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Hindawi Publishing Corporation Journal of Chemistry Volume 2015, Article ID 860381, 9 pages http://dx.doi.org/10.1155/2015/860381

Research Article Nutritional Content and Elemental and Analyses of oleifera Grown in Mexico

Mónica A. Valdez-Solana,1 Verónica Y. Mejía-García,1 Alfredo Téllez-Valencia,2 Guadalupe García-Arenas,3 José Salas-Pacheco,4 José J. Alba-Romero,1 and Erick Sierra-Campos1

1 Facultad de Ciencias Qu´ımicas, Universidad Juarez´ del Estado de Durango, Avenida Art´ıculo 123 S/N, Fracc, Filadelfia, 35010 Gomez´ Palacio, DGO, Mexico 2Facultad de Medicina y Nutricion,´ Universidad Juarez´ del Estado de Durango, Avenida Universidad y Fanny Anitua´ S/N, 34000 Durango, DGO, Mexico 3Facultad de Medicina, Universidad Juarez´ del Estado de Durango, Calzada Palmas 1, Colonia Revolucion,´ 35050 Gomez´ Palacio, DGO, Mexico 4Instituto de Investigacion´ Cient´ıfica, Universidad Juarez´ del Estado de Durango, Avenida Universidad S/N, 34000 Durango, DGO, Mexico

Correspondence should be addressed to Erick Sierra-Campos; [email protected]

Received 16 November 2014; Revised 28 March 2015; Accepted 31 March 2015

Academic Editor: Deborah Pacetti

Copyright © 2015 Monica´ A. Valdez-Solana et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Moringa oleifera is a tree distributed in Mexican semiarid and coastal regions. M. oleifera is used in practice in the treatment of various diseases and is available without a medical prescription, often in the form of an herbal infusion for everyday use. The aim of the present study was to evaluate the chemical composition and nutritional values of dried M. oleifera powder collected from two different regions in Mexico. All samples of M. oleifera exhibited moisture levels varying from 3.06 to 3.34%, lipids from 10.21 to 10.31%, fiber from 7.29 to 9.46%, ashes from 10.71 to 11.18%, crude from 10.74 to 11.48%, and from 54.61 to 57.61%. The predominant elements in the leaf powder according to ICP-MS were Ca (2016.5–2620.5 mg/100 g), K (1817–1845 mg/100 g), and Mg (322.5–340.6 mg/100 g). The HPLCnalysis a indicated the presence of phenolic acids (gallic and chlorogenic acids) and flavonoids (rutin, luteolin, quercetin, apigenin, and ). We concluded that Lombardia M. oleifera samples could be employed in edible and commercial applications. Our results showed that the highest mean value of As from the San Pedro samples exceeds the recommended level and may constitute a health hazard to consumers.

1. Introduction has been attributed to the antioxidant properties of their con- stituents, usually associated with a wide range of amphipathic have been an important source of medicine for thou- moleculesthatarebroadlyreferredtoaspolyphenoliccom- sands of years. Even today, the World Health Organization pounds [2]. There is a growing interest in the development (WHO) estimates that up to 80% of people still rely primarily and evaluation of natural antioxidants from medicinal on traditional remedies such as herbs for their medicines [1]. materialsinthefoodindustryandthefieldofpreventive The medicinal value of these plants is due to the presence of health care. Among those herbs, one promising species is a variety of and their elemental composition. Moringa oleifera Lam. (Moringa or drumstick tree), which The role of medicinal plants in disease prevention or control is native to the sub-Himalayan regions of Northwest India. 2 Journal of Chemistry

