Duromide is an effective urease inhibitor when combined with vinasse and blended with ammonium sulphate

ABSTRACT

A recent trend in sugarcane production is to apply urea fertilizers in combination with vinasse (V) to balance nitrogen (N) and potassium (K) supply. This practice promotes a circular economy and a possible reduction in N losses through ammonia (NH₃) volatilization due to the acidic nature of V. However, it is unclear whether V reduces NH₃ volatilization, and whether the addition of urease inhibitors further reduces NH₃ losses. Furthermore, the efficacy of formulations containing urea and urease inhibitors may decrease over time when blended with acidic fertilizers such as ammonium sulfate (AS). This study aimed to determine the efficacy of urease inhibitors N-(n-butyl) thiophosphoric triamide (NBPT) and Duromide in reducing NH₃ volatilization losses from urea applied with V and from urea blended with AS and stored for 10 months in a warehouse. Two different experiments were conducted under controlled conditions. Seven days after fertilizer application, urea addition to V resulted in a 35 % NH₃ loss (a reduction of only 36 % compared to urea), whereas NBPT-treated urea + V and Duromide-treated urea + V resulted in reductions of 51 and 74 %, respectively. The efficacy of NBPT-treated urea blended with AS to control volatilization decreased from 41 to 9 % after 10 months of storage. In contrast, the efficacy of Duromide-treated urea in reducing the NH₃ volatilization remained relatively constant. Duromide is a more stable and effective urease inhibitor than NBPT, supporting agronomic practices that reduce NH₃ volatilization and maintain inhibitory effects when blended with acidic fertilizers such as AS or combined with acidic organo-mineral fertilizers like V.

Keywords
NBPT; nitrogen; sugarcane; fertilizer; urease inhibitor

INTRODUCTION

Brazil is the largest producer of sugarcane (Saccharum spp.) worldwide (USDA, 2024). Sugarcane biomass is used to produce ethanol, sugar, electricity, and other biorenewables (Bordonal et al., 2018). The majority of this biomass is used in ethanol production, with Brazil producing 29.7 billion liters of ethanol in 2023 (Conab, 2024). Vinasse (V) is a by-product generated during ethanol rectification and distillation (Parsaee et al., 2019). About 13 L of V are generated for every 1 L of ethanol produced from sugarcane (Gallucci et al., 2019). Owing to its high concentrations of organic matter and potassium (K), V is commonly applied to sugarcane fields (Rossetto et al., 2018). With the increasing availability of V and the ongoing need to reduce application costs and balance the nitrogen (N):K ratio, sugarcane producers enrich V with N fertilizers. In addition to this, there is some evidence that V reduces N losses through ammonia (NH₃) volatilization (Gallucci et al., 2019; Oliveira et al., 2023), however losses remain high (Otto et al., 2017).

Urea is the most widely used N fertilizer worldwide, accounting for more than 50 % of current N demand (IFA, 2024). Its popularity is explained by its high N concentration, widespread availability, and low production costs (Cantarella et al., 2018). However, when applied to the soil surface or at shallow depths, urea undergoes hydrolysis by the urease enzyme, resulting in the loss of NH₃ to the atmosphere. In sugarcane fields mulched with straw, NH₃ volatilization can range from 24 to 61 % of the applied N (Cantarella et al., 2008; Mariano et al., 2012; Mira et al., 2017; Pinheiro et al., 2018). The deleterious effects of NH₃ volatilization include reduced yield (Torralbo et al., 2022), economic losses (Pan et al., 2016), air pollution, and contamination of terrestrial and aquatic ecosystems (Behera et al., 2013; Wyer et al., 2022).

The most cost-effective method for reducing NH₃ volatilization is applying urease inhibitors to urea. The most widely used urease inhibitor worldwide is N-(n-butyl) thiophosphoric triamide (NBPT) (Cantarella et al., 2018; Modolo et al., 2018; Klimczyk et al., 2021). Silva et al. (2017) found that NBPT-treated urea resulted in a 52 % reduction in NH₃ volatilization compared with conventional urea. However, the stability of NBPT decreases when it is added to organic matrices (Gioacchini et al., 2000), suggesting that combining V with NBPT-treated urea may not be as effective at managing NH₃ volatilization.

