Research Article Nutritional Content and Elemental and ...

Research ArticleNutritional Content and Elemental and PhytochemicalAnalyses of Moringa oleifera Grown in Mexico

Moacutenica A Valdez-Solana1 Veroacutenica Y Mejiacutea-Garciacutea1 Alfredo Teacutellez-Valencia2

Guadalupe Garciacutea-Arenas3 Joseacute Salas-Pacheco4 Joseacute J Alba-Romero1

and Erick Sierra-Campos1

1Facultad de Ciencias Quımicas Universidad Juarez del Estado de Durango Avenida Artıculo 123 SN Fracc Filadelfia35010 Gomez Palacio DGO Mexico2Facultad de Medicina y Nutricion Universidad Juarez del Estado de Durango Avenida Universidad y Fanny Anitua SN34000 Durango DGO Mexico3Facultad de Medicina Universidad Juarez del Estado de Durango Calzada Palmas 1 Colonia Revolucion35050 Gomez Palacio DGO Mexico4Instituto de Investigacion Cientıfica Universidad Juarez del Estado de Durango Avenida Universidad SN34000 Durango DGO Mexico

Correspondence should be addressed to Erick Sierra-Campos ericksiergmailcom

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

Academic Editor Deborah Pacetti

Copyright copy 2015 Monica A Valdez-Solana et al This is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

Moringa oleifera is a tree distributed in Mexican semiarid and coastal regions M oleifera is used in practice in the treatment ofvarious diseases and is available without a medical prescription often in the form of an herbal infusion for everyday use The aimof the present study was to evaluate the chemical composition and nutritional values of dried M oleifera leaf powder collectedfrom two different regions in Mexico All samples ofM oleifera exhibited moisture levels varying from 306 to 334 lipids from1021 to 1031 fiber from 729 to 946 ashes from 1071 to 1118 crude protein from 1074 to 1148 and carbohydrates from5461 to 5761 The predominant mineral elements in the leaf powder according to ICP-MS were Ca (20165ndash26205mg100 g)K (1817ndash1845mg100 g) and Mg (3225ndash3406mg100 g) The HPLC analysis indicated the presence of phenolic acids (gallic andchlorogenic acids) and flavonoids (rutin luteolin quercetin apigenin and kaempferol) We concluded that LombardiaM oleiferasamples could be employed in edible and commercial applications Our results showed that the highest mean value of As from theSan Pedro samples exceeds the recommended level and may constitute a health hazard to consumers

1 Introduction

Plants have been an important source of medicine for thou-sands of years Even today the World Health Organization(WHO) estimates that up to 80 of people still rely primarilyon traditional remedies such as herbs for their medicines [1]The medicinal value of these plants is due to the presence ofa variety of phytochemicals and their elemental compositionThe role of medicinal plants in disease prevention or control

has been attributed to the antioxidant properties of their con-stituents usually associated with a wide range of amphipathicmolecules that are broadly referred to as polyphenolic com-pounds [2] There is a growing interest in the developmentand evaluation of natural antioxidants from medicinal plantmaterials in the food industry and the field of preventivehealth care Among those herbs one promising species isMoringa oleifera Lam (Moringa or drumstick tree) whichis native to the sub-Himalayan regions of Northwest India

Hindawi Publishing CorporationJournal of ChemistryVolume 2015 Article ID 860381 9 pageshttpdxdoiorg1011552015860381

brought to you by COREView metadata citation and similar papers at coreacuk

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2 Journal of Chemistry

It is widely distributed throughout Africa Saudi ArabiaSoutheast Asia the Caribbean Islands and South AmericaEvery part of M oleifera has medicinal properties and iscommercially exploitable for the development of medicinaland industrial byproducts [3]

Traditionally the leaves fruits flowers and immaturepods of this tree are edible they are used as a highly nutritivevegetable in many countries particularly in India Pakistanthe Philippines Hawaii and some African nations [4ndash6] Indeveloping nations M oleifera is used as an alternative toimported food supplements to treat and combat malnutri-tion especially among infants and nursingmothers by virtueof its chemical constituents [7]

Several valuable reviews of the ethnobotanical usesof M oleifera are available [8ndash11] Moringa has beenfound to be a good source of polyphenols and antiox-idants [11] Phytochemicals such as vanillin omega fattyacids carotenoids ascorbates tocopherols beta-sitosterolmoringine kaempferol and quercetin have been reported inits flowers roots fruits and seeds The leaves in particularhave been found to contain phenolics and flavonoids [12 13]these compounds have various biological activities includingantioxidant anticarcinogenic immunomodulatory antidia-betic antiatherogenic and hepatoprotective functions andthe regulation of thyroid status [14ndash16] Moreover leavescontain trace elements that are essential to human health Forinstancemagnesium iron selenium and zinc play an impor-tant role in metabolism and interest in these elements isincreasing together with reports relating trace element statusand oxidative diseases [17 18] However a recent study hasshown that dried M oleifera leaves contain lead at very highvalues of 3520mgL [19] Therefore it is very important toidentify the mineral composition ofM oleifera leaves that arewidely consumed by humans and animals

In Mexico M oleifera is widely cultivated in differentzones of the country and is found inmore than ten states fromSonora to Oaxaca on the Pacific side Few studies have beenconducted on nutritional and phytochemical composition[20 21] however to date a detailed composition of the leavesof M oleifera that is native to Mexico has not been reportedyet In addition it is important to bear in mind that themineral and phenolics contents present in leaves depend onseveral factors such as geographical area where the plant iscultivated type of soil water and fertilizers industrializationprocess and storage conditions Taking these precedents intoconsideration the aim of this study was to evaluate thephytochemical constituents of methanolic extracts and traceelements and nutritional values present in M oleifera leavesgrown in the northeast and west regions of Mexico