It is widely distributed throughout Africa, Saudi Arabia, Milli-Q Plus system (Millipore, Bedford, Southeast Asia, the Islands, and South America. USA). The multielement standard mixture solutions con- Every part of M. oleifera has medicinal properties and is tainedBa,Bi,LiCu,Co,In,Li,Ni,Pb,andV;theothers commercially exploitable for the development of medicinal contained Ba, Ca, Mg, and Sr. Analytical grade 65% w/v nitric and industrial byproducts [3]. acid (HNO3) (JT Baker) was used in the decomposition. Traditionally, the , fruits, flowers, and immature pods of this tree are edible; they are used as a highly nutritive 2.2. Samples Preparation. M. oleifera leaves were collected in many countries, particularly in India, Pakistan, from Mexican cultivars (Lombardia, Michoacan,´ located thePhilippines,,andsomeAfricannations[4–6]. In ∘ 󸀠 󸀠󸀠 ∘ 󸀠 󸀠󸀠 on 19 01 30 Nand10205 39 WandSanPedro,Coahuila, developing nations, M. oleifera is used as an alternative to ∘ 󸀠 󸀠󸀠 ∘ 󸀠 󸀠󸀠 locatedon2545 35 Nand10258 54 W,Mexico, April 2011) imported food supplements to treat and combat malnutri- and a voucher of the specimen was deposited in the CIIDIR- tion, especially among infants and nursing mothers, by virtue IPN herbarium, Durango, Mexico, for future references. The of its chemical constituents [7]. plants were identified and authenticated by M. Sc. Alberto Several valuable reviews of the ethnobotanical uses Gonzalez´ Zamora, Faculty of Biology, UJED. The leaves were of M. oleifera are available [8–11]. Moringa has been cleaned and processed as described by previous researchers found to be a good source of and antiox- [22, 23]. A set of five samples from each geographical origin idants [11]. Phytochemicals such as vanillin, omega fatty were randomly collected and analyzed. All materials were air- acids, carotenoids, ascorbates, tocopherols, beta-sitosterol, dried and powdered and protected from light until further moringine, kaempferol, and quercetin have been reported in analysis. its flowers, , fruits, and . The leaves, in particular, have been found to contain phenolics and flavonoids [12, 13]; these compounds have various biological activities, including 2.3. Chemical Analysis. Representative subsamples were ∘ antioxidant, anticarcinogenic, immunomodulatory, antidia- dried in a forced oven at 105–110 Ctoaconstant betic, antiatherogenic, and hepatoprotective functions and weight for moisture determination. Crude protein, crude the regulation of thyroid status [14–16]. Moreover, leaves lipid, crude fiber, and ash were analyzed by triplicate accord- contain trace elements that are essential to human health. For ing to AOAC [24]. The total carbohydrates were determined instance, , iron, selenium, and play an impor- by the difference method [100 − ( + + moisture tant role in metabolism, and interest in these elements is + ash in percentage)] [25, 26]. The energy values (kcal/100 g) increasing together with reports relating trace element status were determined by multiplying the values of carbohydrates, and oxidative diseases [17, 18]. However, a recent study has lipids,andproteinsbyfactorsof4,9,and4,respectively,and shown that dried M. oleifera leaves contain lead at very high taking the sum expressed in kilocalories [27]. values of 352.0 mg/L [19]. Therefore, it is very important to identify the mineral composition of M. oleifera leaves that are 2.4. Extraction of Phenolic Compounds. The extracts were widely consumed by humans and animals. prepared using 23 g of dry-ground sample and 260 mL of 80% In Mexico, M. oleifera is widely cultivated in different methanolic aqueous solution by successive maceration. The zones of the country and is found in more than ten states from mixtures were shaken in a magnetic grid at room temperature Sonora to Oaxaca on the Pacific side. Few studies have been for 21 h. They were then filtered through Whatman filter conducted on nutritional and phytochemical composition paper number 1. The final extract was concentrated ona [20, 21]; however, to date, a detailed composition of the leaves rotator evaporator, and the filtrate was placed in a deep of M. oleifera that is native to Mexico has not been reported freezer for 24 h and lyophilized to obtain a powdered extract. yet. In addition, it is important to bear in mind that the All of the extractions were performed in triplicate, and the mineral and phenolics contents present in leaves depend on ∘ supernatants were kept at −40 C until further analysis. several factors such as geographical area where the plant is cultivated, type of soil, water and , industrialization process, and storage conditions. Taking these precedents into 2.5. ICP-MS Analysis. The instrumentation used for diges- consideration, the aim of this study was to evaluate the tion consisted of a closed microwave oven (MARS-Xpress, phytochemical constituents of methanolic extracts and trace CEM). The multielemental analysis was performed by induc- elements and nutritional values present in M. oleifera leaves tively coupled plasma mass spectrometry (ICP-MS) with a grown in the northeast and west regions of Mexico. reaction cell (XSeries 2, Thermo Scientific). Prior to analysis, thesampleswerehomogenizedinamortar;approximately 2. Materials and Methods 0.5gofthesamplewasaccuratelyweighedinanacid-washed polytetrafluoroethylene (PTFE) digestion tube, and 5 mL of 2.1. Reagents and Standards. The methanol and acetonitrile HNO3 (65% w/v) was added. The tube was heated in a ∘ used in this study were of HPLC grade and were purchased microwaveovenatapowersettingof50%and150Cfor ∘ ∘ from JT Baker, USA, and Caledon Lab., Canada. HPLC-grade 10 min, 70% and 220 C for 20 min, and 10% and 100 Cfor phenolic standards (gallic and chlorogenic acids, rutin, lute- 15 min. The digest was transferred into a 50 mL acid-washed olin, quercetin, apigenin, and kaempferol) were of analytical volumetric flask, which was filled with demineralized water reagent grade and were purchased from Sigma-Aldrich (St. andstoredinapolypropylenecontainer.Onewaterblankwas Louis, MO, USA). Demineralized water was taken from a run with each batch of samples. Journal of Chemistry 3