Blending urea with ammonium sulfate (AS) is a common practice to supply nitrogen and sulfur (S) to crops. Ammonium sulfate is less prone to substantial ammonia (NH₃) volatilization losses in non-alkaline soils. (Minato et al., 2020; Santos et al., 2020; Corrêa et al., 2021), but losses can be significant in alkaline soils (Rao and Batra, 1983; Schwenke et al., 2014; Powlson and Dawson, 2022). In contrast, NH₃ volatilization losses from urea are significant regardless of soil pH. An alternative to using more concentrated fertilizers and reducing N losses from volatilization is to treat urea with NBPT. However, as can occur with organic mixtures, NBPT, when mixed with inorganic matrices, can also have its efficacy reduced by up to 34 % (Cassim et al., 2024).

Duromide is a novel urease inhibitor (CAS # 2093385-47-6; reaction product of N-(n-butyl) thiophosphoric triamide, urea, and formaldehyde) that acts as a urease inhibitor (Slater, 2020), slowing urea hydrolysis and thereby preventing ammonia volatilization. Duromide is a more complex molecule than NBPT, resulting in greater stability. Recent studies showed that Duromide-treated urea was more effective in reducing NH₃ losses (Cassim et al., 2021) and increasing Brachiaria (Urochloa spp.) yield (Cassimiro et al., 2023) than NBPT-treated urea. Since the Duromide molecule is more stable when compared with NBPT, it can provide greater efficacy in reducing NH₃ volatilization when mixed with V or AS. However, no studies have tested this hypothesis.

Recycling agricultural waste and reducing nutrient losses in sugarcane plantations are key strategies to promote a circular economy and enhance agronomic performance. To the best of our knowledge, no study has yet assessed the effect of the urease inhibitor Duromide on NH₃ volatilization losses when urea is dissolved in V, nor changes in its efficacy during storage when blended with AS. Our hypotheses are that urea + V + Duromide provides the greatest reduction in NH₃ volatilization than urea + NBPT; and that Duromide-treated urea maintains its efficacy in reducing N losses over a longer storage period than NBPT when blended with AS. This study aimed to (i) determine the efficiency of urease inhibitors NBPT and Duromide in reducing N losses through NH₃ volatilization when urea is dissolved in V; and (ii) assess the efficacy of Duromide and NBPT-treated urea blended with AS in reducing NH₃ volatilization after ten months of storage in a warehouse.

MATERIALS AND METHODS

Soil description

The soil (0.00–0.10 m layer) used in the experiments was an Argissolo Vermelho Distrófico arênico of sandy texture, according to the Brazilian Soil Classification System (SiBCS) (Santos et al., 2018), corresponding to an Ultisol in the USDA Soil Taxonomy (Soil Survey Staff, 2014). After visible organic residues had been removed, the soil was sieved to 4 mm and stored at room temperature until incubation. The main soil chemical properties were: pH(H₂O) of 5.2 and cation exchange capacity (CEC) at pH 7.0 of 10.5 cmolc dm^-3 (Tedesco et al., 1995); organic carbon, 6.96 g kg^-1 (measured by dry combustion using a Flash EA 1112, Thermo Finnigan, Milan, Italy); and clay content, 102 g kg^-1 (Claessen, 1997).