2 Materials and Methods

21 Reagents and Standards The methanol and acetonitrileused in this study were of HPLC grade and were purchasedfrom JT Baker USA andCaledon Lab Canada HPLC-gradephenolic standards (gallic and chlorogenic acids rutin lute-olin quercetin apigenin and kaempferol) were of analyticalreagent grade and were purchased from Sigma-Aldrich (StLouis MO USA) Demineralized water was taken from a

Milli-Q Plus water purification system (Millipore BedfordUSA) The multielement standard mixture solutions con-tained Ba Bi Li Cu Co In Li Ni Pb and V the otherscontained Ba CaMg and Sr Analytical grade 65wv nitricacid (HNO

3) (JT Baker) was used in the decomposition

22 Samples Preparation M oleifera leaves were collectedfrom Mexican cultivars (Lombardia Michoacan locatedon 19∘0110158403010158401015840N and 102∘0510158403910158401015840W and San Pedro Coahuilalocated on 25∘4510158403510158401015840N and 102∘5810158405410158401015840WMexico April 2011)and a voucher of the specimen was deposited in the CIIDIR-IPN herbarium Durango Mexico for future references Theplants were identified and authenticated by M Sc AlbertoGonzalez Zamora Faculty of Biology UJEDThe leaves werecleaned and processed as described by previous researchers[22 23] A set of five samples from each geographical originwere randomly collected and analyzed All materials were air-dried and powdered and protected from light until furtheranalysis

23 Chemical Analysis Representative subsamples weredried in a forced drought oven at 105ndash110∘C to a constantweight for moisture determination Crude protein crudelipid crude fiber and ash were analyzed by triplicate accord-ing to AOAC [24] The total carbohydrates were determinedby the difference method [100 minus (proteins + fats + moisture+ ash in percentage)] [25 26] The energy values (kcal100 g)were determined by multiplying the values of carbohydrateslipids and proteins by factors of 4 9 and 4 respectively andtaking the sum expressed in kilocalories [27]

24 Extraction of Phenolic Compounds The extracts wereprepared using 23 g of dry-ground sample and 260mL of 80methanolic aqueous solution by successive maceration Themixtures were shaken in amagnetic grid at room temperaturefor 21 h They were then filtered through Whatman filterpaper number 1 The final extract was concentrated on arotator evaporator and the filtrate was placed in a deepfreezer for 24 h and lyophilized to obtain a powdered extractAll of the extractions were performed in triplicate and thesupernatants were kept at minus40∘C until further analysis

25 ICP-MS Analysis The instrumentation used for diges-tion consisted of a closed microwave oven (MARS-XpressCEM)Themultielemental analysis was performed by induc-tively coupled plasma mass spectrometry (ICP-MS) with areaction cell (XSeries 2 Thermo Scientific) Prior to analysisthe samples were homogenized in a mortar approximately05 g of the sample was accurately weighed in an acid-washedpolytetrafluoroethylene (PTFE) digestion tube and 5mL ofHNO

3(65wv) was added The tube was heated in a

microwave oven at a power setting of 50 and 150∘C for10min 70 and 220∘C for 20min and 10 and 100∘C for15min The digest was transferred into a 50mL acid-washedvolumetric flask which was filled with demineralized waterand stored in a polypropylene container Onewater blankwasrun with each batch of samples

Journal of Chemistry 3

26 HPLC-DAD Analysis Seven phenolic standards andsamples were dissolved in the mobile phase yielding con-centrations of 25120583gmL The solutions were filtered througha 045120583m membrane filter and the evaluation of eachcalibration curve was fitted by linear regression An Agilent1200HPLCSeries (Agilent PaloAlto CAUSA)was operatedat 30∘C equipped with a degasser a diode array detector(DAD) a quaternary gradient pump and an Eternity C18reversed-phase analytical column of 150mm times 46mm anda 35 120583m particle (Sigma-Aldrich USA) The mobile phasewas a binary gradient methanol and buffer solution Thebuffer contained TCA 01 solution in water and its pH wasadjusted to 21 The linear gradient began with 20 to 50methanol over the first 30min followed by 70 methanolover the next 30minThe flow rate was 06mLmin and datawere collected at 270 and 352 nm The injection volume was20120583L Data acquisition peak integration and calibrationswere performed with Agilent 1200 Chemstation softwareThe results are reported as means plusmn standard deviations oftriplicate independent analyses

27 Statistical Analysis The data were expressed as means plusmnSD and the results were statistically analyzed using a Holm-Sidak test and one-way analysis of variance (ANOVA) on aSigmaPlot program v 110 forWindowsThe differences wereconsidered statistically significant if 119901 lt 005

3 Results

31 Chemical Analysis The proximate and nutrient analysesof M oleifera play a crucial role in assessing its nutritionalsignificance (119901 lt 005) The proximate analysis for bothcultivars is presented in Table 1 along with the values foundin the literature for identical M oleifera for comparativepurposes Nonsignificant differences were observed betweenthe proximate analyses of the samples studied in Lombardiaand San Pedro The mean moisture lipid fiber and ashvalues found in the present study are in agreement withthe values reported [28] The result shows that Mexicancultivars contain crude protein (106) crude fiber (about8) ash (about 11) carbohydrates (about 56) moisture(about 32) and lipid (about 102) Carbohydrates are theprincipal sources of energy The ash content of about 11indicates that the leaves are rich in mineral elements Themean protein content found in the Mexican samples rangedfrom 1074 to 1148 This is a lower protein level than the272 reported by other authors [21 29ndash31] though it maystill be valuable as a protein sourceThe variation in chemicalanalyses between our cultivars and the reference may dependon seasonal variations the plantsrsquo stages of developmentand the techniques employed to collect leaf samples beforethe experimental analysis The chemical composition valuesconfirmed thatM oleifera leaves are an excellent food sourcejustifying its direct use in human nutrition or development ofbalanced diets for animal nutrition

