Agave Sap Research Articles

Health-Related Composition and Bioactivity of an Agave Sap/Prickly Pear Juice Beverage

In this study, a beverage made from a combination of Agave sap (AS) and prickly pear juice (PPJ) was analyzed for its nutrients and bioactive and potentially health-promoting compounds. The beverage was evaluated for its ability to act as an antioxidant, regulate glycemic properties, and undergo gut bacterial fermentation in vitro. The major mono- and oligosaccharides present in the beverage were galacturonic acid (217.74 ± 13.46 mg/100 mL), rhamnose (227.00 ± 1.58 mg/100 mL), and fructose (158.16 ± 8.86 mg/mL). The main phenolic compounds identified were protocatechuic acid (440.31 ± 3.06 mg/100 mL) and catechin (359.72 ± 7.56 mg/100 mL). It was observed that the beverage had a low glycemic index (<40) and could inhibit digestive carbohydrases. The combination of ingredients also helped to reduce gas production during AS fermentation from 56.77 cm3 to 15.67 cm3. The major SCFAs produced during fermentation were butyrate, acetate, and propionate, with valerate being produced only during the late fermentation of the AS. This beverage is rich in bioactive compounds, such as polyphenols and dietary fiber, which will bring health benefits when consumed.

Fermentation of Agave americana L. sap produced in Cayambe – Ecuador

Fermentation of agave sap, also known as exudate, has become an ancestral practice throughout Ecuadorian Andean. In Cayambe, located in this region, grows Agave americana L., which is recollected, and its sap is fermented. The agave-based fermented beverage, locally named "tzawar mishki", exhibits variable features, mainly ethanol concentration. In this work, fermentation conditions of agave sap were studied to enhance ethanol yield. Two thermal treatments for raw exudate were evaluated, pasteurization at boiling point for 30 minutes and sterilization at 121°C for 15 minutes; fermentation temperature, 30°C and room (around 18°C); and two yeast strains. Thermal pretreatments have a positive impact on reducing sugars and sucrose concentration. In the first case, an increase of 76 % and 30 % has been reported, while sucrose concentration quadrupled and doubled in pasteurized and sterilized samples, respectively. The highest ethanol concentration (63,31 g/L) and the best yield (66,21 %) were accomplished through agave sap pasteurized and fermented for 96 hours at 30°C. Negligible differences have been evidenced in ethanol and other volatile compounds content between the two yeast strains evaluated. Keywords: agave sap, ethanol, fermentation, fermentable sugars, yeast

Microencapsulation of steroidal saponins from agave sap concentrate using different carriers in spray drying.

Concentrated agave sap is a product with in vivo proven hypocholesterolemic and hypoglycemic activities, as well as in vitro anticancer potential. In the present work, a factorial design was used to determine the suitable drying conditions of concentrated agave by studying the effect of inlet temperature (150 °C, 180 °C and 210 °C) and the type of carrier agent (maltodextrin, hydroxypropyl methylcellulose, guar gum and xanthan gum). The response variables for each treatment were the product recovery and microencapsulated saponins. Further characterization of concentrated agave powders was performed: solubility in water, hygroscopicity, moisture content, tap density, bulk density, Carr's index followability and morphology by scanning electron microscopy analysis. The hydroxypropyl methylcellulose proved to improve physicochemical properties and enhance product yield, using 210 °C inlet temperature and a mix of carrier agents of maltodextrin/hydroxypropyl methylcellulose/xanthan gum at 50/48.5/1.5 (w/w/w) proportion exhibited the highest saponin recovery of 53.81%. Moreover, different carrier agents in powders revealed two shapes, regular spherical shape with smooth surface and collapsed shapes. The use of polymers excipients helped to decrease the stickiness of the desired product and enhanced the powder stability and microencapsulation of the steroidal saponins.

Morphological and genetic diversity of traditional varieties of agave in Hidalgo State, Mexico.

