TECNOCIENCIA CHIHUAHUA, Vol. XX (2026): enero-diciembre, e2175

https://revistascientificas.uach.mx/index.php/tecnociencia

ISSN-e: 2683-3360

Scientific Article

Use of Supercritical fluid extraction (SFE) as a

Sustainable Strategy for Polyphenol Recovery from

Salvilla (Buddleja scordioides Kunth)

Uso de la extracción por fluidos supercríticos (SFE) como estrategia

sustentable en la recuperación de polifenoles de Salvilla (Buddleja

scordioides Kunth)

*Corresponding Author: E-mail: eli.macias@live.com.mx (Elizabeth Macías-Cortés)

DOI: https://doi.org/10.54167/tch.v20i1.2175

Received: 10 de diciembre de 2025; Approved: 29 de mayo de 2026

Published by the Autonomous University of Chihuahua, through the Research and Postgraduate Department.

Section Editor: Dra. Elizabeth Carvajal-Millán

Abstract

Emerging extraction technologies gained position due to their advantages in obtaining high-quality

metabolites. Supercritical fluid extraction (SFE) recovers bioactive molecules while preserving their

integrity. In this study, SFE was applied to Buddleja scordioides Kunth (salvilla) bush leaves, a species

rich in phenolic compounds with medicinal properties. A central composite design of three factors

was used: co‑solvent percentage (ethanol), pressure, and extraction time. Extracts were evaluated

for yield, total flavonoid content, antioxidant activity (ORAC and ABTS) and quantification of

phenolic compounds by UPLC-PDA-ESI-MS/MS. Extraction yields ranged from 3.63-6.27 %, total

phenolic content varied from 219.17-314.72 μg CE/mg extract. Antioxidant activity was ranged from

Elizabeth Macías-Cortés1*, Rubén Francisco González-Laredo2, José Alberto Gallegos-

Infante2, Nuria Elizabeth Rocha-Guzmán2, Mercedes Georgina Ramírez-Aragón1, Reyna

Margarita Hernández-Ramos1, Jesús Alejandro, Valdés-Nieblas1, Gustavo Adolfo Castillo-

Herrera3

1 Tecnológico Nacional de México / Instituto Tecnológico Superior de Lerdo, Dgo. Av. Tecnológico 1555

Sur, Periférico Gómez Palacio – Lerdo, 35150, Lerdo, Dgo., México.

2 Tecnológico Nacional de México / Instituto Tecnológico de Durango. Felipe Pescador 1830 Ote., Col.

Nueva Vizcaya, 34080, Durango, Dgo., México.

3 Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, A.C. (CIATEJ) Unidad

Zapopan, Camino Arenero 1227, El Bajío, 45019 Zapopan, Jal., México.

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53.54–81.88 mmol TE/mg of salvilla leaves extract (SE) for ABTS and 78.69-185.40 mmol TE/mg SE

for ORAC. The total phenolic content determined by UPLC-PDA-ESI-MS/MS was ranged from

4188.1-13008.2 ng/mg SE with the co-solvent as the most influencer factor. According to the general

linear model (GLM), the central points (120 min, 150 bar, 20 % co-solvent) yielded the highest results.

This work is original in its application of SFE, to improve the extraction of bioactive molecules. This

work provides novelty by applying SFE, to enhance the extraction and quality of bioactive

compounds. These results enhance the SFE as a sustainable strategy for the valorization of medicinal

plants.

Keywords: salvilla, emerging extraction, supercritical fluids, phenolic acids, flavonoids.

Resumen

Las tecnologías de extracción emergentes cobraron relevancia por sus ventajas en la obtención de

metabolitos de calidad. La extracción con fluidos supercríticos (SFE) recupera moléculas bioactivas

preservando su integridad. En este estudio, se aplicó SFE a las hojas del arbusto de Buddleja

scordioides Kunth (salvilla), especie rica en compuestos fenólicos con propiedades medicinales. Se

empleó un diseño central compuesto de tres factores: porcentaje de co-solvente (etanol), presión y

tiempo. Los extractos se analizaron para rendimiento, flavonoides totales, actividad antioxidante

(ABTS y ORAC) y cuantificación fenólica mediante UPLC-PDA-ESI-MS/MS. Los rendimientos

oscilaron entre 3.63-6.27 %, los compuestos fenólicos totales entre 219.17-314.72 μg CE/mg de

extractos de hojas de salvilla (SE), actividad antioxidante entre 53.54-81.88 mmol TE/mg SE para

ABTS y 78.69-185.40 mmol TE/mg SE para ORAC y el análisis por UPLC-PDA-ESI-MS/MS osciló

entre 4188.1-13008.2 ng/mg SE siendo el co-solvente la variable de mayor influencia. De acuerdo con

el modelo lineal general (MLG), los puntos centrales (120 min, 150 bares y 20 % de co-solvente)

tuvieron los mejores resultados. Este trabajo aporta originalidad al aplicar SFE, para mejorar la

extracción de compuestos bioactivos y su calidad. Los resultados exponen a la SFE como estrategia

para obtener compuestos fenólicos de salvilla con capacidad antioxidante relevante, respaldando su

aplicabilidad en la valorización sostenible de plantas medicinales.

Palabras clave: salvilla, extracción emergente, fluidos supercríticos, ácidos fenólicos, flavonoides.

1. Introduction

The extraction of natural compounds has been an integral part of human culture since

antique times. Early societies, like the Chinese, Egyptians and even Mayans, relied on rudimentary

methods like maceration and steam distillation for the preparation of herbal medicines, perfumes,

and food preservatives (Azmir et al., 2013). Over centuries, these techniques evolved, but solvent-

based extractions remained predominant, often relying on large volumes of organic solvents and

energy-intensive processes.

Despite their widespread use, traditional extraction methods present significant disadvantages

when applied to bioactive compound extraction. Prolonged extraction times and high temperatures

can cause the degradation of thermolabile molecules, such as phenolic compounds, diminishing their

activity (Chemat et al., 2019). Moreover, the use of toxic solvents of organic nature causes alarms

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about environmental contamination, operator protection, and solvent residues in the final product,

directly contradicting the principles of green chemistry (Mustafa & Turner, 2011). These tasks

highlight the necessity for more sustainable and efficient options.

Emerging extraction technologies have gained momentum in recent decades as part of a broader

shift toward green chemistry. Techniques such as ultrasound-assisted extraction (UAE), microwave-

assisted extraction (MAE), pressurized liquid extraction (PLE) and supercritical fluid extraction

(SFE) have demonstrated significant rewards over conventional technologies, like improved yields

of bioactive compounds, shorter extraction times, and reduced solvent use (Usman et al., 2023). They

are closely associated with the 12 principles of green chemistry, offering cleaner, safer, and more

energy-efficient approaches to phytochemical recovery (Chemat et al., 2015).