Experiment 1: Urea combined with V and urease inhibitor

The first experiment was conducted in a greenhouse using a completely randomized design with five treatments and four replications. Treatments consisted of a control (without N application), urea (Ur) (46 % N), Ur + V, NBPT-treated Ur + V (Ur-NBPT + V), and Duromide-treated Ur + V (Ur-Duromide + V). The pH of V was 4.0. Each experimental unit consisted of a 2.8 L polyethylene pot containing 2.8 kg of soil moistened to 80 % of field capacity, equivalent to 2.5 kg of dry soil. Nitrogen fertilizers were applied by broadcasting at a rate of 0.8 mg cm^-2, equivalent to 80 kg N ha^-1. The N supplied by V was disregarded, as the amendment had low contents of ammonium N (NH₄⁺-N, 68.5 g m^-3) and nitrate N (NO₃^--N, 1.0 g m^-3). The V rate was 30 m³ ha^-1 (Cetesb, 2020).

In this study, Anvol^TM was used as a source of Duromide to treat urea in both experiments. Anvol is a commercially available stabilizer containing two active ingredients: Duromide (the main active ingredient) and NBPT. For simplicity, the treatments in which urea was treated with Anvol will be referred to in this paper as Ur-Duromide.

For treatments including V and urea (with or without inhibitor), the fertilizer formulation was diluted in V and then applied to the soil surface using a directed jet application method, with a total volume of 71 mL per pot (28.4 g water kg^-1 of dry soil). For treatments with inhibitors, granular urea was treated with the urease inhibitors, prior to dissolution in V. For Ur (without V), the fertilizer was diluted in an equivalent volume of distilled water. The treatments were applied to partially moistened soil (60 % of field capacity). The total volume of solution applied per treatment increased the gravimetric soil moisture to 80 % of field capacity. During the experimental period, soil moisture was adjusted to 80 % of field capacity every two days by weighing the pots and adding distilled water as needed. The greenhouse was equipped with temperature and humidity control, with the temperature maintained at 27 °C ± 2.5 °C and the humidity at 70 % ± 5 %.

The quantification of N losses via NH₃ volatilization was carried out within pots using the semi-open static chamber method (Araújo et al., 2009). A 2 L (Ø = 10.0 cm) static chamber was used. A piece of polyurethane sponge (2.5 cm wide and 25 cm long) was placed inside the chamber, suspended on a flask containing 50 mL of 1 mol L⁻¹ sulfuric acid (H₂SO₄) and 4 % (v/v) glycerin. The NH₃-N collection device was placed in the central area of pots immediately after treatments were applied. The evaluation lasted 30 days, and NH₃-N collections were made on days 1, 2, 3, 4, 5, 7, 9, 11, 13, 15, 17, 20, 24, and 30.

The sponge was replaced at each collection with a new sponge containing the acid + glycerin solution. Used sponges and excess acid solution were then transferred to flasks with snap caps containing 20 mL of distilled water. The flasks were shaken on a horizontal shaker at 2 Hz for 30 min to extract ammonium sulfate from the sponge via the reaction between H₂SO₄ and captured NH₃. The NH₃-N content of the extract was determined by the semi-micro Kjeldahl method. Briefly, 20 mL of the solution from snap-capped flasks was distilled by adding 5 mol L^-1 NaOH, followed by titration with 0.01 mol L^-1 H₂SO₄. The measured amounts of NH₃-N were multiplied by a factor of 1.66 to correct the NH₃ capture efficiency of the semi-open static chamber method used (Martins et al., 2021).

Experiment 2: Urea treated with urease inhibitors blended with AS and stored for 10 months

The second experiment was conducted in the laboratory using a completely randomized design with six treatments and four replications. Treatments consisted of five blends of urea or urea + urease inhibitor with AS and a control without N application. The blends used were: urea (Ur) + ammonium sulfate (AS) (Ur + AS); NBPT-treated Ur 72 h before the start of the experiment + AS (Ur-NBPT + AS Fresh), Duromide-treated Ur 72 h before the start of the experiment + AS (Ur-Duromide + AS Fresh), NBPT-treated Ur + AS stored for 10 months (Ur-NBPT + AS Stored), and Duromide-treated Ur + AS stored for 10 months (Ur-Duromide + AS Stored). The proportion of N fertilizers in the blends was 76 % Ur (treated or untreated with inhibitor) and 24 % AS. Urea was treated with NBPT and Duromide prior to blending with ammonium sulfate, as in the process used for commercial formulations. The AS used had a pH of 4.24. The blends were stored for 10 months in 50-kg bags at room temperature in a warehouse before the start of the NH₃ volatilization trial. During storage, the temperature of the blends inside the bags and the room air temperature were measured using T-type sensors coupled to a data logger (CR1000, Campbell Scientific) (Figure 1). Subsamples were collected from stored bags to prepare the replications of Ur-NBPT + AS (Stored) and Ur-Duromide + AS (Stored) treatments and Ur + AS.

