Journal of Applied Botany and Food Quality 92, 15 - 23 (2019), DOI:10.5073/JABFQ.2019.092.003 1 Recursos Genéticos y Productividad-Genética, Colegio de Postgraduados, Mexico 2 Departamento de Fitotecnia-Universidad Autónoma Chapingo, Edo. México, Mexico 3 Laboratorio de Análisis Bioquímico e Instrumental, CINVESTAV, Guanajuato, Mexico Metabolomic study of volatile compounds in the pigmented fruit from Mexico Crataegus genotypes María Dolores Pérez-Lainez1, Tarsicio Corona-Torres1, María del Rosario García-Mateos2*, Robert Winkler3, Alejandro F. Barrientos-Priego2, Raúl Nieto-Ángel2, Víctor Hebert Aguilar-Rincón1, José Armando García-Velázquez1 (Submitted: July 12, 2018; Accepted: December 30, 2018) * Corresponding author Summary Crataegus is distributed worldwide and presents a phenotypic diversity in size, shape, color and aroma of the fruit. The objective of this study was to identify genotypes of Crataegus with a similar profile of volatile compounds by means of a metabolomic study. In addition, the content of pigment was evaluated to contribute to the agronomic, medicinal and chemotaxonomic value. Color determination, total carotenoids (TC) and total anthocyanins (TA) were determined in the exocarp and mesocarp of fresh fruits by means of spectrophotometry. The volatile compounds were determined by Low Temperature Plasma coupled to Mass Spectrometry (LTP- MS). A total of 75 volatile compounds were detected, according to abundance and mass-to-charge ratio, which by means of Principal Component Analysis (PCA) and selection of variables; genotypes were grouped according to size and origin. The pigment content was related to the physical color of the fruit. The highest concentration of carotenoids was 42.35 μg·g-1 FW. in the genotype PO5, and 992.34 μg·g-1 FW for anthocyanins in the genotype CH18, concen- trations of both compounds found in the exocarp of the fruit. Keywords: Anthocyanins, carotenoids, Crataegus sp., volatile com- pounds. Introduction The genus Crataegus (Family Rosaceae) displays wide phenotypic and genetic diversity grouping approximately 150 species, of which 55 are located in the Euro-Asian continent (Europe, the Middle East, East Asia), and 95 in the American continent (PhiPPs, 1997; PhiPPs et al., 2003). In Mexico there are 15 species reported (EgglEston, 1909; PhiPPs, 1997; núñEz-Colín et al., 2011), which are called “tejocote” (Mexican hawthorn), a word that derives from Nahuatl “te-xocotl” in reference to the hardness and acidity of the fruit (CabrEra, 1992). The phenotypic diversity of this genus is appreciated in the different organs of the tree, among them the fruit, which vary in shape, size and color of the exocarp (skin) (yellow, orange, red and black), with different tonality and intensity. The mesocarp (pulp) goes from yellow, orange, greenish white, reddish white to non-homogeneous diffuse red (niEto-angEl and borys, 1992; PhiPPs et al., 2003). The fruit is also characterized by its flavor (flavor and taste) particular property of this species; in China, it has an intense and unique aroma that influences the acceptability and taste of food products, such as juices and jams due to the presence of some volatile compounds (zhao et al., 2015). Attributes such as color, aroma, taste, texture, appearance, food safety and the nutritional value of fresh fruit are factors critical to consumer (barrEt et al., 2010). In tejocote, specifically the color and aroma influence the quality of the fruit and importance of its agro-industrial use (preparation of beverages with and without liquor, fruit paste and syrups); as well as, its consumption as fresh fruit. In Mexico, since prehispanic times, different parts of the Mexican hawthorn tree have been used in traditional medicine (Edwards et al., 2012), current- ly, this fruit is used preferably in traditional offerings, religious ceremonies and Christmas (borys and lEszCzyñska-borys, 1994). Morphological and molecular studies (niEto, 2007; bEtanCourt- olvEra et al., 2017) of some genotypes and species of the genus Crataegus have contributed to the taxonomic characterization and conservation of this under-utilized resource. Many studies of some species located in China, Turkey, Europe and the United States report the presence of bioactive compounds (simple phenols, polyphenols, carotenoids, anthocyanins and flavonoids, among others) (Caliskan et al., 2012; Kostić et al., 2012; vEbEriC et al., 2015; liu et al., 2016); which justify its medicinal properties (hypertension, angina pectoris, atherosclerosis, indigestion and ab- dominal distension) (Chang et al., 2002) and pharmacological properties of the cardiovascular system, as well as its antioxidant activity (Cui et al.,2006). Despite the great genetic and phenotypic diversity, few metabolites are identified in Mexican species (bandEras-tarabay et al., 2015; garCía et al., 2012). The variation of volatile compounds and pig- ment content (anthocyanins and carotenoids) among these species is hitherto unknown; the profile of these elements could be considered the chemical fingerprint of each species, keys for the understanding of their genetic variation. On the other hand, metabolomic studies of bioactive compounds have been used to describe the phenotypic variation of a wide range of phytochemicals or changes in plant chemical composition. Therefore, the objective of this study was to identify genotypes of Crataegus that presented a similar profile of volatile compounds by means of a metabolomic study, and to evaluate the pigment content to contribute to the agronomic, medicinal and mainly chemotaxonomic value. Materials and methods Plant material