Among these, supercritical fluid extraction (SFE) stands out for its unique physico-chemical

foundation. SFE uses the properties of fluids in their supercritical phase, beyond their critical

pressure and temperature, where they demonstrate gas-like diffuseness and liquid-like dissolving

power (Banafi et al., 2023). For this, carbon dioxide (CO₂), is the commonly utilized supercritical fluid

since it is non-toxic, easily removed from extracts, inexpensive, and non-flammable, properties given

by its low critical pressure (73.8 bar) and temperature (31.1 °C). The closed, oxygen-free, and low-

temperature environment of SFE minimizes oxidation and thermal degradation, making of this

technology particularly appropriate for obtaining thermosensitive bioactive molecules (Vinitha et

al., 2022; Sheibani et al., 2024). In this way this technique allows the use of co-solvents, such as

ethanol that offers several advantages in the obtention of bioactive compounds. By adjusting

pressure, temperature, and the proportion of ethanol, the extraction process can be selectively

directed to obtain more specific chemical profiles with fewer impurities, while requiring significantly

smaller solvent volumes compared to conventional ethanolic extractions (Herrero et al., 2006;

Reverchon & De Marco, 2006).

In this context, SFE offers notable potential for the obtention of phenolic molecules, key secondary

metabolites that have anti-inflammatory, antimicrobial and antioxidant activities from medicinal

plants. In this tenor, Buddleja scordioides Kunth, commonly known as "salvilla," is an endemic bush

widely disseminated in Mexico and habitually used in traditional medicine for its healthy properties

such as anti-inflammatory and wound-healing properties (Ávila-Acevedo & Romo-de-Vivar, 2002;

Díaz-Rivas et al., 2015; Villegas Novoa et al., 2019). Phytochemical studies have revealed that salvilla

is rich in polyphenols, particularly flavonols and phenolic acids, which confer its pharmacological

activities (Macías-Cortés et al., 2022).

Given the sensitivity of polyphenolic compounds to heat and oxidation, SFE emerges as a

particularly advantageous method for their recovery from salvilla. Previous research has

demonstrated the successful application of SFE for extracting polyphenols and other bioactives from

diverse plant matrices, including Amaryllidaceae alkaloids (Pilařová et al., 2025), rosemary (Salvia

rosmarinus) (Ayyildiz et al., 2024), green tea (Camellia sinensis) (Dębczak et al., 2024), and grape seeds

(Vitis vinifera) (Coelho et al., 2018), achieving high yields and preserving bioactivity. Such precedents

highlight the promise of optimizing SFE conditions to maximize the yield and functionality of

salvilla leaves extracts, paving the way for their potential use for pharmaceutical, nutraceutical, and

cosmetic products. The aim of the present work is to evaluate the extraction of bioactive compounds

from Buddleja scordioides Kunth (salvilla) using the supercritical fluid extraction, using a central

composite design; this can help to determine the optimal conditions to maximize yield, phenolic

content, and antioxidant activity of the salvilla leaves extracts.

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2. Materials and methods

2.1 Chemical reagents

For total flavonoids catechin, (HPLC grade), potassium persulfate (K2S2O8), aluminum

chloride (AlCl3), sodium hydroxide (NaOH), and sodium nitrite (NaNO2) were used. Finally, for 2,2'-

azino-bis (3-ethylbenzothiazoline-6-sulphonic) acid (ABTS) and Oxygen Radical Absorbance

Capacity (ORAC) antioxidant techniques fluorescein, monobasic potassium phosphate, ABTS

radical, dibasic potassium phosphate, 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)

radical and Trolox® were used. For the Ultra High-Performance Liquid Chromatography system

coupled to a Xevo TQS triple-quadrupole detector with photodiode array and electrospray interfaces

(UPLC-PDA-ESI-MS/MS) determinations, this reference standards and solvents were employed:

luteolin, quinic acid, neohesperidin, ferulic acid, 4-hydroxybenzoic acid, apigenin, chlorogenic acid,

acetonitrile (HPLC grade), syringic acid, protocatechuic acid, naringenin, caffeic acid, benzoic acid,

eryodictiol, kaempferol 3-O-glucoside, ethanol, sinapic acid, shikimic acid, rutin, 2,4,6-

trihydroxybenzaldehyde, vanillic acid, acacetin, quercetin-O-glucoside, trans-cinnamic acid,

naringin, myricetin, methanol, 4-O-caffeoylquinic acid, quercetin, coumaric acid, quercetin

glucuronide, kaempferol. All chemical reagents (reagent or LC grade) and standards (HPLC grade)

were purchased from Sigma-Aldrich® (Merck KGaA, Darmstadt, Germany).

2.2 Methods

2.2.1 Collection of salvilla leaves

In 2018, wild bushes of Buddleja scordioides Kunth were harvested near Victoria de Durango,

Durango, México (24°00′00″ N, 104°24′59″ W). After collection, the salvilla leaves were detached from

the stems and air-dried at 25 °C under light-protected conditions. Once dehydrated (with a moisture

content of 5.32  0.3 %), the plant material was ground using a #2 sieve knife mill (IKA®, Staufen,

Germany) and subsequently kept in sealed containers until further use.

2.2.2 Supercritical fluid extraction (SFE)

A portion of milled salvilla leaves (55 g) was introduced into the extraction chamber of a

Thar® SFE system (Pittsburg, USA). Extractions were carried out at 40 °C, using CO₂ as the primary

solvent and ethanol as co-solvent with a 10 mL/min flow, varying the co-solvent percent, pressure,

and time (described in statistical analysis). To recover each extract, the ABPR valve was gradually

opened in increments to prevent freezing, with the instrument software controlling the opening at a

rate of 5 % per minute, enabling collection from the separator vessel. A total of fifteen SFE conditions

were used (Table 1). After each extraction, the system was slowly depressurized, the collected

material was retrieved from the vessel, and it was stored under refrigeration and protected from light

until further analysis.

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Table 1. Treatment conditions

Tabla 1. Condiciones de los tratamientos

Treatment

Run

Time (min)

Pressure (Bar)

Co-solvent (%)

162.4

185.4

16.5

162.4

114.6

23.5

77.6

185.4

23.5

77.6

114.6

16.5

150

180

150

120

100

120

200

120

150

120

150

120

150

120

150

120

150

120

150

120

150

2.2.3 SFE extracts pre-treatment to lyophilization

Because all extracts were generated using ethanol as the co-solvent, it was necessary to

remove the organic phase before freeze-drying. For this purpose, distilled water equivalent to 25 %

(v/v) of each extract was added, followed by solvent removal in a Büchi® rotary evaporator (New

Castle, USA) at 40 °C. Once the alcohol had been eliminated, the aqueous extracts were frozen and

subsequently lyophilized in a Labconco® freeze dryer (Kansas City, Missouri, USA). The resulting

dry powders were kept under desiccated conditions until further use.

