Abstract
The Kato–Katz method is the standard technique for diagnosing Soil-Transmitted Helminth (STH) infections; however, the conventional malachite green stain poses potential environmental and health hazards. Therefore, the development of natural, biodegradable staining agents is needed to support sustainable laboratory practices. This study aimed to evaluate the effectiveness of natural cellophane stains developed from fragrant pandan leaf (Pandanus amaryllifolius) and mangosteen peel (Garcinia mangostana) extracts as alternatives to malachite green in the Kato–Katz method for STH diagnosis. Methods: A true experimental laboratory study with a post-test only control group design was conducted. Malachite green served as the control, while pandan leaf and mangosteen peel extracts were prepared at concentrations of 75%, 85%, and 95%. Qualitative phytochemical screening was performed, followed by microscopic evaluation of Ascaris lumbricoides, Trichuris trichiura, and hookworm eggs. Staining quality was assessed based on transparency, contrast, morphological clarity, and ease of identification. Data were analyzed using the Shapiro–Wilk, Kruskal–Wallis, and Mann–Whitney U tests. Results: Phytochemical analysis confirmed the presence of chlorophyll, flavonoids, tannins, and phenolic compounds in pandan leaf extract, whereas mangosteen peel extract contained anthocyanins, xanthones, flavonoids, tannins, and phenolic compounds. Both extracts produced homogeneous and stable staining solutions that preserved the characteristic morphology of all STH eggs. The 95% pandan leaf extract demonstrated the highest staining quality among the natural stains, followed by the 95% mangosteen peel extract. No statistically significant difference was observed between the high-concentration natural stains and conventional malachite green (p > 0.05). Conclusion: Natural cellophane stains derived from fragrant pandan leaf and mangosteen peel extracts have strong potential as safe, biodegradable, and sustainable alternatives to malachite green for routine STH diagnosis using the Kato–Katz method.