Hawthorn fruit was randomly collected at a commercial maturity stage in September-October 2016 and 2017 from the ex situ germ- plasm bank at the University of Chapingo (UACH), Estado de Mexico, Mexico; located at 19° 29’N and 98° 53’ W, at 2249 m height. The climate is classified as C (Wo) (w)b (i’) g, corresponds to temperate sub-humid, with a mean annual precipitation of 645 mm and an average annual temperature of 15 ± 2 °C (garCía, 1988). The material was placed in paper bags previously labeled, in a coo- ler at 4 °C. The samples harvested in 2016 were stored at -4 °C until analysis. The material harvested in 2017 was stored at -20 ± 2 °C until study. Tab. 1 shows the site of origin of the genotypes studied, identified and grouped according to their morphological characte- ristics (niEto, 2007; bEtanCourt-olvEra et al., 2017). Before the analysis of metabolites, the equatorial diameter of 10 fruits was 16 M.D. Pérez-Lainez, T. Corona-Torres, M. del Rosario García-Mateos, R. Winkler, A.F. Barrientos-Priego, Raúl Nieto-Ángel, V.H. Aguilar-Rincón, J.A. García-Velázquez measured using a vernier, an interval was obtained from the mea- surements; based on that fruits were grouped by size as follows: small (12.60 - 18.40 mm), medium (18.50 - 28.00 mm) and large (29.00 - 34.00 mm). Fig. 1 shows the size and color of each of the genotypes studied. Preparation of the sample Three fruits fruit of each genotype (EU) were analyzed separately, without pre-treatment, each fruit was cut longitudinally, immediately the surface of the cut was exposed directly to the mass spectrometry team for analysis. For this study we used the methodology pro- posed by MartínEz-Jarquín and winklEr (2013), with a Low- Temperature Plasma ionization source coupled to Mass Spectrometer (LTP-MS) LCQ Fleet (Thermo Scientific, Waltham, Massachusetts, USA). The advantage of this technique allows to analyze the volatile compounds of a sample without the previous extraction, avoiding low yields and losses of some compounds during the extraction, as well as the detection of these metabolites present in low concentrations (MartínEz-Jarquín and winklEr, 2017). Recently, the method has been applied for the analysis of volatile compounds in coffee, tequila and mezcal (gaMboa-bECErra et al., 2017; MartínEz-Jarquín et al., 2017). Detection of volatile compounds The fresh fruit cut was exposed directly to the plasma beam coupled to the mass spectrometer until obtaining 20 mass spectra in a range of 50 to 500 m/z per cut in a fruit. Three fruits were analyzed separately, finally 60 spectra were obtained by genotype. Helium was used as discharge gas, the capillary temperature was 80 ± 2 °C, voltage of 10 Kv and a frequency of 10 Khz. The equipment’s previous calibra- tion was previously performed as reported by MartínEz-Jarquín and winklEr (2013). Measurement of color parameters For the color analysis (physical color measurement), 23 genotypes were used because the genotype CH69 was not fruitful in the year of collection (2017). The color of the exocarp and mesocarp of fresh fruit was determined by means of the evaluation of L* (luminosity), Hue angle (Hue) and color purity or chromaticity index (Chroma) with a Miniscan® EZ 4500L spectrophotometer (Hunterlab, Vir- ginia, USA). The readings of a* and b* were obtained to identify the color differences between tissue and genotypes in numerical form. Variables were estimated with the following equations: Hue = tan-1 (a/b); Chroma = (a2+ b2) ½ (MC. guirE, 1992). Three fruits of each genotype with similar commercial maturity stage were selected; for each fruit three different points of the exocarp were studied. Subsequently, for the measurement of pulp color, the fruit was cut into three portions longitudinally, each measurement was made in triplicate. Measurement of total carotenoids The total quantification of carotenoids (TC) was carried out separately in the exocarp and mesocarp of the fresh fruit as indicated by MéndEz-iturbidE et al. (2013) with minor modifications. To obtain the sample, between three and twenty fruits were used per genotype, depending on the size of the fruit. The tissue (2 g) Tab. 1: Geographical characteristics of 24 genotypes of the Mexican hawthorn (Crataegus sp.). Genotype Species* Statea Originb Lat Long H(m) MDc CH04 sulfurea Chiapas S C de las Casas 16.75 92.67 2300 Oct-Dec CH08 tracyi Chiapas S C de las Casas 16.75 92.67 2300 Oct-Nov CH10 tracyi Chiapas S C de las Casas 16.75 92.67 2300 Sept-Nov CH13 tracyi Chiapas Rancho Nuevo 16.67 92.57 2400 Dec-Jan CH15 aurescens Chiapas Mitzitan 16.65 92.55 2380 Jan-Feb CH16 tracyi Chiapas Mitzitan 16.65 92.55 2380 Nov-Jan CH18 tracyi Chiapas Rancho Robelo 16.67 92.45 2250 Nov-Feb CH19 baroussana Chiapas Mitzitan 16.65 92.55 2380 Dec-Jan CH22 rosei Chiapas Mitzitan 16.65 92.55 2380 Oct-Nov CH27 gracillior Chiapas Rancho Robelo 16.67 92.45 2250 Jan-Feb CH43 greggiana Chiapas Rancho Robelo 16.67 92.45 2250 Jan-Feb CH44 gracillior Chiapas Mitzitan 16.65 92.55 2380 Oct-Nov CH51 mexicana Chiapas Mitzitan 16.65 92.55 2380 Nov-Feb CH69 cuprina Chiapas Candelaria 16.70 92.53 2320 Oct-Nov CH72 baroussana Chiapas S J Yashitinin 16.65 92.45 2350 Nov-Dec CH83 tracyi Chiapas S C de las Casas 16.75 92.67 2300 Sept-Nov P02 tracyi Puebla Rancho Nuevo 19.14 98.57 2620 Sept-Oct P05 aurescens Puebla Mitzitan 19.14 98.50 2628 Sept-Oct P06 tracyi Puebla Mitzitan 19.14 98.50 2625 Sept-Oct P25 cuprina Puebla Origen 19.10 98.47 2420 Dec-Feb P55 sulfurea Puebla S C de las Casas 19.17 98.40 2280 Dec-Feb P86 mexicana Puebla Huejotzingo 19.17 98.40 2280 Nov-Dec P100 mexicana Puebla Huejotzingo 19.17 98.40 2280 Oct-Dec