The agaves are plants of cultural importance which have been used by humans for about 10,000 years and about 40 specific uses. The most culturally and economically important of those uses are for the production of fermented (pulque) and distilled beverages (mescal). Pulque continues to be produced in nearly all of Mexico, and the agaves used for this purpose have shown domestication syndrome. We carry out an ethnobotanical, morphological, and genetic analysis of the traditional varieties of pulque agave used in the production of aguamiel (agave sap) and pulque in the state of Hidalgo. We did semi-structured interviews, free listings, and tours with 11 agave managers. We analyzed morphology and studied genetic diversity and structure using nuclear microsatellites. We found wild-collected, tolerated, transplanted, and cultivated varieties of agave. This comprised 19 traditional varieties of pulque agave, 12 of them in production during the study, which corresponded to the species Agave americana, A. salmiana y A. mapisaga and five intraspecific entities. The varieties were grouped morphologically according to a management gradient; the wild-collected varieties were the smallest, with more lateral teeth and a larger terminal spine. The cultivated varieties clearly exhibited domestication syndrome, with larger plants and smaller dentition. The expected heterozygosity (He) of the varieties ranged from 0.204 to 0.721. Bayesian clustering suggested the existence of three genetic groups, both at the level of traditional varieties of pulque agaves and for management categories, a result that matches multivariate clustering. Pulque producers in the studied localities maintain high agrobiodiversity. The cultivated varieties exhibit domestication syndrome, as has been reported for other species of the genus with the same selection purposes. Our results support the hypothesis of a decrease in genetic diversity in crops compared to wild-growing agaves, which seems to be due to vegetative propagation, among other factors.

Fructosyltransferase production by Aspergillus oryzae BM-DIA using solid-state fermentation and the properties of its nucleotide and protein sequences.

Fructosyltransferase (FTase) catalyzes the transfer of a fructosyl group to a sucrose molecule or a fructooligosaccharide (FOS) when a FOS with a longer chain is formed. Production of FTase by two Aspergillus species and its mixture was exploited using solid-state fermentation (SSF) and employing agave sap as substrate. The maximum FTase activity (1.59 U/mL) by Aspergillus oryzae was obtained after 24h, using a temperature of 30°C, with an inoculum of 2 × 107 spores/mL. The nucleotide sequence coding for the fructosyltransferase showed 1494bp and encodes for a protein of 498 amino acids. The hypothetical molecular tertiary structure of Aspergillus oryzae BM-DIA FTase showed the presence of structural domains, such as a five-bladed beta-propeller domain characteristic of GH (glycoside hydrolase) and C terminal, which forms a beta-sandwich module. This study contributes to the knowledge of stability, compatibility, and genetic expression of Aspergillus oryzae BM-DIA under SSF bioprocess conditions for industrial production of fructosyltransferase.

Assessment of the bacterial diversity of agave sap concentrate, resistance to in vitro gastrointestinal conditions and short-chain fatty acids production

Low bacterial diversity in the gut has been associated with the development of several diseases. Agave sap concentrate (ASC) is obtained from the thermal treatment of the fresh sap called “aguamiel”, an artisanal Mexican food. In this study, we assessed the microbial diversity from three different ASC producing regions in Mexico using high-throughput sequencing of the 16S rRNA gene and evaluated their resistance to an in vitro gastrointestinal process as well as their ability to produce short-chain fatty acids (SCFA). Seven phyla and 120 genera were detected in ASC samples; Firmicutes had the highest relative read abundance at the phylum level, whereas Bacillus was the most abundant genus. Bacterial diversity at phylum and genus levels was highly dependent on the region where ASC was produced. The microbiota from a selected sample was resistant to low pH conditions, bile salts and intestinal enzymes. Moreover, bacteria were able to survive and grow in the colonic environment. SCFA production was comparable with that observed for a well-known probiotic, Lactobacillus plantarum 299v, that was used as control. These findings demonstrate that ASC contains a bacterial ecosystem with potential probiotic benefits.

