ABSTRACT
Objective: The objective of this research was to determine the physicochemical composition of honey produced in the Sierra de Flores Magón region, Oaxaca, Mexico, to characterize its physicochemical parameters and verify Whether it meets the specifications established in national and international regulations.
Theoretical Framework: Honey produced in Mexico has a varied composition and is well accepted in the international market. However, evidence is lacking to support the appropriate composition of honey, free of signs of adulteration.
Method: A total of 29 honey samples from three municipalities in the Sierra de Flores Magón were collected directly from apiaries. The protocols of the Official Mexican Standard and the Codex Alimentarius were followed to determine: moisture, total soluble solids, electrical conductivity, ash, pH, acidity (free, lactone, and total), hydroxymethylfurfural (НМ), diastase index, and color.
Results and Discussion: The honeys met both national and international regulations. A significant difference (P < 0.05) was found in conductivity and ash content. Some parameters indicated that the honeys were fresh, with no signs of adulteration or fermentation.
Research Implications: Compliance with regulations could improve honey marketing conditions and generate better income for beekeepers.
Originality/Value: This research contributes to the understanding of the functional biological properties of honeys from the aforementioned region, improving the marketing process.
Keywords: Apis mellifera, Hydroxymethylfurfural, Diastase index, mmPfund.
RESUMO
Objetivo: O objetivo desta pesquisa foi determinar a composição fisico-quimica dos méis produzidos na região de Sierra de Flores Magón, Oaxaca, México, para caracterizar seus parámetros fisico-quimicos e verificar se atendem as especificações estabelecidas nas regulamentações nacionais e internacionais.
Referencial teórico: O mel produzido no México apresenta composição variada e boa aceitação no mercado internacional. No entanto, são necessárias evidências que confirmem que esse alimento tenha a composição adequada, sem sinais de adulteração.
Método: Foram coletadas diretamente dos apiários 29 amostras de mel de três municípios da Serra de Flores Magón. Foram seguidos os protocolos da Norma Oficial Mexicana e do Codex Alimentarius para determinar: umidade, sólidos solúveis totais, condutividade elétrica, cinzas, pH, acidez (livre, lactônica e total), Hidroximetilfurfural (HMD), índice de diastase e cor.
Resultados e Discussão: Os méis atendem às regulamentações nacionais e internacionais. Foi encontrada diferença significativa (P < 0,05) na condutividade e no teor de cinzas. Alguns parâmetros indicam que os méis eram frescos, sem sinais de adulteração ou fermentação.
Implicações da pesquisa: A conformidade com as regulamentações pode melhorar as condições de comercialização do mel e gerar maiores rendas para os apicultores.
Originalidade/Valor: Esta pesquisa contribui para o conhecimento das propriedades biológicas funcionais do mel da região citada, melhorando o processo de comercialização.
Palavras-chave: Apis mellifera, Hidroximetilfurfural, Índice de diastase, mmPfund.
RESUMEN
Objetivo: El objetivo de esta investigación fue determinar la composición fisicoquímica de mieles producidas en la región Sierra de Flores Magón, Oaxaca, México, para caracterizar sus parámetros fisicoquímicos, y verificar si cumplen con las especificaciones establecidas en la normatividad nacional e internacional.
Marco Teórico: La miel producida en México es de composición variada, com buena aceptación en el mercado internacional, sin embargo, hace falta evidencia que avale que este alimento presenta la composición adecuada, sin signos de adulteraciones.
Método: Un total de 29 muestras de miel de tres municipios de la Sierra de Flores Magón fueron recolectadas directamente en los apiários. Se siguieron los protocolos de la Norma Oficial Mexicana y del Codex alimentarius para determinar: humedad, sólidos solubles totales, conductividad eléctrica, cenizas, pH, acidez (libre, lactona y total), Hidroximetilfurfural (HMD), indice de la diastasa y color.
Resultados y Discusión: Las mieles cumplen tanto con la normatividad nacional e internacional. Se encontró diferencia significativa (P <0.05) en la conductividad y contenido de cenizas. Algunos parámetros indican que las mieles eran frescas, sin signos de adulteración o fermentación.
Implicaciones de la investigación: El cumplimiento de la normatividad podría favorecer las condiciones de comercialización de la miel y generar mejores ingresos económicos para los apicultores.
