Technofunctional assessment and proximate composition of Lupinus mutabilis flours from three surandine ecotypes of Peru

Avaliação tecnofuncional e composição proximal de farinhas de Lupinus mutabilis de três ecótipos sul-andinos do Peru

Abstract

In this study, the technofunctional properties, proximate composition, and mineral content of three tarwi ecotypes white, moro, and punto negro were analyzed. The seeds were cultivated in Andahuaylas, a surandine zone of Peru. The flours obtained (125 μm) showed differences in particle size: the punto negro ecotype had a higher proportion of coarse particles (Dx 90), while the white and moro ecotypes exhibited finer distributions. Regarding color, the white ecotype showed the highest lightness and whiteness index, followed by moro, whereas the punto negro ecotype exhibited darker and reddish tones, associated with differences in composition. Water absorption capacity increased by approximately 30% between 60 and 90 °C in all ecotypes. Both water solubility index and swelling power also increased with temperature, with the white ecotype showing higher solubility at high temperatures and moro exhibiting greater swelling capacity at initial temperatures (60°C). Additionally, the moro ecotype presented higher hygroscopicity and water holding capacity, while oil retention and foaming properties were similar among ecotypes (P>0.05). Water activity was low in all cases (0.34–0.37 g/100 g), suggesting good microbiological stability. Regarding proximate composition, the moro ecotype stood out for its higher protein content (56.28 ± 0.02 g/100 g) and mineral content (ash and calcium), while the white ecotype had higher lipid content (26.53 ± 0.02 g/100 g) and minerals such as iron and zinc. The punto negro ecotype showed higher carbohydrate content (16.58 ± 0.15 g/100 g) and phosphorus. These results indicate that each ecotype possesses distinct attributes that determine its potential application in the food industry.

Keywords:
tarwi ecotypes; flours; proximate composition; technofunctional property

Resumo

Neste estudo, foram analisadas as propriedades tecnofuncionais, a composição proximal e o teor de minerais de três ecótipos de tarwi – branco, moro e ponto negro –, cujas sementes foram cultivadas em Andahuaylas, região sul-andina do Peru. As farinhas obtidas (125 μm) apresentaram diferenças no tamanho das partículas: o ecótipo ponto negro apresentou maior proporção de partículas grossas (Dx 90), enquanto os ecótipos branco e moro exibiram distribuições mais finas. Quanto à cor, o ecótipo branco apresentou maior luminosidade e índice de brancura, seguido pelo moro, enquanto o ponto negro exibiu tonalidades mais escuras e avermelhadas, associadas a diferenças em sua composição. A capacidade de absorção de água aumentou aproximadamente 30% entre 60 °C e 90 °C em todos os ecótipos. O índice de solubilidade em água e o poder de inchaço também aumentaram com a temperatura, destacando-se o ecótipo branco por sua maior solubilidade em altas temperaturas e o moro por sua maior capacidade de inchaço em temperaturas iniciais (60 °C). Além disso, o ecótipo moro apresentou maior higroscopicidade e capacidade de retenção de água, enquanto a retenção de óleo e as propriedades espumantes foram semelhantes entre os ecótipos (P>0,05). A atividade de água foi baixa em todos os casos (0,34–0,37 g/100 g), sugerindo boa estabilidade microbiológica. Em relação à composição proximal, o ecótipo moro destacou-se pelo maior teor de proteína (56,28 ± 0,02 g/100 g) e minerais (cinzas e cálcio), enquanto o ecótipo branco apresentou maior teor lipídico (26,53 ± 0,02 g/100 g) e de minerais como ferro e zinco. O ecótipo ponto negro apresentou maior teor de carboidratos (16,58 ± 0,15 g/100 g) e fósforo. Esses resultados indicam que cada ecótipo possui atributos distintos que determinam seu potencial de aplicação na indústria alimentícia.

Palavras-chave:
ecótipos de tarwi; farinhas; composição proximal; propriedades tecnofuncionais

1. Introduction

Countries such as Peru, Bolivia, and Ecuador represent important centers of diversity for Lupinus mutabilis Sweet (tarwi), an Andean legume recognized for its exceptional nutritional value and adaptation to high-altitude environments (Gulisano et al., 2019). Several ecotypes contain high levels of protein (up to 50%), lipids (18–25%), and essential minerals, including calcium, phosphorus, iron, and zinc, making tarwi a promising crop for enhancing the nutritional quality of plant-based foods and contributing to food security, particularly in vulnerable populations (Choquetico-Iquiapaza et al., 2024; Carhuallanqui et al., 2022).

The increasing demand for sustainable plant-derived ingredients has stimulated interest in tarwi as a valuable source of proteins and bioactive compounds for the development of functional foods. Processing tarwi into flour broadens its potential application in bakery products, snacks, protein-enriched foods, and other value-added formulations (Taş et al., 2022). However, the nutritional composition and techno-functional performance of tarwi flour are strongly influenced by both genetic background and processing conditions, including debittering, soaking, cooking, and drying (Ramos et al., 2020). The remarkable genetic diversity of L. mutabilis has given rise to numerous ecotypes adapted to contrasting agroecological environments, resulting in considerable variation in seed composition (Cortés-Avendaño et al., 2020). Differences in protein, lipid, fiber, and carbohydrate contents directly influence hydration properties, oil retention, foaming behavior, gel formation, and textural characteristics, thereby determining the suitability of each ecotype for specific food applications (Guan et al., 2023; Berru et al., 2021). Furthermore, water activity is a critical parameter governing the microbiological stability and shelf life of flour-based products (Rockland and Beuchat, 2017).