Figure 1
Daily temperature data during fertilizer storage. Arrow A indicates the start of fertilizer storage on March 1, 2023. Arrow B indicates the start of the ammonia (NH₃) volatilization experiment on January 8, 2024, after 10 months of fertilizer storage.

Each experimental unit consisted of a 1,750 mL glass jar containing moist soil at 80 % field capacity, equivalent to 300 g of dry soil. Nitrogen fertilizers were applied by broadcasting. The N rate was 1.0 mg cm^-2, equivalent to 100 kg ha^-1. All experimental units were stored in an incubator in the absence of light at constant temperature (28 ± 0.3 °C). Soil used in experiment 2 was the same soil as in experiment 1.

Quantification of N losses via NH₃-N volatilization was carried out in each glass jar using an adapted method proposed by Nômmik (1973). A polyurethane sponge was suspended 0.10 m above the soil to capture the volatilized NH₃-N. The sponge, with a density of 0.017 g cm^-3, diameter of 0.07 m, and a thickness of 1.2 cm, was soaked with 40 mL of a 1 mol L^-1 H₂SO₄ solution containing 4 % glycerin (v/v) immediately before being placed in the glass jar. At each collection time, the sponge was replaced with a new sponge. The evaluation lasted 20 days, and collections were made on days 1, 2, 3, 4, 5, 7, 11, 13, 15, 17, and 20.

The collected sponges were transferred to glass flasks (900 mL) containing 250 mL of distilled water. The glass flasks were shaken in a horizontal shaker at 2 Hz for 30 min to extract ammonium sulfate. The NH₃-N content of the extracts was determined by distillation using the semi-micro Kjeldahl method previously described in Experiment 1.

Statistical analysis

Ammonia volatilization data from both experiments were subjected to non-linear regression analysis using the logistic model (Equation 1). The Akaike information criterion (AIC) was used for model selection. The one with the lowest AIC was chosen (Akaike, 1974). After selecting the model, the data were subjected to non-linear regression using a three-parameter (α, β, and γ) logistic model, as described by Seber and Wild (2003). The model is commonly used to estimate plant growth (Lisboa et al., 2018) and nutrient uptake patterns (Barth et al., 2018). More recently, it has been used to estimate NH₃-N volatilization losses over time (Minato et al., 2020; Oliveira et al., 2024; Lisboa et al., 2024) and nitrous oxide emissions (Besen et al., 2021).

Eq. 1 $\hat{Y}=\frac{\alpha }{1+exp\left[-\right(t-\beta \left)/\gamma \right]}$

in which: $\hat{Y}$ is the amount of N volatilized in the form of NH₃-N (kg ha^-1) at time t (days); α is the maximum cumulative volatilization; β is the day at which the maximum daily loss (MDL) of NH₃-N occurs, corresponding to the inflection point of the curve; and γ is a model parameter used to calculate the MDL of NH₃-N, as shown in equation 2.

Eq. 2 $MDL=\frac{\alpha }{4\gamma }$

Following the estimation of model parameters α, β, and γ, data from days 7 and 30 of Experiment 1 and days 7 and 20 of Experiment 2 were subjected to tests of homogeneity of variances (Bartlett) and normality of errors (Shapiro–Wilk) at p<0.05. Given that the assumptions of data normality were met, data were then subjected to analysis of variance (ANOVA) at p<0.05 (Eisenhart, 1947). Means were compared using the Tukey test at the 5 % significance level in SISVAR.