Originalidad/Valor: Esta investigación conribuye al conocimiento de las propiedades biológicas funcionales de las mieles de la región antes mencionada, mejorando el processo de comercialización.
Palabras clave: Apis mellifera, Hidroximetilfurfural, Índice de la diastasa, mmP fund.
1 INTRODUCTION
Honey from the bee Apis mellifera is defined as "the natural sweet substance produced by bees from the nectar of flowers or from secretions of living parts of plants or from excretions of plant-sucking insects that remain on living parts of the same and that the bees collect, transform and combine with their own specific substances, and deposit, dehydrate, store and leave in the honeycomb so that it matures or can age" (NOM, 2018; Codex, 2019). In 2022, world honey production was 2,149,256 tons (FAOSTAT, 2022). In 2021, honey production in Mexico was 63,362 tons (SIAP, 2019).
In recent decades, honey consumption has increased due to its beneficial health properties, such as: anti-inflammatory, antimicrobial, antimutagenic, antioxidant, antiproliferative, and antithrombotic (Hau-Y ama et al ., 2020). However, these properties could be modified if the physicochemical composition of honey is not adequate (Rysha et al ., 2022). Therefore, studies related to the physicochemical composition, functional properties, botanical and geographical origin are of utmost importance during the characterization and commercialization of honey, in addition to guaranteeing its safety and authenticity (Escudero and Saijo, 2022).
Around 200 chemical compounds present in honey have been identified (Lanjwani and Channa, 2019; Sakib et al ., 2017), with carbohydrates being the main constituents (approximately 95% of dry matter), followed by water, as well as a wide variety of substances, such as; proteins, enzymes, amino acids, organic acids, pigments, vitamins, minerals, phenolic compounds, flavonoids, carotenoids, aromatic substances and water-insoluble particles from the collection and/or handling of honey (Seraglio ef al ., 2019; Machado et al ., 2018). Pauliuc et al . (2020) mention that the color, aroma and flavor of honey is directly related to the type of flowering, geographical origin, climatic conditions of the region and the bee species. However, these characteristics can be affected by climatic conditions, poor hygiene during the harvesting, handling, packaging, and storage processes (Sakib et al ., 2017). Therefore, the objective of this study was to characterize honeys from three municipalities in the Sierra de Flores Magón region of the state of Oaxaca, in terms of their physicochemical parameters, and to verify whether they comply with the specifications established in national and international regulations.
2 MATERIALS AND METHODS
2.1 STUDY AREA
The samples analyzed came from the Sierra de Flores Magón region, located in the state of Oaxaca, in the Cañada and Papaloapan-Tuxtepec regions, comprising more than 20 municipalities. Seventy percent of the area is covered by high mountain ranges, while the rest is crisscrossed by small mountain ranges and hills. Furthermore, it is an area with a great diversity of ecosystems. The predominant vegetation is low deciduous forest, and it is one of the most important areas in terms of the richness and number of endemic flora and fauna species in Mexico (INPI, 2017).
2.2 COLLECTION OF HONEY SAMPLES
A total of 29 honey samples were analyzed (spring-summer 2020 harvest), San Jerónimo Tecóatl (n=12) (18°10'00"N, 96°55'00"W), San Pedro Ocopetatillo (n=6) (18%12'00"N, 96°55'00"W) and San Antonio Eloxochitlán (n=11) (18°10'35"N, 96 · 5230"W). With the support of the beekeeping organizations: "Miel Néctar Mazateco" and "Sociedad de Apicultores de Eloxochitlán". The samples were collected directly from the apiary, placed in previously sanitized containers of approximately 1.4 kg, labeled, hermetically sealed, and stored at room temperature until analysis.