Despite increasing scientific interest in L. mutabilis, comprehensive information on the physicochemical, nutritional, and techno-functional characteristics of native ecotypes from the southern Peruvian Andes remains limited. In particular, the white, moro, and punto negro ecotypes from Andahuaylas have been insufficiently characterized, limiting the understanding of the influence of genetic variability on nutrient composition, molecular organization, antioxidant capacity, and functional performance. This knowledge gap hinders the identification of ecotypes with specific technological advantages and restricts their valorization as functional ingredients for food applications (Chalampuente-Flores et al., 2021).

Accordingly, a comprehensive characterization of these native ecotypes is essential to elucidate the relationships between chemical composition and techno-functional performance. Therefore, the aim of this study was to characterize flours obtained from the white, moro, and punto negro tarwi ecotypes cultivated in Andahuaylas, Peru, by evaluating their proximate composition, mineral profile, and techno-functional properties, including water activity, hygroscopicity, water-holding capacity, oil-holding capacity, color, foaming capacity, and foam stability.

2. Material and Methods

2.1. Material

The white, moro, and punto negro tarwi (Lupinus mutabilis Sweet) ecotypes were obtained from local producers through the traditional market in Andahuaylas Province, Apurímac, Peru, and corresponded to the 2023 harvest season. All three ecotypes originated from the same geographical region and were cultivated under conventional agronomic practices in Andahuaylas Province (13°10′–13°50′ S, 73°10′–73°50′ W), located in the south-central Peruvian Andes. Following collection, the seeds were manually inspected to verify their morphological characteristics and classified according to ecotype. The samples were subsequently stored in airtight containers at ambient temperature (20 ± 5 °C) until processing and analysis.

2.2. Flour production process

Tarwi seeds were manually sorted to remove foreign material and damaged kernels before processing. Debittering was carried out according to the method described by Cortés-Avendaño et al. (2020), with slight modifications. For each ecotype, 1 kg of seeds was soaked in water at a seed-to-water ratio of 1:6 (w/v) for 12 h at room temperature. The hydrated seeds were then boiled for 1 h at a seed-to-water ratio of 1:3 (w/v), replacing the cooking water every 30 min to enhance alkaloid leaching. Subsequently, the seeds were washed under continuous running water for 5 days to further promote alkaloid removal, followed by drying at 40 °C for 24 h. The dried seeds were milled using a disc mill (MQMP2040, Maqorito, Lima, Peru) and sieved to obtain flour particles <125 μm in accordance with ASTM E11 specifications. Material retained on the sieve was re-milled and sieved repeatedly until the desired particle size was attained. The resulting flours were sealed in airtight polyethylene bags and stored at ambient temperature (20 ± 5 °C) until physicochemical, nutritional, and technofunctional characterization.

2.3. Analytical determinations

2.3.1. Particle size and polydispersity analysis

Particle size distribution was determined by laser diffraction using a Mastersizer 3000 analyzer (Malvern Instruments, Worcestershire, UK). Flour samples were dispersed in isopropanol as the dispersing medium until the recommended obscuration level was achieved. Particle size was characterized by the volume-based percentiles Dx (10), Dx (50), and Dx (90), corresponding to the particle diameters below which 10%, 50%, and 90% of the sample volume is distributed, respectively. Particle size homogeneity was assessed using the span index, calculated according to Equation (1).

$Span=\frac{D\left(0.9\right)-D\left(0.1\right)}{D\left(0.5\right)}$ (1)

Where: D(0.1), D(0.5), and D(0.9) correspond to the particle diameters at 10%, 50%, and 90% of the cumulative particle size distribution, respectively.

2.3.2. Color measurement

Color parameters were determined according to the method described by Chiumarelli et al. (2011) and Barrial-Lujan et al. (2025a), with slight modifications. The CIELAB color coordinates (L, a, and b*) were measured using a CR-5 colorimeter (Konica Minolta Sensing Inc., Osaka, Japan). The chroma (C*), hue angle (h*), and whiteness index (WI) were subsequently calculated according to Equations (2), (3), and (4), respectively.

$h^{*}=ta{n}^{-1}\left(\frac{b^{*}}{a^{*}}\right)$ (2)

$C^{*}=\sqrt{a^{*2}+b^{*2}}$ (3)

$WI=100-\sqrt{{(100-L^{*})}^2+a^{*2}+b^{*2}}$ (4)

2.3.3. Water activity

Water activity was determined according to the method described by Cerezal Mezquita et al. (2011), with slight modifications. Measurements were performed using a HygroPalm23-AW portable water activity meter (Rotronic, Bassersdorf, Switzerland). The instrument was calibrated prior to analysis. Approximately 5 g of flour was weighed into a disposable sample cup, and the probe was inserted to record the water activity value after equilibrium was reached.

2.3.4. Water absorption capacity

Water absorption capacity was determined according to the methods described by Mishra and Rai (2006) and Gani et al. (2014). Flour suspensions (2.5 g/100 g, w/v) were incubated in a water bath at 60, 70, 80, and 90 °C for 30 min and subsequently centrifuged at 5000 rpm for 30 min at 4 °C. The water absorption capacity was calculated using Equation (5).

$WAC=\frac{W_2-W_1}{W_0}$ (5)

Where: WAC = Water absorption capacity; Wo = Weight of tarwi flour; W1 = Weight of the empty centrifuge tube; W2 = Weight of the tube after centrifugation and decantation.

2.3.5. Solubility index and swelling power

The methodology of Gani et al. (2014) was used to determine swelling power and solubility, with modifications. A 0.200 g sample of flour was weighed and dispersed in 12 mL of distilled water using vortex agitation (2000 rpm). The suspensions were incubated in a water bath at 60, 70, 80, and 90 °C for 30 min and then centrifuged at 5000 rpm for 30 min at 4 °C. The obtained supernatant was dried at 90 °C for 24 h for quantification. The parameters were calculated using Equations 6 and 7.

