Research has demonstrated the significance of urine in the area of production of BioH 2 gas for the purpose of energy utilization using wood saw dust as the raw material. The bio-nutrients of the urine was characterized by considering the following parameter and its value determined as demonstrated in the area of pH (6.80), Total Solids (TS) (6050mg/l), Total Dissolved Solids (TDS) (4250mg/l), Total Suspended Solid (TSS) (1800mg/l), Chemical Oxygen Demand (COD) (8535mg/l) Biochemical Oxygen Demand (BOD 5 ) (2300mg/l) and other parameters such as Total Kjeldahl Nitrogen (TKN) (4800mg/l), Ammonium (NH 4 + -N) (3200mg/l), phosphate (PO 4 3- -P) (230mg/l), Sulphate (SO 4 2- ) (156mg/l), Potassium (K + ) (905mg/l), Chloride (Cl - ) (1850mg/l), Sodium (Na + ) (2450mg/l), Calcium (Ca 2+ ) (156mg/l), Magnesium (Mg 2+ ) (120mg/l) and Urea 18900mg/l. The yield of biohydrogen from the different samples revealed that HSP1 > HMP3 > HSM1 > HSHM1 > HSHP1 > HHP1 > HHM1 and this process was operated with bio nutrient in the production of hydrogen gas. However, for the hydrogen gas production with the aid of bio- nutrient revealed the order of magnitude of UHP2 > USP2 > USHP2 > UHM2 > UMP3 > USM2 > USHM2. This research has illustrated the significance of the urine production with the mechanism of hydrolyzed and unhydrolyzed process as well as the role of bio-nutrient in the production of BioH 2 gas.

Key words: Characterization, urine, production, bio-Hydrogen, energy, utilization, wood

1. Ahmed SF, Rafa N, Mofijur M, Badruddin IA, Inayat A, Ali MS, Farrok O, Yunus Khan TM. Biohydrogen production from biomass sources: metabolic pathways and economic analysis. Frontiers in Energy Research. 2021 Sep 10;9:753878. 2. Fangkum A, Reungsang A. Biohydrogen production from sugarcane bagasse hydrolysate by elephant dung: Effects of initial pH and substrate concentration. International Journal of Hydrogen Energy. 2011 Jul 1;36(14):8687-96. 3. Budiman PM, Wu TY. Role of chemicals addition in affecting biohydrogen production through photofermentation. Energy Conversion and Management. 2018 Jun 1;165:509- 27. 4. Багрій ІД, Рєпкін ОО, Паюк СО, Криль ЯМ, Гафич ІП. Scientific justification of spatial distribution and mapping of anomalous manifestations of white hydrogen–energy raw material of the XXI century. Мінеральні ресурси України. 2023 Apr 10(1):4-10. 5. Li YC, Chu CY, Wu SY, Tsai CY, Wang CC, Hung CH, Lin CY. Feasible pretreatment of textile wastewater for dark fermentative hydrogen production. International journal of hydrogen energy. 2012 Oct 1;37(20):15511-7. 6. Lin CY, Chiang CC, Nguyen ML, Lay CH. Enhancement of fermentative biohydrogen production from textile desizing wastewater via coagulation-pretreatment. international journal of hydrogen energy. 2017 Apr 27;42(17):12153-8. 7. Lin YH, Zheng HX, Juan ML. Biohydrogen production using waste activated sludge as a substrate from fructose-processing wastewater treatment. Process Safety and Environmental Protection. 2012 May 1;90(3):221-30. 8. Lin PY, Whang LM, Wu YR, Ren WJ, Hsiao CJ, Li SL, Chang JS. Biological hydrogen production of the genus Clostridium: metabolic study and mathematical model simulation. International Journal of Hydrogen Energy. 2007 Aug 1;32(12):1728-35. 9. Ahmad T, Aadil RM, Ahmed H, ur Rahman U, Soares BC, Souza SL, Pimentel TC, Scudino H, Guimarães JT, Esmerino EA, Freitas MQ. Treatment and utilization of dairy industrial waste: A review. Trends in Food Science & Technology. 2019 Jun 1;88:361-72. 10. Kothari R, Tyagi V, Pathak AK. Renewable and sustainable energy reviews Renewab Sustainab. ResearchGate. 2010;14:1744–51. Available from: https://www.researchgate.net/publication/285197707_Renewable_and_sustainable_energ y_reviews_Renewab_Sustainab 11. Kothari R, Tyagi VV, Pathak A. Waste-to-energy: A way from renewable energy sources to sustainable development. Renewable and Sustainable Energy Reviews. 2010 Dec 1;14(9):3164-70. 12. Kotsopoulos TA, Fotidis IA, Tsolakis N, Martzopoulos GG. Biohydrogen production from pig slurry in a CSTR reactor system with mixed cultures under hyper-thermophilic temperature (70 C). biomass and bioenergy. 2009 Sep 1;33(9):1168-74. 