Effect of Solvent Composition on Ultrasound-Generated Intensity and Its Influence on the Ultrasonically Assisted Extraction of Bioactives from Agave Bagasse (Agave salmiana)

Agave bagasse is a waste material generated during agave sap collection for “pulque” production. This residue is a potential feedstock for steroidal saponins and other phytochemicals. Ultrasonically assisted extraction (UAE) allows the use of green solvents such as water or ethanol for phytochemicals recovery. The influence of solvent composition and its effect on the intensity of the ultrasonic field during the UAE of agave bagasse saponins were evaluated. The best extraction conditions were a temperature of 60 °C (T) and a solvent-to-mass ratio of 20 (S/M), for conventional and UAE. In contrast to conventional extraction, where mixtures of ethanol-water were more efficient, water was the best extraction solvent in UAE. The amount of saponins obtained without ultrasound in 58% ethanol (22.48 ± 1.34 mg PE/g dw) was similar to the amount obtained by UAE in water (24.41 ± 0.84 mg PE/g dw). When only water was used, the ultrasound intensity was higher (271.40 ± 11.91 W/L) than in ethanol mixtures (144.81–202.30 W/L) due to cavitation intensity, demonstrated in a test on aluminum foil and SEM images of exhausted bagasse. Despite lower chemical affinity of water for saponins, its lower vapor pressure and viscosity, as well as higher surface tension, compared with ethanol, provided a greater cavitation intensity. Therefore, UAE using water is an excellent alternative to recover bioactive compounds from agave bagasse.

Evolution of Fructans in Aguamiel (Agave Sap) During the Plant Production Lifetime.

Aguamiel is the sap collected from agave, while pulque is the result of the natural fermentation of aguamiel. Despite its ancestral origin and numerous publications on pulque production, little is known about the evolution and concentration of sugars and fructo-oligosaccharides in aguamiel, either during its daily accumulation or through the agave production lifetime. In this study, we examined aguamiel composition in three agave plants during their productive lifetime (4 to 9 months). After each collection, the agave pine is scraped to induce aguamiel to flow into an internally created cavity (cajete), producing a residual bagasse (metzal). We found that the concentration of agave fructans and sucrose, as well as the fructan profile, change during the aguamiel production process. During the daily collection, a small amount of agave fructans released from the pine by scraping is drawn into the cajete with the first milliliters of sap where it is then diluted with the inflow of aguamiel. The main component of aguamiel is the sucrose produced in high concentration in the leaves through photosynthesis and then hydrolyzed in the cajete as aguamiel accumulates. We also describe how the fructan profile changes during the accumulation of aguamiel in the cajete. In addition to the varying amount of sucrose that is hydrolyzed in the aguamiel accumulated, we found that fructo-oligosaccharides are either diluted, consumed, or hydrolyzed, depending on the plant and its production stage, thus yielding different fructan profiles. New fructo-oligosaccharides are, in some cases, synthesized by bacteria present in aguamiel. These profiles were also observed in aguamiel collected from ten different plants in the same production region. We also found that a considerable amount of agave fructans is lost in metzal (bagasse), the agave material that is scraped and thrown away twice a day during the production process.

Genomic profiling of bacterial and fungal communities and their predictive functionality during pulque fermentation by whole-genome shotgun sequencing