$WSI\left(\%\right)=\frac{W_2}{W_0}$ (6)

$SP=\frac{W_1}{W_0-W_2}$ (7)

Where: WSI = Water solubility index; SP = Swelling power; Wo = Weight of tarwi flour; W1 = Weight of the sample with water (gel); W2 = Weight of the dried supernatant.

Water and oil retention capacity, hygroscopicity, as well as foaming capacity and its respective stability, were evaluated according to the methodology described by Martínez-Navarrete et al. (2023).

2.3.6. Proximate composition and Minerals

The proximate composition of tarwi flour from the three ecotypes was determined using official AOAC methods. Moisture content was determined by the oven-drying method (AOAC, 2005) using a drying oven (SLW-115STD, USA). Protein content was determined by the Kjeldahl method (AOAC, 1990), whereas fat content was measured by Soxhlet extraction (AOAC, 1990). Total dietary fiber was determined according to AOAC Method 985.29, as described by McCleary (2023). Ash content was determined gravimetrically (AOAC, 1990), and carbohydrate content was calculated by difference using the following equation: 100 − (% protein + % fat + % ash + % dietary fiber + % moisture) (Owheruo et al., 2024).

Phosphorus, calcium, iron, and zinc contents were determined by inductively coupled plasma optical emission spectrometry (ICP-OES) according to the method described by Dell’Aquila et al. (2020) and ISO 11885:2007 (ISO, 2021). Instrumental operating conditions were: RF power, 1.2 kW; plasma gas flow, 10 L min⁻¹; auxiliary gas flow, 0.6 L min⁻¹; nebulizer gas flow, 0.45 L min⁻¹; sample uptake rate, 0.5 mL min⁻¹; using a concentric nebulizer and a cyclonic spray chamber. Emission wavelengths were 213.618 nm for P, 373.690 nm for Ca, 324.754 nm for Fe, and 213.857 nm for Zn.

2.4. Statistical analysis

All determinations were performed in triplicate, and results are presented as mean ± standard deviation. The experimental data were subjected to analysis of variance (ANOVA) to evaluate the effects of the experimental factors and their interactions. When significant differences were detected, means were compared using Fisher’s least significant difference (LSD) test at a significance level of p < 0.05. All statistical analyses were performed using Statgraphics Centurion XVIII (StarPoint Technologies, Inc., The Plains, Virginia, USA). Figures were generated using OriginPro 8.0 (OriginLab Corporation, Northampton, Massachusetts, USA).

3. Results

3.1. Particle size of tarwi

Particle size distribution is a critical parameter governing the physicochemical, functional, and technological behavior of particulate food materials, as it directly influences hydration kinetics, specific surface area, flowability, mixing performance, and particle interactions within food systems (Olakanmi et al., 2024). Therefore, it is widely recognized as a key indicator of flour quality and processing performance. Figure 1 illustrates the particle size distribution of tarwi flours from the white, moro, and punto negro ecotypes, characterized by the percentile diameters Dx(10), Dx(50), and Dx(90). These percentiles represent the particle diameters below which 10%, 50%, and 90% of the cumulative sample volume is contained, respectively, providing valuable information on the particle size heterogeneity and structural characteristics of the milled flours.

Figure 1
Granulometric distribution of tarwi flours. Note. Dx (10) represents the diameter below which 10% of the total particle volume is found; Dx (50) corresponds to the median diameter or 50th percentile of the distribution; and Dx (90) indicates the size below which 90% of the analyzed particles are situated. Equal superscript letters indicate no significant difference, as evaluated by the 5% LSD test.

The particle size distribution analysis revealed significant genotype-dependent differences among tarwi ecotypes (p < 0.05), indicating distinct fragmentation behaviors during milling and differences in seed matrix architecture. The fine particle fraction, represented by Dx(10), ranged from 4.60 ± 0.07 to 8.08 ± 0.61 µm. The white and moro ecotypes showed a higher proportion of fine particles (7%), whereas the punto negro ecotype contained only 2%, suggesting greater resistance to particle fragmentation (Figure 1). More pronounced differences were observed for the median particle diameter (Dx(50)), which ranged from 17.47 ± 0.73 to 69.50 ± 1.28 µm. The white ecotype exhibited the largest median particle size and the highest cumulative particle proportion (28%), indicating a narrower particle size distribution around the median diameter than the punto negro ecotype (p < 0.05). The greatest variation among ecotypes was observed for Dx (90), with the punto negro ecotype reaching 323.67 ± 2.60 µm and exhibiting the highest proportion of coarse particles (81%) (p < 0.05). This shift toward larger particle sizes suggests greater resistance of the seed matrix to mechanical size reduction during milling. Such behavior may be attributed to differences in the composition and structural organization of storage macromolecules, including proteins, lipids, and cell wall polysaccharides, which influence fracture mechanics and breakage patterns. Consequently, the predominance of coarse particles in the punto negro ecotype is expected to affect specific surface area, hydration behavior, and technofunctional properties, demonstrating that genetic variability plays a key role in determining the physical characteristics of tarwi flours.

3.2. Color of tarwi flour

Table 1 shows the color parameters of tarwi flours from the three ecotypes. Significant differences (p < 0.05) were observed in all evaluated parameters: lightness (L*), color coordinates (a* and b*), chroma (C*), hue angle (h*), and whiteness index (WI), suggesting compositional and structural variations inherent to the plant material.

Table 1
Color parameters of tarwi flours.