EM66 tracyi Edo. Mex. S C de las Casas 19.48 98.77 2700 Sept-Oct a S C de las Casas = San Cristóbal de las Casas; S J Yashitinin = San José Yashitinin; S C del Monte = Santa Catarina del Monte. b Edo. Mex. = Estado de México. c MD = Commercial maturity date (niEto, 2007). Metabolomic study of volatile compounds in Mexican Crataegus genotypes 17 was ground in a mortar with 20 mL of hexane, acetone and ethanol (50:25:25 v/v), the solution was kept at 4 °C in the dark for 30 min, then it was filtered. Each extraction was made in triplicate. The absorbance of the organic phase was measured at 450 nm in a Multiscan® GO spectrophotometer (Thermo Scientific, Waltham, Massachusetts, USA). Hexane was used as a blank solution. To calculate the concentration (μg) of total carotenoids the following equation was used: μg = A × V (mL) × 106/ ε × 100; where ε = extinction coefficient of β-carotenoid (2505); V = volume of sample; A = Absorbance obtained (MéndEz-iturbidE et al., 2013). Results are expressed in μg·g1 of fresh weight (FW): μg·g-1 = μg / weight of sample (g). Measurement of total anthocyanins The concentration of total anthocyanins (TA) was determined by the differential pH method (giusti and wrolstad, 2001). The extrac- tion of anthocyanins was carried out in the exocarp and mesocarp of the fresh fruit, in triplicate. For each replication, 2.0 g of fresh tissue was ground in a mortar with 20 mL of methanol acidified with HCL at 0.01 (v/v %) in a 10:1 ratio. Then the extract was allowed to rest for 24 h in the dark at 4 °C. Two test tubes were prepared with 0.5 mL of the extract, for each sample; then 25 mL of KCL and 25 mL of CH3COONa·3H2O solution were added to one tube. The tubes were allowed to rest for 15 min in the dark to stabilize the reactions. The absorbance of each sample was measured at 520 and 700 nm in a Multiscan® GO spectrophotometer (Thermo Scientific, Waltham, Massachusetts, USA). The blank solution used was acidified metha- nol. The concentration of anthocyanins was expressed as cyanidin- 3-glucoside with the equation: TA (mg·L-1) = A × MW × DF × 1000 ε-1; where TA (mg·L-1) is the concentration of total anthocya- nins, MW = molecular weight of cyanidin-3-glucoside (445.2), DF = dilution factor, ε = extinction coefficient (26 900) and A = absor- bance obtained with the following equation: Abs = (A520-A700)pH1- (A520-A700)pH4.5 (wrolstad et al., 2005). Statistic analysis The R Program Version 3.4.3 (http://www.rproject.org) with the Rstudio interface (http://www.rstudio.com) was used for the data analysis (volatile compounds and physical-chemical color variables). The mass spectra obtained in the analysis of volatile compounds in a range of 50 to 500 m/z in format .raw were changed to the numerical format .mzML using the Program MSConver. The package MALDI- quant was used to process the mass data of the 60 spectra obtained by genotype, to obtain the final mass spectrum. Subsequently, the package pheatmap generated the heat map with the abundance of the volatile compounds and the principal components analysis (PCA) was obtained by means of the prcomp, to integrate the metabolomic analysis (graCE and hudson, 2016). The color variables (L, hue, chroma, TA and TC) studied in the exo- carp and mesocarp of the fruit were subjected to a one-way ANOVA and to the analysis of the ANOVA assumptions and a comparison of means (P < 0.05). Due to the fact that not all the variables fulfilled the data normality test, for the variable hue in the exocarp, and TA and TC in the mesocarp, a data transformation was carried out using the logarithm method. The TC variables in the exocarp and hue in the mesocarp were analyzed by means of the Kruskal-Wallis test (P < 0.05). The packages used in this study were: agricolae, to obtain Tukey’s mean comparison (P < 0.05); car, to perform the Levene test in the analysis of assumptions and pgirmes to carry out the Kruskal- Wallis test. Results and discussion Volatile compounds The analysis of volatile compounds by ionization with low-Ttem- perature plasma coupled to mass sSpectrometry (LTP-MS) allowed to detect 75 compounds with increased relative intensity (%). Their low molecular weight, 50 to 500 m/z, suggests that those volatile compounds are associated with the aroma of the fruit. Fig. 2 shows only the mass spectra of two genotypes (P86, spectrum A and CH44, spectrum B) due to reasons of space. A Principal Components Analysis (PCA) was carried out with the 75 metabolites detected (compounds of the fruit aroma). The results were interpreted based on their eigenvalues and eigenvectors. The eigenvalues and the variance explained for each of the compounds are shown in Tab. 2, where it is observed that five compounds jus- tify 81.96% of the total variance. It was also found that the vola- tile compounds that provided 34.89% of the total variance in the first component (PC1) were those with m/z ratio: 144.98, 173.01, 270.8, 344.74, 117.03, 187.01, 200.99, 284.8, 159.0, 298.77, 256.79, 278.97, 358.74, 330.73, 235.1, 218.48, 103.06, 345.67, 204.12, 223.04 and 400.76. The volatile compounds of m/z: 288.74, 260.78, 98.99, 274.77, 117.03, 232,86, 302.68, 56.95, 81.01, 246.76, 153.12, 92.92, 127.06 and 250.91 provided 23.61% of the total variance of the se- cond component (PC2). In the third component (PC3) the volatile compounds 228.87, 214.90, 219.0, 262.87, 258.86 and 230.79 pro- vided 22.18% of the total variance. From the PCA, the dispersion of the study genotypes in the first two principal components was obtained graphically (Fig. 3), where a cluster was observed with the genotypes CH08, CH10 and CH51, from the state of Chiapas. The second group corresponded to the genotypes PA06, PA05, PA02 and P100 from Atexcac and Huejotz- Fig. 1: Fruit image of 24 hawthorn (Crataegus sp.) genotypes. 