. 2022 Aug 28;12(9):806. doi: 10.3390/metabo12090806

Table 2.

Modification of the composition and/or bioactivity determined in alfalfa (Medicago sativa L.) because of different treatments.

Treatments	Conditions	Effects	References
Alfalfa sprout supplementation in a rabbit diet.	Ninety mixed white rabbits were fed for 50 days, divided into 3 groups: Standard (S) diet, Standard diet + 20 g d⁻¹ of alfalfa sprouts (A), Standard diet + 20 g d⁻¹ of flax sprouts (F).	The alfalfa sprouts presents increase in the total content of fatty acids (PUFA) (linoleic acid by 38.46% and linolenic acid by 70.05%), isoflavones (daidzein) in diets. The linolenic acid content in muscle of the alfalfa group was three times higher than control group. α-tocopherol content and α-tocotrienol, were up.	[12]
Dietary supplementation with alfalfa sprouts.	Dietary supplementation with alfalfa sprouts (40 g d⁻¹) and quantification of bioactive compounds and cholesterol in chicken and chicken eggs.	Decreased cholesterol in chicken plasma from 79.2 to 65.2 mg dL⁻¹ and in egg yolk from 11.5 to 10.4 mg g⁻¹.	[13]
Sprouts alfalfa exposed to sound wave	Frequencies (250, 500, 800, 1000, and 1500 Hz) for two 1-h periods until 6 days.	Increase (24–50%) in the expression of genes that promote the production of L-ascorbic acid in sprouts (to 500 and 1000 Hz). The treatments increased the concentration of ascorbic acid and the antioxidant enzyme superoxide dismutase.	[14]
Substitution with alfalfa seed flour.	Adding alfalfa seed flour (0, 15, 30, 45% w/w) in rice flour biscuits (gluten free).	Increased linearly in: crude protein, total dietary fiber, total polyunsaturated, hardness, total phenolic content (22.9 to 112.9 mg GAE 100 g⁻¹ DW for control and 45% substituted flour), and resistant starch. The antioxidant capacity increased proportionally from 14.7 to 194.6 μmol GAE 100 g⁻¹ DW (FRAP) and from 739.3 to 3627.7 μmol TE 100 g⁻¹ DW (ORAC), for control and 45% of alfalfa flour, respectively.	[11]
Different types of LED lighting in alfalfa sprout composition (FMC: fresh mass of cotyledons).	Four variants using cold white (10,032 K), warm white (3279 K), red green blue (RGB) LEDs with two chips activated: red and blue, which combined gave a violet colour.	Chlorophyll (a) up to 998.9 mg kg⁻¹ in FMC with cold LED. β-carotene up to 44.6 mg kg⁻¹ in FMC with red-green-blue LED (RGB). Chlorophyll a up to 843.3 mg kg⁻¹ FMC, chlorophyll b up to 256.7 mg kg⁻¹ FMC, β-carotene up to 21.6 mg kg⁻¹ FMC, lutein up to 82.6 mg kg⁻¹ FMC, neoxanthin up to 15 mg kg⁻¹ FMC and violaxanthin up to 43.7 mg kg⁻¹ FMC in shoots with sunlight. Total phenols up to 697 mg GAE kg⁻¹ in FMC with blue LED (RGB). Ascorbic acid up to 155 mg kg⁻¹ with sunlight.	[15]
Dry alfalfa sprouts (dry heat, freeze-dried).	Heat-dried samples (HD): stove at 60 °C for 24 h, and the freeze-dried samples (FD) for 24 h under vacuum (50 mTorr).	Greater decrease in isoflavone composition with the application of oven heat drying than with lyophilization. The lyophilisate increased the sterols concentration (41.82% for stigmasterol). The presence of carotenoids (zeaxanthin, β-carotene, retinol, lutein) was only detected after drying processes (not fresh).	[10]
Soak in water of seeds alfalfa.	Seeds were disinfected with hot water: soak at 85 °C, for 10 s; at 85 °C, for 40 s; at 90 °C, for 10 s; and at 100 °C for 10 s.	No effect on: lutein (23.4–26.6 mg kg⁻¹), violaxanthin (16.0–17.2 mg kg⁻¹), neoxanthin (3.5–4.1 mg kg⁻¹), β-carotene (10.1–11.7 mg kg⁻¹), total phenols (486.5–599.4 mg kg⁻¹) and chlorophyll b (64.7–72.8 mg kg⁻¹). The application of 100 °C caused a decrease in the content of ascorbic acid (from 84.5 a 67.5 mg kg⁻¹) and a increased phenolic content (from 537.1 to 599.4 mg kg⁻¹).	[16]
Supplementation, digestion and in vitro fermentation.	Adding alfalfa seed flour (0, 30, 45% w/w) in rice flour biscuits (gluten free). Simulated in vitro digestion and fermentation process.	Cookies with 30 and 45% of alfalfa seed flour presented the highest total phenolic content (0.42 and 0.56 mg g⁻¹, respectively) (control 0.15 mg g⁻¹). The in vitro fermentation of 8–48 h increased the concentration of lignans and phenolic acids, whose bioaccessibility at 24 h of in vitro fermentation were 16.2 and 12.2%, respectively.	[17]
Incorporation of selenium to alfalfa crop.	Inorganic selenium was used in two chemical forms: selenite (Na₂SeO₃) or selenate (Na₂SeO₄) (0, 20, or 200 μmol L⁻¹).	Increase in anthocyanins in alfalfa (29%) after of 20 µmol L⁻¹ selenite solution (≈8% reduction of DPPH).	[18]
Ultrasound in fresh alfalfa leaves.	Study factors and ranges: Solvent/raw material ratio (mL g⁻¹): 5, 10, 15. Time (h): 1, 2, 3. Temperature (°C): 50, 65, 80. Power (W): 50, 100, 150. Ethanol concentration (%): 60, 75, 90.	Better yield (up to 1.61%) and bioaccessibility (up to 19.7%) of saponins. Conditions: solvent/raw material (9.5 mL g⁻¹), extraction time (2.90 h), extraction temperature (79.1 °C), ultrasound power (111.0 W), ethanol concentration (88.2%).	[8]