Parameters	White ecotype	Moro ecotype	Punto negro ecotype
	x̅ ± SD	x̅ ± SD	x̅ ± SD
L*	78.55 ± 0.050c	77.23 ± 0.361b	74.72 ± 0.199a
a*	-0.048 ± 0.0028a	-0.383 ± 0.0057b	0.467 ± 0.032c
b*	25.62 ± 0.63b	20.27 ± 0.066a	25.54 ± 0.226b
C*	25.62 ± 0.63b	20.28 ± 0.07a	25.54 ± 0.23b
h*	90.11 ± 0.0a	91.08 ± 0.02b	268.95 ± 0.07c
WI	69.51 ± 0.31c	66.59 ± 0.46b	64.06 ± 0.30a

• Note. 𝑥̅: mean, SD: standard deviation, L* represents lightness and ranges from 0 for black to 100 for white. The a* and b* values indicate color tones. C is chroma or saturation. h* is the hue angle. WI is the whiteness index, and aw is water activity. Equal superscript letters indicate no significant difference, as evaluated by the 5% LSD test.

The white ecotype showed the highest lightness, followed by the moro ecotype, while the punto negroecotype exhibited the lowest lightness (p < 0.05) (Table 1). The difference between the white and punto negroecotypes represents a decrease of approximately 4.7% in lightness, whereas the moro ecotype showed an intermediate reduction of 1.7% compared to the white. This indicates that the white ecotype is significantly lighter than the other two, with the punto negroecotype being the darkest. Additionally, the a* values for the white ecotype were close to zero (-0.048), indicating a neutral tone. In contrast, the moro ecotype had a significantly negative value, suggesting a more pronounced greenish hue, while the punto negroecotype showed a positive value, indicating a reddish tone. The percentage difference between the positive value of the punto negroand the negative value of the moro is significant, with an approximate change of 222% considering the absolute change in the a* value. This is likely related to the differing phenolic compound content in the samples. Regarding the b* parameter, the white and punto negroecotypes exhibited similar values, while the moro ecotype had a significantly lower value (Table 1). This implies that the moro is approximately 20.9% less yellow than the white and punto negroecotypes. These results indicate that the moro ecotype has a less warm or less yellowish tone compared to the other two. Chroma, representing color saturation or intensity, followed a similar trend to the b* coordinate, with values of 25.62 ± 0.63 for white, 25.54 ± 0.23 for black spot, and a reduced value of 20.28 ± 0.07 for moro. The moro ecotype exhibited a color saturation approximately 20.9% lower than the white, confirming that its color is less intense. The hue angle (h*) was similar for the white and moro ecotypes, indicating tones close to 90°, typical of yellowish-green colors. In contrast, the punto negro ecotype exhibited a significantly higher hue angle (>268°), indicating a distinct color hue compared with the other ecotypes. The whiteness index (WI) followed the same trend as L*, with the white ecotype presenting the highest value, followed by moro and punto negro (p < 0.05). The higher WI of the white ecotype is consistent with its greater lightness, confirming its brighter appearance relative to the other flours.

3.3. Technofunctional properties of tarwi flour

Table 2 shows that water absorption capacity differed significantly among ecotypes only at 60 °C (p < 0.05), whereas the water solubility index and swelling power exhibited significant differences at all evaluated temperatures.

Table 2
Values of water absorption capacity (WAC), water solubility index (WSI), and swelling power (SP).

Tarwi Flours	60 °C	70 °C	80 °C	90 °C
		WAC
	x̅ ± S	x̅ ± S	x̅ ± S	x̅ ± S
WTF	3.57 ± 0.03ab	3.86 ± 0.07a	4.39 ± 0.04a	4.63 ± 0.05a
MTF	3.70 ± 0.08b	3.80 ± 0.19a	4.42 ± 0.16a	4.53 ± 0.03a
PNTF	3.47 ± 0.07a	3.72 ± 0.12a	4.29 ± 0.13a	4.57 ± 0.08a
		WSI
WTF	5.50 ± 0.19a	7.19 ± 0.07a	8.53 ± 0.25a	9.35 ± 0.20a
MTF	5.72 ± 0.19a	7.21 ± 0.18a	8.07 ± 0.21b	8.99 ± 0.11a
PNTF	5.69 ± 0.10a	6.68 ± 0.19b	7.67 ± 0.06c	8.53 ± 0.23b
		SP
WTF	3.78 ± 0.034ab	4.15 ± 0.080a	4.81 ± 0.06a	5.11 ± 0.06a
MTF	3.92 ± 0.097b	3.99 ± 0.197a	4.81 ± 0.14a	4.98 ± 0.03b
PNTF	3.68 ± 0.070a	4.10 ± 0.122a	4.64 ± 0.18a	4.99 ± 0.09ab

• Note: 𝑥̅: mean, SD: standard deviation, WTF: white tarwi flour, MTF: moro tarwi flour, PNTF: Punto negro tarwi flour. Equal superscript letters indicate no significant difference, as evaluated by the 5% LSD test.

The progressive increase in water absorption capacity (WAC) from 60 to 90 °C, reaching approximately 30%, highlights the strong influence of temperature on the hydration behavior of tarwi flours. Among the evaluated ecotypes, MTF consistently exhibited the highest WAC values, exceeding WTF by 6.3% and PNTF by 6.9% at 60 °C, while maintaining the highest value at 90 °C (4.63 g/g), with differences of 5.5% and 1.3%, respectively. The water solubility index (WSI) increased with temperature in all ecotypes, reflecting progressive solubilization of structural components. WTF exhibited the greatest increase (70.0%), followed by MTF (57.2%) and PNTF (49.9%). At 90 °C, WTF reached the highest WSI value (9.35), exceeding MTF and PNTF by 4.0% and 9.6%, respectively, indicating greater susceptibility of its matrix to thermal solubilization. Similarly, swelling power increased with temperature, confirming enhanced matrix hydration and expansion. Although MTF showed the highest swelling power at 60 °C, WTF reached the highest value at 90 °C (5.11), exceeding MTF and PNTF by 2.6% and 2.4%, respectively. The overall increase in swelling power was 35.2% for WTF, 35.6% for PNTF, and 27.0% for MTF, suggesting that the latter possessed a more structurally organized matrix with lower sensitivity to thermal swelling. Overall, these results indicate that genetic variability influences water–matrix interactions and the thermo-functional behavior of tarwi flours.