2.6. HPLC-DAD Analysis. Seven phenolic standards and Table 1: Nutritional composition of dried M. oleifera leaf powder. samples were dissolved in the mobile phase, yielding con- centrations of 25 𝜇g/mL. The solutions were filtered through Parameter Lombardia San Pedro [28] a0.45𝜇m membrane filter, and the evaluation of each Moisture (%) 3.34 ± 1.36 3.06 ± 1.38 7.4 ± 2.89 calibration curve was fitted by linear regression. An Agilent Lipid (%) 10.31 ± 1.2 10.21 ± 1.83 6 ± 2.5 1200 HPLC Series (Agilent, Palo Alto, CA, USA) was operated ± ± ± ∘ Fiber (%) 7. 2 9 0.84 9.46 1.14 9 7. 4 5 at 30 C, equipped with a degasser, a diode array detector Ash (%) 10.71 ± 0.81 11.18 ± 0.19 17.6b (DAD), a quaternary gradient pump, and an Eternity C18 ∗ 10.74 ± 1.3 11.48 ± 1.4 24 ± 5.8 reversed-phase analytical column of 150 mm × 4.6 mm and Protein (%) a ± ± ± ∗ a 3.5 𝜇m particle (Sigma-Aldrich, USA). The mobile phase Carbohydrates (%) 57.61 2.19 54.61 0.6 36 9.2 was a binary gradient: methanol and buffer solution. The Energy value 366.2 ± 4.23 356.25 ± 6.48 304 ± 87 buffercontainedTCA0.1%solutioninwater,anditspHwas (Kcal/100 g) adjusted to 2.1. The linear gradient began with 20% to 50% Values are mean ± SD, analyzed individually in triplicate, and are expressed methanol over the first 30 min, followed by 70% methanol as g/100 g leaf powder. a, bCalculated by difference. over the next 30 min. The flow rate was 0.6 mL/min, and data ∗ Difference significative between Mexican samples against a reference 𝑝< were collected at 270 and 352 nm. The injection volume was 0.018. 20 𝜇L. Data acquisition, peak integration, and calibrations wereperformedwithAgilent1200Chemstationsoftware. The results are reported as means ± standard deviations of Table 2: Measured concentration in (mg/100 g) of trace element in the M. oleifera leaf powder. triplicate independent analyses. Elements Lombardia San Pedro [28] ± ∗∗ 2.7. Statistical Analysis. The data were expressed as means Calcium (Ca) 2016.5 ± 22.6 2620.5 ± 5.6 1897 ± 748.4 ∗∗ SD, and the results were statistically analyzed using a Holm- Magnesium (Mg) 322.5 ± 0.0 340.6 ± 2.8 473 ± 429.4 Sidak test and one-way analysis of variance (ANOVA) on a (K) 1845 ± 7. 0 1 8 1 7 ± 14.1 1467 ± 636.7 SigmaPlot program v. 11.0 for Windows. The differences were ∗∗ 𝑝 < 0.05 Sodium (Na) 8.13 ± 0.6 40.78 ± 0.7 220 ± 180 considered statistically significant if . ∗ Iron (Fe) 19.37 ± 6.6 7.07 ± 0.4 32.5 ± 10.78 ± ± ± 3. Results Zinc (Zn) 1.0 0.7 1.6 0.6 2.4 1.12 Copper (Cu) 1.03 ± 0.47 0.41 ± 0.0 0.9 ± 0.48 −3 ∗∗ 3.1. Chemical Analysis. The proximate and nutrient analyses Selenium (Se) 9.55 × 10 ± 0.0 0.107 ± 0.0 ND ∗∗ of M. oleifera play a crucial role in assessing its nutritional Lead (Pb) 0.355 ± 0.0 0.2 ± 0.0 ND 𝑝 < 0.05 −3 ∗∗ significance ( ). The proximate analysis for both Arsenic (As) 5.5 × 10 ± 0.0 0.28 ± 0.0 ND cultivars is presented in Table 1, along with the values found Results are mean ± SD, analyzed individually in triplicate. in the literature for identical M. oleifera for comparative ND: nondetermined. purposes. Nonsignificant differences were observed between Statistically significant differences between the means of both cultivars are ∗∗ ∗ the proximate analyses of the samples studied in Lombardia denoted as p < 0.001; p < 0.05. and San Pedro. The mean moisture, lipid, fiber, and ash values found in the present study are in agreement with thevaluesreported[28]. The result shows that Mexican and mineralogy) [32]. Ten elements, including toxic ones cultivars contain crude protein (10.6%), crude fiber (about such as lead and arsenic, were determined to be present in 8%), ash (about 11%), carbohydrates (about 56%), moisture two crops of M. oleifera collected from Lombardia and San (about 3.2%), and lipid (about 10.2%). Carbohydrates