RESULTS

Ammonia volatilization increased sharply after Ur and Ur + V application in Experiment 1, reaching 48 and 26 %, respectively, three days after fertilizers application (Figure 2). In contrast, NH₃ volatilization was less than 2 % for Ur-NBPT + V and Ur-Duromide + V. Seven days after fertilizer application, cumulative NH₃ losses were 55, 35, 27 and 14 %, respectively, for Ur, Ur + V, Ur-NBPT + V and Ur-Duromide + V (Table 1), which represented a 36, 51 and 75 % reduction in NH₃ losses for Ur + V, Ur-NBPT + V and Ur-Duromide + V compared with Ur.

Figure 2
Cumulative ammonia (NH₃-N) volatilization after broadcast applications of urea (Ur), Ur + vinasse (V), Ur-NBPT + V, and Ur-Duromide + V at a nitrogen (N) rate of 80 kg ha^-1. Vertical bars represent the standard deviation.

Table 1
Cumulative loss of ammonia nitrogen (NH₃-N) by volatilization on the 7th and 30th days of Experiment 1 and the 7th and 20th days of Experiment 2

Treatments	Experiment 1
	%N lost until day 7	%N lost until day 30
Urea (Ur)	55 a	57 a
Ur + vinasse (V)	35 b	40 b
Ur-NBPT + V	27 c	37 c
Ur-Duromide + V	14 d	24 d
CV (%)	1.02	1.05
Tukey's HSD	0.69	0.87
p-value	0.000*	0.000*
Treatments	Experiment 2
	%N lost until day 7	%N lost until day 20
Urea (Ur) + ammonium sulfate (AS)	21 a	22 a
Ur-NBPT + AS (Stored)	10 b	20 b
Ur-Duromide + AS (Stored)	2 d	8 d
Ur-NBPT + AS (Fresh)	6 c	13 c
Ur-Duromide + AS (Fresh)	1 e	7 e
CV (%)	2.33	1.81
Tukey's HSD	0.42	0.56
p-value	0.000*	0.000*

• Means followed by the same lowercase letter do not differ by Tukey test.
• ^*
  Significant at p<0.05. CV (%): Coefficient of variation.

The cumulative losses (α), peak volatilization (β), and MDL of NH₃-N of treatments in Experiment 1 are described in table 2. Considering cumulative loss (α), Ur-Duromide + V was the most effective in reducing NH₃-N volatilization losses compared with urea alone (60 % reduction), followed by Ur-NBPT + V (36 % reduction) and Ur + V (31 % reduction) (Table 2). The efficacy of Ur-Duromide + V was consistent, as this treatment afforded the lowest NH₃-N losses at both 7 and 30 days after N application (Table 1).

Table 2
Non-linear logistic model parameters for cumulative ammonia (NH₃-N) volatilization losses, coefficient of determination (R²) and NH₃-N reduction relative to urea from a sandy soil

Experiment	Treatments	Parameters					Reduction of NH3-N
		α	γ	β	R2	MDL	loss in relation to Ur or Ur + AS
		%		days		%	%
1	Urea (Ur)	57	0.41	2	0.99	35	-
	Ur + vinasse (V)	40	0.72	3	0.96	14	31
	Ur-NBPT + V	37	1.18	6	0.98	8	36
	Ur-Duromide + V	24	1.42	7	0.99	4	60
2	Ur + ammonium sulfate (AS)	22	0.44	2	0.99	13	-
	Ur-NBPT + AS (Stored)	20	1.16	5	0.99	4	9
	Ur-Duromide + AS (Stored)	8	0.82	6	0.99	2	67
	Ur-NBPT + AS (Fresh)	13	0.89	5	0.99	4	43
	Ur-Duromide + AS (Fresh)	7	0.84	6	0.99	2	71

• α: cumulative NH₃-N loss (% of applied N); β: time taken for the cumulative NH₃-N loss to reach 50 % (days); γ: model parameter used to calculate the maximum daily loss of NH₃-N (MDL).