13. Kourougianni F, Arsalis A, Olympios AV, Yiasoumas G, Konstantinou C, Papanastasiou P, Georghiou GE. A comprehensive review of green hydrogen energy systems. Renewable Energy. 2024 Sep 1;231:120911. 14. Kovalev AA, Kovalev DA, Zhuravleva EA, Laikova AA, Shekhurdina SV, Vivekanand V, Litti YV. Biochemical hydrogen potential assay for predicting the patterns of the kinetics of semi-continuous dark fermentation. Bioresource Technology. 2023 May 1;376:128919. 15. Kuehn M. Transitioning from a Grey to Clean Hydrogen Economy: Considerations for Scaling Production and Making Use of Existing Transport Infrastructure. InAbu Dhabi International Petroleum Exhibition and Conference 2023 Oct 2 (p. D031S106R007). SPE. 16. Kumar G, Bakonyi P, Periyasamy S, Kim SH, Nemestóthy N, Bélafi-Bakó K. Lignocellulose biohydrogen: practical challenges and recent progress. Renewable and Sustainable Energy Reviews. 2015 Apr 1;44:728-37. 17. Kumar N, Das D. Enhancement of hydrogen production by Enterobacter cloacae IIT-BT 08. Process Biochemistry. 2000 Jan 1;35(6):589-93. 18. De Vrije T, Bakker RR, Budde MA, Lai MH, Mars AE, Claassen PA. Efficient hydrogen production from the lignocellulosic energy crop Miscanthus by the extreme thermophilic bacteria Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana. Biotechnology for biofuels. 2009 Jun 17;2(1):12. 19. De Vrije T, De Haas GG, Tan GB, Keijsers ER, Claassen PA. Pretreatment of Miscanthus for hydrogen production by Thermotoga elfii. International journal of hydrogen energy. 2002 Nov 1;27(11-12):1381-90. 20. Sigurbjornsdottir MA, Orlygsson J. Combined hydrogen and ethanol production from sugars and lignocellulosic biomass by Thermoanaerobacterium AK54, isolated from hot spring. Applied Energy. 2012 Sep 1;97:785-91. 21. Singh A, Sevda S, Abu Reesh IM, Vanbroekhoven K, Rathore D, Pant D. Biohydrogen production from lignocellulosic biomass: technology and sustainability. Energies. 2015 Nov 17;8(11):13062-80. 22. Lakshmidevi R, Muthukumar K. Enzymatic saccharification and fermentation of paper and pulp industry effluent for biohydrogen production. International Journal of Hydrogen Energy. 2010 Apr 1;35(8):3389-400. 23. Levin DB, Chahine R. Challenges for renewable hydrogen production from biomass. International journal of hydrogen energy. 2010 May 1;35(10):4962-9. 24. Li J, Wei YM, Liu L, Li X, Yan R. The carbon footprint and cost of coal-based hydrogen production with and without carbon capture and storage technology in China. Journal of Cleaner Production. 2022 Aug 15;362:132514. 25. Li Q, Liu CZ. Co-culture of Clostridium thermocellum and Clostridium thermosaccharolyticum for enhancing hydrogen production via thermophilic fermentation of cornstalk waste. International Journal of Hydrogen Energy. 2012 Jul 1;37(14):10648- 54. 26. Li YC, Liu YF, Chu CY, Chang PL, Hsu CW, Lin PJ, Wu SY. Techno-economic evaluation of biohydrogen production from wastewater and agricultural waste. International journal of hydrogen energy. 2012 Oct 1;37(20):15704-10. 27. Liu F, Fang B. Optimization of bio‐hydrogen production from biodiesel wastes by Klebsiella pneumoniae. Biotechnology Journal: Healthcare Nutrition Technology. 2007 Mar;2(3):374-80. 28. Liu H, Zhang T, Fang HH. Thermophilic H2 production from a cellulose-containing wastewater. Biotechnology Letters. 2003 Feb;25(4):365-9. 29. Liu S, Li W, Zheng G, Yang H, Li L. Optimization of cattle manure and food waste co- digestion for biohydrogen production in a mesophilic semi-continuous process. Energies. 2020 Jul 28;13(15):3848. 30. Palmqvist E, Hahn-Hägerdal B. Fermentation of lignocellulosic hydrolysates. II: inhibitors and mechanisms of inhibition. Bioresource Technology. 2000 Aug;74(1):25–33. Available from:https://www.researchgate.net/publication/222245618_Fermentation_of_lignocellulo sic_hydrolysates_II_Inhibitors_and_mechanisms_of_inhibition 31. Koroglu EO, Ozdemir OK, Ozkaya B, Demir A. An integrated system development including PEM fuel cell/biogas purification during acidogenic biohydrogen production from dairy wastewater. International Journal of Hydrogen Energy. 2019 Jun 28;44(32):17297-303. 