32 Elemental Analysis Different soils contain a particular ofmineral elements qualities and quantities whose bioavailabil-ity depends on soil properties (pH clay and humid complex

Table 1 Nutritional composition of driedM oleifera leaf powder

Parameter Lombardia San Pedro [28]Moisture () 334 plusmn 136 306 plusmn 138 74 plusmn 289Lipid () 1031 plusmn 12 1021 plusmn 183 6 plusmn 25Fiber () 729 plusmn 084 946 plusmn 114 9 plusmn 745Ash () 1071 plusmn 081 1118 plusmn 019 176b

Protein () 1074 plusmn 13 1148 plusmn 14 24 plusmn 58lowast

Carbohydrates ()a 5761 plusmn 219 5461 plusmn 06 36 plusmn 92lowast

Energy value(Kcal100 g) 3662 plusmn 423 35625 plusmn 648 304 plusmn 87

Values are mean plusmn SD analyzed individually in triplicate and are expressedas g100 g leaf powdera bCalculated by differencelowastDifference significative between Mexican samples against a reference 119901 lt0018

Table 2 Measured concentration in (mg100 g) of trace element intheM oleifera leaf powder

Elements Lombardia San Pedro [28]Calcium (Ca) 20165 plusmn 226 26205 plusmn 56lowastlowast 1897 plusmn 7484Magnesium (Mg) 3225 plusmn 00 3406 plusmn 28lowastlowast 473 plusmn 4294Potassium (K) 1845 plusmn 70 1817 plusmn 141 1467 plusmn 6367Sodium (Na) 813 plusmn 06 4078 plusmn 07lowastlowast 220 plusmn 180Iron (Fe) 1937 plusmn 66 707 plusmn 04lowast 325 plusmn 1078Zinc (Zn) 10 plusmn 07 16 plusmn 06 24 plusmn 112Copper (Cu) 103 plusmn 047 041 plusmn 00 09 plusmn 048Selenium (Se) 955 times 10minus3 plusmn 00 0107 plusmn 00lowastlowast NDLead (Pb) 0355 plusmn 00 02 plusmn 00lowastlowast NDArsenic (As) 55 times 10minus3 plusmn 00 028 plusmn 00lowastlowast NDResults are mean plusmn SD analyzed individually in triplicateND nondeterminedStatistically significant differences between the means of both cultivars aredenoted as lowastlowastp lt 0001 lowastp lt 005

and mineralogy) [32] Ten elements including toxic onessuch as lead and arsenic were determined to be present intwo crops of M oleifera collected from Lombardia and SanPedro locations The elements were determined by ICP-MSafter wet digestion of the dried sample with concentratednitric acid in closed PTFE vessels using a microwave ovenCa Mg K Fe and Na were present at levels of mg100 gdry matter whereas Zn Cu and Se were present at 120583g100 glevels in close agreementwith levels previously reported [30]and these values fall within the ranges reported (Table 2reference column) The elemental analysis of our samplesrevealed high contents of Ca (2016 to 2620mg100 g) Mg(322 to 3406mg100 g) and K (1817 to 1845mg100 g) whileZn Cu and Se were 1 1 and 01mg100 g respectively Ironwas abundant in Lombardia cultivar whereas the Ca Mgand Na levels were higher in San Pedro cultivar (Table 2)These values of M oleifera leaves from Mexico agree withthose found in Burkina Faso and India [29 30]The Fe variedamong the leaf samples collected from Lombardia and SanPedro ranging from 1937 to 707 respectively However thislevel is low when compared to the contents of some results


from India [30] The Pb level in Lombardia and San Pedrocultivars was below the limit set by the WHO acceptabledaily intake of Pb for adults of 021ndash025mgday [33] Thelowest As content was observed in the Lombardia cultivar(00055mg100 g) whereas the content of As in the samplescollected from San Pedro was the highest (028mg100 g)which correlates with the extremely high As level in SanPedro soil Because the levels in the Lombardia samplewere still within the prescribed limit for food at 1mgkgaccording to theWHOFAO standard [34] this sample couldbe recommended as a source of essential elements Howeverbecause the As levels in San Pedro samples were found tobe higher than the limit set by the WHO it is necessary tostudy them in further detail to avoid overconsumption andcumulative toxicity from long-term use

33 Quantification of Phenolics RP-HPLC with DAD detec-tion was employed for the identification and quantificationof main phenolic compounds present in the methanolicextracts The quantifications of phenolics (mgg dry matter)were accomplished by comparing retention times and peakareas between the standards and the samples Curves of thestandard compounds were made using serial dilutions ofstandards dissolved inHPLC-grademethanolThe separationconditions of the phenolic compounds were taken froma previous study [35] A representative chromatogram ofa phenolic standard mixture under optimal conditions ispresented in Figure 1 demonstrating that the peaks of thephenolic standardswere separatedwith satisfactory efficiencyand resolution in 70min of analysis time The eluting peakswere monitored with a diode array detector at 270 nm and352 nm and the spectra were recorded between 0 and 62minDetection was accomplished by using these two wavelengthsto observe a wide range of phenolic classes phenolic acidsand isoflavonoids at 270 nm and flavonoids at 352 nm Ahigh degree of linearity was observed in all of the phenoliccompounds (Table 3) the square of correlation coefficientfor the calibration curve estimates the reproducibility of themethod All of the M oleifera methanolic extracts from thedifferent varieties were injected under these conditions

