All students provided written feedback on the activity. Representative quotations from multiple participants were used to determine common themes. Quotations used within were sourced from six separate individuals.

Data Collection and Analysis

Paired sample t tests were conducted to evaluate changes in student understanding. Survey data was analyzed using descriptive statistics for quantitative items, while open-ended responses were coded thematically to identify patterns in student reflections and learning outcomes.

For between-group comparison, final exam scores from students who completed the laboratory activity were compared to those who completed only the lecture portion of the course. Analysis focused specifically on questions related to oxygen delivery to assess whether participation in the activity correlated with improved long-term understanding.

Ethical Considerations

All procedures were reviewed and approved by the university’s Institutional Review Board and deemed exempt under federal educational research guidelines. Participation in physical activity was strictly voluntary and accompanied by clearly communicated safety protocols. Students could withdraw at any time without penalty. All identifying information was anonymized to ensure confidentiality prior to data analysis.

Glossary

ABbreviations

RER

respiratory exchange ratio

PUI

primarily undergraduate institution

WFT

wearable fitness tracker

IRB

institutional review board

METs

metabolic equivalents

HR

heart rate

The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acsomega.5c07272.

• Complete laboratory protocol; informed consent form; pre- and postlab quiz questions; postlab survey; postlab reflection questions; rubric for Bohr effect narrative response (Table S1); rubric for hemoglobin affinity response (Table S2) (PDF)

The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript.

This work was partially funded by the Canisius Earning Excellence Program and Canisius University Chemistry and Biochemistry Research Endowment.

The authors declare no competing financial interest.

Published as part of ACS Omega special issue “Undergraduate Research as the Stimulus for Scientific Progress in the USA”.