18 M.D. Pérez-Lainez, T. Corona-Torres, M. del Rosario García-Mateos, R. Winkler, A.F. Barrientos-Priego, Raúl Nieto-Ángel, V.H. Aguilar-Rincón, J.A. García-Velázquez exocarp of some genotypes. Group I grouped the genotypes of small size (12.60 - 18.40 mm), more than 50% with red exocarp and 92% are from Chiapas, with the exception of the genotype P25 (yellow exocarp from Puebla). Group II has genotypes from three states (Chiapas, Puebla and Estado de Mexico) that were characterized by medium to large fruits (18.50 - 34.00 mm), with yellow, yellow- orange and red exocarp. The variation in the size and in some cases the origin explain the differences between groups, the coloration of the exocarp of the fruit, as the species were not determining fac- tors. The clusters showing high similarity (P100-P02, CH08-CH10, CH44-P86, CH13-CH72, CH15-CH27) in their volatile profile had in common the origin, species and/or color of the fruit, the exception was the cluster with the genotypes CH83 and P05. On the other hand, the volatile compounds of greater abundance are shown on the heat map (Fig. 4), this is represented by the color scale where yellow corresponds to the lowest abundance up to red which corresponds to greater abundance. The five most abundant volatiles were only present in some genotypes (Tab. 3) of the two groups of the dendrogram, e. g. the compound 260.78 m/z was found only in the genotypes of Group II, with different abundance. In contrast, the compound 144.98 m/z with different abundance was identified in all genotypes of Groups I and II. In this regard, two structural metabo- lites (hexyl acetate and butyl butanoate) with the same m/z (144,21) have been reported in different species (C. aestivalis, C. opaca and Fig. 2: Metabolomic profile of the volatile compounds obtained by LTP-MS. A) Genotype P86 (C. mexicana); B) genotype CH44 (C. gracillior). Tab. 2: Eigenvalues and variance proportion explained by the principal component analysis of volatile compounds from the genotypes of Crataegus. Compound Eigen values VPC (%) CV (%) 1 26.170 34.89 34.89 2 17.707 23.61 58.50 3 8.385 22.18 69.68 4 5.930 7.91 77.59 5 3.279 4.37 81.96 VPC: Variance per compound. CV: Cumulative variance. Fig. 3: Principal components of Crataegus sp. genotypes by volatile compounds. ingo, Puebla. In this same group, we found the genotypes CH83 and CH18 from two different regions of the state of Chiapas (Fig. 3). In the dendrogram located in the upper part of Fig. 4 there are two groups (I and II) with the 24 genotypes evaluated; in the lower part, the heat map is shown containing the abundance (relative intensity %) and the presence of the 37 volatile compounds (m/z ratio) selected by the principal component analysis. The color row of the dendrogram represents the color of each geno- type, e.g. the red boxes are associated with the red coloration of the Metabolomic study of volatile compounds in Mexican Crataegus genotypes 19 C. rufula) of Crataegus from the United States by horvat and ChaPMan (1991) and Cha et al. (2011), however the last metabolite was found in the aroma of a different species (C. pinnatifida) culti- vated in China (lingyun and biJun, 1997). Both compounds have also been identified in the aroma of apple (EsPino-díaz et al., 2016). There are few studies that report the volatile profile in the Crataegus fruit; of the 37 compounds reported in this study only seven coincide with those reported in the fruit (Tab. 4), however volatile compounds have been reported in flowers (kovalEva et al., 2009) and leaves (lakaChE et al., 2014). In this study, even though the detection of volatiles with the LTP- MS technique did not allow the structural identification of the com- pounds identified by their m/z ratio, the heat map allowed to prove a different chemical profile for each genotype. Color parameters The color parameters evaluated provide quality attributes that are important in consumer preferences. The lower values (< 52) of chro- ma (purity of color) obtained in the exocarp of the red fruits showed great variability in the intensity of color in comparison with yellow fruits which showed higher values (> 66) (Tab. 4). The fruits with yellow exocarp (P55 and P05) and mesocarp (CH72 and CH83) had the highest values (> 61 and > 71, respectively) of luminosity (tis- sues with brighter colors) and hue (yellow exocarp and mesocarp), compared to red exocarp fruits with a luminosity of 32.46 to 47. 49 h 26 to 32 ° (Tab. 5). It is important to mention that yellow and larger fruits (P55, P02 and P100), grown in Puebla (first producer state at a national level with a production of 3500 t per year), are the most demanded in Mexico due to their agroindustrial and cultural use (SAGARPA, 2015). Content of carotenoids Carotenoids are natural pigments metabolized by plants, are re- sponsible for the colors yellow, orange and red in some fruits and vegetables. The apocarotenoids (derivatives) are the result of the breaking of the carotenoids present in the aroma of flowers and fruits (naMitha and nEgi, 2010). The exocarp of the genotypes P55 and P05 showed the highest concentrations of carotenoids (42.09 and 42.35 μg·g-1 FW, respectively), which coincides with the higher val- ues found for hue in the present study. However, despite of having small fruits, the mesocarp of genotype P25 had the highest concen- tration of carotenoids (20.92 μg·g-1 FW), but also a considerable con- centration in the exocarp (38.17 μg·g-1 de FW), this non-commercial genotype from Puebla was the only one in Group I, where most of the Fig. 4: The heat map shows the abundance (color intensity) of each of the volatile compounds listed in the right column (m/z ratio). The dendrogram (upper part) shows the clusters of the genotypes by coloration of the hawthorn (Crataegus sp.) fruit. 