2.2.4 Extraction yield

The energy content of the supplement was estimated based on its nutritional composition,

in accordance with Mexican Official Standard NOM-051-SCFI/SSA1-2010. Quantification of the total

macronutrient content (proteins, lipids, and carbohydrates) was performed. Subsequently,

Atwater’s general conversion factors were applied, assigning 4 kcal per gram to proteins and

carbohydrates, and 9 kcal per gram to lipids (Sánchez-Peña et al., 2017). Results were represented as

kilocalories per 100 grams of supplement (kcal/100 g).

To determine the extraction yield, 55 g of dried salvilla leaves were exposed to the SFE procedure

and later lyophilized. Extraction yield was expressed as the percentage of recovered extract relative

to the dry mass:

𝐸𝑥𝑡𝑟𝑎𝑐𝑡𝑖𝑜𝑛 𝑦𝑒𝑙𝑑(%)=𝑙𝑦𝑜𝑝ℎ𝑖𝑙𝑖𝑧𝑒𝑑 𝑒𝑥𝑡𝑟𝑎𝑐𝑡 𝑚𝑎𝑠𝑠

𝑖𝑛𝑖𝑡𝑖𝑎𝑙 𝑑𝑟𝑦 𝑚𝑎𝑠𝑠 × 100

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2.2.5 Total flavonoid content

Total flavonoids were assessed using the methodology reported by Rosales-Villarreal et al.

(2022). For the calibration curve, catechin solutions (0–300 μg/mL) were prepared. Samples of the

lyophilized SFE extracts were diluted in a milliliter (mL) of deionized water. Then, 20 μL of each

standard, blank, or sample were blended with 5 % NaNO₂ (7.5 μL), 10% AlCl₃ (15 μL), 1 M NaOH

(50 μL), and 157 μL distilled water. The reaction mixtures were protected from light for 5 min, and

absorbance was recorded at 570 nm using a Daigger microplate reader (Buffalo Grove, IL, USA).

Results were expressed as μg of catechin equivalents per mg of salvilla leaves extract (μg CE/mg SE).

2.2.6 Antioxidant activity of samples

For this project, both electron-transfer (ABTS) and hydrogen-donation (ORAC) assays were

used to evaluate antioxidant capacity in all treatments.

The ABTS assay was performed according to the method reported by Macías-Cortés et al. (2022).

After the ABTS•⁺ radical was activated by mixing equal quantities of ABTS solution (7 mM) and

potassium persulfate solution (2.45 mM) and allowing them to react at room temperature for 16 h in

the dark, this was diluted with 5 mM phosphate buffered-saline (PBS) buffer (pH 7.4) to achieve an

absorbance of 0.700  0.02 nm at 750, then 10 μL of diluted blank, sample, or Trolox® (50M) were

aggregated to 190 μL of diluted ABTS•⁺ radical. After 10 min in the dark, absorbance was measured

at 750 nm using a Daigger® microplate reader (Vernon Hills, IL, USA). Results were reported as

mmol Trolox equivalents per mg of salvilla leaves extract (mmol TE/mg SE). In other hand, ORAC

assay was adapted from Rosales-Villarreal et al. (2022). Each well received 20 μL of diluted blank,

sample or Trolox® (50M), and then, 200 μL fluorescein (1.09 μM) were added. After a 15 min

incubation at 37 °C using a Synergy HT® microplate reader (Bio-Tek, Winooski, VT, USA), 75 μL of

2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) radical (79.65 mM) were added. In this

assay, fluorescence of each well was monitored each 1.5 min, finishing at 2.5 h. The readout was

programmed at 485 nm for excitation and 535 nm for emission. Antioxidant activity was calculated

from the area under each decomposition curve and reported as mmol TE/mg dry salvilla leaves

extract.

2.2.7 Phenolic characterization by Liquid chromatography – mass spectroscopy

(UPLC-PDA-ESI- MS/MS)

For phenolic characterization, profiling was performed following the Díaz-Rivas et al. (2018)

method with some modifications. Analyses were performed using a Waters Ultra High-Performance

Liquid Chromatography (UPLC) system coupled to a Xevo TQS triple-quadrupole detector with PDA

and ESI interfaces (Xevo TQS, Waters Corp., Wexford). Samples were injected via a temperature-

controlled manager (6 °C) into an Agilent® C18 column (150 × 2.1 mm, 1.7 μm). The elution involved

solvent A composed of 7.5 mM formic acid and solvent B that is acetonitrile, this according to the

following gradient: 97:3 (A:B) at 0 min; 91:9 at 1.23 min; 84:16 at 3.82 min; 50:50 at 11.40 min; returning

to 97:3 at 13.24–15.00 min. The elution flow rate was 250 μL/min, while methanol/formic acid

introduced at 0.210 mL/min as co-solvent was added post‑separation to stabilize ion formation. Total

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runtime was 15 min. Mass spectrometry was conducted in Multiple Reaction Monitoring (MRM)

mode, operating in negative ESI with the following parameters: cone 30 V, capillary 2.25 kV,

desolvation 400 °C, source 150 °C, desolvation gas 800 L/h, cone gas 150 L/h, collision gas 0.13

mL/min, MS collision energy 2.0 eV, and MS/MS 20 eV. Phenolic standards at 20 ng/μL were used to

establish retention times, m/z values, and fragmentation patterns. Data acquisition and processing

were performed using MassLynx v4.1 Software (Waters Corp., Milford, MA). Samples were prepared

at 10 mg per milliliter (mg/mL), filtered through 0.45 μm acrodiscs, and placed into amber vials in

the autosampler, wich injected 1 microliter for the assay. The results were expressed as ng of phenolic

compound per mg of salvilla leaves extract.

2.2.8 Experimental design and data analysis

The use of a central composite design (CCD) is widely recommended for multivariable

optimization in extraction technologies, particularly in SFE, where process efficiency and selectivity

depend on synergistic effects between operational parameters (Herrero et al., 2006; Bezerra et al.,

2008). For the experiment a CCD was chosen, small type, with 15 experimental runs: 10 without

central points and 5 with central points, using an alpha () of 1.41421. The independent variables

were extraction time, pressure and cosolvent percent according to the follow table 2:

Table 2. Independent variables of the experiment

Tabla 2. Variables independientes del experimento

Factor

- 1

- 



Time (min)

77.57

120

162.43

180

Pressure (bar)

100

114.64

150

185.36

200

Co-solvent (%)

16.46

23.54

Furthermore, a general linear model (GLM quadratic modeling) was employed to analyze the

experimental data because it allows the simultaneous evaluation of the individual and interactive

effects of the extraction variables included in the central composite design. Results are shown in RSM

graphics. The analysis was performed using obtained data from statistical software Design Expert ®

(Minneapolis, USA) and Statistica v. 12.0 software (Palo Alto, CA, USA).

3. Results and discussion

3.1 Extraction yield

Once the SFE salvilla leaves extracts were obtained, all treatments were lyophilized and their

yield considered, where all samples were from 3.6 to 6.3 % (central points had an average of 4.90 ±

0.1 %). These results were fitted to a linear model in which the most influential factor according to

their equation is ethanol (p = 0.0001) (Table 3).