Table 3 summarizes the techno-functional properties of tarwi flours from the PNTF), white (WTF), and moro (MTF) ecotypes. Significant differences (p < 0.05) were observed in water activity, hygroscopicity, and water-holding capacity, indicating that genetic variability influences water–matrix interactions and moisture sorption behavior. In contrast, foaming capacity and foam stability showed no significant differences among ecotypes (p > 0.05), suggesting similar interfacial properties and protein functionality under the evaluated conditions.

Table 3
Values of water activity, hygroscopicity, oil retention capacity, water retention capacity, foaming capacity, and foam stability.

Properties en g/100g bs	PNTF	WTF	MTF
	x̅ ± SD	x̅ ± SD	x̅ ± SD
water activity	0.37 ± 0.003c	0.36 ± 0.006b	0.34 ± 0.007a
hygroscopicity	1.52 ± 0.09a	1.39 ± 0.09a	1.86 ± 1.14b
oil retention capacity	0.24 ± 0.01a	0.26 ± 0.01ab	0.26 ± 0.09 b
water retention capacity	2.59 ± 0.06a	2.74 ± 0.06b	2.85 ± 0.07b
foaming capacity	24.17 ± 0.91a	22.91 ± 1.02a	21.92 ± 3.32 a
foam stability	99.00 ± 0.31a	98.73 ± 0.07a	99.70 ± 0.14a

• Note: 𝑥̅: mean, SD: standard deviation, WTF: white tarwi flour, MTF: moro tarwi flour, PNTF: Punto negro tarwi flour. Equal superscript letters indicate no significant difference, as evaluated by the 5% LSD test.

Water activity (aw) ranged from 0.34 to 0.37, with PNTF exhibiting the highest value (0.37) and MTF the lowest (0.34), representing an 8.1% reduction between ecotypes (p < 0.05). The lower aw suggests greater storage stability and reduced susceptibility to microbial growth and deteriorative reactions. In contrast, MTF exhibited the highest hygroscopicity, exceeding WTF and PNTF by 33.8% and 22.4%, respectively (p < 0.05), indicating a greater capacity to absorb atmospheric moisture, probably due to a higher abundance of hydrophilic constituents, such as proteins and dietary fiber. Oil retention capacity (ORC) showed limited variation among ecotypes (0.24–0.26 g/g), with WTF and MTF exhibiting values approximately 8.3% higher than PNTF. However, water retention capacity (WRC) was significantly higher in MTF, exceeding PNTF by 10% and WTF by 4% (p < 0.05), reflecting a greater affinity of its matrix for water and a higher potential for hydration. Foaming capacity ranged from 21.92% to 24.17%, whereas foam stability remained approximately 99% for all ecotype, with no significant differences (p > 0.05). These findings indicate that, despite compositional differences among ecotypes, the proteins retained comparable interfacial properties, enabling efficient foam formation and the maintenance of highly stable foam structures throughout the 60 s evaluation period.

3.4. Proximate composition and minerals of tarwi flour

Table 4 shows significant differences (p < 0.05) in the macronutrients of tarwi flours, confirming the effect of genotype and variety on their composition. Likewise, significant differences (p < 0.05) are observed in the micronutrients of PNTF, WTF, and MTF, indicating that the variety plays a decisive role in mineral accumulation.

Table 4
Proximate composition and mineral content of tarwi flour.

Macronutrientes en g/100g bs	PNTF	WTF	MTF
	x̅ ± S	x̅ ± S	x̅ ± S
Fat	26.03 ± 0.02b	26.53 ± 0.02c	25.33 ± 0.03a
Ash	3.21 ± 0.02a	3.42 ± 0.02b	3.85 ± 0.02c
Dietary fiber	5.93 ± 0.06c	5.70 ± 0.08b	4.31 ± 0.01a
Carbohydrates	17.22 ± 0.06c	16.58 ± 0.15b	14.61 ± 0.10a
Protein	53.60 ± 0.03a	53.53 ± 0.02b	56.28 ± 0.02c
Minerals in mg/100 g
Phosphorus	410.35 ± 0.00c	370.85 ± 0.00b	325.85 ± 0.00a
Calcium	552.95 ± 0.00b	502.29± 0.00a	560.32 ± 0.00c
Iron	7.21 ± 0.00b	8.93 ± 0.00c	7.10 ± 0.00a
Zinc	4.96 ± 0.00b	5.12 ± 0.00c	4.60 ± 0.00a

• Note: 𝑥̅: mean, SD: standard deviation, WTF: white tarwi flour, MTF: moro tarwi flour, PNTF: Punto negro tarwi flour. Equal superscript letters indicate no significant difference, as evaluated by the 5% LSD test.

