Mexican Propolis Research Articles

Effect of Mexican Propolis on Wound Healing in a Murine Model of Diabetes Mellitus.

Diabetes mellitus (DM) affects the wound healing process, resulting in impaired healing or aberrant scarring. DM increases reactive oxygen species (ROS) production, fibroblast senescence and angiogenesis abnormalities, causing exacerbated inflammation accompanied by low levels of TGF-β and an increase in Matrix metalloproteinases (MMPs). Propolis has been proposed as a healing alternative for diabetic patients because it has antimicrobial, anti-inflammatory, antioxidant and proliferative effects and important properties in the healing process. An ethanolic extract of Chihuahua propolis (ChEEP) was obtained and fractionated, and the fractions were subjected to High-Performance Liquid Chromatography with diode-array (HPLC-DAD), High-Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS) and Gas Chromatography-Mass Spectrometry (GC-MS) analyses and 46 compounds were detected. Deep wounds were made in a murine DM model induced by streptozotocin, and the speed of closure and the wound tensile strength were evaluated by the tensiometric method, which showed that ChEEP had similar activity to Recoveron, improving the speed of healing and increasing the wound tensile strength needed to open the wound again. A histological analysis of the wounds was performed using H&E staining, and when Matrix metalloproteinase 9 (MMP9) and α-actin were quantified by immunohistochemistry, ChEEP was shown to be associated with improved histological healing, as indicated by the reduced MMP9 and α-actin expression. In conclusion, topical ChEEP application enhances wound healing in diabetic mice.

Potential Antidiabetic Activity of Apis mellifera Propolis Extraction Obtained with Ultrasound.

Recent studies have linked phenolic compounds to the inhibition of digestive enzymes. Propolis extract is consumed or applied as a traditional treatment for some diseases. More than 500 chemical compounds have been identified in propolis composition worldwide. This research aimed to determine Mexican propolis extracts' total phenolic content, total flavonoid content, antioxidant activity, and digestive enzyme inhibitory activity (ɑ-amylase and ɑ-glucosidase). In vitro assays measured the possible effect on bioactive compounds after digestion. Four samples of propolis from different regions of the state of Oaxaca (Mexico) were tested (Eloxochitlán (PE), Teotitlán (PT), San Pedro (PSP), and San Jerónimo (PSJ)). Ethanol extractions were performed using ultrasound. The extract with the highest phenolic content was PE with 15,362.4 ± 225 mg GAE/100 g. Regarding the flavonoid content, the highest amount was found in PT with 8084.6 ± 19 mg QE/100 g. ABTS•+ and DPPH• radicals were evaluated. The extract with the best inhibition concentration was PE with 33,307.1 ± 567 mg ET/100 g. After simulated digestion, phenolics, flavonoids, and antioxidant activity decreased by 96%. In contrast, antidiabetic activity, quantified as inhibition of ɑ-amylase and ɑ-glucosidase, showed a mean decrease in enzyme activity of approximately 50% after the intestinal phase. Therefore, it is concluded that propolis extracts could be a natural alternative for treating diabetes, and it would be necessary to develop a protective mechanism to incorporate them into foods.

Activity of propolis from Mexico on the proliferation and virulence factors of Candida albicans

BackgroundThis research evaluated the anti-Candida albicans effect of Mexican propolis from Chihuahua.Chemical composition of the ethanolic extract of propolis was determined by GC-MS, HPLC-DAD, and HPLC-MS. The presence of anthraquinone, aromatic acid, fatty acids, flavonoids, and carbohydrates was revealed.ResultsThe anti-Candida activity of propolis was determined. The inhibitions halos were between 10.0 to 11.8 mm; 25% minimum inhibitory concentration (0.5 mg/ml) was fungistatic, and 50% minimum inhibitory concentration (1.0 mg/ml) was fungicidal.The effect of propolis on the capability of C. albicans to change its morphology was evaluated. 25% minimum inhibitory concentration inhibited to 50% of germ tube formation.Staining with calcofluor-white and propidium iodide was performed, showing that the propolis affected the integrity of the cell membrane.INT1 gene expression was evaluated by qRT-PCR. Propolis significantly inhibited the expression of the INT1 gene encodes an adhesin (Int1p).Chihuahua propolis extract inhibited the proliferation of Candida albicans, the development of the germ tube, and the synthesis of adhesin INT1.ConclusionsGiven the properties demonstrated for Chihuahua propolis, we propose that it is a candidate to be considered as an ideal antifungal agent to help treat this infection since it would not have the toxic effects of conventional antifungals.