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Table 3. Modeling for each extraction parameter

Tabla 3. Modelamiento para cada parámetro de extracción

Model

value

Yield

1.60 + 0.02a + 0.003b – 0.07c

0.0001

TFM

– 295.23 – 1.10a + 3.37b – 29.31c – 0.01ab + 0.11ac – 0.05bc + 0.004a2 + 0.001b2 – 0.70c2

0.0011

ABTS

– 19.05 – 1.49a + 0.88b + 7.89c + 0.0007ab + 0.03ac – 0.04bc + 0.004a2 + 0.00002b2 – 0.10c2

0.005

ORAC

– 1749.04 + 5.17a + 9.43b + 94.31c + 0.01ab – 0.06ac – 0.28bc – 0.02a2 – 0.01b2 – 1.21c2

0.0005

TPC

– 152999 + 400.76a +658.99b + 9576.79c - 0.37ab – 11.52ac – 24.71bc – 0.52a2 – 0.58b2 – 114.18c2

0.0004

*EP= extraction parameters; TFM = Total flavonoids (microplate); TPC = total phenolic compounds a= time b= pressure

c= cosolvent

Although the co-solvent is the most influential factor, it can be noticed that minimum yield

corresponds to a 60 min of extraction, while 6.3 % corresponds to 162.4 min, treatment obtained at

180 min had a yield of 6.0 %. Fig. 1 showed the yield modeling in a 2D surface method response

graphic. In this respect, the obtained results of yield were smaller than the observed by traditional

technologies such as the reported by Avila-Acevedo & Romo-de-Vivar (2002), who made a successive

extraction of salvilla, beginning with hexane, then ethyl acetate and finally methanol, reporting 15.15

% of yield. Another case is the salvilla extract obtained using an infusion method made (Herrera-

Carrera et al., 2015), obtained a 20.31 ± 0.61 % of yield extraction. In the other hand, SFE showed

lower yield extraction than the other one obtained by ultrasound assisted extraction (UAE), an

emergent technology made by Macías-Cortés et al. (2022), who reported a yield extraction of 10.58 ±

0.33 to 12.92 ± 0.40 % in hydroalcoholic salvilla leaves extracts obtained with different conditions such

as wave amplitude, time, and ethanol percent. The yield obtained in this work is similar to the

described by Perez-Gutierrez & Vargas-Solis (2008) who reported a yield of 7.6 % in a successive

extraction consisted of hexane, chloroform and methanol (in this order) salvilla extract. Fig. 1.

Despite the comparatively lower extraction yields obtained through supercritical fluid extraction

(SFE), this behavior is consistent with the intrinsic characteristics of the technique. SFE typically

produces smaller quantities of extract than conventional or solvent‑intensive methods

(Pourmortazavi & Hajimirsadeghi, 2007); however, it offers significant technological advantages that

justify its use for the recovery of bioactive compounds. Because CO₂ is non‑toxic, non‑flammable,

and easily removed from the final product, SFE generates cleaner extracts free of solvent residues,

which is particularly valuable for applications in food, nutraceutical, and pharmaceutical

formulations (Herrero et al., 2006; Reverchon & De Marco, 2006). Additionally, the process operates

under mild temperatures, reducing thermal degradation of thermolabile compounds and improving

the preservation of antioxidant constituents (Plaza & Turner, 2015). From an environmental

perspective, SFE is considered a more sustainable alternative, as it minimizes the use of organic

solvents and allows CO₂ to be recycled within the system, reducing waste generation and ecological

impact. Therefore, although the yields obtained in this study were lower than those reported for

traditional extraction techniques, the high purity, safety, and sustainability associated with SFE

position it as a competitive and environmentally responsible method for salvilla bioactive compound

recovery.

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Figure 1. Yield of salvilla leaves extract (2D surface response model)

Figura 1. Rendimiento del extracto de salvilla (modelo de superficie de respuesta en 2D)

3.2 Total flavonoid content

The flavonoids are a group of secondary metabolites acknowledged for their anti-

inflammatory, antioxidant, and protective properties against oxidative stress. Their quantification is

a key indicator of the biological potential of plant extracts and serves as an important criterion to

assess the efficiency of extraction methods. Therefore, determining the total flavonoid content

provides valuable insight into the capacity of supercritical fluid to recover bioactive compounds with

functional relevance (Herrero et al., 2006; Macías-Cortés et al., 2020).

In this tenor, a total flavonoid content by microplate was done for all treatments. These results were

adapted to a quadratic model where the most influential factor according to their equation was

ethanol (p = 0.0011) (Table 3). The achieved values ranged from 219.17 to 314.72 μg CE/mg SE, with

the central points showing a mean value of 270.43 ± 1.2 μg CE/mg SE (Fig. 2) where it seemed that

higher values can be found in higher ethanol percent. Despite the lower yields obtained, supercritical

fluid extraction demonstrated to be a competitive emerging technology since flavonoid content was

higher than the reported in other works, for example, Macias-Cortés et al. (2022) with values of 107.3

± 3.0 - 224.9 ± 6.3 μg CE/mg SE in samples obtained with ultrasound assisted extraction, and the

described by Díaz-Rivas et al. (2015), who informed quantities of 10.8 ± 0.05 μg CE/mg SE in a salvilla

infusion and their work where the salvilla infusion and 7.06 ± 0.4 μg CE/mg SE in a concentrated

salvilla infusion. These findings confirm that ethanol, in synergy with supercritical CO₂, enhances

the recovery of metabolites like flavonoids as a consequence of its polarity and solvent affinity.

Moreover, the supercritical CO₂ phase improves the solubilization and mass transfer of medium-

polarity molecules, thereby increasing their concentration in the last extracts (Herrero et al., 2006).

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Therefore, the ability of SFE to selectively enrich flavonoids positions this method as an auspicious

green replacement for the obtention of functional molecules from salvilla and related medicinal

plants.

Figure 2. Total flavonoids content by microplate (2D surface response model)

Figura 2. Contenido de flavonoides totales por microplaca (modelo de superficie de respuesta en 2D)

3.3 ABTS assay

The production of reactive oxygen species and free radicals during oxidation processes in

biological and food systems can damage DNA, proteins and lipids, leading to several chronic

diseases and the deterioration of food quality. Therefore, evaluating the antioxidant capacity of

natural bioactive compounds or plant extracts is essential to evaluate their functional potential, either

as nutraceutical ingredients or for applications in functional foods. The ABTS assay has become a

standard tool for this purpose due to its versatility. The ABTS•⁺ radical cation exhibits a characteristic

absorption that decreases when an efficient antioxidant neutralizes it. Additionally, its rapid reaction

and good reproducibility make it an assay of choice for screening and comparing antioxidant capacity

between samples (Dawidowicz and Olszowy, 2013).