2.3 PHYSICOCHEMICAL PARAMETERS
The total soluble solids content was expressed in degrees °Brix, according to NOM (2018), it was determined using a digital refractometer (Atago, PAL-3, USA), 1 to 2 g of honey were placed at 20 °C. The moisture content at 20 °C was determined according to AOAC (2012), by means of the refractive index, using a refractometer (HANNA, HI96800, USA), 1 to 2 g of honey were placed at 20 °C. Ash quantification was carried out according to AOAC (2012), 5 g of honey were placed in a crucible. Subsequently, a pre-calcination of the honey was carried out, then the crucible was placed in a muffle (Thermo Lyne, F6018, USA), at 550 °C for five and a half hours. The results were expressed as ash percentage. The pH was determined according to the AOAC (2012), 10 g of honey and 75 mL of distilled water were placed in a beaker, stirred until completely homogenized, and then the potentiometer electrode (Orion, STAR A2115, USA) was placed. The free, lactone and total acidity were determined according to the NOM (2018), 10 g of sample and 75 mL of distilled water were weighed in a beaker. The pH was measured by titration with 0.05 N sodium hydroxide, until reaching pH 8.5, subsequently titrated with 0.05 N hydrochloric acid until obtaining a pH 8.3. The results of free, lactone and total acidity were expressed in milliequivalents/kg of honey (meq acid/kg). To determine electrical conductivity, a 20% w/v honey solution was prepared, then the electrical conductivity meter (HANNA, HI99163) was placed inside the glass and the sample reading was taken (NOM, 2018). The results were expressed in mS/cm. To determine the hydroxymethylfurfural (HMF) content in honey, a UV/V spectrophotometer (Perkin Elmer, LAMBDA35, USA) was used. The methodology is based on the absorbance of hydroxymethylfurfural in the ultraviolet and visible regions of the spectrum at 284 nm (NOM, 2018). The results were expressed in milligrams per kilogram (mg/kg). Diastase activity was determined according to the amount of starch hydrolyzed by a honey solution (NOM, 2018). The absorbance was measured in a UV/V spectrophotometer (Perkin Elmer, LAMBDA35, USA) at 660 nm, at different times until an absorbance equal to or less than 0.235 was obtained. The diastase activity was calculated as the diastase index (DI) or grams of starch hydrolyzed at 40 °C per hour per 100 g of honey.
2.4 DETERMINATION OF THE COLOR OF HONEY
The honey color was determined by two methodologies: 1) By the CIELAB method using a spectrophotocolorimeter (3NH, NR110, China) with an aperture of 8 mm and illuminant of D65. 2) According to Montenegro et al. (2005), the honey was heated to 50 °C in order to dissolve any sugar crystals, then 10 mL of a 50% (w/v) honey solution was prepared with distilled water, the solution was homogenized with a vortex for 2 minutes, then centrifuged at 3200 rpm for 5 min, the supernatant was placed in 3 mL quartz cells and the reading was taken on the spectrophotometer at 635 nm (Perkin Elmer, LAMBDA 35, USA), using distilled water as a blank. The results were expressed on the mm Pfund scale (mmPfund= -38.70 + 371.39 · Abs).
2.5 STATISTICAL ANALYSIS
Results were expressed as the mean and standard deviation per municipality for each of the parameters evaluated. An ANOVA was performed for each of the study variables (by municipality), with a significance level of P <0.05; when a significant difference was found, a comparison of means was performed using the Tukey test. All analyses were performed using Statgraphics Centurion XVI software version 16.1.11 (Stat Point Technologies, Inc., The Plains, VA, USA).
3 RESULTS AND DISCUSSIONS
Table 1 shows the results of the physicochemical parameters of 29 honey samples from the Sierra de Flores Magón Region, Oaxaca, Mexico. On average, the moisture content for SJ honeys was 16.99 + 0.83, SP 17.14 + 0.42, and SA 17.28 + 0.83, with no significant differences (P <0.05). Moisture content is an important quality parameter in honey, since its stability and shelf life depend on it (Damto et al ., 2024). Honeys with high moisture content could be fermented, which could affect flavor and quality. However, honeys with low moisture content could granulate and crystallize (Chen, 2019). The moisture content of honey depends mainly on the botanical origin, time of year, level of maturity in the hive, climatic conditions of harvest, handling conditions during harvest and storage (Thrasyvoulou et al ., 2018). According to the Mexican Official Standard (NOM, 2018), the moisture content in honey should not exceed 20%, all samples analyzed presented values lower than 20% (Figure 2a). The results of this research are similar to honeys from northern Kenya, Kosovo, Greece, Romania and Turkey, the climatic conditions of these countries are very varied (Muhati and Warui, 2022; Rysha et al ., 2022; Tsavea et al ., 2022; Albu et al ., 2021; Akgün et al ., 2021).