Anti-Inflammatory and Histological Analysis of Skin Wound Healing through Topical Application of Mexican Propolis.

Skin wound healing is a complex biochemical process of tissue repair and remodeling in response to injury. Currently, the drugs used to improve the healing process are inaccessible to the population, are costly, and have side effects, making the search for new treatment alternatives necessary. Propolis is a natural product produced by bees that is widely recognized and used in folk medicine for its multiple biomedical activities. However, therapeutic information regarding Mexican propolis is limited. This study aimed to evaluate the wound-healing effect of the Chihuahua ethanolic extract of propolis (ChEEP). Macroscopic and histological analyses were performed using a mouse wound-healing model. The topic acute toxicity assay showed that propolis at 10% w/v had no toxic effects. ChEEP has antibacterial activity against the Gram-positive bacteria Staphylococcus aureus and Staphylococcus epidermidis. Moreover, it exhibited good anti-inflammatory activity evaluated through mouse ear edema induced by 12-O-tetradeca-noylphorbol-13-acetate (TPA). A full-thickness incision lesion was created in mice and treated topically with 10% ChEEP. At Day 14 post-treatment, it was observed that propolis increased wound contraction and reduced healing time and wound length; furthermore, propolis increased the tensile strength of the wound, as determined with the tensiometric method, and promoted the formation of type I collagen at the site of injury, as evaluated with Herovici stain. These findings suggest that the topical administration of ChEEP can improve skin wound healing, probably due to the synergistic effect of its components, mainly polyphenols, in different steps of the wound-healing process. It should be noted this is the first time that the wound-healing activity of a Mexican propolis has been evaluated.

Antifungal Activity of Mexican Propolis on Clinical Isolates of Candida Species.

Infections caused by micro-organisms of the genus Candida are becoming a growing health problem worldwide. These fungi are opportunistic commensals that can produce infections—clinically known as candidiasis—in immunocompromised individuals. The indiscriminate use of different anti-fungal treatments has triggered the resistance of Candida species to currently used therapies. In this sense, propolis has been shown to have potent antimicrobial properties and thus can be used as an approach for the inhibition of Candida species. Therefore, this work aims to evaluate the anti-Candida effects of a propolis extract obtained from the north of Mexico on clinical isolates of Candida species. Candida species were specifically identified from oral lesions, and both the qualitative and quantitative anti-Candida effects of the Mexican propolis were evaluated, as well as its inhibitory effect on C. albicans isolate’s germ tube growth and chemical composition. Three Candida species were identified, and our results indicated that the inhibition halos of the propolis ranged from 7.6 to 21.43 mm, while that of the MFC and FC50 ranged from 0.312 to 1.25 and 0.014 to 0.244 mg/mL, respectively. Moreover, the propolis was found to inhibit germ tube formation (IC50 ranging from 0.030 to 1.291 mg/mL). Chemical composition analysis indicated the presence of flavonoids, including pinocembrin, baicalein, pinobanksin chalcone, rhamnetin, and biochanin A, in the Mexican propolis extract. In summary, our work shows that Mexican propolis presents significant anti-Candida effects related to its chemical composition, and also inhibits germ tube growth. Other Candida species virulence factors should be investigated in future research in order to determine the mechanisms associated with antifungal effects against them.

In vitro antiviral effect of Mexican and Brazilian propolis and phenolic compounds against human coronavirus 229E

ABSTRACT Propolis is a resinous substance collected by bees (Apis mellifera). It is used for its biological properties. This natural product is available as a safe therapeutic option. Herein, we report the antiviral effects of brown propolis extract from Mexico and green and red propolis extracts from Brazil, as well as their phenolic compounds (quercetin, caffeic acid, and rutin) in preventing infection of MRC-5 cells by HCoV-229E. Normal human fibroblast lung cells (MRC-5) were used to determine the cytotoxicity of the compounds. All samples studied showed antiviral activity. Green and brown propolis extracts, and quercetin exhibited the best EC50 values with values of 19.080, 11.240, and 77.208 µg/mL against HCoV-229E, respectively, and with TC50 of 62.19, 29.192, and 298 µg/mL on MRC-5 cells, respectively. These results are the first in vitro study of the effects of propolis on HCoV-229E and provide the basis for the development of natural formulations against other coronavirus strains.