are the Pedro locations. The elements were determined by ICP-MS principal sources of energy. The ash content of about 11% after wet digestion of the dried sample with concentrated indicates that the leaves are rich in mineral elements. The nitric acid in closed PTFE vessels using a microwave oven. mean protein content found in the Mexican samples ranged Ca,Mg,K,Fe,andNawerepresentatlevelsofmg/100g from 10.74 to 11.48%. This is a lower protein level than the dry matter, whereas Zn, Cu, and Se were present at 𝜇g/100 g 27.2% reported by other authors [21, 29–31], though it may levels, in close agreement with levels previously reported [30], still be valuable as a protein source. The variation in chemical and these values fall within the ranges reported (Table 2, analyses between our cultivars and the reference may depend reference column). The elemental analysis of our samples on seasonal variations, the plants’ stages of development, revealed high contents of Ca (2016 to 2620 mg/100 g), Mg and the techniques employed to collect leaf samples before (322 to 340.6 mg/100 g), and K (1817 to 1845 mg/100 g), while the experimental analysis. The chemical composition values Zn,Cu,andSewere1,1,and0.1mg/100g,respectively.Iron confirmed that M. oleifera leaves are an excellent food source, was abundant in Lombardia cultivar, whereas the Ca, Mg, justifying its direct use in human nutrition or development of andNalevelswerehigherinSanPedrocultivar(Table 2). balanced diets for animal nutrition. These values of M. oleifera leaves from Mexico agree with those found in Burkina Faso and India [29, 30]. The Fe varied 3.2. Elemental Analysis. Different soils contain a particular of among the leaf samples collected from Lombardia and San mineral elements qualities and quantities whose bioavailabil- Pedro, ranging from 19.37 to 7.07, respectively. However, this ity depends on soil properties (pH, clay and humid complex, level is low when compared to the contents of some results 4 Journal of Chemistry from India [30].ThePblevelinLombardiaandSanPedro Table 3: Elution order of polyphenolic compounds separated by cultivars was below the limit set by the WHO acceptable HPLC-DAD. daily intake of Pb for adults of 0.21–0.25mg/day [33]. The Reference Retention Linear range 𝑟2 lowest As content was observed in the Lombardia cultivar compound time (min) (𝜇g/mL) (0.0055 mg/100 g), whereas the content of As in the samples 270 nm collected from San Pedro was the highest (0.28 mg/100 g), ± which correlates with the extremely high As level in San Gallic acid 4.53 0.05 1.0–9.9 0.9999 Pedrosoil.BecausethelevelsintheLombardiasample 352 nm Chlorogenic were still within the prescribed limit for food at 1 mg/kg 15.52 ± 0.38 0.6–23.1 0.9995 according to the WHO/FAO standard [34], this sample could acid be recommended as a source of essential elements. However, Luteolin 46.89 ± 0.29 0.5–4.0 0.9993 because the As levels in San Pedro samples were found to Rutin 48.25 ± 0.25 9.9–49.5 0.9996 be higher than the limit set by the WHO, it is necessary to Quercetin 56.07 ± 0.21 0.99–3.0 0.9999 study them in further detail to avoid overconsumption and Kaempferol 59.79 ± 0.81 0.66–3.3 0.9998 cumulative toxicity from long-term use. Apigenin 60.59 ± 0.15 0.1–1.0 0.9995 Each value is the mean ± SD of triplicate determinations. 3.3. Quantification of Phenolics. RP-HPLC with DAD detec- tion was employed for the identification and quantification Table 4: Characterization (𝜇g/g dry matter) of polyphenolic com- of main phenolic compounds present in the methanolic pounds separated by HPLC-DAD. extracts. The quantifications of phenolics (mg/g dry matter) Compound San Pedro Lombardia were accomplished by comparing retention times and peak Gallic acid 49.07 ± 4.53 43.28 ± 1.83 areas between the standards and the samples. Curves of the ∗∗ Chlorogenic acid 286.13 ± 15.09 479.53 ± 16.24 standard compounds were made using serial dilutions of ∗∗ standards dissolved in HPLC-grade methanol. The separation Luteolin 44.56 ± 2.03 94.27 ± 7. 