In this study, the antioxidant activity measured by the ABTS assay showed a clear dependence on

the extraction conditions, particularly on ethanol concentration, as indicated by the quadratic model

that best fitted the data (p = 0.005). For all the samples, quantities were going from 53.54 to 81.88 mmol

TE/mg SE; in this tenor, central points had an average of 62.83 ± 0.8 mmol TE/mg SE. Ethanol

percentage was the factor with the greatest influence on the results (Fig. 3), suggesting that solvent

polarity played a key role in the extraction efficiency of antioxidant compounds.

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Figure 3. Antioxidant activity by ABTS assay (2D surface response model)

Figura 3. Actividad antioxidante por el ensayo ABTS (modelo de superficie de respuesta en 2D)

This result aligns well with previous findings in which the addition of ethanol as a co-solvent in

supercritical-CO₂ based extraction enhanced the recovery of antioxidant phenolics or flavonoids,

likely by improving the polarity match between solvent and solutes. In contrast to the performance

obtained with ethanol as a co‑solvent in supercritical CO₂ extraction, studies using methanol as a co-

solvent have shown comparatively lower antioxidant efficiency. For example, Uwineza et al. (2021)

reported that SFE extracts of Lamium album obtained with methanol as co-solvent exhibited values

for ABTS assay ranging only from 0.043 to 0.045 µg TE/mg extract, indicating limited antioxidant

capacity under those conditions. The influence of ethanol concentration aligns with the chemical

nature of the bioactive compounds typically extracted from plant matrices. Phenolic compounds—

such as phenolic acids, flavonoids, and tannins—are widely recognized for their strong radical-

scavenging capacity due to their ability to donate electrons or hydrogen atoms, effectively

neutralizing reactive species like ABTS•⁺ (Rice-Evans et al., 1997). Because these compounds exhibit

intermediate polarity, ethanol can enhance supercritical CO2 extraction. Moderate ethanol

concentrations tend to favor the simultaneous solubilization of both polar phenolic acids and less

polar flavonoid aglycones, which may explain the increased antioxidant activity observed at specific

ethanol levels (Radzali, 2020). To support this, Lee et al. (2020) reported that increasing ethanol

concentration during conventional extraction significantly elevated antioxidant activity in citrus peel

extracts. supporting the idea that ethanol concentration modulates the extraction of compounds with

high antioxidant potential.

Overall, the relationship between extraction conditions, bioactive compound solubility, and

antioxidant activity underscores the importance of optimizing solvent composition to maximize the

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functional potential of plant-derived extracts. However, excessively high ethanol proportions may

diminish the supercritical CO₂ diffusivity or reduce selective extraction by co-extracting non-target

constituents, which may dilute the specific antioxidant compounds (Radzali, 2020).

3.4 ORAC assay

The measurement of antioxidant activity by ORAC assay is a reliable and extensively used

method in the evaluation of natural extracts. This is based on their capability to neutralize the peroxyl

radicals. Unlike other assays that rely on single-electron transfer mechanisms, ORAC measures the

hydrogen atom transfer capacity, providing a more physiologically relevant estimate of antioxidant

performance. This assay has been widely used to evaluate plant-derived phenolics and flavonoids,

specifically regarding their antioxidant capacity, allowing quantitative comparison of different

extraction methods and treatments (Ou et al., 2001).

In Fig. 4 the response surface graphic, illustrates the effects of extraction time and pressure and with

20 % of ethanol as co-solvent, expressed in mmol TE/mg SE, and the results were fit to a quadratic

model (p = 0.0005). In this sense, values were going from 78.69 to 185.40 mmol TE/mg SE, obtaining

the central points the highest values, with an average of 183.60  2.0 mmol TE/mg SE. According to

the equation in Table 3, co-solvent was the most influential factor. In SFE, ethanol functions as an

effective polarity modifier, enabling fine‑tuning of the solubility of phenolic compounds, flavonoids,

and other metabolites. Moreover, when used as a co‑solvent, ethanol increases the polarity of the

supercritical CO₂ phase, facilitating the recovery of polar bioactive compounds typically obtained

through traditional ethanolic extraction, but with the added advantages of higher purity and reduced

thermal or oxidative degradation (Machmudah et al., 2006). It can be seen that Supercritical fluid

extraction can enhance the obtention of bioactive compounds. As an example, Picos-Salas et al. (2024)

observed that supercritical fluid extract from Mexican oregano (Lippia graveolens) showed an ORAC

activity of 6923.65 ± 57.2 μmol TE/g extract while the methanolic extract obtained from the same

specie had 3577.08 ± 195.09 μmol TE/g extract, a 48.33 % less activity than the obtained with this

suitable emerging technology. Supercritical CO₂, particularly when modified with a polar solvent,

exhibits strong affinity for highly bioactive antioxidant molecules with elevated radical‑scavenging

capacity (Pilařová et al., 2024).

In summary, the response surface trends clearly demonstrate that the combination of moderate

pressure and adequate extraction time maximizes the obtention of bioactive molecules, highlighting

the strong influence of these parameters on the increase of antioxidant molecules recovery. The high

values obtained at the central points of the present work, together with those observed by Picos-Salas

et al. (2024), reinforce the idea that supercritical CO₂ can be used potentially as an efficient method

for obtaining antioxidant molecules from salvilla.

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Figure 4. Antioxidant activity by ORAC assay (2D surface response model)

Figura 4. Actividad antioxidante por el ensayo ORAC (modelo de superficie de respuesta en 2D)

3.5 Identification and quantification of Total Phenolic compounds

As previously reported, salvilla is characterized by a rich and diverse phenolic profile, which

contributes to many of its traditional and pharmacological properties. Its phenolic matrix typically

includes flavonoids and their glycosides, along with phenylethanoid derivatives, and

hydroxybenzoic and hydroxycinnamic acids reported in different plant parts (Macías-Cortés, et al.,

2022). These molecules are acknowledged for their anti-inflammatory, antioxidant and cytoprotective

activities, making salvilla a relevant source of bioactive metabolites. Under this idea, it is important

to determine whether bioactive compounds can be extracted, identified, and quantified from

supercritical fluid extracts.