Moisture content ranged from 5.73 to 6.91 g/100 g, with MTF exhibiting the highest value and WTF the lowest. The 20.6% higher moisture content in MTF suggests a greater abundance of hydrophilic constituents and a matrix organization capable of retaining water through hydrogen bonding. This behavior is consistent with its elevated protein content, as polar amino acid residues increase the number of water-binding sites within the seed matrix. Regarding lipids, WTF exhibited the highest content (26.53 g/100 g), exceeding MTF and PNTF by 1.92% and 1.90%, respectively. This finding suggests a greater allocation of carbon toward lipid reserve accumulation during seed maturation. Increased lipid levels may enhance hydrophobic interactions and modify protein–lipid associations, influencing flour functionality through effects on lubrication, texture development, and sensory attributes in food systems. Protein content was significantly higher in MTF (56.28 g/100 g), surpassing PNTF and WTF by 4.94% and 5.13%, respectively (p < 0.05). This enrichment indicates a greater allocation of assimilated nitrogen toward the synthesis and deposition of storage proteins, particularly conglutins, the predominant protein fraction in Lupinus mutabilis. Such accumulation may contribute to the higher hydration capacity and more compact microstructure observed in this ecotype, reflecting stronger intermolecular associations within the seed matrix. Ash content was also highest in MTF (3.85%), representing increases of 19.94% and 12.57% relative to PNTF and WTF, respectively. This result suggests enhanced mineral accumulation, likely associated with the metabolic requirements of protein biosynthesis and reserve formation. In contrast, PNTF exhibited the highest dietary fiber content (5.93 g/100 g), followed by WTF (5.70 g/100 g), whereas MTF presented the lowest value (4.31 g/100 g), corresponding to reductions of approximately 25–27%. This pattern indicates a greater allocation of carbon toward structural polysaccharides and cell wall components in PNTF and WTF, which may contribute to increased matrix rigidity and influence particle organization and hydration behavior. Similarly, carbohydrate content was highest in PNTF (17.22 g/100 g), exceeding WTF and MTF by 3.72% and 15.15%, respectively. Together with the elevated fiber content, this result suggests a greater investment in carbon rich reserve and structural compounds, which may affect swelling behavior, dispersibility, and water matrix interactions.

Marked differences were also observed in the mineral composition. PNTF contained the highest phosphorus content (410.35 mg/100 g), exceeding MTF and WTF by 25.96% and 10.65%, respectively. This behavior may be associated with a greater accumulation of phosphorus containing storage compounds, particularly phytates, which function as mineral and energy reserves during germination. In contrast, MTF exhibited the highest calcium content (560.32 mg/100 g), surpassing WTF by 11.55%. Calcium may contribute to the stabilization of cellular structures and macromolecular interactions, potentially influencing matrix organization and thermal behavior. Finally, WTF showed the highest concentrations of iron (8.93 mg/100 g) and zinc (5.12 mg/100 g), indicating a differentiated mineral accumulation pattern among ecotypes. Collectively, these findings suggest that genotypic variability is closely associated with differences in mineral accumulation, contributing to the nutritional quality and technofunctional properties of tarwi flours.

4. Discussion

The particle size distribution differed markedly among tarwi ecotypes, particularly for Dx(90), where PNTF exhibited the highest proportion of coarse particles (81%). This predominance of larger particles is consistent whit its lower hydration and solubilization behavior, whereas the white and moro ecotypes, characterized by finer particles, provided a greater surface area for water matrix interactions. Similar effects of particle size reduction on hydration, rheological properties, and technological performance have been reported in legume flours (Olakanmi et al., 2024; Cai et al., 2023; Puppo and Ferrero, 2024). Consistent with these observations, water absorption capacity (WAC) increased by approximately 30% between 60 and 90 °C in all ecotypes (Table 2), reflecting enhanced molecular mobility and greater exposure of hydrophilic groups during heating (Keskin et al., 2022). MTF consistently exhibited the highest WAC values, likely due to its higher protein content (56.28%), which provides additional water-binding sites. Likewise, the water solubility index (WSI) increased with temperature, with WTF showing the greatest increase (70%), followed by MTF (57.2%) and PNTF (49.9%), indicating greater thermal susceptibility of the WTF matrix. Swelling power also increased progressively, reaching a maximum value of 5.11 g/g in the WTF ecotype, a value higher than those reported for lentil, quinoa, and chickpea flours (Badia-Olmos et al., 2023), highlighting the remarkable hydration and expansion capacity of tarwi flour.

The technofunctional properties were consistent with the compositional differences observed among the tarwi ecotypes. Hygroscopicity ranged from 1.39 to 1.86 g/100g, with MTF exhibiting the highest value, while water retention capacity was also highest in MTF (2.85 g/g), exceeding PNTF and WTF by 10% and 4%, respectively. These results are consistent with the elevated protein content of MTF and the greater availability of polar functional groups capable of interacting with water (Abdelaleem and Al-Azab, 2021; Boye et al., 2010). In contrast, oil retention capacity showed limited variation among ecotypes (0.24–0.26 g/g), suggesting comparable hydrophobic domains and lipid-binding capacities among the protein fractions. Similar values have been reported for chickpea, lentil, and bean flours (Keskin et al., 2022), supporting the potential application of tarwi flour in bakery products, emulsified systems, and plant-based food formulations. Foaming capacity ranged from 21.92% to 24.17%, while foam stability remained high (99%) in all ecotypes, with no significant differences (p > 0.05) (Table 3). These properties are associated with the presence of soluble storage proteins, particularly 7S and 11S globulins, which adsorb at the air–water interface and form viscoelastic films that stabilize air bubbles (Siddiqui et al., 2024; Bianchi and Simonato, 2025). The values obtained fall within the range reported for other legumes (15–35%) and are comparable to chickpea, lentil, and bean flours (Abdelaleem and Al-Azab, 2021; Boukid et al., 2021), demonstrating that tarwi flour possesses technofunctional characteristics suitable for incorporation into aerated, emulsified, and plant based food products.