Figure 2 presents a comparison between Lombardiaextract and San Pedro extract regarding the presence ofbioactive compounds by HPLC-DAD analysis The resultsshow that their chromatographic separations were similarhowever the peak sizes were different which determinesthe amount and proportion of each compound Only sevenphenolic compounds were identified gallic acid chlorogenicacid luteolin rutin quercetin kaempferol and apigeninThe most abundant phenolic acid was chlorogenic acidand rutin was the most abundant flavonoid in all analyzedsamples of both cultivars The quantitative determinationof these compounds is shown in Table 4 An effect ofcultivation location on five phenolics content was observedfor chlorogenic acid luteolin and apigenin but not forgallic acid and quercetin (Figure 2 and Table 4) The plantcultivated at Lombardia contained higher phenolics thatthose at San Pedro In addition Lombardia cultivar showedlarger quantities of chlorogenic acid and luteolin that those

Table 3 Elution order of polyphenolic compounds separated byHPLC-DAD

Referencecompound

Retentiontime (min)

Linear range(120583gmL) 119903

2

270 nmGallic acid 453 plusmn 005 10ndash99 09999

352 nmChlorogenicacid 1552 plusmn 038 06ndash231 09995

Luteolin 4689 plusmn 029 05ndash40 09993Rutin 4825 plusmn 025 99ndash495 09996Quercetin 5607 plusmn 021 099ndash30 09999Kaempferol 5979 plusmn 081 066ndash33 09998Apigenin 6059 plusmn 015 01ndash10 09995

Each value is the mean plusmn SD of triplicate determinations

Table 4 Characterization (120583gg dry matter) of polyphenolic com-pounds separated by HPLC-DAD

Compound San Pedro LombardiaGallic acid 4907 plusmn 453 4328 plusmn 183Chlorogenic acid 28613 plusmn 1509 47953 plusmn 1624lowastlowast

Luteolin 4456 plusmn 203 9427 plusmn 76lowastlowast

Rutin 60335 plusmn 1348 84525 plusmn 1883lowastlowast

Quercetin 4618 plusmn 06 4989 plusmn 698Kaempferol 4643 plusmn 214 6736 plusmn 786lowast

Apigenin 2441 plusmn 216 874 plusmn 095lowastlowast

The concentration is given in 120583gg of the dry plant material for triplicateinjectionsStatistically significant differences between the means of both cultivars aredenoted as lowastlowastp lt 0001 lowastp lt 005

of San Pedro cultivar The average mean of apigenin in SanPedro cultivar was about 3 times greater than that found inLombardia cultivar Overall both the phenolic compoundsand their concentrations are generally consistent with thosefound inM oleifera leaves by other plant researchers [13 36]The differences observed could be attributed to the influenceof several factors such as different climate irrigation systemand location of the cultivars These phenolic compoundsact as potent metal chelators and free-radical scavengerstherefore our results indicate that the methanolic leavesextracts of M oleifera grown in Mexico may be used intreating diseases related to free-radical reactions

4 Discussion

M oleifera is perhaps the most useful traditional medicinalplant found in several African and Asian countries Anecdo-tal evidence of the benefits of M oleifera has fueled a recentincrease in the exploitation and attention to its many healingbenefits specifically the high nutrient composition of theleaves and seeds

Although various studies have been conducted on theMoringa species in India Africa and Pakistan gaps andinconsistency between the information on the nutrient of this


(min)10 20 30 40 50 60

(mAU

)

020406080

268

32

884

157

56

469

3148

261

519

22

560

5757

962

598

19 606

22

655

2665

736

C

LR

QK

ADAD1 D sig = 352 16 ref = 500100 (AgilentACGALICOSTD0441D)

Figure 1 Chromatogram of a mixture of standardsrsquo polyphenols DAD detection is at 352 nmThe chromatographic conditions are describedin the text Peak identification C chlorogenic acid L luteolin R rutin Q quercetin K kaempferol A apigenin

10 20 30 40 50 60

020406080

100

264

7

585

6 670

3

149

00 390

03

424

7643

469

443

11

464

2147

291

480

5849

283

516

5849

663

533

7352

001

541

8153

672

626

08510

69

607

9760

506

553

6955

918

562

7757

805

583

58

545

66

(min)

(mAU

)

10 20 30 40 50 60

020406080

100120140

(min)

(mAU

)

146

70666

0

265

3 388

63

434

1544

284

464

2448

077

493

1649

682

625

7960

777

605

00

577

8658

301

558

8456

227

533

5954

162

536

95

510

7851

668

520

10

San Pedro

Lombardia

DAD1 D sig = 352 16 ref = 500100 (AgilentACGALICOSTD0409D)


Figure 2 Comparison of gradient HPLC analyses of two varieties ofM oleifera flavonols Detection was performed at 352 nm

plant remain [28] and there are few experimental studiesregarding the potential nutritional value of the plant grownin Central and South America [23 37 38] and some reportson M oleifera cultivated in Mexico [20 21] However inrecent years the pharmaceutical properties associated withM oleifera have garnered increasing attention in theMexicanmarket in part because the dietary supplements are naturalcompounds