6 ∗∗ conditions of the phenolic compounds were taken from Rutin 603.35 ± 13.48 845.25 ± 18.83 apreviousstudy[35]. A representative chromatogram of Quercetin 46.18 ± 0.6 49.89 ± 6.98 ∗ a phenolic standard mixture under optimal conditions is Kaempferol 46.43 ± 2.14 67.36 ± 7. 8 6 ∗∗ presented in Figure 1, demonstrating that the peaks of the Apigenin 24.41 ± 2.16 8.74 ± 0.95 phenolic standards were separated with satisfactory efficiency The concentration is given in 𝜇g/g of the dry plant material for triplicate and resolution in 70 min of analysis time. The eluting peaks injections. were monitored with a diode array detector at 270 nm and Statistically significant differences between the means of both cultivars are ∗∗ ∗ 352 nm, and the spectra were recorded between 0 and 62 min. denoted as p < 0.001; p < 0.05. Detection was accomplished by using these two wavelengths to observe a wide range of phenolic classes: phenolic acids and isoflavonoids at 270 nm and flavonoids at 352 nm.A of San Pedro cultivar. The average mean of apigenin in San high degree of linearity was observed in all of the phenolic Pedro cultivar was about 3 times greater than that found in compounds (Table 3);thesquareofcorrelationcoefficient Lombardia cultivar. Overall, both the phenolic compounds for the calibration curve estimates the reproducibility of the and their concentrations are generally consistent with those method. All of the M. oleifera methanolic extracts from the found in M. oleifera leaves by other plant researchers [13, 36]. different varieties were injected under these conditions. The differences observed could be attributed to the influence Figure 2 presents a comparison between Lombardia of several factors, such as different climate, irrigation system, extract and San Pedro extract regarding the presence of and location of the cultivars. These phenolic compounds bioactive compounds by HPLC-DAD analysis. The results act as potent metal chelators and free-radical scavengers; show that their chromatographic separations were similar; therefore, our results indicate that the methanolic leaves however, the peak sizes were different, which determines extracts of M. oleifera growninMexicomaybeusedin the amount and proportion of each compound. Only seven treating diseases related to free-radical reactions. phenolic compounds were identified: gallic acid, chlorogenic acid, luteolin, rutin, quercetin, kaempferol, and apigenin. 4. Discussion The most abundant phenolic acid was chlorogenic acid, and rutin was the most abundant flavonoid in all analyzed M. oleifera is perhaps the most useful traditional medicinal samples of both cultivars. The quantitative determination plant found in several African and Asian countries. Anecdo- of these compounds is shown in Table 4.Aneffectof tal evidence of the benefits of M. oleifera has fueled a recent cultivation location on five phenolics content was observed increase in the exploitation and attention to its many healing for chlorogenic acid, luteolin, and apigenin, but not for benefits, specifically the high nutrient composition of the gallic acid and quercetin (Figure 2 and Table 4). The plant leaves and seeds. cultivated at Lombardia contained higher phenolics that Although various studies have been conducted on the those at San Pedro. In addition, Lombardia cultivar showed Moringa species in India, Africa, and Pakistan, gaps and larger quantities of chlorogenic acid and luteolin that those inconsistency between the information on the nutrient of this Journal of Chemistry 5