20 M.D. Pérez-Lainez, T. Corona-Torres, M. del Rosario García-Mateos, R. Winkler, A.F. Barrientos-Priego, Raúl Nieto-Ángel, V.H. Aguilar-Rincón, J.A. García-Velázquez genotypes from Chiapas were grouped. Concentrations in the exo- carp of the two genotypes (P100 and P02) from C. mexicana were higher (38.68 and 33.61 μg·g-1 FW) than that reported (26.4 μg·g-1 FW) by MéndEz-iturbidE et al. (2013), but in lyophilized exocarp of the same species from the state of Tlaxcala, Mexico (Tab. 5). The differences in the contents of both studies could be explained mainly by environmental factors, different stage of fruit ripening and post- harvest management (rEilly, 2013), as has been reported in other fruits (lóPEz-vidal et al., 2014; MEsEJo et al., 2011). The content of carotenes in hawthorn was lower than that reported for melon Tab. 3: The five volatile compounds with increased relative intensity (%) by genotype. Genotype m/z (% Abundance) CH4 173.01 (4.31) 270.80 (4.11) 144.98 (4.01) 242.81 (3.99) 98.99 (3.94) CH8 270.80 (5.18) 173.01 (5.12) 344.74 (2.74) 98.99 (2.87) 144.98 (2.47) CH10 173.01 (5.02) 270.80 (4.39) 344.74 (2.87) 144.98 (2.57) 187.01 (2.51) CH13 173.01 (5.03) 144.98 (4.14) 270.80 (3.44) 200.99 (2.37) 288.74 (2.34) CH15 144.98 (4.95) 270.8 0 (4.22) 173.01 (3.70) 98.99 (3.34) 288.74 (3.21) CH16 173.01 (6.53) 144.98 (4.53) 270.80 (2.73) 344.74 (2.63) 288.74 (2.36) CH18 144.98 (4.11) 173.01 (3.79) 270.80 (2.16) 98.99 (2.11) 288.74 (2.02) CH19 98.99 (3.19) 173.01 (3.15) 144.98 (2.87) 270.80 (2.83) 200.99 ( 2.44) CH22 144.98 (6.91) 288.74 (6.82) 274.77 (2.68) 260.78 (2.54) 173.01 (2.13) P25 144.98 (5.45) 173.01 (4.62) 288.74 (3.32) 270.80 (2.99) 98.99 (2.44) CH27 144.98 (4.62) 173.01 (4.35) 270.80 (4.01) 288.74 (3.08) 98.99 (2.56) CH43 144.98 (4.28) 173.01 (3.87) 270.80 (3.75) 98.99 (3.14) 288.74 (2.11) CH44 288.74 (12.25) 260.78 (8.12) 144.98 (7.85) 274.77 (3.72) 117.03 (3.10) CH51 173.01 (4.46) 270.80 (4.42) 187.01 (2.97) 98.99 (2.81) 144.98 (2.75) P55 288.74 (6.45) 144.98 (6.07) 260.78 (4.11) 274.77 (3.82) 117.03 (2.46) EM66 144.98 (5.62) 288.74 (4.70) 173.01 (2.99) 270.80 (2.52) 260.78 (2.46) CH69 173.01 (4.56) 144.98 (4.02) 270.80 (2.29) 200.99 (2.24) 117.03 (2.07) CH72 173.01 (5.95) 270.80 (3.98) 144.98 (3.72) 344.74 (3.11) 200.99 (2.30) CH83 144.98 (5.95) 98.99 (3.19) 28.84 (3.02) 173.01 (2.19) 270.80 (1.56) P86 288.74 (8.03) 144.98 (6.74) 260.78 (5.21) 117.03 (2.73) 274.77 (2.65) P100 144.98 (6.55) 288.74 (4.56) 98.99 (2.30) 260.78 (1.85) 117.03 (1.77) P02 144.98 (5.52) 288.74 (3.81) 98.99 (2.25) 173.01 (1.62) 117.03 (1.61) P05 144.98 (6.46) 288.74 (3.52) 98.99 (2.59) 173.01 (1.69) 117.03 (1.45) P06 144.98 (4.58) 98.99 (4.52) 173.01 (2.06) 117.03 (2.00) 270.8 (1.72) Tab. 4: Volatile compounds identified by GC-MS in the Crataegus fruit Name Formula m/z M W Species Source 2-Hexenal C6H10O 98.99 98.145 C. aestivalis, lingyun and biJun, 1997 C. opaca, horvat and ChaPMan, 1991 C. rufula y C. piinnatifida Benzaldehyde C7H6O 106.99 106.124 C. aestivalis, lingyun and biJun, 1997 C. opaca, robErtson et al., 1993 C. rufula y C. piinnatifida Hexyl acetate C8H16O2 144.98 144.214 C. aestivalis, lingyun and biJun, 1997 C. opaca, horvat and ChaPMan, 1991 C. rufula y C. piinnatifida Butyl butanoate C8H16O2 144.98 144.214 C. aestivalis y Cha et al., 2011 C. opaca Citral C10H16O 152.06 152.237 C. piinnatifida lingyun and biJun, 1997 Hexyl hexanoate C12H24O2 200.99 200.322 C. aestivalis, horvat and ChaPMan, 1991 C. opaca y C. rufula β-caryophyllene C15H24 204.12 204.357 C. monogyna robErtson et al., 1993 m/z: Mass-to-charge ratio; MW: Molecular weight. Metabolomic study of volatile compounds in Mexican Crataegus genotypes 21 (185.0 μg·g-1 FW) by MartínEz-valdiviEso et al. (2014), but higher than that found in peach (12.0 μg·g-1FW) (CaMbEll and Padilla, 2013). Therefore, the consumption of hawthorn fruits represent a source of carotenoids in the human diet, important in the prevention of vitamin A deficiencies, cancer, cardiovascular diseases, age-rela- ted macular degeneration and cataract formation; beside this fruit has an important antioxidant activity (arathi et al., 2015). Anthocyanin content The anthocyanins comprise another type of pigments responsible for the coloring of fruits from red to purple, are phenolic with an- tioxidant properties, which justify their nutraceutical properties (dElgado-vargas et al., 2000). It was found that the genotypes with intense and dark red exocarp (CH18, CH51 and CH19) had the highest concentrations (992.34, 844.69 and 747.69 μg·g-1FW, re- spectively) (Tab. 4) which corresponds to the smallest values of hue (< 28 °) and chroma (< 52), concentrations higher than those reported by FroEhliChEr et al. (2009) (58.0 mg ·100 g-1 FW) in the exocarp of C. monogyna; however, lower than those reported for cranberries (206.0 mg·100 g FW) (ribEra et al., 2010) and raspberry (28.7 - 55.6 mg·100 g-1 FW) (PEña-varEla et al., 2006). In contrast, yellow exocarp fruits showed very small concentrations (5.56 - 30.3 μg·g-1 FW). In the mesocarp of all genotypes a lower concentration of these pigments was observed (6.95 - 29.22 μg·g-1 FW), the exceptions were the genotypes CH10 and CH18, which had the highest anthocyanin values (29.22 and 28.38 μg·g-1) and higher hue values (> 51°) (Tab. 5). It should be noted that in most of the