The results of our study suggest that the concentrationof moisture ash fiber lipid and carbohydrates exhibitedno changes in the proximate analysis because there wasno statistical difference (at the level of 95) from the twodifferent M oleifera cultivars and their amounts were com-parable to the concentration ranges found in the literature[28] However numerous studies have shown thatM oleiferaleaves offer high protein content contrary to these findingsour study did not find a high level of protein This variationmay be explained by several factors such as climate and thegeography of development of the cropDespite this differencethe proximate analysis shows thatM oleifera leaves are a goodsource of proteins lipids and carbohydrates In addition the

high ash content indicates that the leaves are a good source ofinorganic minerals

The previous data on the characterization and quantita-tion of trace element studies present inM oleifera leaves arerather limited Important studies in this area were presentedby [39] who characterized eleven different elements in SouthIndia leaf samples and by [29] who identified and quantifiedseven diverse elements inM oleifera cultivated in Africa

The most abundant macroelements found in our analysiswere Ca Mg and K The San Pedro cultivar investigated inthis study exhibited higher concentrations of Ca and Mg Incontrast the K content was higher in the Lombardia cultivarThe concentrations of these macronutrients overlapped withthose reported for leaves ofM oleifera grown in the literatureFurthermore the concentration of Na in our cultivars waslower than the levels reported in several studies [40 41]Therefore the minerals found in M oleifera play both acurative and preventive role in combating human diseaseFor example Ca is a multifunctional nutrient essentialto the body metabolism [42] and Ca deficiency leads toosteoporosis Thus M oleifera is considered to be a natural


cure for osteoporosis [43] Furthermore there is strongbiological plausibility for the direct impact of Mg intake oncardiovascular disease prevention insulin sensitivity anddiabetes [44]

The most abundant microelements found in this studywere Fe Zn Cu and Se In the Philippines M oleifera isknown as ldquothe motherrsquos best friendrdquo due to its use to increasenursing mothersrsquo milk production [45] This effect can beattributed to Cu and Zn which are essential in increasingthe rate of pregnant female milk production [46] Zn is alsoimportant in the healing of wounds and functions as anantioxidantM oleifera extract has shown significant wound-healing activity against excision restored incision and dead-space wounds This effect could be due to M oleifera highzinc content [47] Fe has several essential functions in thebody such as its roles in oxygen transport and oxidativemetabolism [48] Se is an essential element in both animaland human nutrition In addition the antitumorigenic effectsof Se compounds have been described in a variety of invitro and animal models suggesting that supplemental Sein human diets may reduce the risk of cancer [49] Theanticarcinogenic activity of M oleifera was described [50]and their results suggest that the minerals present in Moleifera leaves may contribute to its therapeutic properties

The use of herbal medicines is increasing in both devel-oping and developed countries due to their reasonable pricesand in particular to the assumption that natural products aresafe [51] However most herbal products are not validatedaccording to the recommended pharmaceutical guidelinesand often contain toxic and lethal concentrations of toxicheavy metals [52]

Arsenic is a metalloid that acts on cells through a varietyof mechanisms influencing numerous signal transductionpathways and resulting in cellular effects such as apoptosisinduction growth inhibition and angiogenesis inhibition[53] Pb is a metallic element that emanates into the envi-ronment fromvarious sources including the industrial wastecombustion of fossil fuels and the use of agrochemicalsHigher levels of Pb cause a variety of acute and chronic healthproblems including cancer kidney damage heart problemsand even death [54]

San Pedro is part of the region known as the ldquoComarcaLagunerardquo It is located in the Coahuila and Durango states inMexico a region in which the aquifers are severely contami-natedwithAs [55] High levels of As have been detected in thegroundwater that is used as both drinking water for humansand dairy cattle and for agricultural irrigation [56]

Plants vary widely in their tolerance to toxic metalsM oleifera from San Pedro can grow normally while accu-mulating arsenic in its roots (data not shown) Among theM oleifera samples considered in this study the lowest Ascontent was observed in the sample grown in Lombardia(00055mg100 g) whereas the As content in the samplescollected from San Pedro was much higher (028mg100 g)showing obvious signs of environmental contamination

The suggested concentration of Pb in plant species is 2to 6mgL [57] The concentration of Pb was minimum at02ndash035mg100 g in our samples These results were similarto those described for plants grown in other parts of the

world Various studies have found Pb at similar levels (lt0001to 26 120583g Pbg) in medicinal plants in Italy Egypt and theUnited States [58ndash60] whereas the highest concentration isfound in herbal medicine used in Brazil [61] A recent studyreported lower Pb and As levels in M oleifera leaves [62]M oleifera varieties in this study contain lower Pb levelwhich further supports their use as food supplement and theirmedicinal benefits

Because it is associated with several metabolic diseasesand age-related degenerative disorders are closely related tothe bodyrsquos oxidative processes the use of M oleifera as asource of antioxidants to combat oxidation warrants furtherattention Some authors reported that methanolic extractof M oleifera leaves had a high antioxidant activity whichmay be attributed to the presence of polyphenolics andother antioxidant substances In addition the data obtainedemphasize the free-radical scavenging effect of an aqueousextract ofM oleifera over DPPH free radical superoxide andnitric oxide radicals and the inhibition of lipid peroxidation[63 64]