Comparison of the Biological Potential and Chemical Composition of Brazilian and Mexican Propolis

Propolis is a resinous substance collected by bees from plants and its natural product is available as a safe therapeutic option easily administered orally and readily available as a natural supplement and functional food. In this work, we review the most recent scientific evidence involving propolis from two countries (Brazil and Mexico) located in different hemispheres and with varied biomes. Brazil has a scientifically well documented classification of different types of propolis. Although propolis from Brazil and Mexico present varied compositions, they share compounds with recognized biological activities in different extraction processes. Gram-negative bacteria growth is inhibited with lower concentrations of different types of propolis extracts, regardless of origin. Prominent biological activities against cancer cells and fungi were verified in the different types of extracts evaluated. Antiprotozoal activity needs to be further evaluated for propolis of both origins. Regarding the contamination of propolis (e.g., pesticides, toxic metals), few studies have been carried out. However, there is evidence of chemical contamination in propolis by anthropological action. Studies demonstrate the versatility of using propolis in its different forms (extracts, products, etc.), but several potential applications that might improve the value of Brazilian and Mexican propolis should still be investigated.

Evaluation of the gastroprotective effects of Chihuahua propolis on indomethacin- induced gastric ulcers in mouse

The aim of this work was to evaluate the gastroprotective activity of a Mexican propolis on indomethacin-induced gastric ulcers in a mouse model. The following contents of the ethanolic extract of propolis of Chihuahua (EEPCh) were determined: antioxidant activity (SA50), total phenolic content (TPC), total flavonoid content (TFC), and chemical composition by HPLC-DAD and HPLC-MS, as well as acute toxicity by OECD Guideline 423. Gastric lesions were induced by intragastric indomethacin treatment in male ICR mice. As the positive control, omeprazole was administered, and three doses of EEPCh were evaluated (50, 150 and 300 mg/kg). Gastric mucosal injury, histological changes and mucosal content were evaluated by means of H&E and PAS staining. For homogenized gastric tissues, the following were evaluated: TBARS, MPO, and PGE2 levels; SOD and GPx antioxidant enzymatic activity; and the concentrations of the proinflammatory cytokines, TNF-α, IL-1β and IL-6. EEPCh had a significant SA50 of 41.55 µg/mL. The TPC of EEPCh was 860 mg GAE/g, and its TFC was 49.58 mg QE/g. Different phenolic compounds were identified in the extract and were not toxic. The EEPCh doses decreased mucosal damage and histological injuries, maintained the mucosal content and reduced the TBARS, MPO and concentrations of proinflammatory cytokines in gastric ulcer tissues. The 150 and 300 mg/kg doses increased the SOD activity and maintained the PGE2 content. Only the 300 mg/kg dose increased the GPx activity. The results of this study suggest that EEPCh displays gastroprotective effects by means of its antioxidant activity and anti-inflammatory effects and promotes ulcer protection through the maintenance of mucosal content and PGE2 levels.

Daño estructural en Cryptococcus neoformans producido por un propóleo mexicano

El propóleo es un producto de las abejas que recolectan de la vegetación y que muestra acción antifúngica por la presencia de compuestos fenólicos y flavonoides. Cryptococcus neoformans es una levadura encapsulada que produce una infección micótica oportunista en hospederos inmunocompetentes como inmunocomprometidos. Debido a la resistencia que C. neoformans ha presentado a los antimicóticos comerciales, es necesario estudiar otras sustancias de origen natural con acción fungicida.En este estudio se evaluó el daño a la estructura de C. neoformans, proveniente de un aislamiento clínico, producido por un extracto etanólico de propóleo mexicano mediante Microscopía Electrónica de Barrido (MEB). Se cuantificaron los compuestos fenólicos y flavonoides de la resina; la evaluación de la actividad antifúngica se realizó mediante la prueba de inhibición del crecimiento radial y para determinar la Concentración Mínima Inhibitoria y Fungicida (CMI y CMF) se utilizó el método de macrodilución.El extracto de propóleo mexicano demostró tener un contenido óptimo de fenoles y flavonoides; una concentración de 2.5 mg/mL del extracto de propóleo provoca la formación de invaginaciones e irregularidades en la superficie del hongo produciendo una disminución del tamaño.Se demuestra la actividad anticriptococica de un extracto de propóleo al evidenciar el daño en la estructura celular, lo cual puede proporcionar la perspectiva de alternativa terapéutica para pacientes con diversas enfermedades producidas por este hongo, considerando ampliar la investigación del mecanismo de acción del propóleo como antifúngico.