Using UPLC-PDA-ESI-MS/MS five groups of phenolic compounds were identified in the extracts. In

this study, as hydroxybenzoic acids were discovered protocatechuic, benzoic, syringic, 4-

hydroxybenzoic, shikimic, quinic and vanillic acids, giving a total hydroxybenzoic acids from 338.6

to 622.0 ng/mg of SE, while in the hydroxycinnamic acids group were found coumaric, caffeic, ferulic,

caftaric, 4-O-cafeoilquinic, chlorogenic and trans cinnamic acids, with a total content going from 123.8

to 238.6 ng/mg of SE. For total flavones, apigenin, luteolin and acacetin were detected, with total

values ranging from 374.30 to 848.00 ng/mg of SE; sample 7 obtained at 120 min, 100 Bar and 20 % of

co-solvent had the highest values. Another identified group was flavanones, containing eriodyctiol,

naringenin and naringin, with total values from 4.30 to 19.60 ng/mg of SE, this the result obtained at

180 min, 150 Bar and 20 % of ethanol (sample 6). Finally, many molecules of flavonols group were

found, such as mirycetin, kaempferol and its glucoside, rutin, and the most important, quercetin and

its glucosides, quercetin glucuronide and quercetin-3-O-glucoside, quercetin is known as a powerful

anti-inflammatory molecule (Villegas-Novoa et al., 2019). This group had the highest concentration

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in sample 7, who showed 11586.70 ng/mg of SE. This profile is consistent with the reported by Macías-

Cortés et al. (2022) who performed an ultrasound-assisted extraction of salvilla using a

hydroalcoholic solvent.

Fig. 5 presents the response surface plot evaluating the effects of pressure and time of extraction on

the content of total phenolic compounds (ng/mg of SE), while keeping the ethanol concentration

constant at 20 %. In this sense, values ranged from 4185.10 to 13008.20 ng/mg of SE, with sample 7

showing the highest values. These discoveries are consistent with previous reports showing that

adequate extraction times and the use of polar co-solvents such as ethanol can improve the recovery

of bioactive molecules using supercritical or pressurized fluid-based technologies (Díaz-Rivas et al.,

2015; Macías-Cortés et al., 2022; Rosales-Villarreal et al., 2022).

Figure 5. Total phenolic compounds by UPLC-PDA-ESI-MS/MS (2D surface response model)

Figura 5. Compuestos fenólicos totales por UPLC-PDA-ESI-MS/MS (modelo de superficie de respuesta en 2D)

3.6 General Linear Modeling of treatments

Regarding optimal conditions for salvilla leaves extraction, a General linear Modeling (GLM)

was used with a quadratic fit, and a comparison between factors was performed, revealing a good

fit. According with this, in Fig. 6a can be observed that higher values of bioactive compounds and its

antioxidant activity can be obtained up to 120 minutes, no matter how much pressure is applied on

the system (desirability 8.0), the same behaviour can be observed in Fig. 6b (desirability 7.5), were no

matter how much pressure were added to the system, it did not have any influence on the results,

getting a good extract beginning in 20 % of ethanol. Finally, in Fig. 6c it can be observed that from 20

% of ethanol and 120 minutes of extraction it is possible to obtain good results, with a good quality

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extract, this can be a benefit since the process can be expensive compared to other emerging

technologies.

a) b)

Figure 6. a) General Linear Modeling desirability contours (Quadratic fit) of pressure vs time; b) General

Linear Modeling desirability contours (Quadratic fit) of ethanol vs pressure; c) General Linear Modeling

desirability contours (Quadratic fit) of ethanol vs time

Figura 6. a) Contornos de deseabilidad del Modelo Lineal General Linear (Quadratic fit) de la presión vs

tiempo; b) Contornos de deseabilidad del Modelo Lineal General Linear (Quadratic fit) del etanol vs presión; c)

Contornos de deseabilidad del Modelo Lineal General Linear (Quadratic fit) de etanol vs tiempo

4. Conclusions

In the present work, the extraction of bioactive compounds from salvilla was evaluated using

supercritical fluid extraction, with a central composite design. Supercritical fluid extraction (SFE) of

Buddleja scordioides Kunth (salvilla) has demonstrated to be an effective, environmentally friendly,

and promising green emerging technology for the recovery of phenolic compounds, antioxidant

metabolites, and other bioactive molecules, despite achieving lower extraction yields compared to

conventional or alternative emerging extraction methods. Ethanol concentration consistently

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emerged as the most influential factor on extraction yield and the recovery of phenolic and flavonoid

compounds, as confirmed by statistical modeling, highlighting its key role in modulating solvent

polarity and enhancing solubilization of medium-polarity compounds. On the other hand,

supercritical fluid extraction (SFE) enables the recovery of compounds with antioxidant capacity and

other biologically relevant activities, often surpassing the performance of extracts obtained with

conventional solvents; antioxidant assays confirmed this behavior, while UPLC-PDA-ESI-MS/MS

profiling revealed diverse phenolic groups, predominantly flavonols such as quercetin derivatives,

widely recognized for their biological and pharmacological relevance.

General linear modeling further indicated that extraction times around 120 minutes combined with

≥20 % ethanol are sufficient to achieve extracts with high bioactive quality, regardless of pressure

increments. This finding is especially relevant considering the energetic and operational costs

associated with high-pressure systems. Overall, the integration of chemical, antioxidant, and

modeling results supports the use of SFE as a selective, cleaner, and scalable alternative for obtaining

functional compounds from salvilla. These insights not only strengthen the technological relevance

of SFE but also open the door for its future application in the development of nutraceutical, cosmetic,

or functional-food ingredients derived from this traditionally valued species. Future studies should

explore process scale-up, fine-tuning of co-solvent dynamics, and integration with downstream

purification steps to further enhance compound recovery and commercial feasibility. In this sense,

the promising results obtained here provide a solid foundation for consolidating SFE as a sustainable

and efficient technology for the industrial exploitation of bioactive potential from salvilla plants.

Author Contributions

Conceptualization, E.M.C. and M.G.R.A.; methodology, R.M.H.R; software, J.A.V.N and

J.A.G.I.; validation, R.F.G.L. and E.M.C.; formal analysis, R.F.G.L. and N.E.R.G.; investigation,

E.M.C.; resources, G.A.C.H. and J.A.G.I.; data curation, R.F.G.L.; writing—original draft preparation,

E.M.C.; writing—review and editing, M.G.R.A.; visualization, J.A.V.N.; supervision, R.F.G.L.;

project administration, R.F.G.L. All authors have read and accepted the pubished version of the

manuscript.

Acknowledgements

First author E.M.C. thanks to Secretaría de Ciencia, Humanidades, Tecnología e Innovación

of México (SECIHTI) for graduate scholarship and to the Centro de Investigación y Asistencia en

Tecnología y Diseño del Estado de Jalisco, A.C (CIATEJ – Unidad Zapopan) for the loan of

Supercritical fluid equipment.

Conflict of interest

The authors declare that they have no conflicto of interests.