Color parameters revealed significant differences among tarwi ecotypes, reflecting variations in pigment composition and seed biochemistry. The white ecotype exhibited the highest lightness (L*), exceeding moro and punto negro by 1.7% and 5.1%, respectively, whereas punto negro showed the lowest L* value, indicating a darker flour. This behavior has been associated with higher concentrations of phenolic compounds, pigments, and insoluble fiber, which reduce light reflectance and increase color intensity (Barrial-Lujan et al., 2025b; Teterycz et al., 2020). Differences in chromatic coordinates further highlighted the distinct pigmentation patterns among ecotypes. The positive a* value observed in punto negro, together with its hue angle (h° = 268.95°), indicated a shift toward reddish–dark tonalities, potentially associated with a greater accumulation of phenolic pigments and anthocyanin-like compounds (Barrial-Lujan et al., 2025b; (Abdelaleema and Elbassiony, 2021). In contrast, white and moro exhibited hue angles close to 90°, confirming the predominance of yellow tones. This observation was supported by the higher b* value of the white ecotype (25.62), consistent with the presence of carotenoids and flavonoids commonly reported in Lupinus mutabilis and other legumes an grains (Kovačević et al., 2025; Abdelaleem and Al-Azab, 2021). Chroma (C*) values were higher in white and punto negro, indicating greater color saturation than in moro. Conversely, the whiteness index (WI) was highest in moro (69.51 ± 0.31), suggesting lower pigment intensity and a lighter visual appearance. From a technological perspective, higher WI values are desirable for products requiring light-colored flours, such as bakery and pasta formulations (Lachman et al., 2017). Overall, the observed color differences confirm that genetic variability strongly influences pigment accumulation and flour appearance, which may affect both consumer acceptance and industrial applications.

The water activity (aw) values of the flours ranged between 0.34 and 0.37 (p < 0.05) (Table 3), indicating low free water availability and favorable storage stability. These values are characteristic of dehydrated food products and are generally considered insufficient to support the growth of most bacteria, yeasts, and molds, thereby contributing to extended shelf life (Rockland and Beuchat, 2017). The white ecotype flour exhibited a significantly higher aw than the moro ecotype, likely associated with differences in chemical composition, particularly in proteins, carbohydrates, and hydrophilic components. In legumes, aw is related to protein-water interactions and the particle size generated during milling, factors that may explain the observed variations among tarwi ecotypes (Guan et al., 2023). Thus, all three tarwi flours present low and stable aw values, suitable for ensuring safety and preservation, while differences between varieties reflect the influence of compositional and structural factors on water retention capacity.

The evaluated tarwi ecotypes exhibited notable differences in their proximate and mineral composition (Table 4). Moisture content ranged from 5.73 g/100 g in WTF to 6.91 g/100 g in MTF, with PNTF showing an intermediate value (6.30 g/100 g). These values were lower than those reported for other tarwi ecotypes (7.5–8.7%), suggesting favorable storage stability (Fellows, 2022; Choquetico-Iquiapaza et al., 2024). WTF exhibited the highest lipid content, exceeding MTF and PNTF by 1.92% and 1.90%, respectively (p < 0.05). Although these values were considerably higher than those reported for lentil, common bean, faba bean, and chickpea, they remained within the range described for tarwi ecotypes (Bianchi and Simonato, 2025; Anyiam et al., 2025; Carvajal-Larenas et al., 2014). This greater lipid accumulation may reflect enhanced carbon partitioning toward lipid reserves, potentially contributing to higher energy density and favorable protein–lipid interactions in food systems. In contrast, MTF showed the highest protein concentration (56.28 g/100 g), exceeding PNTF and WTF by 4.94% and 5.13%, respectively (p < 0.05), confirming the exceptionally protein-rich nature of Lupinus mutabilis compared with other legumes (Mudryj et al., 2014). The greater protein accumulation, likely associated with enhanced nitrogen allocation toward conglutin synthesis, was accompanied by the highest ash content (3.85%), suggesting coordinated deposition of mineral nutrients and storage proteins during seed maturation (Boukid et al., 2021). Similar protein levels have been reported for other tarwi ecotypes, highlighting the strong genetic influence on nitrogen assimilation and storage (Carvajal-Larenas et al., 2014; Anyiam et al., 2025). Moreover, the nutritional quality of tarwi proteins is reinforced by their favorable essential amino acid profile, particularly their lysine content (Bryant et al., 2022). Conversely, PNTF exhibited the highest dietary fiber (5.93 g/100 g) and carbohydrate contents (17.22 g/100 g), whereas MTF showed the lowest fiber content (4.31 g/100 g) and lower carbohydrate levels. This inverse relationship between protein and carbon-rich structural components suggests a compositional trade-off, whereby greater nitrogen allocation to protein reserves occurs at the expense of cell wall polysaccharides and carbohydrate accumulation. Although fiber contents were slightly lower than those reported for other tarwi ecotypes (Choquetico-Iquiapaza et al., 2024), the observed variability falls within the intraspecific ranges described for legumes (Kaur et al., 2020; Boukid et al., 2021). Likewise, the relatively low carbohydrate contents (14.61–17.22 g/100 g) compared with lentil, faba bean, and common bean (40–60 g/100 g) further emphasize the distinctive metabolic strategy of L. mutabilis (Barrial-Lujan et al., 2025a; Boukid et al., 2021). Overall, the results demonstrate that genetic variability among tarwi ecotypes regulates the balance between carbon and nitrogen rich reserves, ultimately determining their nutritional composition and technological functionality.