Pulque is a culturally important 4,000-year-old traditional Mexican fermented drink. Pulque is produced by adding fresh aguamiel (agave sap) to mature pulque, resulting in a mixture of microbial communities and chemical compositions. We performed shotgun metagenomic sequencing of five stages of pulque fermentation to characterize organismal and functional diversity. We identified 6 genera (Acinetobacter, Lactobacillus, Lactococcus, Leuconostoc, Saccharomyces and Zymomonas) and 10 species (Acinetobacter boissieri, Acinetobacter nectaris, Lactobacillus sanfranciscensis, Lactococcus lactis, Lactococcus piscium, Lactococcus plantarum, Leuconostoc citreum, Leuconostoc gelidum, Zymomonas mobilis and Saccharomyces cerevisiae) that were present ≥ 1% in at least one stage of pulque fermentation. The abundance of genera and species changed during fermentation and was associated with a decrease in sucrose and increases in ethanol and lactic acid, suggesting that resource competition shapes organismal diversity. We also predicted functional profiles, based on organismal gene content, for each fermentation stage and identified an abundance of genes associated with the biosynthesis of folate, an essential B-vitamin. Additionally, we investigated the evolutionary relationships of S. cerevisiae and Z. mobilis, two of the major microbial species found in pulque. For S. cerevisiae, we used a metagenomics assembly approach to identify S. cerevisiae scaffolds from pulque, and performed phylogenetic analysis of these sequences along with a collection of 158 S. cerevisiae strains. This analysis suggests that S. cerevisiae from pulque is most closely related to Asian strains isolated from sake and bioethanol. Lastly, we isolated and sequenced the whole-genomes of three strains of Z. mobilis from pulque and compared their relationship to seven previously sequenced isolates. Our results suggest pulque strains may represent a distinct lineage of Z. mobilis.

Novel process to obtain agave sap (aguamiel) by directed enzymatic hydrolysis of agave juice fructans

Kluyveromyces marxianus and Saccharomyces paradoxus, which are the native microorganisms found in aguamiel, were used successfully for exo- and endo-fructanase synthesis to hydrolyze agave fructans important in agave sap production. We studied culture media for enzyme synthesis, as well as the physicochemical properties and enzyme activity, and we optimized the bioprocess parameters and the enzyme extract mixture to maximize agave fructan hydrolysis. The results showed that the best culture medium for fructanase synthesis contained 14.0, 20.0, and 1.0 (g-L−1) of sucrose, peptone, and K2HPO4, respectively. The best enzyme-to-substrate ratio for agave juice containing 90 g-L−1 of fructans was 10:1. Under constant operating conditions of 45.25 °C, 65.47 rpm, and pH 4.28, a product with 93.62 g-L−1of total reducing sugars (TRS) and fructooligosaccharides (FOS) with a polymerization degree (PD) from 3 to 10 hexose units was obtained using a mixture of 98.4% K. marxianus and 1.6% S. paradoxus enzyme extracts. The resulting agave sap preserves the natural flavor and tastes closest to natural agave sap (aguamiel); hence, the results show, for the first time, a resolution of the technical challenge for stable agave sap production and confirm the importance of using native strains to evolve from spontaneous to directed fermentation.

Physical, Chemical, and Microbiological Characteristics of Pulque: Management of a Fermented Beverage in Michoacán, Mexico.

Pulque is a beverage that has been prepared in Mexico since pre-Hispanic times from the fermented sap of more than 30 species of wild and domesticated agaves. We conducted studies in two communities of the state of Michoacán, in central-western Mexico, where we documented its traditional preparation and analyzed the relationship between preparation conditions and the composition and dynamics of microbiological communities, as well as the physical and chemical characteristics of the beverage. In one of the communities, Santiago Undameo (SU), people boil the sap before inoculating it with pulque inoculum; this action causes this local pulque to be sweeter, less acidic, and poorer in bacteria and yeast diversity than in the other community, Tarimbaro (T), where the agave sap is not boiled and where the pulque has more diversity of microorganisms than in SU. Fermentation management, particularly boiling of the agave sap, influences the dynamics and diversity of microbial communities in the beverage.

Processing temperature effect on the chemical content of concentrated aguamiel syrups obtained from two different Agave species