In Experiment 2, NH₃ volatilization also increased sharply for Ur + AS, reaching 18 % only three days after fertilizer application (Figure 3). Seven days after fertilizer application, cumulative NH₃ losses were 21 % for Ur + AS, 10 and 6 % for Ur-NBPT + AS (stored and fresh, respectively), and 2 and 1 % for Ur-Duromide + AS (stored and fresh, respectively) (Table 1). Consequently, NH₃ losses reduction was 52 and 71 % for Ur-NBPT + AS (stored and fresh, respectively) and 90 and 95 % for Ur-Duromide + AS (stored and fresh, respectively).

The model parameters describing the efficacy of treatments in Experiment 2 are described in table 2. The Ur-NBPT + AS (Fresh) and Ur-Duromide + AS (Fresh) reduced N loss via volatilization by 43 and 71 %, respectively, compared with Ur. Therefore, although both inhibitors were effective in reducing NH₃ volatilization when combined with urea shortly before application, Ur-Duromide was more effective. After 10 months of storage, the efficacy of Ur-NBPT + AS (Stored) in reducing NH₃ losses decreased significantly from 43 to 9 % compared with urea (Table 2). In contrast, Ur-Duromide + AS (Stored) maintained its efficacy, achieving a 67 % reduction in NH₃ losses, close to the 71 % obtained with the fresh blend without storage (7 and 8 % N loss at 20 days; Table 2). Overall, Duromide, fresh or stored, was more effective than NBPT in reducing NH₃-N losses, as indicated by the model parameters (Table 2) and cumulative NH₃ losses on days 7 and 20 of the trial (Table 1).

Figure 3
Cumulative ammonia (NH₃-N) volatilization after broadcast applications of urea treated or not with urease inhibitors (NBPT or Duromide) at a nitrogen (N) rate of 100 kg ha^-1. Fresh: unstored fertilizer recently treated with NBPT or Duromide. Stored: fertilizer treated with NBPT or Duromide and stored for 10 months in a warehouse. Vertical bars represent the standard deviation.

DISCUSSION

Effect of urease inhibitors on NH₃ volatilization in urea-vinasse mixture

The greatest losses by NH₃-N volatilization were observed in urea treatments, given its susceptibility to hydrolysis by urease. Hydrolysis consumes H⁺ protons, producing ammonium (NH₄⁺) and bicarbonate (HCO₃^-) (Ferguson et al., 1984; Sunderlage and Cook, 2018). The HCO₃^- ions increase the pH of the soil surrounding urea to 7.5–9.0, reflecting the soil buffering capacity (Pelster et al., 2018). This process favors the transformation of NH₄⁺ to NH₃, which is quickly lost to the atmosphere in gaseous form (Rochette et al., 2009). The hydrolysis reaction is rapid, with the pH increasing by 0.19 pH h^-1 during the first 4.7 h. The soil pH can reach 9.0, 8.8, and 8.4 at 0.5–1 cm, 1–1.5 cm, and 1.5–2 cm, respectively, from the urea granule after 64.7 h (Merl et al., 2024). This pattern explains why the half-maximal loss occurred on the second day after untreated urea application (Table 2).

The reduction in N loss via volatilization with urea dissolution in V indicates that V acidity (pH 4.0) lowered pH at the microsites where liquid fertilizer was applied. Oliveira et al. (2023) demonstrated the potential of combining urea with V to mitigate N-NH₃ losses, with reductions of 50 and 91 % in the dry and rainy seasons, respectively, compared with urea alone. The acidity of V also contributed to reducing NH₃-N loss in experiments carried out by Trivelin et al. (1997, 1998). However, our results show that N losses by volatilization of Ur + V remain high (circa 40 % of N applied). In a study by Otto et al. (2017), NH₃ losses from the urea-V mixture remained high, ranging from 29 to 35 % of the applied N. For this reason, Otto et al. (2017) evaluated other mixtures, such as urea + V + monoammonium phosphate (MAP) + boric acid (H₃BO₃), to enhance N-loss reduction; however, these mixtures showed limited improvement in efficacy compared to urea + V.

