Abstract
To address the challenges of complex workflows, limited student engagement, and insufficient integration of green and interdisciplinary skills in traditional laboratory teaching, this study introduces a multi-day experimental teaching framework with a staged instructional design for the extraction, purification, and identification of berberine hydrochloride (BH) in undergraduate natural product chemistry courses. The framework is implemented as a multi-day practical, pedagogically organized into three instructional stages (extraction, separation and purification, and identification), each of which is aligned with specific learning objectives. This stage-based instructional structure reduces cognitive load and enhances learning outcomes. Green chemistry principles were incorporated into the G-RPWAM (green-reagents, procedures, waste, awareness, methodology) framework to guide solvent selection and procedural design, optimize reagent use, minimize waste, and reduce energy consumption in undergraduate laboratory teaching while fostering environmental awareness. A comprehensive evaluation system, including technical metrics (extraction yield, purity), student performance (experimental skills, data analysis ability, creative thinking, problem-solving ability), and student feedback (content clarity, structured organization, green principles integration, overall effectiveness), was employed to assess the teaching innovation. The results demonstrated that the staged instructional design improved the accessibility and scalability of the experiment, enhanced student satisfaction, and successfully cultivated both sustainability awareness and laboratory skills. This study provides a structured, pedagogy-oriented approach aligned with green chemistry principles for undergraduate natural product chemistry education.
Data availability
The data supporting the findings of this study are available from the corresponding author upon reasonable request.
References
Egambaram, O. et al. The future of laboratory chemistry learning and teaching must be Accessible. J. Chem. Educ. 99 (12), 3814–3821 (2022).
Guntero, V. A., Mancini, P. M. E. & Kneeteman, M. Introducing organic chemistry students to the extraction of natural products found in vegetal Species. World J. Chem. Educ. 4 (5), 142–147 (2017).
Onuoha, C. P., Garner, M. P. & Fenn, N. E. R. Student perceptions of co-curricular activities on pharmacy education: A review. Curr. Pharm. Teach. Learn. 13 (5), 576–584 (2021).
Cooper, M. M. & Stowe, R. L. Chemistry education Research-From personal empiricism to Evidence, Theory, and informed Practice. Chem. Rev. 118 (12), 6053–6087 (2018).
Wu, M. M. & Yezierski, E. J. Investigating Teacher-Teacher feedback: Uncovering useful Socio-pedagogical norms for Reform-Based chemistry Instruction. J. Chem. Educ. 100 (11), 4224–4236 (2023).
Nugraheni, A. R. E. & Srisawasdi, N. Development of pre-service chemistry teachers’ knowledge of technological integration in inquiry-based learning to promote chemistry core competencies. Chem. Educ. Res. Pract. 26 (2), 398–419 (2025).
Zhang, L. et al. The strategies and techniques of drug discovery from natural products. Pharmacol. Ther. 216, 107686 (2020).
Zhao, J. et al. Berberine hydrochloride ameliorates PM2.5-induced pulmonary fibrosis in mice through inhibiting oxidative stress and inflammatory. Chem. Biol. Interact. 386, 110731 (2023).
Xiao, Y. et al. Berberine hydrochloride enhances innate immunity to protect against pathogen infection via p38 MAPK pathway. Front. Immunol. 16, 1536143 (2025).
Yang, Y. et al. Determination of alkaloid contents in various tissues of Coptis chinensis Franch. By reversed Phase-High performance liquid chromatography and ultraviolet Spectrophotometry. J. Chromatogr. Sci. 55 (5), 556–563 (2017).
Wang, Y. et al. Simultaneous determination of eight active compounds in Baitouweng decotion and its single Herbs. J. Chromatogr. Sci. 57 (6), 502–510 (2019).
Jakubčinová, J., Feszterová, M. & Silliková, V. Active learning in the extraction of organic compounds: A study of undergraduate chemistry Students. Educ. Sci. 14 (10), 1051 (2024).
Wackerly, J. W., Dunne, J. F. & Shriver, J. A. Multistep synthesis of diphenhydramine: A modular laboratory for First-Semester organic chemistry Students. J. Chem. Educ. 102 (7), 2859–2865 (2025).
Fleischer, T. et al. Digital sequential scaffolding during experimentation in chemistry Education-Scrutinizing influences and effects on Learning. Educ. Sci. 13 (8), 811 (2023).
Young, J. Q. et al. Cognitive load theory: implications for medical education: AMEE guide 86. Med. Teach. 36 (5), 371–384 (2014).