Mexico has an enormous diversity of Agave species. The agave sap (called aguamiel) is concentrated to obtain a syrup. In this study, the aguamiel was obtained from two different Agave species: A. salmiana and A. atrovirens, then it was concentrated using four different temperature treatments (70 °C, 80 °C, 90 °C, and 95 °C). During thermal processing, the chemical, proximal, and sweetness analyses were determined. The temperature affected the color, samples treated at 70 °C and 80 °C were the darkest. The pH of the syrups was acidic (4.15 to 5.47). The moisture of the samples remains constant (ca. 40%). The fat value in the samples was under 1.3%, and the fiber was not detected in any of the samples. The A. salmiana syrup had the highest ash content (8%), A. atrovirens contain 50% less. In protein content, A. atrovirens syrups presented 8.5% and A. salmiana nearly 3%. The 5-hydroxymethyl-furfural was not affected by any of the used temperatures. A. atrovirens syrups contained more glucose (ca. 23%). The temperature affected the sucrose content; it was higher at 70 °C. The samples revealed the content of fructooligosaccharides, such as 1-kestose, 1-nystose, and 1-fructofuranosyl nystose. 1-nystose was detected only in A. salmiana syrups. The food analysis of these syrups was essential to understand the effect of the Agave species and temperature treatments used during the production of agave syrup.

Influence of fermentation by different microflora consortia on pulque and pulque bread properties.

Pulque bread is a traditional Mexican product obtained by fermentation using microflora present only in pulque. In this study, the possibility of creating a pulque microbial consortium under laboratory conditions and its applications were evaluated. A laboratory-made consortium was compared with a consortium originating in Mexico in bread and pulque production. They were tested in various growth medium systems: pulque made from agave sap and malt extract, Mexican wheat and rye pulque bread, and European wheat and rye bread. Depending on the growth medium, consortiums showed differing influence on many factors, such as specific volume, weight loss after baking, soluble proteins, and crust and crumb color. Indigenous starters increased sensorial acceptance of pulque and Mexican rye bread, decreased pH, and increased titratable acidity of the breads at the highest level whereas laboratory consortia improved sensory acceptance of wheat breads. The laboratory-prepared starter in some cases improved antiradical activity. All pulques received similar consumer evaluations. However, malt pulque was the least appreciated beverage. The results show the possibility of creating a pulque microbial consortium under laboratory conditions. Depending on the flour type and the breadmaking technique, the use of a particular microbial consortium allowed modification of certain physicochemical parameters. In conclusion, it is feasible to modify bread parameters to obtain features corresponding to consumer demands by using an appropriate microflora, pulque, or flour type. Moreover, this research describes, for the first time, the use of rye malt for pulque and rye flour for pulque bread preparation as raw materials. © 2019 Society of Chemical Industry.

Occupational asthma due to tampico fiber bystander exposure in a brush production company—case report and literature review

A female machine operator in a brush factory presented for an expert medical examination due to suspected occupational obstructive pulmonary disease. She reported the occurrence of severe respiratory distress whenever colleagues in the same working area processed tampico fibers or a mixture of tampico fibers and bassine. The challenge posed in terms of the expert appraisal was to either prove or rule out occupational causality and IgE-mediated (IgE, immunglobulin E) sensitization in relation to the reported workplace-related symptoms. As part of the further diagnostic work-up, tampico fibers were isolated from the workplace and the proteins biotinylated and coupled to a streptavidin ImmunoCAP for specific IgE testing. The material taken from the workplace was checked for mite allergen contamination. In addition, a systematic literature search was conducted in PubMed on “agave-/tampico fibers and diseases.” Tampico-specific IgE antibodies (10.5 kU/l and immunoblot labeling of protein bands in the 25-kDa range) were detected in the patient’s serum. The inhibition assay showed that mite-allergen contamination of the tampico materials was not responsible for the IgE reactivity. Tampico fibers are robust fibers obtained from the leaves of the Mexican Agave lechugilla. The literature describes several cases of skin reactions in the form of irritant contact dermatitis due to occupational exposure to agave leaves or contact with agave sap. In the present case, the asthma symptoms of a female worker in a brush factory were attributed to inhalant bystander exposure to tampico fibers in the workplace. The unequivocal evidence of sensitization to tampico fibers met the medical requirements for the recognition of an occupational disease (BK 4301).