The results in Experiment 1 show that V should be combined with Ur treated with urease inhibitors to promote further reductions in NH₃ volatilization. The most effective treatment for reducing NH₃ volatilization was Ur-Duromide + V (Table 2), resulting in agronomically significant additional reductions in NH₃-N losses compared with Ur + V. The lower efficacy of Ur-NBPT + V compared to Ur-Duromide + V can be attributed to the sensitivity of NBPT to the acidity of V. Acidic conditions are known to reduce the chemical stability of NBPT, decreasing its effectiveness as a urease inhibitor (Engel et al., 2013; Soares and Cantarella, 2023). Conversely, Duromide has a different chemical structure from NBPT, comprising radicals that increase molecular stability under adverse conditions, such as low pH (Cassim et al., 2021). It should be mentioned that, in this study, Ur and V solutions were applied uniformly over the entire soil surface to ensure comparability among treatments under controlled conditions. In commercial sugarcane fields, however, V enriched with N fertilizers is typically applied in narrow bands (0.20-0.30 m wide) along the crop rows (Oliveira et al., 2023), thereby increasing the local solution volume and promoting deeper infiltration. Therefore, the magnitude of NH₃ volatilization losses observed under our experimental conditions may be higher than those observed under banded applications, thereby enhancing the apparent efficiency of urease inhibitors. Additional trials should be conducted to validate these results under field conditions.

The greater stability of urea treated with Duromide resulted in the largest β value (7 days) and, consequently, the smallest MDL (4 %) (Table 2). A delay in the time to reach half-maximal N loss (β) can lead to an increase in N use efficiency. The longer urea N is kept in the field, the greater the chance of the nutrient being incorporated into the soil profile by precipitation. This minimizes urea-N losses and improves N availability for plant uptake (Lisboa et al., 2024).

Efficiency of urease inhibitors on NH₃ volatilization when mixed with AS and stored

Both Ur-NBPT + AS (Fresh) and Ur-Duromide + AS (Fresh) were effective in reducing NH₃ volatilization losses compared with Ur + AS. However, Ur-Duromide + AS Fresh achieved a 71 % reduction, whereas Ur-NBPT + AS Fresh resulted in a 43 % reduction. These results highlight Duromide superior efficacy in mitigating NH₃ volatilization losses. For instance, the cumulative NH₃-N loss on day 7 was only 1 % in the Ur-Duromide + AS (Fresh) treatment (Table 1), whereas in the Ur-NBPT + AS (Fresh) treatment, it was 6 %. In the field, the longer the period of urease inhibition, the greater the likelihood that N fertilizer will be incorporated into the soil by rainfall or irrigation water (Holcomb III et al., 2011).

Storage for 10 months affected the efficacy of Ur-NBPT + AS (Stored) in reducing NH₃-N losses by volatilization (9 % compared with conventional Ur + AS; Table 2). Watson et al. (2008) found that the half-life of NBPT-treated urea was approximately 5 months at 25 °C. However, the loss of NBPT efficacy cannot be attributed solely to storage time. Other conditions may accelerate NBPT degradation, such as high storage temperature (Gioacchini et al., 2000; Watson et al., 2008) and soil acidity (Sunderlage and Cook, 2018; Soares and Cantarella, 2023). One of the main causes of NBPT molecule degradation is contact with acidity. For example, Engel et al. (2015) showed that the NBPT half-life is longer in alkaline soils, with 0.07, 0.59, 2.70, and 3.43 days at pH 5.1, 6.1, 7.6, and 8.2, respectively. Although the study by Engel and co-workers addressed soil acidity, it provides evidence that mixing urea treated with NBPT with acidic fertilizers such as AS may also initiate NBPT degradation before the fertilizer is even applied to the soil. Fonseca et al. (2023) observed that the degradation of NBPT was drastically accelerated when stored in mixtures of NBPT-treated urea and phosphate fertilizers. These authors attributed the degradation of NBPT to the high free acidity of phosphate fertilizers, concluding that the mixture of urea with NBPT and P-fertilizers is incompatible. In contrast to the NBPT-treated urea blend, results from this study showed that the Ur-Duromide + AS (Stored) maintained its efficacy in reducing NH₃ losses regardless of storage time, demonstrating that it is a more stable urease inhibitor and that retains its efficacy even when blended with acidic fertilizers.