genotypes the highest concen- tration of both pigments predominated in the exocarp, the exception was the genotypes P25 and CH18, where the concentrations in the fruit (exocarp + mesocarp) were important from the nutraceutical and medicinal point of view, because their consumption could pro- vide high content of antioxidant pigments, for health benefits. Conclusions The profiles of chemical volatile compounds were different among the 24 genotypes of the Mexican Crataegus. The metabolomic study allowed to cluster the genotypes according to their chemical profile. The differences in the volatile profile were related to the size of the fruit, and in some genotypes to their origin (state of Chiapas). No relationship of volatile compounds by color or by species was found. The highest concentration of carotenoids was observed in the exo- carp in comparison with the mesocarp of yellow fruits, in contrast the red fruits showed the highest concentrations in the exocarp. The LTP-MS technique allowed to detect volatile compounds of low abundance when analyzing the sample without pretreatment. The results of the present research could be used for the development of products derived from hawthorn with high levels of bioactive com- pounds that would justify their medicinal properties. Acknowledgments We thank Dr. Maria Teresa Carrillo Rayas (CINVESTAV Unidad Irapuato, CUI), and Maria Isabel Cristina Elizarraraz Anaya (CUI), Thermo and Waters Mexico for technical support. The project was funded by the CONACyT Fronteras project 2015-2/814 and the bi- lateral grant CONACyT-DFG 2016/277850. MDPL acknowledges her CONACyT postgraduate scholarship. References Arathi, b.P., sowMya, P.r., viJay, kbaskaran, v., lakshMinarayana, r., 2015: Metabolomics of carotenoids: The challenges and prospects – A review. Trends Food Sci. Tech. 45, 105-117. DOI: 10.1016/j.tifs.2015.06.003 barrEtt, d.M., bEauliEu, J.C., shEwFElt, r., 2010: Color, flavor, texture, and nutritional quality of fresh-cut fruits and vegetables: Desirable le- vels, instrumental and sensory measurement, and the effects of process- ing. Crit. Rev. Food Sci. Nutr. 50, 369-389. DOI: 10.1080/10408391003626322. bandEras-tarabay, J.a., CErvantEs-rodríguEz, M., grada-sánChEz, M., EsPindola-lozano, M., CuEvas-roMEro, E., navarro-oCaña, a., MEndEz-iturbidE, d., 2015: Antioxidant-mediated protective effect of hawthorn (Crataegus mexicana) peel extract in erythrocytes agains oxidative damage. Afr. J. Food Sci. 9, 208-222. DOI: 10.5897/AJFS2015.1269 bEtanCourt-olvEra, M., niEto-ángEl, r., urbano, b., gonzálEz- andrés, F., 2018: Analysis of the biodiversity of hawthorn (Crataegus spp.) from the morphological, molecular, and ethnobotanical approach- es, and implications for genetic resource conservation in scenery of in- creasing cultivation: the case of Mexico. Genet. Resour. Crop. Evol. 65, 897-916. DOI: 10.1007/s10722-017-0583-4. borys, M., lEszCzyñska-borys, w.y.h., 1994: Tejocote (Crataegus spp.) planta para solares, macetas e interiores. Rev. Chapingo Serie Hort. 2, 95-107. CabrEra l.g., 1992: Diccionario de Aztequismos. Ediciones Colofón. Distrito Federal, México. Caliskan, o., gündüz, K., serce, s., toplu, c., Kamiloğlu, o., sengül, M., ErCisli, s., 2012: Phytochemical characterization of several haw- thorn (Crataegus spp.) species sampled from the Eastern Mediterranean region of Turkey. Pharmacogn. Mag. 8, 16-21. DOI: 10.4103/0973-1296.93305 CaMPbEll, o.E., Padilla-zakour, o.i., 2013: Phenolic and carotenoid composition of canned peaches (Prunus persica) and apricots (Prunus armeniaca) as affected by variety and peeling. Food Res. Int. 54, 448- 455. DOI: 10.1016/j.foodres.2013.07.016 Cui, n., nakaMura, k., tian, s., kayahara, h., tian, y., 2006: Poly- phenolic content and physiological activities of chinese hawthorn ex- tracts. Biosci. Biotech. Bioch. 70, 2948-2956. Cha, d.h., PowEll, t.h.q., FEdEr, J.l., linn, C.E. Jr., 2011: Identification of host fruit volatiles from three mayhaw species (Crataegus series es- tivales) attractive to mayhaw-origin Rhagoletis pomonella flies in the Southern United States. J. Chem. Ecol. 37, 961-973. DOI 10.1007/s10886-011-0013-6 Chang, q., zuo, z., harrison, F., Chow, M.s., 2002: Hawthorn. J. Clin. Pharmacol 42, 605-612. dElgado-vargas, F., JiMénEz, a.r., ParEdEs-lóPEz, o., 2000: Natural pigments: Carotenoids, anthocyanins, and betalains − characteristics, biosynthesis, processing, and stability. Crit. Rev. Food Sci. Nutr. 40, 173- 289. DOI: 10.1080/10408690091189257 Edwards, J.E., brown, P.n., talEnt, n., diCkinson, t.a., shiPlEy, P.r., 2012: A review of the chemistry of the genus Crataegus. Phytochemistry 79, 5-26. DOI: 10.1016/j.phytochem.2012.04.006 EChEvErría, g., FuEntEs, M.t., graEll, J., lóPEz, M.l., 2003: Rela- tionships between volatile production, fruit quality and sensory evalu- ation of Fuji apples stored in different atmospheres by means of multi- variate analysis. J. Sci. Food Agric. 84, 5-20. DOI: 10.1002/jsfa.1554 EsPino-díaz, M., robErto sEPúlvEda, d., gonzálEz-aguilar, g., olivas, g.i., 2016: Biochemistry of apple aroma: A Review. Food Technol. Biotechnol. 