DAD1 D, sig. = 352, 16, ref. =500,100 (Agilent/ACGALICOSTD0441.D) A K L Q 80 R 60.622 59.819 60 46.931 C 56.057 48.261

(mAU) 40

20 15.756 57.962 51.922 65.526 65.736 2.884 0 2.683 10 20 30 40 50 60 (min)

Figure 1: Chromatogram of a mixture of standards’ polyphenols; DAD detection is at 352 nm. The chromatographic conditions are described in the text. Peak identification: C: chlorogenic acid; L: luteolin; R: rutin; Q: quercetin; K: kaempferol; A: apigenin.

DAD1 D, sig. = 352, 16, ref. =500,100 (Agilent/ACGALICOSTD0409.D)

San Pedro 48.058

100 49.283 51.658 80 53.373 60 49.663 39.003

(mAU) 40 6.703 46.421 51.069 47.291 53.672 52.001 44.311 14.900 56.277

20 54.181 57.805 58.358 55.918 43.469 60.506 60.797 55.369 42.476 5.856 62.608 54.566 0 2.647 10 20 30 40 50 60 (min)

DAD1 D, sig. = 352, 16, ref. =500,100 (Agilent/ACGALICOSTD0410.D)

140 Lombardia 48.077 120 100 49.316 80 60 (mAU) 6.660

40 46.424 49.682 38.863 51.668 44.284 52.010 14.670 51.078 53.359 54.162 56.227 57.786 55.884 60.500 60.777 53.695 20 43.415 58.301 62.579 0 2.653 10 20 30 40 50 60 (min)