The urease inhibitor NBPT has been on the market for almost 30 years. Its efficacy in reducing NH₃ volatilization is well documented in review and meta-analysis studies (Pan et al., 2016; Silva et al., 2017; Cantarella et al., 2018; Klimczyk et al., 2021). However, limitations, such as molecular degradation when blended with acidic fertilizers, reduce NBPT efficacy. Therefore, there is space for new technologies that can improve urease inhibition and further reduce volatilization losses. Duromide represents a next generation of enhanced-efficiency urease inhibitors, consisting of a new substance with superior stability, promoting reduction of losses by NH₃ volatilization, when blended with acidic fertilizers such as AS or when combined with acidic organo-mineral fertilizers like V. This greater stability and efficiency of Duromide should be further investigated, considering different soil pH levels and temperature, to evaluate its performance in different agricultural scenarios.

CONCLUSIONS

The addition of urea to vinasse (V) reduced nitrogen (N) losses via volatilization, but remained high, about 40 % of applied N. Our results show that mixing urea with V is not sufficient to significantly reduce N losses through volatilization, and that urease inhibitors, especially Duromide, can provide additional reductions in ammonia (NH₃) volatilization, even 30 days after fertilizer application. These results highlight the greater Duromide stability compared to N-(n-butyl) thiophosphoric triamide (NBPT) under acidic conditions, such as when mixed with V (pH 4.0). This conclusion was supported by lower NH₃ losses observed when urea was blended with ammonium sulfate (AS) (pH = 4.24) and treated with Duromide rather than NBPT. Furthermore, Duromide-treated urea blended with AS maintained its efficacy in reducing NH₃ losses after 10 months of storage, whereas NBPT under the same conditions showed a 4-fold decrease in efficacy. The greater stability of Duromide compared to NBPT allows for superior fertilizer efficacy for farmers, flexibility in the storage and transportation of urea treated with urease inhibitors, when blended with acidic fertilizers such as AS or when combined with acidic organo-mineral fertilizers like V. When applied to the soil, the prolonged urease inhibition efficacy of the Duromide promotes a greater reduction in NH₃ volatilization, thereby enhancing N use efficiency in crops.

ACKNOWLEDGEMENTS

We thank the Federal University of Santa Maria (UFSM) and Koch Agronomic Services for their participation in this research. We thank LABCEN’s laboratory staff and students who participated in the collection and processing of data.

• How to cite:
  Pilecco GE, Pujol SB, Cassim BMAR, Alves DA, Dias MR, Giacomini SJ. Duromide is an effective urease inhibitor when combined with vinasse and blended with ammonium sulphate. Rev Bras Cienc Solo. 2026;50:e0250093. https://doi.org/10.36783/18069657rbcs20250093
• FUNDING
  This study was supported by Carbon and Nitrogen Biogeochemistry Laboratory (LABCEN), Federal University of Santa Maria, Brazil.

DATA AVAILABILITY

The data will be provided upon request.

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Edited by

• Editor:
  José Miguel Reichert https://orcid.org/0000-0001-9943-2898 and Marco Aurélio Carbone Carneiro https://orcid.org/0000-0003-4349-3071

Publication Dates

• Publication in this collection
  27 July 2026
• Date of issue
  2026

History

• Received
  30 Apr 2025
• Accepted
  02 Jan 2025