Sweller, J. Cognitive load during problem solving: effects on learning. Cogn. Sci. 12 (2), 257–285 (1988).
Yin, D. S. & Yin, X. Scaffolding learning: from specific to generic with large Language models. PloS One. 19 (9), e310409 (2024).
Wood, D., Bruner, J. S. & Ross, G. The role of tutoring in problem solving. J. Child Psychol. Psychiatry Allied Discip. 17 (2), 89–100 (1976).
Long, E. M. & Gummelt, G. Experiential service learning: Building skills and sensitivity with kolb’s learning theory. Gerontol. Geriatr. Educ. 41 (2), 219–232 (2020).
Kolb, D. A. Experiential Learning: Experience as the Source of Learning and Development. (Prentice-Hall, 1984).
Worrall, A. F. et al. University teaching of mass spectrometry as a key practical technique within the context of a fully integrated, spiral curriculum. Rapid Commun. Mass Spectrom. RCM. 30, e9851 (2024).
Fauzi Ah, L. et al. Modified spiral organic curriculum on organic chemistry courses for chemistry education undergraduate students. AIP Conf. Proc. 1911(1), (2017).
Schmidkunz, H. & Büttner, D. Teaching Chemistry According To a Spiral curriculum (European Journal of Science Education, 1986).
Freese, T. et al. The relevance of sustainable laboratory practices. RSC Sustain. 2 (5), 1300–1336 (2024).
de Marco, B. A. et al. Evolution of green chemistry and its multidimensional impacts: A review. Saudi Pharm. J. 27 (1), 1–8 (2019).
Tobiszewski, M., Mechlińska, A. & Namieśnik, J. Green analytical chemistry-theory and practice. Chem. Soc. Rev. 39 (8), 2869–2878 (2010).
Etzkorn, F. A. & Ferguson, J. L. Integrating green chemistry into chemistry Education. Angew Chem. Int. Ed. Engl. 62 (2), e202209768 (2023).
de la Parra, J. et al. Crafting a More Environmentally Benign Extraction and Analysis of Pharmaceutical Precursors from a Medicinal Plant: A Student-Led Innovation (ChemRxiv Preprint, 2019).
Plotka-Wasylka, J. et al. Green chemistry in higher education: State of the Art, Challenges, and future Trends. ChemSusChem 11 (17), 2845–2858 (2018).
Krasnodębski, M. The bumpy road to sustainability: Reassessing the history of the twelve principles of green chemistry. Stud. Hist. Philos. Sci. 103, 85–94 (2024).
Lu, J. et al. Separation of Berberine hydrochloride and Tetrahydropalmatine and their quantitative analysis with thin layer chromatography involved with ionic Liquids. J. Anal. Methods Chem. 2015, 642401 (2015).
Zhen, H. S., Chen, Y. & Zhong, X. J. Quality standards of Huanglian Jiedu pills. Zhongguo Zhong Yao Za zhi = Zhongguo Zhongyao Zazhi = China. J. Chin. Materia Med. 19 (7), 414–447 (1994).
Jebb, A. T., Ng, V. & Tay, L. A review of key likert scale development advances: 1995–2019. Front. Psychol. 12, 637547 (2021).
Korrel, M. et al. Framework for training in minimally invasive pancreatic surgery: An international Delphi consensus Study. J. Am. Coll. Surg. 235 (3), 383–390 (2022).
Braun, V. & Clarke, V. Using thematic analysis in psychology. Qual. Res. Psychol. 3 (2), 77–101 (2006).
Lynch, B. L. A behaviorally anchored rating scale for the evaluation of student performance. Am. J. Med. Technol. 43 (1), 54–63 (1977).
Miladinović, S. M. Green analytical chemistry: Integrating sustainability into undergraduate education. Anal. Bioanal. Chem. 417 (4), 665–673 (2025).
Liu, Z. et al. Comprehensive undergraduate experiment: Determination of phenylurea herbicide in tea by magnetic solid-phase extraction-high performance liquid chromatography. Se pu = Chin. J. Chromatogr. 42 (11), 1094–1100 (2024).
Ho, C. C. et al. Employing pressurized hot water extraction (PHWE) to explore natural products chemistry in the undergraduate Laboratory. J. Vis. Exp. 7 (141), 10–3791 (2018).
Murcia, J. E. et al. Risk assessment and green chemistry applied to waste generated in university laboratories. Heliyon 9 (5), e15900 (2023).