To better understand the association of flavonoids intakeand health outcomes analyses of flavonoids in plant foodsan intense area of research and clinical and epidemiologicalstudies are required [65 66] A phytochemical analysiswas performed to determine the major class of compoundspresent in the leaf extracts The quantitative estimations ofthe polyphenols have shown that the Lombardia samplescontained a higher amount of chlorogenic acid (47953 plusmn1624 120583gg) rutin (84525 plusmn 1883 120583gg) and luteolin (9427 plusmn76 120583gg) whereas the San Pedro samples had higher amountsof apigenin (2441 plusmn 216 120583gg) Our results are consistentwith those from previous studies which strongly suggestthat M oleifera may be an important source of naturalantioxidants [67 68] Previous phytochemical investigationshave identified five flavonols in M oleifera including gal-lic acid chlorogenic acid rutin ellagic acid ferulic acidquercetin and kaempferol [13 36] however only quercetinkaempferol gallic acid and chlorogenic acid were foundin all the studies However our values of quercetin andkaempferol are out of the range of those reported by differentauthors for methanolic extract of M oleifera leaves [13 68ndash70] these two compounds have been reported as the mostabundant in the leaves of M oleifera from India Pakistanand African nations It has been reported that the leavesof this plant have very low levels of luteolin (62120583gg) [71]This value corresponds to 15 times less than the obtainedin this study (see Table 4) A possible explanation for theseresults is that the flavonols are the most widespread ofthe flavonoids in plant food and unlike flavonols apigeninand luteolin are not widely distributed with significant con-centrations In addition the extraction yield of antioxidantcompounds from plant materials is influenced primarilyby the conditions under which the process of liquid-solidextraction is achieved the type of solvent used to separate thesoluble fraction from the permeable solid and the degree ofpolymerization of the phenolics and their interactions withthe other components [72 73]

As mentioned earlier the abundance and diversity offlavonoids present inM oleiferamay be responsible for their


therapeutic effectiveness against various diseases [11]ThoughM oleifera is known to contain quercetin and kaempferoltraceable amounts of chlorogenic acid and derivatives havebeen detected within the leaves from Ghana Senegal andZambia [74] Chlorogenic acid and its isomers are esters ofquinic and caffeic acids that have abilities to inhibit oxidationand also promote various pharmacological activities suchas antiobesity reduction of plasma and liver lipids andinhibition of acute lung injury [75ndash77] On the other handrutin is present in substantial amounts in our M oleiferaleaves and some investigations showed that this compoundhas a broad range of physiological activities [78]

It is interesting to mention that when flavones aremethoxylated metabolic stability and membrane transportin the intestineliver dramatically increase thus improvingoral bioavailability In addition methoxyflavones also showincreased cancer chemopreventive properties when com-pared to more common unmethylated flavones [79]

Although the bioactivity of the individual compoundsmay be known their effects on health may not be assignificant as the combination of the entire class of bioactivesworking through multiple mechanisms of action The use ofdried leaf powder is recommended to achieve the health ben-efits of the additive and synergistic effects of the constituentspresent in the whole leaves Obviously it is not yet clear howpolyphenolic compounds and minerals are associated withthe reduction of human diseases Therefore it is necessary toconduct further studies on their in vivo activity bioavailabil-ity and toxicity Actually we are conducting studies on thetherapeutic properties of ourM oleifera leaves extracts

5 Conclusion

Our results showed that the Mexican species of M oleiferahave nutritional potential because their leaves contain a highconcentration of energy nutrients minerals and phenolicconstituents mainly flavonoids and phenolic acids whichrepresent a good source of natural antioxidants Thereforethe therapeutic potential of M oleifera may be due to thepresence of these constituents The presence of harmfulelements such as arsenic and lead in Lombardia M oleiferaleaf powder appears to be within permissible limits butwe discovered that a maximum permissible level of As isexceeded in San Pedro samples Our results suggest thatquality assurance and monitoring of toxic metals are neededfor plants intended for human consumption

Conflict of Interests

The authors declare no conflict of interests

Acknowledgments

The authors thank Angel Trejo for providing us with amicrowave oven They thank Jorge Humberto Marın for histechnical assistanceThey are grateful to Berenice HernandezRamos Coordinator of the Chemical Analysis Laboratory ofCIATEC Guanajuato Mexico who enabled them to conduct

some ICP-MS analyses The authors are also grateful tothe Mexican Moringa producers for the supply of samplesused in this study A special acknowledgement is due toJose Ruvalcaba Quinones for his technical assistance in thecourse of this research Financial support was obtained frompromeP-SEP (IDCA-9365) Mexico

References

[1] M Ekor ldquoThe growing use of herbal medicines issues relatingto adverse reactions and challenges in monitoring safetyrdquoFrontiers in Pharmacology vol 4 pp 1ndash10 2014

[2] S Demiray M E Pintado and P M L Castro ldquoEvaluation ofphenolic profiles and antioxidant activities of Turkishmedicinalplant Tilia argentea Crataegi folium leaves and Polygonumbistorta rootsrdquo World Academy of Science Engineering andTechnology vol 54 pp 312ndash317 2009

[3] Miracle Trees September 2014 httpmiracletreesorg[4] B A Anhwange V O Ajibola and S J Oniye ldquoChemical

studies of seeds ofMoringa oleifera and Detarium microcarpumseedsrdquo Journal of Biological Sciences vol 4 no 6 pp 711ndash7152004

[5] F Anwar M Ashraf and M I Bhanger ldquoInterprovenancevariation in the composition of Moringa oleifera oilseeds fromPakistanrdquo Journal of the American Oil Chemistsrsquo Society vol 82no 1 pp 45ndash51 2005

[6] A O Oluduro ldquoEvaluation of antimicrobial properties andnutritional potentials of Moringa oleifera Lam leaf in South-Western NigeriardquoMalaysian Journal of Microbiology vol 8 no2 pp 59ndash67 2012

[7] RDhakar B PooniyaMGupta et al ldquoMoringa the herbal goldto combat malnutritionrdquo Chronicles of Young Scientists vol 2no 3 pp 119ndash125 2011

[8] S Hussain F Malik and S Mahmood ldquoReview an expositionof medicinal preponderance of Moringa oleifera (Lank)rdquo Pak-istan Journal of Pharmaceutical Sciences vol 27 no 2 pp 397ndash403 2014