Mass Spectrometry-Based Metabolomics of Agave Sap (Agave salmiana) after Its Inoculation with Microorganisms Isolated from Agave Sap Concentrate Selected to Enhance Anticancer Activity

Saponins have been correlated with the reduction of cancer cell growth and the apoptotic effect of agave sap concentrate. Empirical observations of this artisanal Mexican food have shown that fermentation occurs after agave sap is concentrated, but little is known about the microorganisms that survive after cooking, or their effects on saponins and other metabolites. The aim of this study was to evaluate the changes in metabolites found in agave (A. salmiana) sap after its fermentation with microorganisms isolated from agave sap concentrate, and demonstrate its potential use to enhance anticancer activity. Microorganisms were isolated by dilution plating and identified by 16S rRNA analysis. Isolates were used to ferment agave sap, and their corresponding butanolic extracts were compared with those that enhanced the cytotoxic activity on colon (Caco-2) and liver (Hep-G2) cancer cells. Metabolite changes were investigated by mass spectrometry-based metabolomics. Among 69 isolated microorganisms, the actinomycetes Arthrobacter globiformis and Gordonia sp. were used to analyze the metabolites, along with bioactivity changes. From the 939 ions that were mainly responsible for variation among fermented samples at 48 h, 96 h, and 192 h, four were correlated to anticancer activity. It was shown that magueyoside B, a kammogenin glycoside, was found at higher intensities in the samples fermented with Gordonia sp. that reduced Hep-G2 viability better than controls. These findings showed that microorganisms from agave sap concentrate change agave sap metabolites such as saponins. Butanolic extracts obtained after agave sap fermentation with Arthrobacter globiformis or Gordonia sp. increased the cancer cell growth inhibitory effect on colon or liver cancer cells, respectively.

Variability in Saponin Content, Cancer Antiproliferative Activity and Physicochemical Properties of Concentrated Agave Sap.

Concentrated agave sap (CAS) has gained popularity as an unrefined sweetener. It is obtained by boiling "aguamiel" that contains phytochemicals with diverse bioactivities. Saponins have been the most widely studied agave phytochemicals due to their cancer antiproliferative effect but their concentration may vary due to maturity of the agave plant and collection site. In this study, 18 CAS samples produced in different states of Mexico were analyzed using multivariate methods to determine which physicochemical or phytochemical parameters were responsible for variation. Additionally, extracts with different saponin profiles were tested to determine possible correlations with antiproliferative activity. Total soluble solids, pH, and water activity were similar to those reported for other agave sweeteners. Antioxidant capacity of samples was correlated to browning index. Eleven steroidal saponins were found in CAS samples and they were the main source of variability. Magueyoside B, a kammogenin tetraglycoside, was the most abundant saponin in all samples. With respect to bioactivity, multivariate analysis indicated that magueyoside B and a gentrogenin tetraglycoside were compounds strongly related with bioactivity. CAS from Hidalgo, Puebla, and Veracruz had higher concentration of magueyoside B than from the other kamogenin tetraglycoside found in the samples from other Mexican states. These results could be used as a first approach to characterize and standardize CAS to validate the potential health benefits derived from its consumption.

The Logistic-Exponential Weibull Model as a Tool to Predict Natural Microflora Inactivation of Agave Mapsiaga Aguamiel (Agave Sap) by High Pressure Treatments