Figure 2: Comparison of gradient HPLC analyses of two varieties of M. oleifera flavonols. Detection was performed at 352 nm. plant remain [28], and there are few experimental studies high ash content indicates that the leaves are a good source of regarding the potential nutritional value of the plant grown inorganic minerals. in Central and South America [23, 37, 38] and some reports The previous data on the characterization and quantita- on M. oleifera cultivated in Mexico [20, 21]. However, in tion of trace element studies present in M. oleifera leaves are recent years, the pharmaceutical properties associated with rather limited. Important studies in this area were presented M. oleifera have garnered increasing attention in the Mexican by [39], who characterized eleven different elements in South market,inpartbecausethedietarysupplementsarenatural India leaf samples, and by [29], who identified and quantified compounds. seven diverse elements in M. oleifera cultivated in Africa. The results of our study suggest that the concentration The most abundant macroelements found in our analysis of moisture, ash, fiber, lipid, and carbohydrates exhibited wereCa,Mg,andK.TheSanPedrocultivarinvestigatedin no changes in the proximate analysis because there was this study exhibited higher concentrations of Ca and Mg. In no statistical difference (at the level of 95%) from the two contrast, the K content was higher in the Lombardia cultivar. different M. oleifera cultivars, and their amounts were com- The concentrations of these macronutrients overlapped with parable to the concentration ranges found in the literature those reported for leaves of M. oleifera grown in the literature. [28]. However, numerous studies have shown that M. oleifera Furthermore, the concentration of Na in our cultivars was leaves offer high protein content; contrary to these findings, lower than the levels reported in several studies [40, 41]. our study did not find a high level of protein. This variation Therefore, the minerals found in M. oleifera play both a may be explained by several factors, such as climate and the curative and preventive role in combating human disease. geography of development of the crop. Despite this difference, For example, Ca is a multifunctional nutrient essential the proximate analysis shows that M. oleifera leaves are a good to the body metabolism [42], and Ca deficiency leads to source of proteins, lipids, and carbohydrates. In addition, the osteoporosis. Thus, M. oleifera is considered to be a natural 6 Journal of Chemistry cure for osteoporosis [43]. Furthermore, there is strong world. Various studies have found Pb at similar levels (<0.001 biological plausibility for the direct impact of Mg intake on to 2.6 𝜇g Pb/g) in medicinal plants in Italy, Egypt, and the cardiovascular disease prevention, sensitivity, and United States [58–60], whereas the highest concentration is diabetes [44]. found in used in Brazil [61]. A recent study The most abundant microelements found in this study reported lower Pb and As levels in M. oleifera leaves [62]. wereFe,Zn,Cu,andSe.InthePhilippines,M. oleifera is M. oleifera varieties in this study contain lower Pb level, known as “the mother’s best friend” due to its use to increase which further supports their use as food supplement and their nursing mothers’ milk production [45]. This effect can be medicinal benefits. attributed to Cu and Zn, which are essential in increasing Becauseitisassociatedwithseveralmetabolicdiseases therateofpregnantfemalemilkproduction[46]. Zn is also and age-related degenerative disorders are closely related to important in the healing of wounds and functions as an the body’s oxidative processes, the use of M. oleifera as a antioxidant. M. oleifera extract has shown significant wound- source of antioxidants to combat oxidation warrants further healing activity against excision, restored incision, and dead- attention. Some authors reported that methanolic extract space wounds. This effect could be due to M. oleifera high of M. oleifera leaves had a high antioxidant activity, which zinc content [47]. Fe has several essential functions in the may be attributed to the presence of polyphenolics and body,suchasitsrolesinoxygentransportandoxidative other antioxidant substances. In addition, the data obtained metabolism [48]. Se is an essential element in both animal emphasize the free-radical scavenging effect of an aqueous and human nutrition. In addition, the antitumorigenic effects extract of M. oleifera over DPPH free radical, superoxide, and of Se compounds have been described in a variety of in nitric oxide radicals and the inhibition of lipid peroxidation vitro and animal models, suggesting that supplemental Se [63, 64]. in human diets may reduce the risk of cancer [49]. The To better understand the association of flavonoids intake anticarcinogenic activity of M. oleifera was described [50], and health outcomes, analyses of flavonoids in plant foods, and their results suggest that the minerals present in M. an intense area of research, and clinical and epidemiological oleifera leaves may contribute to its therapeutic properties. studies are required [65, 66]. A phytochemical analysis The use of herbal medicines is increasing in both devel- was performed to determine the major class of compounds oping and developed countries due to their reasonable prices present in the leaf extracts. The quantitative estimations of and, in particular, to the assumption that natural products are the polyphenols have shown that the Lombardia samples safe [51]. However, most herbal products are not validated contained a higher amount of chlorogenic acid (479.53 ± according to the recommended pharmaceutical guidelines 16.24 𝜇g/g), rutin (845.25 ± 18.83 𝜇g/g), and luteolin (94.27 ± and often contain toxic and lethal concentrations of toxic 7. 