[9] J S Aney T Rasmi KMaushumi and B Kiran ldquoPharmacolog-ical and pharmaceutical potential ofMoringa oleifera a reviewrdquoJournal of Farmacy Research vol 2 no 9 pp 1424ndash1426 2009

[10] F Anwar S Latif M Ashraf and A H Gilani ldquoMoringaoleifera a food plant with multiple medicinal usesrdquo Phytother-apy Research vol 21 no 1 pp 17ndash25 2007

[11] G Mishra P Singh R Verma et al ldquoTraditional uses phyto-chemistry and pharmacological properties of Moringa oleiferaplant an overviewrdquo Der Pharmacia Lettre vol 3 no 2 pp 141ndash164 2011

[12] M Mbikay ldquoTherapeutic potential ofMoringa oleifera leaves inchronic hyperglycemia and dyslipidemia a reviewrdquo Frontiers inPharmacology vol 3 article 24 12 pages 2012

[13] A R Verma M Vijayakumar C S Mathela and C V Rao ldquoInvitro and in vivo antioxidant properties of different fractions ofMoringa oleifera leavesrdquo Food and Chemical Toxicology vol 47no 9 pp 2196ndash2201 2009

[14] L A Cajuday and G L Pocsidio ldquoEffects of Moringa oleiferaLam (Moringaceae) on the reproduction of male mice (Musmusculus)rdquo Journal of Medicinal Plants Research vol 4 no 12pp 1115ndash1121 2010

[15] I L Jung ldquoSoluble extract from Moringa oleifera leaves witha new anticancer activityrdquo PLoS ONE vol 9 no 4 Article IDe95492 10 pages 2014


[16] M M Khalafalla E Abdellatef H M Dafalla et al ldquoActiveprinciple from Moringa oleifera Lam leaves effective againsttwo leukemias and a hepatocarcinomardquo African Journal ofBiotechnology vol 9 no 49 pp 8467ndash8471 2010

[17] E Pelus J Arnaud V Ducros H Faure A Favier and A MRoussel ldquoTrace element (Cu Zn Fe Mn Se) intakes of a groupof Frenchmen using the duplicate diet techniquerdquo InternationalJournal of Food Sciences and Nutrition vol 45 no 1 pp 63ndash701994

[18] Q Shazia Z H Mohammad T Rahman and H U ShekharldquoCorrelation of oxidative stress with serum trace elementlevels and antioxidant enzyme status in beta thalassemia majorpatients a review of the literaturerdquoAnemia vol 2012 Article ID270923 7 pages 2012

[19] I Offor R Ehiri and C Njoku ldquoProximate nutritional analysisand heavy metal composition of dried Moringa oleifera leavesfrom Oshiri Onicha LGA Ebonyi State Nigeriardquo IOSR Journalof Environmental Science Toxicology and Food Technology vol8 no 1 pp 57ndash62 2014

[20] D I Sanchez-Machado J Lopez-Cervantes and N J RVazquez ldquoHigh-performance liquid chromatography methodto measure 120572- and 120574-tocopherol in leaves flowers and freshbeans fromMoringa oleiferardquo Journal of Chromatography A vol1105 no 1 pp 111ndash114 2006

[21] D I Sanchez-Machado J A Nunez-Gastelum C Reyes-Moreno B Ramırez-Wong and J Lopez-Cervantes ldquoNutri-tional quality of edible parts ofMoringa oleiferardquo Food Analyti-cal Methods vol 3 no 3 pp 175ndash180 2010

[22] S E Atawodi J C Atawodi G A Idakwo et al ldquoEvaluation ofthe polyphenol content and antioxidant properties of methanolextracts of the leaves stem and root barks of Moringa oleiferaLamrdquo Journal of Medicinal Food vol 13 no 3 pp 710ndash716 2010

[23] P Siddhuraju and K Becker ldquoAntioxidant properties of varioussolvent extracts of total phenolic constituents from three dif-ferent agroclimatic origins of drumstick tree (Moringa oleiferaLam) leavesrdquo Journal of Agricultural and Food Chemistry vol51 no 8 pp 2144ndash2155 2003

[24] AOAC International Official Methods of Analysis Associationof Official Analytical Chemists Rockville Md USA 15thedition 1990

[25] H G Muller and G Tobin Nutrition and Food ProcessingCroom Helm 1980

[26] H Javid U K Farman R Ullah et al ldquoNutrient evaluation andelemental analysis of four selected medicinal plants of khyberpakhtoon khwa Pakistanrdquo Pakistan Journal of Botany vol 43no 1 pp 427ndash434 2011

[27] M Imran H Khan S S Hassan and R Khan ldquoPhysicochemi-cal characteristics of variousmilk samples available in PakistanrdquoJournal of Zhejiang University Science B vol 9 no 7 pp 546ndash551 2008

[28] K A Witt The Nutrient Content of Moringa oleifera LeavesMessiah College Department of Nutrition and Dietetics 2013

[29] C W Yameogo M D Bengaly A Savadogo P A Nikiemaand S A Traore ldquoDetermination of chemical composition andnutritional values of Moringa oleifera leavesrdquo Pakistan Journalof Nutrition vol 10 no 3 pp 264ndash268 2011

[30] L J FuglieTheMiracle TreeTheMultiple Attributes ofMoringaTechnical Centre for Agricultural and Rural CooperationWageningen The Netherlands 2001

[31] D Mukunzi J Nsor-Atindana Z Xiaoming A Gahungu EKarangwa and GMukamurezi ldquoComparison of volatile profile

of Moringa oleifera leaves from Rwanda and china using HS-SPMErdquo Pakistan Journal of Nutrition vol 10 no 7 pp 602ndash6082011