Advances in Chemical Composition and Biological Activity of Mexican Propolis

Propolis is resinous material produced by honeybees, which collect secretions from plants organs and then they are mixed...

Mexican Propolis: A Source of Antioxidants and Anti-Inflammatory Compounds, and Isolation of a Novel Chalcone and ε-Caprolactone Derivative.

The propolis produced by bees are used in alternative medicine for treating inflammation, and infections, presumably due to its antioxidant properties. In this context, five propolis from México were investigated to determine their inhibitory lipid peroxidation properties. The ethyl acetate extract from a red propolis from Chiapas State (4-EAEP) was the most potent (IC50 = 1.42 ± 0.07 μg/mL) in the TBARS assay, and selected for further studies. This extract afforded two new compounds, epoxypinocembrin chalcone (6), and an ε-caprolactone derivative (10), as well as pinostrobin (1), izalpinin (2), cinnamic acid (3), pinocembrin (4), kaempherol (5), 3,3-dimethylallyl caffeate in mixture with isopent-3-enyl caffeate (7a + 7b), 3,4-dimethoxycinnamic acid (8), rhamnetin (9) and caffeic acid (11). The HPLC profile, anti-mycobacterial, and antioxidant properties of this extract was also determined. Most of the isolated compounds were also tested by inhibition of reactive oxygen species (ROS) in challenged mouse bone marrow-derived mast cells (BMMCs), and DPPH. Their anti-inflammatory activity was evaluated by TPA, and MPO (myeloperoxidase) activity by ear edema test in mice. The most potent compounds were 7a + 7b in the TBARS assay (IC50 = 0.49 ± 0.06 μM), and 2 which restored the ROS baseline (3.5 μM). Our results indicate that 4-EAEP has anti-oxidant, and anti-inflammatory properties due to its active compounds, suggesting it has anti-allergy and anti-asthma potential.

Hypoglycaemic and Antioxidant Effects of Propolis of Chihuahua in a Model of Experimental Diabetes.

Propolis is a bee-collected natural product that has been proven to have various bioactivities. This study tested the effects of a Mexican propolis on streptozotocin-induced diabetes mellitus in a murine model. The results showed that an ethanolic extract of propolis of Chihuahua (EEPCh) significantly inhibited increases in blood glucose and the loss of body weight in diabetic mice. EEPCh increased plasma insulin levels in STZ-diabetic mice, whereas, in untreated diabetic mice, there was no detection of insulin. EEPCh had a high antioxidant capacity (SA50 = 15.75 μg/mL), which was directly related to the concentrations of total phenols (314 mg GAE/g of extract) and flavonoids (6.25 mg QE/g of extract). In addition, increased activities of the enzymes superoxide dismutase, catalase, and glutathione peroxidase were observed in diabetic mice treated with EEPCh. Compounds such as pinocembrin, quercetin, naringin, naringenin, kaempferol, acacetin, luteolin, and chrysin were identified by HPLC-MS analysis. This investigation demonstrated that propolis of Chihuahua possesses hypoglycaemic and antioxidant activities and can alleviate symptoms of diabetes mellitus in mice. These effects may be directly related to the chemical composition of propolis, as most of the compounds identified in propolis are reportedly active in terms of the different parameters evaluated in this work.

Comparison between In Vitro Antiviral Effect of Mexican Propolis and Three Commercial Flavonoids against Canine Distemper Virus.