54, 375-394. DOI: 10.17113/ftb.54.04.16.4248 EgglEston, w.w., 1909: The Crataegi of Mexico and Central America. Bulletin of the Torrey Botanical Club 36, 501-514. FroEhliChEr, t., hEnnEbEllE, t., Martin-nizard, F., ClEEnEwErCk, P., hilbErt, J.l., trotin, F., grEC, s., 2009: Phenolic profiles and anti- oxidative effects of hawthorn cell suspensions, fresh fruits, and medici- nal dried parts. Food Chem. 115, 897-903. DOI: 10.1016/j.foodchem.2009.01.004 gaMboa-bECErra, r., MontEro-vargas, J.M., MartínEz-Jarquín, s., gálvEz-PonCE, E., MorEno-PEdraza, a., winklEr, r., 2017: Rapid 22 M.D. Pérez-Lainez, T. Corona-Torres, M. del Rosario García-Mateos, R. Winkler, A.F. Barrientos-Priego, Raúl Nieto-Ángel, V.H. Aguilar-Rincón, J.A. García-Velázquez Ta b. 5 : C ol or p ar am et er s, to ta l c ar ot en oi d co nt en t ( T C ) a nd to ta l a nt ho cy an in s (T A ) i n th e ex oc ar p an d m es oc ar p of 2 3 ge no ty pe s of th e ge nu s C ra ta eg us . E xo ca rp M es oc ar p G en ot yp e L um in os it y H ue C hr om a T C * TA * L um in os it y H ue C hr om a T C * TA * (μ g· g- 1 ) (μ g· g- 1 ) (μ g· g- 1 ) (μ g· g- 1 ) C H 4 60 .7 3 a 64 .1 4 a b 66 .2 1 a bc 23 .8 0 ab 5. 56 e 65 .4 3 d ef g 65 .7 5 a b 59 .0 2 a 15 .6 1 a b 22 .2 6 a bc C H 8 35 .9 5 e f 29 .9 1 e fg 54 .8 8 g hj 16 .4 1 a b 56 3. 59 c 64 .0 7 e fg 62 .4 2 a b 49 .1 7 b cd 9. 85 b cd 16 .7 0 a bc C H 10 37 .1 0 d ef 28 .7 3 fg 54 .6 0 h i 11 .8 4 a b 36 8. 77 d 61 .1 5 fg 51 .0 8 b c 51 .8 3 a bc 4. 51 f g 29 .2 2 a C H 13 47 .4 9 b c 37 .3 1 d 62 .1 5 b cd ef 28 .7 0 a b 23 9. 35 d 68 .4 7 b cd ef 66 .8 8 a b 50 .1 4 b c 7. 39 c de f 16 .7 0 a bc C H 15 64 .3 5 a 67 .2 5 a 68 .2 7 a 13 .7 7 a b 6. 95 e 65 .8 7 c de fg 68 .5 4 a b 54 .3 0 a b 9. 78 b cd 10 .4 3 a bc C H 16 42 .8 0 c d 32 .4 1 e 57 .5 1 fg hi 11 .9 9 a b 28 5. 27 d 67 .9 1 b cd ef 67 .6 0 a b 46 .5 1 b cd 10 .5 7 b cd 14 .6 1 a bc C H 18 32 .4 6 f 26 .9 9 g 45 .0 5 j 19 .5 4 a b 99 2. 34 a 70 .0 8 a bc de 71 .9 3 a b 48 .1 5 b cd 5. 04 e fg 28 .3 8 a b C H 19 34 .6 8 e f 28 .0 6 fg 51 .9 4 i 5. 71 b 74 7. 69 b 67 .7 4 bc de f 70 .2 2 a b 47 .5 4 b cd 3. 95 g 18 .1 7 a bc C H 22 50 .7 3 b 55 .3 1 c 57 .7 8 e fg h 22 .2 6 a b 16 .1 9 e 61 .4 9 fg 69 .9 5 a b 45 .6 9 b cd 7. 28 d ef 12 .0 1 a bc P2 5 52 .4 8 b 59 .3 8 b c 60 .9 4 a bc 38 .1 7 a b 23 .6 6 e 52 .9 8 h 64 .6 9 ab 52 .0 2 a bc 20 .9 2 a 16 .7 0 a bc C H 27 52 .1 3 b 61 .4 7 a bc 60 .5 6 c de fg 14 .4 1 a b 15 .8 3 e 60 .0 0 g h 66 .9 7 a b 49 .2 0 b cd 7. 77 c de 14 .9 6 a bc C H 43 59 .9 7 a 63 .0 9 a b 63 .7 6 a bc d 10 .7 1 a b 23 .6 6 e 66 .2 1 c de fg 70 .9 1 a b 52 .8 6 a b 9. 55 b cd 9. 73 ab c C H 44 64 .4 2 a 63 .4 7 a b 66 .1 6 a bc 16 .9 2 a b 10 .4 3 e 66 .7 2 c de fg 70 .2 1 a b 45 .6 0 b cd 6. 67 d ef g 8. 35 ab c C H 51 34 .8 3 e f 28 .7 7 fg 52 .5 9 h i 15 .3 7 a b 84 4. 69 a b 60 .1 3 g h 57 .3 5 b 43 .9 3 c d 9. 6 b cd 26 .4 4 a bc P5 5 64 .9 3 a 63 .0 8 a b 67 .3 6 a b 42 .0 9 a 12 .5 1 e 71 .9 4 a bc d 69 .5 8 a b 47 .6 2 b cd 8. 98 b cd 6. 95 c E M 66 60 .6 4 a 62 .7 5 a b 65 .1 1 a bc 13 .3 8 a b 6. 96 e 68 .3 9 b cd ef 65 .8 1 a b 53 .6 0 a b 9. 11 b cd 16 .7 0 a bc C H 72 40 .9 3 d e 32 .6 2 e 58 .0 0 d ef gh 38 .1 0 a b 31 5. 89 d 71 .6 0 a bc d 72 .6 5 a b 52 .4 3 a bc 7. 37 d ef 15 .3 0 a bc C H 83 40 .3 1 d e 31 .3 8 e f 55 .7 6 g hi 22 .0 6 a b 52 8. 80 c 73 .2 2 a bc 73 .3 1 a b 46 .9 4 b cd 7. 87 c de 13 .9 2 a bc P8 6 62 .2 3 a 60 .7 5 a bc 65 .5 2 a bc 13 .5 3 a b 12 .5 2 e 69 .6 6 b cd ef 72 .4 8 a b 46 .1 2 b cd 4. 74 e fg 23 .6 6 a bc P1 00 61 .7 9 a 62 .2 3 a b 62 .8 6 a bc de f 33 .6 1 a b 25 .0 4 e 71 .7 1 a bc d 69 .3 4 a b 53 .8 5 a b 12 .8 4 a bc 8. 35 bc P0 2 62 .4 4 a 61 .9 4 a bc 67 .2 8 a b 38 .6 8 ab 18 .0 9 e 70 .1 8 a bc de 69 .3 2 a b 53 .9 3 a b 5. 22 e fg 20 .8 7 a bc P0 5 61 .2 6 a 59 .2 6 b c 67 .4 4 a b 42 .3 5 a 20 .8 7 e 75 .9 9 a 79 .8 0 a 41 .2 3 d 10 .3 4 b cd 11 .1 3 a bc P0 6 60 .8 9 a 62 .0 5 a bc 63 .4 2 a bc de 22 .0 6 a b 30 .3 0 e 74 .7 4 a b 79 .7 1 a 40 .4 1 d 9. 83 b cd 9. 67 ab c H SD 6. 26 5. 81 5. 79 8. 09 15 8. 92 7. 42 8. 89 8. 89 5. 57 24 .0 4 * μg ·g -1 o f f re sh ti ss ue . M ea ns w ith th e sa m e le tte rs a re n ot s ta tis tic al ly d iff er en t ( P < 0. 05 ). H SD : H on es tly s ig ni fic an t d iff er en ce . Metabolomic study of volatile compounds in Mexican Crataegus genotypes 23 classification of coffee products by data mining models from direct electrospray and plasma-based mass spectrometry analyses. Food Anal. Method. 10, 1359-1368. DOI: 10.1007/s12161-016-0696-y garCía, E., 1988: Modificaciones al Sistema de Clasificación Climática de Köppen. Edit. UNAM. D. F., México. garCía-MatEos, r., aguilar-santElisEs, l., soto-hErnándEz, M., niEto-angEl, r., kitE g., 2012: Compuestos fenólicos totales, flavonoi- des y actividad antioxidante en las flores de Crataegus spp. Agrociencia 46, 651-662. giusti, M.M., wrolstad, r.E., 2001: Anthocyanins. Characterization and measurement with UV-visible spectroscopy. In: Wrolstad, R.E. (ed.), Current protocols in food analytical chemistry, unit F1.2.1-13. New York. John Wiley &Sons. graCE, s.C., hudson, d.a., 2016: Processing and visualization of meta- bolomics data using R. In: Prasain, J.K. (ed.), Metabolomics-funda- mentals and applications, 667-694. Croatia. Intechopen. DOI: 10.5772/65405 horvat, r.J., ChaPMan, J.r., 1991: Identification of volatile compounds from ripe mayhaw fruit (Crataegus opaca, C. aestivalis, and C. rufula. J. Food Quality 14, 307-312. DOI: 10.1111/j.1745-4557.1991.tb00071.x Kostić, D.a., VelicKoVić, J.m., mitić, s.s., mitić, m.n., ranDeloVić, s.s., 2012: Phenolic content, and cntioxidant and antimicrobial Activities of Crataegus oxyacantha L. (Rosaceae) fruit extract from Southeast Serbia. Trop. J. Pharm. Res. 11, 117-124. DOI: 10.4314/tjpr.v11i1.15 kovalEva, a.M., gonCharov, n.F., koMissarEnko, a.n., sidora, n.v., kovalEv, s.v., 2009: GC/MS Study of essential oil components from flowers of Crataegus jackii, C. robesoniana, and C. flabellata. Chem. Nat. Compd. 