The purpose of study was to obtain the best combination of natural cryoprotectant (honey solution) and synthetic cryoprotectant (methanol) on the quality of spermatozoa Chromobotia macracanthus Bleeker 1852, after freezing at -80 ℃ for 48 hours. The combination of 10% methanol with various concentrations of honey solution (0%, 0.1%, 0.3%, 0.5%, 0.7% and 0.9%) was tested in this study. Ringer's solution was used as an extender. The diluted sperm was then equilibrated for 25 minutes at 4 ℃, then frozen at -80 ℃ for 48 hours. The sperm was then thawed at 40 ℃ for 13 seconds. Viability, motility and percentage of fertilization were evaluated. The results of the analysis of variance showed that the combination of 10% methanol with several concentrations of honey solution had a significant effect (P <0.05) on the viability of spermatozoa and the percentage of fertilization, but had no effect on sperm motility (P> 0.05). The combination of 0.1% honey solution and 10% methanol showed the highest percentage of both motility (89.4 ± 5.45%), viability (85.75 ± 4.79%), and the percentage of fertilization (98.55 ± 1.69%). %). The conclusion of this study is that 0.1% honey solution combined with 10% methanol in Ringer's solution is the best cryoprotectant for C. macracanthus spermatozoa stored at -80 ℃ for 48 hours. Abstrak Penelitian ini bertujuan untuk mendapatkan kombinasi terbaik antara krioprotektan alami (larutan madu) dan krioprotektan sintetik (metanol) terhadap kualitas spermatozoa Chromobotia macracanthus Bleeker 1852, pasca penyimpanan pada suhu -80 ℃ selama 48 jam. Kombinasi metanol 10% dengan berbagai konsentrasi larutan madu (0%, 0,1%, 0,3%, 0,5%, 0,7% dan 0,9%) diuji dalam penelitian ini. Larutan Ringer digunakan sebagai ekstender. Sperma yang telah diencerkan, kemudian diekuiliberasi selama 25 menit pada suhu 4 ℃, kemudian disimpan beku pada suhu -80 ℃ selama 48 jam. Sperma kemudian dicairkan pada suhu 40 ℃ selama 13 detik. Viabilitas, motilitas dan persentase fertilisasi dievaluasi. Hasil uji analisis sidik ragam menunjukkan bahwa kombinasi metanol 10% dengan beberapa konsentrasi larutan madu berpengaruh secara signifikan (P<0,05) terhadap viabilitas spermatozoa dan persentase fertilisasi, tapi tidak berpengaruh terhadap motilitas spermatozoa (P>0,05). Kombinasi larutan madu 0,1% dan metanol 10% menunjukkan persentase tertinggi baik motilitas (89,4 ± 5,45%), viabilitas (85,75 ± 4,79%), maupun persentase fertilisasi (98,55 ± 1,69%). Kesimpulan penelitian ini adalah bahwa larutan madu 0,1% yang dikombinasikan dengan metanol 10% dalam larutan Ringer merupakan krioprotektan terbaik bagi spermatozoa C. macracanthus yang disimpan pada suhu -80 ℃ selama 48 jam.