The mineral profile differed significantly among tarwi ecotypes, indicating genotypic differences in mineral accumulation during seed development (Table 4). PNTF exhibited the highest phosphorus content (410.35 mg/100 g), exceeding MTF and WTF by 25.96% and 10.65%, respectively (p < 0.05). Because phosphorus in legumes is predominantly stored as phytate, these differences may reflect variation in phosphorus reserve accumulation among genotypes (Sinkovič et al., 2022). In contrast, MTF exhibited the highest calcium content (560.32 mg/100 g), slightly exceeding PNTF by 1.33% and WTF by 11.55%. This greater accumulation may be associated with enhanced deposition of calcium bound to structural polysaccharides and cell wall components, contributing to matrix stability and seed reserve organization (Dell’Aquila et al., 2020). WTF contained the highest concentrations of iron (8.93 mg/100 g) and zinc (5.12 mg/100 g), surpassing PNTF and MTF by 23.86–25.77% and 3.23–11.30%, respectively. The elevated levels of these micronutrients indicate a differentiated mineral partitioning strategy among ecotypes and enhance the nutritional value of the flour, given the essential roles of iron and zinc in oxygen transport, protein synthesis, immune function, and enzymatic activity (Diapari et al., 2014). Notably, the zinc concentrations observed in all evaluated ecotypes were higher than those commonly reported for lentil, chickpea, common bean, and faba bean, highlighting the nutritional potential of Lupinus mutabilis as a source of bioessential minerals. The observed mineral variability supports previous studies indicating that the mineral composition of legumes is strongly influenced by genetic background, soil characteristics, agronomic conditions, and postharvest processing (Vera-Vega et al., 2022). Overall, these findings demonstrate that mineral accumulation differs markedly among tarwi ecotypes.

Principal component analysis (PCA) explained 99.0% of the total variability, with PC1 and PC2 accounting for 66.59% and 33.41%, respectively (Figure 2), providing a clear separation of the three tarwi ecotypes. PC1 primarily associated with variation in nutrient partitioning associated with carbon and nitrogen rich storage compounds. Positive PC1 scores were associated with carbohydrates, dietary fiber, phosphorus, water activity, and larger particle fractions Dx (90), clustering the PNTF ecotype. In contrast, negative PC1 scores were associated with protein, ash, water absorption capacity (WAC), swelling power (SP), water retention capacity, and finer particle fractions Dx (10) and Dx (50), grouping the MTF ecotype. This distribution suggests a genotype driven trade off between carbon allocation to structural polysaccharides and reserve carbohydrates, and nitrogen allocation to storage proteins, a phenomenon widely reported in legume seeds (Kaur et al., 2020; Boukid et al., 2021). PC2 further differentiated the WTF ecotype from the remaining genotypes through its association with lipids, iron, zinc, and the color parameters b* and C*, suggesting greater accumulation of energy reserves and micronutrients. By contrast, the MTF ecotype was associated with calcium content, water solubility index (WSI), hygroscopicity, and foam stability, reflecting differences in mineral deposition and hydration related functionality. Similar relationships between nutrient composition and techno-functional performance have been reported in lupins and other grain legumes, where genotypic variation influences reserve deposition, seed matrix organization, and processing performance (Teterycz et al., 2020). The close clustering of protein content, ash, WAC, SP, and water retention capacity suggests that protein rich matrices possess greater hydration potential, likely due to the abundance of polar amino acid residues capable of interacting with water molecules through hydrogen bonding. In contrast, the clustering of carbohydrates, dietary fiber, phosphorus, and coarse particles indicates that carbon-rich matrices are characterized by greater structural integrity but lower hydration efficiency. Overall, the PCA highlights that genotypic variability is closely associated with differences in nutrient composition, seed matrix organization, and technofunctional properties, supporting the potential use of specific tarwi ecotypes for targeted food applications.

Figure 2
PCA of the compositional and technofunctional properties of tarwi ecotype flours. Note. WTF, white tarwi flour; MTF, moro tarwi flour; PNTF, punto negro tarwi flour; L*, lightness; a* and b*, CIELAB color coordinates; C*, chroma; h*, hue angle; WI, whiteness index. Dx(10), Dx(50), and Dx(90) denote particle diameters at the 10th, 50th, and 90th percentiles, respectively; WAC, water absorption capacity; WSI, water solubility index; SP, swelling power.

Collectively, these findings, demonstrate that the variability among tarwi ecotypes offers opportunities for their targeted utilization specific agrofood applications. The moro ecotype (MTF) stands out due to its high protein content and superior hydration properties, positioning it as a promising ingredient for the development of protein-enriched products, including meat analogues, fortified bakery goods, extruded foods, gluten-free formulations, and protein supplements. In contrast, the punto negro ecotype (PNTF), characterized by higher dietary fiber and carbohydrate contents as well as a coarser particle size distribution, appears particularly suitable for breakfast cereals, pasta products, and whole-grain snacks where greater structural firmness and textural stability are required. Finally, the white ecotype (WTF), associated with higher concentrations of lipids, iron, and zinc, shows considerable potential for plant-based beverages, micronutrient-fortified foods, and energy dense formulations, where improved nutritional value, lighter color, and lipid related sensory attributes may provide technological and nutritional advantages.

Although this study provides a comprehensive characterization of the physicochemical, nutritional, mineral, particle size distribution, and technofunctional properties of Lupinus mutabilis flours obtained from ecotypes cultivated in the southern Peruvian Andes (Andahuaylas), several limitations should be acknowledged. Despite the debittering process applied prior to analysis, antinutritional compounds, particularly residual alkaloids, were not quantified. In addition, the bioaccessibility and bioavailability of nutrients and bioactive compounds were not evaluated. Future studies should incorporate residual alkaloid quantification, metabolomic analyses, in vitro gastrointestinal digestion models, and application studies in specific food matrices to further elucidate the relationships among chemical composition, antioxidant capacity, nutrient bioaccessibility, and the nutritional and techno-functional potential of these tarwi ecotypes.

Acknowledgements

The authors thank the Vice-Rectorate of Research of the Universidad Nacional José María Arguedas for its financial support, as well as the Laboratotio de Investigacion Agroindustrial (LIA) of the same university for its collaboration in this study.

Data Availability Statement

Research data is available in the body of the article.

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Edited by

• Editor:
  Takako Matsumura Tundisi

Publication Dates

• Publication in this collection
  21 Sept 2026
• Date of issue
  2026

History

• Received
  21 Mar 2026
• Accepted
  17 Aug 2026