45, 582-584. DOI: 10.1007/s10600-009-9373-3 lakaChE, z., tigrinE-kordJani, n., tigrinE, C., kaMEll, a., MEklatt, b.y., 2014: Volatile constituents, phenolic compounds, and antioxidant activity of Crataegus azarolus leaves and flowers growing in Algeria. Chem. Nat. Comp. 50, 1132-1135. DOI 10.1007/s10600-014-1183-6 lingyun, C., biJun, X., 1993: Identification of volatile compounds of haw- thorn by Gas Chromatography/Mass Spectrometry (GC/MS). Chinese J. Cromatography 15(3), 219-221. lóPEz-vidal, o., EsCalona-buEndía, h., PElayo-zaldívar, C., Cruz- salazar, J., villa-hErnándEz, J.M., rivEra-CabrEra, F., villEgas- torrEs, o., aliatEJaCal, i., PérEz-FlorEs, l.J., díaz dE lEón- sánChEz F., 2014: Carotenoides, capacidad antioxidante y volátiles del aroma durante la maduración de jitomate. Rev. Int. Bot. Exp. PHYTON 83, 139-146. liu, s., Chang, X., liu, X., shEn, z., 2016: Effects of pretreatments on an- thocyanin composition, phenolics contents and antioxidant capacities during fermentation of hawthorn (Crataegus pinnatifida) drink. Food Chem. 212, 87-95. DOI: 10.1016/j.foodchem.2016.05.146 MartínEz-Jarquín, s., MorEno-PEdraza, a., CázarEz-garCía, d., winklEr, r., 2017: Automated chemical fingerprinting of Mexican spirits derived from Agave (tequila and mezcal) using Direct-Injection Electrospray Ionisation (DIESI) and Low-Temperature Plasma (LTP) Mass Espectrometry. Anal. Methods 9, 5023-5028. DOI: 10.1039/C7AY00793K MartínEz-Jarquín, s., winklEr, r., 2013: Design of a Low-Temperature Plasma (LTP) probe with adjustable output temperature and variable beam diameter for the direct detection of organic molecules. Rapid Commun. Mass Sp. 27, 629-634. DOI: 10.1002/rcm.6494 MartínEz-valdiviEso, d., Font, r., blanCo-díaz, M.t., MorEno-roJas, J.M., góMEz, P., alonso-Moraga, a., dEl río-CElEstino, M., 2014: Application of near-infrared reflectance spectroscopy for predicting ca- rotenoid content in summer squash fruit. Comput. Electron. Agr. 108, 71-79. DOI: 10.1016/j.compag.2014.07.003 MCguirE, g.r., 1992: Reporting of objective color measurements. HortScience 27, 1254-12555. MéndEz-iturbidE, d., bandEras-tarabay, J.a., niEto-CaMaCho, a., roJas-ChávEz, a., garCía-MEza, M.g., 2013: Antioxidant capacity of extracts from hawthorn (Crataegus mexicana) skin. Afr. J. Food Sci. 7, 50-158. DOI: 10.5897/AJFS2013.1006 MEsEJo, C., gaMbEtta, g., gravina, a., MartinEz-FuEntEs, a., rEiga, C., agustia, M., 2012: Relationship between soil temperature and fruit colour development of ‘Clemenpons’ Clementine mandarin (Citrus cle- mentina Hort ex. Tan). J. Sci. Food Agr. 92, 520 -525. naMitha, k.k., nEgi, P.s., 2010: Chemistry and biotechnology of carote- noids. Crit. Rev. Food. Sci. 50, 728-760. DOI: 10.1080/10408398.2010.499811 niEto a.r., 2007: Tejocote. In: Nieto, A.R. (ed.), Frutales Nativos, Un Recurso Fitogénetico de México, 25-118. Universidad Autónoma Chapingo. Chapingo, México. niEto-angEl, r., borys, M.w., 1992: Banco de germoplasma de tejocote (Crataegus spp.) de la República Mexicana. Revista Chapingo 77, 126- 130. núñEz-Colín, C.a., hErnándEz-MartínEz, M.a., 2011: La problemática en la taxonomía de los recursos genéticos de Tejocote (Crataegus sp.) en México. Rev. Mex. Cienc. Agríc. 2, 141-153. PEña-varEla, g., salinas-MorEno, y., ríos-sánChEz. r., 2006: Con- tenido de antocianinas totales y actividad antioxidante en frutos de fram- buesa (Rubus idaeus L.). Rev. Chapingo Serie Hort. 12, 159-163. PhiPPs, J.b., 1997: Monograph of Northern Mexican Crataegus (Rosaceae subfam. Maloideae). SIDA Botanical Miscellany 15, 1-94. PhiPPs, J.b., o’kEnnon, r.J., lanCE, r.w., 2003: Hawthorns and Medlars. Royal horticultural society plant collector guide. Portland, USA.Timber Press. rEilly, k., 2013: On farm and fresh produce managent. In: Tiwari, B.K., Bruton, N.P., Brennan, C.S. (eds.), Handbook of Plant Food Phyto- chemichals: sources, stability and extraction, 201-234. Oxford, UK. John Wiley and Sons, Ltd. DOI: 10.1002/9781118464717.ch9 SAGARPA, 2015: Autoriza EUA la importación de higos y tejocotes mexi- canos. SAGARPA. http://comunicacion socialguanajuato.blogspot.com. es/2015_04_06_archive. html. Accessed 19 June 2017. robErtson, g.w., griFFiths, d.w., woodFord, J.a.t., birCh, a.n.E., PiCkEt, J.a., wadhaMs, l.J., 1993: A comparison of the flower volatiles from hawthorn and four raspberry cultivars. Phytochemistry 33, 1047- 1053. DOI: 10.1016/0031-9422(93)85021-I vEbEriC, r., slatnar, a., bizJak, J., staMPar, J., MikuliC-PEtkovsEk, M., 2015: Anthocyanin composition of different wild and cultivated berry species. LWT - Food Sci. Tech. 60, 509-517. DOI: 10.1016/j.lwt.2014.08.033 wrolstad, r.E., durst, r.w., lEE, J., 2005: Tracking colour and pigment changes in anthocyanins products. Trends Food Sci. Tech. 16, 423-428. DOI: 10.1016/j.tifs.2005.03.019 zhao, y., wang, y., wang, J., wu, z., zuli sun, z., tian, t., niu, h., Jing, l., Fang, z. yang, J., 2015: Characterization of volatile consti- tuents of Chinese hawthorn (Crataegus spp.) Fruit Juices, 533-545. In: Zhang, T.C., Nakajima, M. (eds.), Advances in Applied Biotechnology. © Springer-Verlag Berlin Heidelberg. New York. DOI: 10.1007/978-3-662-46318-5_55 Addresses of authors: María Dolores Pérez-Lainez, Tarsicio Corona-Torres, Victor Hebert Aguilar- Rincón, José Armando García-Velázquez. Colegio de Postgraduados. Texcoco, Estado de Mexico. Postal Code 56230. Mexico. María del Rosario García-Mateos*, Alejandro F. Barrientos-Priego, Raúl Nieto Ángel. Universidad Autónoma Chapingo. Texcoco, Estado de México. Postal Code 56230. Mexico * rosgar08@hotmail.com (Corresponding author) Robert Winkler. Laboratorio de Análisis Bioquímico e Instrumental. CINVESTAV. Irapuato, Guanajuato. Postal Code 36824. Mexico © The Author(s) 2019. This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creative- commons.org/licenses/by/4.0/deed.en).