## 2 Materials and Methods
### Determination of Betalains  
To quantify total betalains, the protocol of [PERSON] et al. (2019) was followed. One gram of each structure was blended in 10 mL of distilled water (1:2) in the dark, then the product was centrifuged (HERMLE Z 232 HKII, Labortechnik GmbH, Wehingen, Germany) at 10,000 rpm for 20 min. Absorbance readings were read on a spectrophotometer (JENWAY Model 6705 Dunmow, United Kingdom) at a wavelength of 483 nm (betaxanthines) and 538 nm (betacyanins). The results were expressed in mg/g of extract. \[\text{Betanins}=\left((\text{a}/1.129)\ \times\ \text{DF}\ \times\ 100\right) \tag{1}\]  
\[\text{Vulgaxantins}=\left((\text{y}/750)\ \times\ \text{DF}\ \times\ 100\right) \tag{2}\]  
where:  
\(\text{a}=1.095\ (\text{A538}\ -\ \text{A600})\)  
\(\text{y}=\text{A476}\ -\ (\text{A538}\ -\ \text{a})-(\text{a}/3.1)\)  
\(\text{DF}=\text{Dilution}\) factor  
Evaluation of the Percentage of Inhibition of the Enzyme Urease as an Indicator of Gastroprotective Effect  
The method described by [PERSON] et al. (2013) with some modifications was followed. The reaction was started by placing 100 \(\mu\)L of urease solution (Jack Bean urease 5 U / mL Sigma-Aldrich, St. Louis, MO, USA), and 100 \(\mu\)L of inhibition extract incubated at 37 \({}^{\circ}\)C in a bath with a horizontal shaker (Scorpion Scientific, CDMX, Mexico) for 10 min; later, 200 \(\mu\)L of phosphate buffer (pH 6.8) with urea (U5128 Sigma-Aldrich, St. Louis, MO, USA) were added (100 mM) and incubated at 37 \({}^{\circ}\)C for 30 min. Subsequently, 500 \(\mu\)L of phenol reagent or reagent A (1% w/v phenol (P1037 Sigma-Aldrich, St. Louis, MO, USA) and 0.005% w/v sodium nitroprusside ([PERSON], Mexico City, Mexico)) and 500 \(\mu\)L of alkaline reagent or reagent B (0.5 % w/v NaOH and 0.1 % NaOCl) was incubated for 30 min at 37 \({}^{\circ}\)C. The reaction was stopped with 600 \(\mu\)L of 4% H\({}_{2}\)SO\({}_{4}\) ([PERSON], Mexico City, Mexico). The absorbance was read at 630 nm using a spectrometer (JENWAY, Model 6705, Dunmow, UK). Thiourea (T8656 Sigma-Aldrich, St. Louis, MO, USA) was used as a standard urease inhibitor. Urease activity was determined by measuring ammonia production using the indophenol method. Indophenol is an intense blue compound that is formed by the reaction of ammonium, hypochlorite, and phenol catalyzed by sodium nitroprusside. An ammonium chloride calibration curve was made to determine the amount of ammonium produced during the tests and thus evaluate the enzymatic activity performed by urease. Evaluation of Percentage of Inhibition of the Elastase Enzyme as an Indicator of Anti-Inflammatory Effect  
The spectrophotometric technique using porcine pancreatic elastase (E1250 Sigma-Aldrich, St. Louis, MO, USA) was applied according to the modified method of [PERSON] et al. (2013) cited by [PERSON] et al. (2014). N-Succinyl-Ala-Ala-Ala-p-nitroanilide (PPE type IV, Sigma-Aldrich, St. Louis, MO, USA) was used at a concentration of 1015 M, dissolved in 0.1 M Tris-HCl (PHG0002 Sigma-Aldrich, St. Louis, MO, USA) at pH 8 as substrate and 0.01 mg of elastase was dissolved with 0.2 M Tris-HCl buffer (pH 8). In order to carry out the reaction, 10 \(\mu\)L of each of the extracts with the different concentrations, 10 \(\mu\)L of elastase and 130 \(\mu\)L of the previously prepared substrate were added to these to obtain a final volume of 150 \(\mu\)L and incubated in a horizontal shaker bath (Scorpion Scientific, CDMX, Mexico) for 40 min. Finally, a reading was taken at 410 nm and the calculations corresponding to the formula were carried out:  
\[\%\ Inhibition=\left[\frac{Abs\ Control\ -\ Abs\ Sample}{abs\ Control} \right]\times 100 \tag{3}\]Evaluation of the Percentage of Inhibition of the Enzyme \(\beta\)-Glucuronidase as an Indicator of Hepatoprotective Effect  
\(\beta\)-Glucuronidase inhibition assay was carried out as per the method of [PERSON] et al. [10] with modifications.