Propolis is a resin that honey bees (Apis mellifera) produce by mixing wax, exudates collected from tree shoots, pollen, and enzymes. It has been used for its biological properties against pathogenic microorganisms including those of viral origin. In the present study, we demonstrate the antiviral effect of Mexican propolis, as well as of the three commercial flavonoids (quercetin, naringenin, and pinocembrin) present in its composition, in cell cultures infected with Canine Distemper Virus. The treatments were carried out with propolis, flavonoids individually, and a mixture of the three flavonoids at three different times. Antiviral activity was evaluated by the inhibition of the relative expression of the virus nucleoprotein gene (Real-Time qPCR) and by the determination of cellular viability (MTT assay). Propolis applied before infection decreased viral expression (0.72 versus 1.0, 1.65, and 1.75 relative expressions) and correlated with increased cell viability (0.314 versus 0.215, 0.259, and 0.237 absorbance units (AU)). The administration of a flavonoid mixture containing the three commercial flavonoids before infection induces a slight decrease in viral expression (0.93 versus 1, 1.42, and 1.82 relative expressions); however, it does not improve cellular viability (0.255 versus 0.247, 0.282, and 0.245 AU). Quercetin administrated at the same time of infection decreases viral expression (0.90 versus 1.0, 3.25, and 1.02 relative expressions) and improves cellular viability (0.294 versus 0.240, 0.250, and 0.245 AU). Pinocembrin and naringenin individually did not show any antiviral activity at the administration times evaluated in this study. The present work is the first in vitro study of the effect of propolis in Canine Distemper Virus and demonstrated the antiviral activity of Mexican propolis, in addition to the synergy that exists between the three flavonoids on cell viability and the expression of the nucleoprotein virus gene.

Prediction of Antimicrobial and Antioxidant Activities of Mexican Propolis by 1H-NMR Spectroscopy and Chemometrics Data Analysis

A feasibility study to predict antimicrobial and antioxidant activity properties of propolis extracts using 700-MHz 1H-NMR spectra and multivariate regression data analysis is presented. The study was conducted with thirty-five propolis samples to develop a rapid and reliable method for the evaluation of their quality. The extracts have been evaluated by measuring phenolic and flavonoid contents; the antioxidant activity; and the antimicrobial activity. The obtained spectral data were submitted to multivariate calibration (partial least squares (PLS) and orthogonal partial least squares (OPLS)) to correlate the relative intensity and position of NMR resonance peaks with the metabolites contents and biological activities. The developed PLS and OPLS model were successfully applied to the determination of the target properties for proof of the concept. The OPLS observed vs. predicted properties plots indicate the absence of systematic errors with determination coefficients between the ranges 0.7207 to 0.9990. Up to 86.1% of explication of variation in the spectral data and 99.9% in the measured properties were attained with 88.6% of prediction capabilities in the best case (S. mutans activity) according to the cross-validation procedure. The figures of merit of the developed PLS and OPLS methods were evaluated and compared as well.

Unusual chemical composition of a Mexican propolis collected in Quintana Roo, Mexico

Propolis is a resinous natural substance collected by honey bees from buds and exudates of various trees and plants. It is widely accepted that the composition of propolis depends on the phytogeographic characteristics of the site of collection. In this study, we have analyzed the chemical composition of a propolis collected in Quintana Roo, Mexico, and evaluated its antioxidant, antifungal, and antibacterial activities. Unexpectedly, the chemical analysis showed that the main components of the ethanolic extract of this Mexican propolis appeared to be pentacyclic triterpenoids, such as α and β-amyrin derivatives, and sterols. The crude extract did not show antioxidant activity when tested using the DPPH-reduction assay, and it also proved inactive when tested for antifungal and antibacterial activities using microdilution and agar diffusion assays, respectively. The fact that the presence of both α and β-amyrins and their derivatives have been reported from the resin of Bursera simaruba, one of the plants used by the bees for propolis production in Quintana Roo, Mexico, confirms the relationship that exists between the flora available to bees in a given region and the chemical composition of the propolis that they produce.

Use of Mexican Propolis for the Topical Treatment of Dermatomycosis in Horses

Five horses with lesions compatible with dermatomycosis were selected and skin scrapings were taken from all affected areas. Samples were then treated with 10% KOH and observed by direct microscopic examination; fungal cultures using Mycobiotic Agar and Borelli Agar were also preformed; Trichophyton mentagrophytes was identified in two cases, whereas Candida albicans was identified in the other three. Skin lesions in cases 1 and 2 were treated topically with ketoconazol ointment, while in the remaining cases a weekly bath with propolis-based shampoo and application of propolis-based ointment on the lesions 2 to 3 times a week were implemented. Treatment in all cases lasted 4 weeks. Pictures were taken before and after treatment and, making use of the Image Pro-Express&#174 software, therapy efficacy was determined by measurement of lesion area diameter and a significant reduction in lesion size was observed by the end of the 4 weeks of treatment with ketoconazole and propolis-based products. Thus proving the effectiveness of such propolis-based products, making them a natural therapeutic alternative for the treatment of cutaneous mycoses in horses, and carrying none of the toxic side effects of conventional pharmaceuticals.