Regarding the total soluble solids content expressed as °Brix, there were no significant differences (P <0.05) between the average honeys of the three municipalities (Table 1). The total soluble solids content in honey must be at least 80 °Brix, this makes the honey more stable during storage (Codex, 2019; NOM, 2018). All samples analyzed presented values higher than 80 °Brix (Figure 2b). The sugar content directly influences the physical characteristics of honey. Likewise, if the honey is heated or stored for a long period of time, simple sugars can undergo enolization, producing furans, and an increase in HMF (da Silva et al ., 2016). These results are similar to those reported in honeys from Kosovo (Rysha et al ., 2022), honeys from Bangladesh (Kamal et al ., 2019), and superior to honeys from northern Thailand (Hempattarasuwan et al , 2019).
The average electrical conductivity for honey samples from SJ, SA and SP was 0.35 + 0.04, 0.43 + 0.04 and 0.5 + 0.06 mS / cm respectively, no significant differences were found (P <0.05). The electrical conductivity of bee honey must be below 0.8 mS / cm (Codex, 2019), all samples comply with the regulations. Electrical conductivity is directly related to the content of minerals, organic acids and proteins (Rysha ег al ., 2022; Guerzou et al ., 2021), even electrical conductivity can be used to know the type of honey, values lower than 0.8 mS / cm belong to flower nectar honeys and values higher than 0.8 mS / cm indicate molasses honey (Pauliuc et al ., 2020). Based on the above, we can say that the honeys analyzed are floral honeys.
The ash content for SJ samples was 0.09 + 0.4, SP 0.11 + 0.05 and SA 0.08 + 0.02%, there was a significant difference (P <0.05) between SP and SA samples. Although national and international regulations do not specify the ash content in honey, Rysha et al . (2022) mention that the ash content in honey is a parameter to determine the geographical and floral origin of honeys. The results of this research are similar to those reported by Guerzou et al ., (2021), and Kamal et al., (2019). Darker honeys have a higher ash content and intense flavor (Craciun et al ., 2020).
Regarding pH, there was no significant difference (P <0.05) between the samples analyzed. The pH of honey is directly related to the texture, stability, and shelf life of honey (Mahnot et al ., 2018). Furthermore, pH is related to the flavor and acceptance of honey, due to the presence of various aromatic acids and aliphatic acids, gluconic acid being the main one, and it is related to the antimicrobial activity of honey (Laaroussi et al., 2020; Garcia et al., 2020). The results of this research are similar to those reported for honeys from Algeria, Romania, and Turkey (Makhloufi et al ., 2021; Albu et al ., 2021; Bayram et al ., 2020).
The average free acidity of SJ honeys was 17.31 + 2.22, SP 16.65 + 2.53 and SA 17.55 + 2.99 тед/Ке and no significant differences (P <0.05) were observed between the honeys of the three municipalities, the lactone content for SJ was 3.76 + 1.05, SP 3.73 + 0.79 and SA 3.78 + 1.14 meq/kg, no significant differences (P <0.05) were found between the honeys of the three municipalities, finally, the total acidity (free acidity + lactone) for SJ honeys was 21.07 + 2.50, SP, 20.39 + 2.57 and SA 21.33 + 3.72 meq/kg, there was no significant difference (P <0.05) between the honeys from the three municipalities. According to Mexican regulations (NOM, 2018), honey must have a maximum total acidity of 50 meg/kg. The total acidity results (Figure 3) are below the maximum recommended value, this could indicate that the honeys analyzed did not show signs of fermentation. These results are similar in honeys from northern Kenya and Morocco (Muhati and Warui, 2022; Laaroussi et al ., 2020), to honeys from India (Mahnot et al ., 2018), and are similar to honeys from bees fed with sugar (Kamal et al ., 2019).