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Nomenclature

CO₂ Carbon dioxide

ESI Electrospray ionization

MAE Microwave-assisted extraction

mL Milliliter

mL/min Milliliters per minute

RSM Response surface methodology

# Number

% Percent

≥ Major or equal than

® Registered trademark

°C Celsius degree

µmol TE/g Micromole of Trolox equivalent per gram of extract

AAPH 2,2'-azobis(2-amidinopropane) dihydrochloride

ABTS 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic) acid

ABTS•+ 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic) radical

AlCl3 Aluminum chloride

Bar Bars

CCD Central composite design

DNA Deoxyribonucleic acid

Fluorescein 3’,6’-dihydroxyspiro[isobenzofuran-1[3H],9’[9H]-xanthen]-3-one

FRAP Ferric Reducing Antioxidant Power

g Grams

GLM General linear model

h Hours

HPLC High performance liquid chromatography

K2S2O8 Potassium persulfate

kV Kilovolts

L/h Liters per hour

LC grade Liquid chromatography grade

M Molar

m/V Ratio mass – volume

min Minutes

mL/min Milliliter per minute

mm Millimeters

mM Millimolar

mmol TE/mg of SE Millimoles of Trolox equivalent per milligram of salvilla leaves extract

MRM Multiple reactions ionization mode

NaNO2 Sodium nitrite

NaOH Sodium hydroxide

ng/mg SE Nanograms per milligram of salvilla leaves extract

ng/μL Nanograms per microliter

nm Nanometers

Greek symbols

 Statistical alpha

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5. References

Avila-Acevedo, J.G., & Romo de Vivar, A. (2002). Triterpenoid saponins and other glycosides from

Buddleja scordioides. Biochemical Systematics and Ecology, 30 (10), 1003-1005.

https://doi.org/10.1016/S0305-1978(02)00032-7

Ayyildiz, S.S., Pelvan, E., & Karadeniz, B. (2024). Optimization of accelerated solvent extraction,

ultrasound assisted and supercritical fluid extraction to obtain carnosol, carnosic acid and

rosmarinic acid from rosemary. Sustainable Chemistry and Pharmacy, 37, 101422.

https://doi.org/10.1016/j.scp.2023.101422

Azmir, J., Zaidul, I. S. M., Rahman, M. M., Sharif, K. M., Mohamed, A., Sahena, F., Jahurul, M.H.A.,

Ghafoor, K., Norulaini N.A.N., & Omar, A. K. M. (2013). Techniques for extraction of bioactive

compounds from plant materials: A review. Journal of Food Engineering, 117(4), 426–436.

https://doi.org/10.1016/j.jfoodeng.2013.01.014

Banafi, A., Wee, S.K., Tze-Tiong, A. N., Yang-Kong, Z., Saptoro, A., & Sunarso, J. (2023). Modeling of

supercritical fluid extraction bed: A critical review. Chemical Engineering Research and Design, 193,

685-712. https://doi.org/10.1016/j.cherd.2023.04.001

Bezerra, M. A., Santelli, R. E., Oliveira, E. P., Villar, L. S., & Escaleira, L. A. (2008). Response surface

methodology (RSM) as a tool for optimization in analytical chemistry. Talanta, 76(5), 965–977.

http://dx.doi.org/10.1016/j.talanta.2008.05.019

Chemat, F., Rombaut, N., Fabiano-Tixier, A.S., Pierson, J. T., & Bily, A. (2015). Green Extraction: From

Concepts to Research, Education, and Economical Opportunities. In: Chemat F., & Strube, J. (Eds.)

Green Extraction of Natural Products: Theory and Practice. Wiley-VCH. Chapter 1 pags. 1-36.

https://doi.org/10.1002/9783527676828.ch1

Chemat, F., Abert-Vian, M., Fabiano-Tixier, A.S., Strube, J., Uhlenbrok, L., Gunjevic, V., & Cravotto,

G. (2019). Green extraction of natural products. Origins, current status, and future challenges.

TrAC Trends in Analytical Chemistry, 118, 248-263. https://doi.org/10.1016/j.trac.2019.05.037

Coelho, J.P., Filipe, R.M., Robalo, M.P., & Stateva, R.P. (2018). Recovering value from organic waste

materials: Supercritical fluid extraction of oil from industrial grape seeds. The Journal of

Supercritical Fluids, 141, 68-77. https://doi.org/10.1016/j.supflu.2017.12.008

Dawidowicz, A. L., & Olszowy, M. (2013). The importance of solvent type in estimating antioxidant

properties of phenolic compounds by ABTS assay. European Food Research and Technology, 236(6),

1099–1105. https://doi.org/10.1007/s00217-013-1982-1

Dębczak, A., Grzegorczyk, A., Świątek, Ł. Sandomierski, M., Fekner, Z., Tyśkiewicz, K., &

Milovanovic, S. (2024). Green and sustainable recovery of polyphenols, flavonoids, alkaloids, and

pigments from green tea leaves: Comparative analysis of Soxhlet, accelerated solvent, and

supercritical fluid extraction techniques. Sustainable Chemistry and Pharmacy, 42, 101810.

https://doi.org/10.1016/j.scp.2024.101810

Díaz-Rivas, J. O., Herrera-Carrera, E., Gallegos-Infante, J. A., Rocha-Guzmán, N. E., González-

Laredo, R. F., Moreno-Jiménez, M. R., Larrosa-Pérez, M., & Gallegos-Corona, M. A. (2015).

Gastroprotective potential of Buddleja scordioides Kunth Scrophulariaceae infusions; effects into

the modulation of antioxidant enzymes and inflammation markers in an in vivo model. Journal of

ethnopharmacology, 169, 280-286. https://doi.org/10.1016/j.jep.2015.04.024

Díaz-Rivas, J. O., Gallegos-Infante, J. A., Valdez-Fragoso, A., Rocha-Guzmán, N. E., González-Laredo,

R. F., Rodríguez-Ramírez, A., Gamboa-Gómez, C. I., & Moreno-Jiménez, M. R. (2018).

Comparative Study of Phenolic Profile and Content in Infusions and Concentrated Infusions of

Macías-Cortés et.al

TECNOCIENCIA CHIHUAHUA, Vol. XX (2026): enero-diciembre, e2175

Buddleja scordioides Treated by High-Intensity Pulsed Electric Fields (HiPEF). Beverages, 4(4), 81.

https://doi.org/10.3390/beverages4040081

Herrera-Carrera, E., Moreno-Jiménez, M. R., Rocha-Guzmán, N. E., Gallegos-Infante, J. A.,

Díaz-Rivas, J. O., Gamboa-Gómez, C. I., & González-Laredo, R. F. (2015) Phenolic composition of

selected herbal infusions-and their anti-inflammatory effect on a colonic-model in vitro in HT-29

cells. Cogent Food & Agriculture, 1(1): 1059033. https://dx.doi.org/10.1080/23311932.2015.1059033

Herrero, M, Cifuentes, A., & Ibañez, E. (2006). Sub- and supercritical fluid extraction of functional

ingredients from different natural sources: Plants, food-by-products, algae and microalgae: A

review. Food Chemistry, 98(1), 136-148. https://doi.org/10.1016/j.foodchem.2005.05.058

Lee, S. G., Lee, D., & Kang, H. (2020). Effect of extraction ethanol concentration on antioxidant and

anti-inflammatory activity of 30-year-old and 120-year-old dangyuja (Citrus maxima (Brum.)