32. Kossalbayev BD, Tomo T, Zayadan BK, Sadvakasova AK, Bolatkhan K, Alwasel S, Allakhverdiev SI. Determination of the potential of cyanobacterial strains for hydrogen production. International Journal of Hydrogen Energy. 2020 Jan 24;45(4):2627-39. 33. Bassey KE, Ibegbulam C. Machine learning for green hydrogen production. Computer Science & IT Research Journal. 2023;4(3):368-85. 34. Bauer C, Treyer K, Antonini C, Bergerson J, Gazzani M, Gencer E, Gibbins J, Mazzotti M, McCoy ST, McKenna R, Pietzcker R. On the climate impacts of blue hydrogen production. Sustainable Energy & Fuels. 2022;6(1):66-75. 35. Dinopoulou G, Sterritt RM, Lester JN. Anaerobic acidogenesis of a complex wastewater: II. Kinetics of growth, inhibition, and product formation. Biotechnology and bioengineering. 1988 Jun 5;31(9):969-78. 36. Guo XM, Trably E, Latrille E, Carrère H, Steyer JP. Hydrogen production from agricultural waste by dark fermentation: a review. International journal of hydrogen energy. 2010 Oct 1;35(19):10660-73. 37. Lal, A., & You, F. (2023). Targeting climate‐neutral hydrogen production: Integrating brown and blue pathways with green hydrogen infrastructure via a novel superstructure and simulation‐based life cycle optimization. AIChE Journal, 69(1), 17-95. 38. Lateef SA, Beneragama N, Yamashiro T, Iwasaki M, Ying C, Umetsu K. Biohydrogen production from co-digestion of cow manure and waste milk under thermophilic temperature. Bioresource Technology. 2012 Apr 1;110:251-7. 39. Manish S, Banerjee R. Comparison of biohydrogen production processes. International Journal of Hydrogen Energy. 2008 Jan 1;33(1):279-86. 40. Majewski P, Salehi F, Xing K. Green hydrogen. AIMS Energy. 2023;11(5):878-95. 41. Maka AO, Mehmood M. Green hydrogen energy production: current status and potential. Clean Energy. 2024 Apr 1;8(2):1-7. 42. Marone A, Varrone C, Fiocchetti F, Giussani B, Izzo G, Mentuccia L, Rosa S, Signorini A. Optimization of substrate composition for biohydrogen production from buffalo slurry co-fermented with cheese whey and crude glycerol, using microbial mixed culture. International Journal of Hydrogen Energy. 2015 Jan 5;40(1):209-18. 43. Mars AE, Veuskens T, Budde MA, Van Doeveren PF, Lips SJ, Bakker RR, De Vrije T, Claassen PA. Biohydrogen production from untreated and hydrolyzed potato steam peels by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana. International journal of hydrogen energy. 2010 Aug 1;35(15):7730-7. 44. Massarweh, Osama;Al-khuzaei, Maha;Al-Shafi, Manal;Bicer, Yusuf;Abushaikha, Ahmad S. Blue hydrogen production from natural gas reservoirs: A review of application and feasibility. Journal of CO2 Utilization. 2023 Apr;70:102438. Available from: https://ui.adsabs.harvard.edu/abs/2023JCOU...7002438M/abstract 45. Lee DJ, Show KY, Su A. Dark fermentation on biohydrogen production: pure culture. Bioresource technology. 2011 Sep 1;102(18):8393-402. 46. Nega Gesese T, Abera Mersha D, Asmare Tezera, Getahun E. Biohydrogen Production Pathways: An Integrated Review of Feedstock Utilization, Technological... BioEnergy Research. 2026 Apr 25;19(1). Available from: https://www.researchgate.net/publication/404190777_Biohydrogen_Production_Pathway s_An_Integrated_Review_of_Feedstock_Utilization_Technological_Developments_Com putational_Modeling_and_Emerging_Prospects 47. ‌Levin DB, Pitt L, Love M. Biohydrogen production: prospects and limitations to practical application. International journal of hydrogen energy. 2004 Feb 1;29(2):173-85. 48. Dong ZY, Yang J, Yu L, Daiyan R, Amal R. A green hydrogen credit framework for international green hydrogen trading towards a carbon neutral future. International Journal of Hydrogen Energy. 2022 Jan 5;47(2):728-34. 49. Bühler K, Bühler B, Klähn S, Krömer JO, Dusny C, Schmid A. 11 biocatalytic production of white hydrogen from water using cyanobacteria. Photosynthesis: Biotechnological applications with microalgae. 2021 Oct 25:279-306. 50. Cai M, Liu J, Wei Y. Enhanced biohydrogen production from sewage sludge with alkaline pretreatment. Environmental science & technology. 2004 Jun 1;38(11):3195-202. 51. Cappelletti BM, Reginatto V, Amante ER, Antônio RV. Fermentative production of hydrogen from cassava processing wastewater by Clostridium acetobutylicum. Renewable Energy. 2011 Dec 1;36(12):3367-72