Mexican propolis flavonoids affect photosynthesis and seedling growth

As a continuous effort to find new natural products with potential herbicide activity, flavonoids acacetin (1), chrysin (2) and 4′,7-dimethylnarangenin (3) were isolated from a propolis sample collected in the rural area of Mexico City and their effects on the photosynthesis light reactions and on the growth of Lolium perenne, Echinochloa crus-galli and Physalis ixocarpa seedlings were investigated. Acacetin (1) acted as an uncoupler by enhancing the electron transport under basal and phosphorylating conditions and the Mg2+-ATPase. Chrysin (2) at low concentrations behaved as an uncoupler and at concentrations up to 100μM its behavior was as a Hill reaction inhibitor. Finally, 4′,7-dimethylnarangenin (3) in a concentration-dependent manner behaved as a Hill reaction inhibitor. Flavonoids 2 and 3 inhibited the uncoupled photosystem II reaction measured from water to 2,5-dichloro-1,4-benzoquinone (DCBQ), and they did not inhibit the uncoupled partial reactions measured from water to sodium silicomolybdate (SiMo) and from diphenylcarbazide (DPC) to diclorophenol indophenol (DCPIP). These results indicated that chrysin and 4′,7-dimethylnarangenin inhibited the acceptor side of PS II. The results were corroborated with fluorescence of chlorophyll a measurements. Flavonoids also showed activity on the growth of seedlings of Lolium perenne and Echinochloa crus-galli.

Protective effect of a mexican propolis on MDBK cells exposed to Aujeszky's disease virus (Pseudorabies virus)

Protective effect of a mexican propolis on MDBK cells exposed to Aujeszky's disease virus (Pseudorabies virus)

PROTECTIVE EFFECT OF MEXICAN PROPOLIS ON MDBK CELLS EXPOSED TO AUJESZKY'S DISEASE VIRUS (PSEUDORABIES VIRUS).

In this work, antiviral effect of Mexican propolis was evaluated in vitro on MDBK (Madin Darby Bovine Kidney) cell cultures infected with pseudorabies virus (PRV). First was carried the cytotoxicity test of the ethanol extract of propolis (EEP) on this cell culture, subsequently, the infective dose of virus Shope strain was determined for the infection of cell cultures and finally the interaction between EPP and virus was done at cell culture level (two hours before, during and two hours later of infection). Also, cell culture samples with the same treatments were processed for transmission electron microscopy (TEM) to observe possible alterations of propolis on the virus. It was determined that using the treatment of 0.5 mg / ml of EEP two hours before being infected, the culture shows a reduction in the number of plaque forming units compared to the other treatments or with the culture infected, it is important to note that the difference found was statistically significant (p

Development and Validation of a RP-HPLC Method for the Simultaneous Quantification of Flavonoids Markers in Mexican Propolis

Acacetin (1), 4′,7-dimethyl naringenin (2), and 4′,7-dimethyl apigenin (3)—three of the main markers active components of propolis—were chosen for the development and validation of a suitable HPLC method for quality control of the crude drug. A 23 factorial design was employed to study the main interactions of independent variables, and the efficient separation was achieved using an XBridge C18 column with an isocratic binary phase consisting of deionized water (TFA; 0.1 % v/v) (eluent A) in methanol (eluent B) (45:65, v/v) at a flow rate of 1.0 mL min−1. The method allows the quantification of 1–3; for each compound, a linear response was evaluated within the range of 100–1,000 μg mL−1 for 1, 25–200 μg mL−1 for 2, and 8–60 μg mL−1 for 3. All the calibration curves showed good linearity within the test ranges (r2 ≥ 0.999). Percentage recoveries of the selected flavonoids 1–3 ranged between 98.3 and 101.2 % (RSD ≤2.0 %). The intra- and interday precision RSDs were no more than 2.0 %, while the repeatability variation was no more than 2.0 %. Finally, the method was applied to establish some seasonal and geographical variations of the markers in propolis obtained from several regions of Mexico. Thus, 4′,7-dimethyl apigenin (3) was selected as a marker compound from Mexican propolis, and, along with 2, could be useful for quality control procedures focused in to establish some geographical variation of Mexican propolis.