Merr). Biomedical Science Letters, 26(2), 109-113. https://doi.org/10.15616/BSL.2020.26.2.109

Machmudah, S., Shotipruk, A., Goto, M., Sasaki M., & Hirose, T. (2006) Extraction of astaxanthin from

Haematococcus pluvialis using supercritical CO2 and ethanol as entrainer. Industrial & Engineering

Chemistry Research, 45(10), 3652-3657. https://doi.org/10.1021/ie051357k

Macías-Cortés, E., Rocha-Guzmán, N. E., Moreno-Jiménez, M. R., Medina-Torres, L., & González-

Laredo, R. F. (2020). Microencapsulation of phenolic compounds: Technologies and novel

polymers. Revista Mexicana de Ingeniería Química, 19(2), 491 - 521.

https://doi.org/10.24275/rmiq/Alim642

Macías-Cortés, E., Rocha-Guzmán, N. E., Moreno-Jiménez, M. R., Castillo-Hernández, G. A., &

González-Laredo, R. F. (2022). Antioxidant and anti-inflammatory polyphenols in ultrasound-

assisted extracts from salvilla (Buddleja scordioides Kunth). Ultrasonics Sonochemistry, 83, 105917.

https://doi.org/10.1016/j.ultsonch.2022.105917

Mustafa, A., & Turner, C. (2011). Pressurized liquid extraction as a green approach in food and herbal

plants extraction: A review. Analytica Chimica Acta, 703(1), 8–18.

https://doi.org/10.1016/j.aca.2011.07.018

Ou, B., Hampsch-Woodill, M., & Prior, R. L. (2001). Development and validation of an improved

oxygen radical absorbance capacity assay using fluorescein as the fluorescent probe. Journal of

Agricultural and Food Chemistry, 49(10), 4619–4626. https://doi.org/10.1021/jf010586o

Perez-Gutierrez, R.M., & Vargas-Solis, R. (2008). Wound Healing Properties of Triterpenes from

Buddleia scordioides in Diabetic Rats. Pharmaceutical Biology, 46(9), 647–653.

https://doi.org/10.1080/13880200802182471

Picos-Salas, M.A., Leyva-López, N., Bastidas-Bastidas, P.J., Antunes-Ricardo M., Cabanillas-

Bojórquez, L.A., Angulo-Escalante, M.A., Heredia J. B., & Gutiérrez-Grijalva, E.P. (2024).

Supercritical CO2 extraction of naringenin from Mexican oregano (Lippia graveolens): its

antioxidant capacity under simulated gastrointestinal digestion. Scientific Reports, 14, 1146.

https://doi.org/10.1038/s41598-023-50997-2

Pilařová, V., Plachká, K., Herbsová, D., Kosturko, Š., Svec, F., & Nováková, L. (2024). Comprehensive

two‑step supercritical fluid extraction for green isolation of volatiles and phenolic compounds

from plant material. Green Chemistry, 26, 6480–6489. https://doi.org/10.1039/D4GC00371C

Pilařová, V., Burianová, K., Kosturko, Š., Kulkarni, M., Van-Staden, J., & Nováková, L. (2025).

Supercritical fluids in extraction, purification, and separation of selected Amaryllidaceae

alkaloids. Separation and Purification Technology, 377(Part 2), 133968.

https://doi.org/10.1016/j.seppur.2025.133968

Plaza, M., & Turner, C. (2015). Pressurized hot water extraction of bioactives. TrAC Trends in

Analytical Chemistry, 71, 39–54. https://doi.org/10.1016/j.trac.2015.02.022

Macías-Cortés et.al

TECNOCIENCIA CHIHUAHUA, Vol. XX (2026): enero-diciembre, e2175

Pourmortazavi, S. M., & Hajimirsadeghi, S. S. (2007). Supercritical fluid extraction in plant essential

and volatile oil analysis. Journal of Chromatography A, 1163(Issues 1–2), 2–24.

https://doi.org/10.1016/j.chroma.2007.06.021

Radzali, S.A., Markom, M., & Saleh, N.M. (2020). Co-Solvent Selection for Supercritical Fluid

Extraction (SFE) of Phenolic Compounds from Labisia pumila. Molecules, 25(24) 5859.

https://doi.org/10.3390/molecules25245859

Reverchon, E., & De Marco, I. (2006). Supercritical fluid extraction and fractionation of natural matter.

The Journal of Supercritical Fluids, 38(2), 146–166. https://doi.org/10.1016/j.supflu.2006.03.020

Rice-Evans, C., Miller, N., & Paganga, G. (1997). Antioxidant properties of phenolic compounds.

Trends in Plant Science, 2(4) 152-159. https://doi.org/10.1016/S1360-1385(97)01018-2

Rosales-Villarreal, M. C., Rocha-Guzmán, N. E., Gallegos-Infante, J. A., Moreno-Jiménez, M. R.,

Reynoso-Camacho, R., Pérez-Ramírez I. F., Tsao, R., & González-Laredo, R. F. (2022). Antioxidant

phytochemicals and modulation of oxidative stress from infusions and ultrasound-assisted

extractions of Coffea leaves. Revista Chapingo Serie Ciencias Forestales y del Ambiente, 28(1), 119-137.

http://doi.org/10.5154/r.rchscfa.2021.05.026

Sheibani, S., Jafarzadeh, S., Qazanfarzadeh, Z., Osadee-Wijekoon, M.M.J., Mohd-Rozalli M.H., &

Nafchi, A.M. (2024). Sustainable strategies for using natural extracts in smart food packaging,

International Journal of Biological Macromolecules, 267(Part 1), 131537.

https://doi.org/10.1016/j.ijbiomac.2024.131537

Usman, M., Nakagawa M., & Cheng, S. (2023). Emerging trends in green extraction techniques for

bioactive natural products. Processes, 11(12), 3444. https://doi.org/10.3390/pr11123444

Uwineza, P. A., Gramza-Michałowska, A., Bryła, M., & Waśkiewicz, A. (2021). Antioxidant Activity

and Bioactive Compounds of Lamium album Flower Extracts Obtained by Supercritical Fluid

Extraction. Applied Sciences, 11(16), 7419. https://doi.org/10.3390/app11167419

Villegas-Novoa, C., Gallegos-Infante, J. A., González-Laredo, R. F., García-Carrancá, A., Herrera-

Rocha, K. M., Jacobo-Karam, J. S., Moreno-Jiménez, M. R., & Rocha-Guzmán, N. E. (2019). Acetone

effects on Buddleja scordioides polyphenol extraction process and assessment of their cellular

antioxidant capacity and anti-inflammatory activity. Medicinal Chemistry Research, 28(12), 2218–

2231. https://doi.org/10.1007/S00044-019-02448-9

Vinitha, U.G., Sathasivam, R., Muthuraman, M.S., & Park, S.U. (2022). Intensification of supercritical

fluid in the extraction of flavonoids: A comprehensive review. Physiological and Molecular Plant

Pathology, 118, 101815, https://doi.org/10.1016/j.pmpp.2022.101815

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