6 𝜇g/g), whereas the San Pedro samples had higher amounts heavy metals [52]. of apigenin (24.41 ± 2.16 𝜇g/g). Our results are consistent Arsenicisametalloidthatactsoncellsthroughavariety with those from previous studies, which strongly suggest of mechanisms, influencing numerous signal transduction that M. oleifera may be an important source of natural pathways and resulting in cellular effects such as apoptosis antioxidants [67, 68]. Previous phytochemical investigations induction, growth inhibition, and angiogenesis inhibition have identified five flavonols in M. oleifera, including gal- [53]. Pb is a metallic element that emanates into the envi- lic acid, chlorogenic acid, rutin, ellagic acid, ferulic acid, ronment from various sources, including the industrial waste, quercetin, and kaempferol [13, 36]; however, only quercetin, combustion of fossil fuels, and the use of agrochemicals. kaempferol, gallic acid, and chlorogenic acid were found Higher levels of Pb cause a variety of acute and chronic health in all the studies. However, our values of quercetin and problems, including cancer, kidney damage, heart problems, kaempferol are out of the range of those reported by different and even death [54]. authors for methanolic extract of M. oleifera leaves [13, 68– San Pedro is part of the region known as the “Comarca 70];thesetwocompoundshavebeenreportedasthemost Lagunera.”ItislocatedintheCoahuilaandDurangostatesin abundant in the leaves of M. oleifera from India, Pakistan, Mexico, a region in which the aquifers are severely contami- and African nations. It has been reported that the leaves nated with As [55]. High levels of As have been detected in the of this plant have very low levels of luteolin (6.2 𝜇g/g) [71]. groundwaterthatisusedasbothdrinkingwaterforhumans This value corresponds to 15 times less than the obtained and dairy cattle and for agricultural irrigation [56]. in this study (see Table 4). A possible explanation for these Plants vary widely in their tolerance to toxic metals. results is that the flavonols are the most widespread of M. oleifera from San Pedro can grow normally while accu- the flavonoids in plant food and, unlike flavonols, apigenin mulating arsenic in its roots (data not shown). Among the andluteolinarenotwidelydistributedwithsignificantcon- M. oleifera samples considered in this study, the lowest As centrations. In addition, the extraction yield of antioxidant content was observed in the sample grown in Lombardia compoundsfromplantmaterialsisinfluencedprimarily (0.0055 mg/100 g), whereas the As content in the samples by the conditions under which the process of liquid-solid collected from San Pedro was much higher (0.28 mg/100 g), extraction is achieved, the type of solvent used to separate the showing obvious signs of environmental contamination. soluble fraction from the permeable solid, and the degree of The suggested concentration of Pb in plant species is 2 polymerization of the phenolics and their interactions with to 6 mg/L [57]. The concentration of Pb was minimum at the other components [72, 73]. 0.2–0.35 mg/100 g in our samples. These results were similar As mentioned earlier, the abundance and diversity of tothosedescribedforplantsgrowninotherpartsofthe flavonoids present in M. oleifera may be responsible for their Journal of Chemistry 7 therapeutic effectiveness against various diseases11 [ ]. Though some ICP-MS analyses. The authors are also grateful to M. oleifera isknowntocontainquercetinandkaempferol, the Mexican Moringa producers for the supply of samples traceable amounts of chlorogenic acid and derivatives have used in this study. A special acknowledgement is due to been detected within the leaves from Ghana, Senegal, and Jose Ruvalcaba Quinones˜ for his technical assistance in the Zambia [74]. Chlorogenic acid and its isomers are esters of course of this research. Financial support was obtained from quinic and caffeic acids that have abilities to inhibit oxidation promeP-SEP (IDCA-9365), Mexico. and also promote various pharmacological activities such as antiobesity, reduction of plasma and liver lipids, and References inhibition of acute lung injury [75–77]. On the other hand, rutin is present in substantial amounts in our M. oleifera [1] M. Ekor, “The growing use of herbal medicines: issues relating leaves and some investigations showed that this compound to adverse reactions and challenges in monitoring safety,” has a broad range of physiological activities [78]. Frontiers in Pharmacology,vol.4,pp.1–10,2014. It is interesting to mention that when flavones are [2] S. Demiray, M. E. Pintado, and P. M. L. 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The use of 2004. dried leaf powder is recommended to achieve the health ben- [5]F.Anwar,M.Ashraf,andM.I.Bhanger,“Interprovenance variation in the composition of Moringa oleifera oilseeds from efits of the additive and synergistic effects of the constituents Pakistan,” Journal of the American Oil Chemists’ Society,vol.82, present in the whole leaves. Obviously, it is not yet clear how no. 1, pp. 45–51, 2005. polyphenolic compounds and minerals are associated with [6] A. O. Oluduro, “Evaluation of antimicrobial properties and the reduction of human diseases. Therefore, it is necessary to nutritional potentials of Moringa oleifera Lam. leaf in South- conduct further studies on their in vivo activity, bioavailabil- Western Nigeria,” Malaysian Journal of Microbiology,vol.8,no. ity, and toxicity. Actually, we are conducting studies on the 2, pp. 59–67, 2012. therapeutic properties of our M. oleifera leaves extracts. [7] R. Dhakar, B. Pooniya, M. 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