Agave sap (aguamiel) is a major source of nutraceutics that may be affected by natural microflora fermentation and thermal processing. High pressure processing (HPP) at 100–400 MPa/25C/1–20 min was investigated as an alternative non-thermal pasteurization by analyzing aerobic mesophiles (AM), coliforms (CL) and yeasts and molds (YM) inactivation. Standard statistics and information theory criteria determined the goodness of fit of first-order kinetics (LKM), Weibull (WBLL) and Gompertz (GMPZ) models. Non-detectable CL were found for 300 MPa/1 min, whereas 400 MPa/4 min HPP treatments reduced AM and YM by 3.86–4.72 log10 units. Statistical analysis showed WBLL best described microbial inactivation. A secondary WBLL model consisting of logistic-exponential b′(P) and exponential decay n(P) functions predicted AM (R2 = 0.835), YM (R2 = 0.867) kinetics for different pressure–time combinations. The secondary WBLL model predicted commercial HPP pasteurization conditions (5-log10 reductions of AM and YM) could be achieved at 400 MPa/5 min or 500 MPa/2 min. Practical Applications High pressure processing (HPP) is a consumer preferred alternative for food pasteurization. Aside from guacamole and some fruit and vegetable juices, there no other significant HPP foods available in México. Our research group believes that the commercialization of food products based on endemic plants such as agave sap (aguamiel), will draw the attention of national and international consumers, researchers and industrial HPP food processors to increase the number of HPP applications. Furthermore, the development of reliable pressure–time predictive models for microbial inactivation would help HPP food processors to ensure food safety and generate better budget estimates for novel products, or redefine current operating conditions since processing costs depend on the applied pressure level and pressure holding time.

Enhancement of fructosyltransferase and fructooligosaccharides production by A. oryzae DIA-MF in Solid-State Fermentation using aguamiel as culture medium

The aim of this work was to improve the production of fructosyltransferase (FTase) by Solid-State Fermentation (SSF) using aguamiel (agave sap) as culture medium and Aspergillus oryzae DIA-MF as producer strain. SSF was carried out evaluating the following parameters: inoculum rate, incubation temperature, initial pH and packing density to determine the most significant factors through Box-Hunter and Hunter design. The significant factors were then further optimized using a Box-Behnken design and response surface methodology. The maximum FTase activity (1347U/L) was obtained at 32°C, using packing density of 0.7g/cm3. Inoculum rate and initial pH had no significant influence on the response. FOS synthesis applying the enzyme produced by A. oryzae DIA-MF was also studied using aguamiel as substrate.

Some Evidences on Effect of Intake Aguamiel (Agave sap)

Honeywater or aguamiel (Agave sap) has been consumed by Mexican population since pre-columbian times. Although, it has been claimed by folk belief that aguamiel possesses some medicinal properties, scientific studies on its effect on human health have not been well documented. The behavior of blood components in nine volunteers (two young males, three adult females and four adult males) after aguamiel consumption (250 mL every three days during a period of 35 days) was analyzed. It was found that, serum red blood cell count, serum white blood cell count, platelet count, minerals (Zn, Mg and Fe) and iron-related proteins (ferritin and transferrin) levels were not negatively affected because of all of these blood indicators ranged within normal reference values. However, this study showed that aguamiel presented a specific functional effect since hypercholesterolemic adult males showed normal levels of serum total cholesterol after aguamiel consumption, whereas total cholesterol levels were kept in normal ranges after aguamiel consumption for normocholesterolemic subjects. Furthermore, aguamiel consumption did not cause hyperglycemia in any of the tested groups.

Fast Centrifugal Partition Chromatography Fractionation of Concentrated Agave (Agave salmiana) Sap to Obtain Saponins with Apoptotic Effect on Colon Cancer Cells.

Separation of potentially bioactive components from foods and plant extracts is one of the main challenges for their study. Centrifugal partition chromatography has been a successful technique for the screening and identification of molecules with bioactive potential, such as steroidal saponins. Agave is a source of steroidal saponins with anticancer potential, though the activity of these compounds in concentrated agave sap has not been yet explored. In this study, fast centrifugal partition chromatography (FCPC) was used coupled with in vitro tests on HT-29 cells as a screening procedure to identify apoptotic saponins from an acetonic extract of concentrated agave sap. The three most bioactive fractions obtained by FCPC at partition coefficients between 0.23 and 0.4 contained steroidal saponins, predominantly magueyoside b. Flow cytometry analysis determined that the fraction rich in kammogenin and manogenin glycosides induced apoptosis, but when gentrogenin and hecogenin glycosides were also found in the fraction, a necrotic effect was observed. In conclusion, this study provides the evidence that steroidal saponins in concentrated agave sap were potential inductors of apoptosis and that it was possible to separate them using fast centrifugal partition chromatography.