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Various pretreatment methods have been combined and employed for maximizing the solubilization of waste-activated sludge (WAS). However, the question "by changing the series of applied combined pretreatments (CPs), can the solubilization efficiency of WAS be affected?" has never been addressed. In this study, firstly, thermal (T), alkaline (A), and ultrasonic (U) pretreatments were individually applied at broad strengths (T = 80–120 °C for 30 min, A = pH 9–12, and U = 5–60 min at 300 W). Then, pretreatment conditions that caused similar solubilization (13.0%) (120 °C, pH 11, and 30 min for T, A, and U, respectively), were adopted for CP with reverse sequences of T&A, U&A, and T&U. A similar disintegration degree was observed in U→A and A→U, while a meaningful difference was found in T&A and T&U: T→A (28.3%), A→T (42.9%), T→U (22.9%), and U→T (27.1%). The difference in pretreatment series also affected the characteristics of soluble matters, which was analyzed by excitation emission matrix and molecular weight distribution. Due to these differences, the highest methane yield of 68.8% (based on (chemical oxygen demand) CODinput) was achieved at A→T, compared to T→A (62.3%). Our results suggested a simple strategy for increasing solubilization, at the same expense of energy, which might be beneficial in the following treatment process, such as dewatering and transportation. [ABSTRACT FROM AUTHOR]

A combined ball milling and ethanol organosolv process is proposed for the pretreatment of three types of herbaceous biomass, giant miscanthus, corn stover, and wheat straw. The combined pretreatment was effective at both removing lignin and increasing the glucan content. After 120 min pretreatment, the glucan content increased to 63.09%, and 55.89% of the acid-insoluble lignin was removed from the giant miscanthus sample. The removal of cellulose, hemicellulose, and acetyl groups were correlated with the removal of lignin. The pretreatment of corn stover showed the highest removal of cellulose, but this was dependent on the removal of acid-insoluble lignin. The slope of the regression lines, which shows the correlation between the removal of lignin and cellulose, was lower than other correlations. The changes in biomass size were analyzed using size distribution graphs. With increasing pretreatment time, the particle size reduction improved in the three types of herbaceous biomass. Because of the combined physicochemical pretreatment, the enzymatic digestibility improved, and a maximum of 91% glucan digestibility was obtained from the pretreated corn stover when 30 FPU/g-glucan enzyme was added. Finally, compositional analysis of the recovered lignin from the remaining black liquor was investigated. [ABSTRACT FROM AUTHOR]