Heni, M. et al. Focused didactic training for skills lab student tutors - which techniques are considered helpful?. GMS Z. Fur Medizinische Ausbildung. 29 (3), Doc41 (2012).
Chen, M., Jeronen, E. & Wang, A. What Lies behind teaching and learning green chemistry to promote sustainability education? A literature review. Int. J. Environ. Res. Public. Health. 17(21), 7876, (2020)
Sahoo, T. et al. Green solvent: green shadow on chemical Synthesis. Curr. Org. Synth. 17 (6), 426–439 (2020).
Rahman, M. U. et al. Green analytical techniques for impurity determination in Pharmaceuticals. Electrophoresis. 47(1):87–105 (2026).
Collins, J. et al. Impact of an international service learning experience in India for DPT students: Short- and Long-Term Benefits. J. Allied Health. 48 (1), 22–30 (2019).
Bowling, A. M. & Underwood, P. W. Effect of simulation on knowledge, self-confidence, and skill performance in the USA: A quasi-experimental study. Nurs. Health Sci. 18 (3), 292–298 (2016).
Xu, B. et al. How does mathematical literacy affect creative thinking? Independent effects and differential impacts across proficiency groups. Acta. Psychol. 260, 105509 (2025).
Smerdel, S. & Zejnilagić Hajrić, M. Demographic characteristics of chemistry teachers in Croatia affecting the use of Pre-laboratory activities in the Classroom. Acta Chim. Slov. 67 (2), 435–444 (2020).
Coleman, A. B. et al. Imagining, designing, and interpreting experiments: Using quantitative assessment to improve instruction in scientific reasoning. Biochem. Mol. Biol. Educ.: Bimon. Public. Int. Union Biochem. Mol. Biol. 51 (3), 286–301 (2023).
Basey, J. M. et al. An evaluation of two hands-on lab styles for plant biodiversity in undergraduate biology. CBE Life Sci. Educ. 13 (3), 493–503 (2014).
Fatemi, F., Dehdashti, A. & Jannati, M. Implementation of chemical Health, Safety, and environmental risk assessment in laboratories: A Case-Series Study. Front. Public. Health. 10, 898826 (2022).
Cooley, C. Z. et al. Design and implementation of a low-cost, tabletop MRI scanner for education and research prototyping. J. Magn. Reson. 310, 106625 (2020).
Reyes, R. L. et al. Enhancing experiential science learning with virtual labs: A narrative account of merits, challenges, and implementation strategies. J. Comput. Assist. Learn. 40 (6), 3167–3186 (2024).
Chiu, W. Pedagogy of emerging technologies in chemical education during the era of digitalization and artificial intelligence: A systematic review. Educ. Sci. 11, 709 (2021).
Funding
This work was supported by the Guangdong Key Laboratory of Functional Substances and Health Products from Medicinal Edible Resources (Grant Number 2021B1212040015), the Guangdong Provincial Department of Education Project (Grant Number 2025KQNCX043), the Hanshan Normal University Quality Engineering Program (Grant Number HSJG-SZ252187, E25070 and e25070), the Third Batch Professor Startup Project of Hanshan Normal University (Grant Number QD2023312), the Hanshan Normal University Scientific Research Mentor Support Program (Grant Number XBF202302), and the Chaozhou Science and Technology Bureau Science and Technology Project (Grant Number 202404GY06).
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Yaqun Liu: Conceptualization, methodology, supervision, writing-original draft preparation, writing-review and editing. Qionglu Huang: Experimental implementation. Zhenxia Zhang: Experimental design, data analysis, visualization. Peikui Yang: Development of the evaluation system, statistical analysis. Yongping Huang: Experimental implementation. Jiayi Huang: Experimental trials, collection of student feedback. Jie Tan: Experimental trials, collection of student feedback. Guohong Xie: Experimental trials, collection of student feedback. Jiechun Chen: Experimental trials, collection of student feedback. Yicun Chen: Analysis of student performance. Chengsong Xie: Microscopy observations, manuscript review. Yuzhong Zheng: Supervision, theoretical framework development, writing-review and editing. All the authors discussed the results and contributed to the final manuscript.
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Liu, Y., Huang, Q., Zhang, Z. et al. A structured multi-day experimental framework integrating green chemistry for the extraction and characterization of Berberine hydrochloride in undergraduate education. Sci Rep (2026). https://doi.org/10.1038/s41598-026-39150-x
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DOI: https://doi.org/10.1038/s41598-026-39150-x