For the hydroxymethylfurfural (HMF) content in honeys from the three municipalities, no significant difference was found (P <0.05). However, SJ honeys presented an average of 8.63 + 4.56 mg/kg, SP 8.07 + 5.79 mg/kg, and SA 12.77 + 4.77 mg/kg. The maximum permissible hydroxymethylfurfural content in honey is 40 mg/kg, with the exception of honeys from tropical climates, where up to 80 mg/kg is permitted (NOM, 2018 and Codex, 2019). We can affirm that the honeys analyzed were fresh and without signs of adulteration. The HMF content in honey can vary widely and is determined by several factors such as geographic location, pH, climate, floral source, honey quality, and especially the processing and preservation method (Al-Farsi ег al ., 2018). The results of this research were similar to those reported for Kenyan honeys (Muhati and Warui, 2022), Estonian honeys (Kivima et al ., 2021), and Turkish honeys (Akgün et al ., 2021).
Regarding the diastase index, national and international regulations recommend a minimum of 8 Schade units in honey from bees ( Apis mellifera ), and a minimum of up to 3 Schade units may be allowed in honeys with values lower than 15 mg / kg of HMF (NOM, 2018). The results show values lower than 8, it should be noted that these results agree with the values obtained for HMF. Ghorab et al . (2021) report values similar to those of this research. Warui et al . (2019) report an average of 10.63 Schade units in honeys from three regions of Kenya, and also emphasize that values close to 0 indicate that they are honeys that have received heat treatment or prolonged storage at high temperatures.
Table 2 shows the colour results of the honeys analysed using the two methods used. On the mmPfund colour scale, the values were: <9 Water White , 9-17 Extra White, 18-34 White, 35-50 Extra Light Amber, 51-85 Light Amber, 86-114 Amber and >114 Dark Amber. The samples were generally soft to light amber in colour.
No significant difference (P <0.05) was found in the color of the honeys. L· (lightness) values ranged from 32 to 42, a· values were in the red tones, b· values in the yellow tones of approximately 10 to 17, and approximately 9 to 22, respectively. The color of honey is a parameter that the consumer considers at the time of purchase, sometimes even the price, and acceptability depends on the color (Albu ef al ., 2021). The color of honey can change during storage or if it has received heat treatment (Guerzou et al ., 2021). Pauliuc et al . (2020) report very similar lightness values in honeys of different floral origin (monofloral and polyfloral), from Romania. Bayram et al . (2020) report lower luminosity values (20 to 29) in honeys from northern Turkey. Ghorab et al . (2021) mention that honeys from Algeria have a light amber to dark amber color, with very good acceptance. Homrani et al . (2020) report color values from 13 to 150 mmPfund, these values correspond to the extra white color to dark amber, they emphasize that the color of honey depends largely on the floral origin, in addition to the content of minerals, terpenes, carotenoids and polyphenols (Laaroussi et al., 2020).
5 CONCLUSION
The results of this research indicate that the 29 honey samples from three municipalities in the Sierra Mazateca (San Jeronimo Тесбай (n=12), San Pedro Ocopetatillo (n=6) and San Antonio Eloxochitlán (n=11), can be considered quality honeys , and comply with the physicochemical specifications of both national and international regulations (Mexican Official Standard and Codex Alimentarium ). Some parameters such as moisture content, total soluble solids, hydroxymethylfurfural and diastase index, indicate that the honeys analyzed were fresh and had not been adulterated. Compliance with regulations could improve the marketing conditions of honey and generate better income for beekeepers.
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Abstract
Objective: The objective of this research was to determine the physicochemical composition of honey produced in the Sierra de Flores Magón region, Oaxaca, Mexico, to characterize its physicochemical parameters and verify Whether it meets the specifications established in national and international regulations. Theoretical Framework: Honey produced in Mexico has a varied composition and is well accepted in the international market. However, evidence is lacking to support the appropriate composition of honey, free of signs of adulteration. Method: A total of 29 honey samples from three municipalities in the Sierra de Flores Magón were collected directly from apiaries. The protocols of the Official Mexican Standard and the Codex Alimentarius were followed to determine: moisture, total soluble solids, electrical conductivity, ash, pH, acidity (free, lactone, and total), hydroxymethylfurfural (НМ), diastase index, and color. Results and Discussion: The honeys met both national and international regulations. A significant difference (P < 0.05) was found in conductivity and ash content. Some parameters indicated that the honeys were fresh, with no signs of adulteration or fermentation. Research Implications: Compliance with regulations could improve honey marketing conditions and generate better income for beekeepers. Originality/Value: This research contributes to the understanding of the functional biological properties of honeys from the aforementioned region, improving the marketing process.