[32] B J Alloway Heavy Metals in Soils Blackie Academic andProfessional 1995

[33] FAOWHO Evaluation of Certain Food Additives and Contam-inants vol 837 World Health Organization Geneva Switzer-land 1993

[34] S B Sarmani I Abugassa A Hamzah and M D Yahya ldquoEle-mental analysis of herbal preparations for traditional medicinesby neutron activation analysis with the k

119900standardization

methodrdquo Biological Trace Element Research vol 71-72 no 1 pp365ndash376 1999

[35] S Fakurazi I Hairuszah and U Nanthini ldquoMoringa oleiferaLam prevents acetaminophen induced liver injury throughrestoration of glutathione levelrdquo Food and Chemical Toxicologyvol 46 no 8 pp 2611ndash2615 2008

[36] B N Singh B R Singh R L Singh et al ldquoOxidative DNAdam-age protective activity antioxidant and anti-quorum sensingpotentials of Moringa oleiferardquo Food and Chemical Toxicologyvol 47 no 6 pp 1109ndash1116 2009

[37] EM B TeixeiraM R B CarvalhoVANevesMA Silva andL Arantes-Pereira ldquoChemical characteristics and fractionationof proteins fromMoringa oleifera Lam leavesrdquo Food Chemistryvol 147 pp 51ndash54 2014

[38] P M P Ferreira D F Farias J T D A Oliveira and A DF U Carvalho ldquoMoringa oleifera bioactive compounds andnutritional potentialrdquoRevista de Nutricao vol 21 no 4 pp 431ndash437 2008

[39] R Gowrishankar M Kumar V Menon et al ldquoTrace elementstudies on Tinospora cordifolia (Menispermaceae) Ocimumsanctum (Lamiaceae) Moringa oleifera (Moringaceae) andPhyllanthus niruri (Euphorbiaceae) using PIXErdquo BiologicalTrace Element Research vol 133 no 3 pp 357ndash363 2010

[40] S O Dania P Akpansubi and O O Eghagara ldquoComparativeeffects of different fertilizer sources on the growth and nutrientcontent of Moringa (Moringa oleifera) seedling in a greenhousetrialrdquo Advances in Agriculture vol 2014 Article ID 726313 6pages 2014

[41] A Sodamode O Bolaji and O Adeboye ldquoProximate analysismineral contents and functional properties ofMoringa oleiferaleaf protein concentraterdquo IOSR Journal of Applied Chemistryvol 4 no 6 pp 47ndash51 2013

[42] F Sizer and E Whitney Nutrition Concepts and ControversiesWadsworth Belmont Calif USA 8th edition 1999

[43] A Howard MoringamdashThe Natural Cure for Osteoporosis FreePress Release 50 Free Press 2014

[44] S Bo and E Pisu ldquoRole of dietary magnesium in cardiovasculardisease prevention insulin sensitivity and diabetesrdquo CurrentOpinion in Lipidology vol 19 no 1 pp 50ndash56 2008

[45] M C P Estrella J B V Mantaring and G Z David ldquoA doubleblind randomized controlled trial on the use of malunggay(Moringa oleifera) for augmentation of volume of breastmilkamong non-nursingmothers of preterm infantsrdquoThePhilippineJournal of Pediatrics vol 49 pp 3ndash6 2000

[46] V V ValkovicTrace Element Analysis vol 5 Taylor and FrancisLondon UK Rice University Houston Tex USA 1975

[47] B S Rathi S L Bodhankar and A M Baheti ldquoEvaluationof aqueous leaves extract of Moringa oleifera Linn for woundhealing in albino ratsrdquo Indian Journal of Experimental Biologyvol 44 no 11 pp 898ndash901 2006


[48] T H Bothwell R V Charlton J D Cook and C A Finch IronMetabolism in Man Blackwell Scientific Publications OxfordUK 1979

[49] H J Jung and Y R Seo ldquoCurrent issues of selenium in cancerchemopreventionrdquo BioFactors vol 36 no 2 pp 153ndash158 2010

[50] L V Costa-Lotufo M T H Khan A Ather et al ldquoStudiesof the anticancer potential of plants used in Bangladeshi folkmedicinerdquo Journal of Ethnopharmacology vol 99 no 1 pp 21ndash30 2005

[51] WHO National Policy on Traditional Medicine and Regulationof Herbal Medicines Report of a WHO Global Survey WorldHealth Organization Geneva Switzerland 2005

[52] M Saeed N Muhammad H Khan and S A Khan ldquoAnalysisof toxic heavy metals in branded Pakistani herbal productsrdquoJournal of the Chemical Society of Pakistan vol 32 no 4 pp471ndash475 2010

[53] P Roy and A Saha ldquoMetabolism and toxicity of arsenic ahuman carcinogenrdquo Current Science vol 82 no 1 pp 38ndash452002

[54] S K Karamala C Sri latha Y Anjaneyulu T S ChandraSekhara Rao D S Vasulu and A P Pidugu ldquoHematobiochem-ical changes of lead poisoning and amelioration with Ocimumsanctum in wistar albino ratsrdquo Veterinary World vol 4 no 6pp 260ndash263 2011

[55] M A Armienta and N Segovia ldquoArsenic and fluoride inthe groundwater of Mexicordquo Environmental Geochemistry andHealth vol 30 no 4 pp 345ndash353 2008

[56] I Rosas R Belmont A Armienta and A Baez ldquoArsenicconcentrations in water soil milk and forage in ComarcaLagunera Mexicordquo Water Air and Soil Pollution vol 112 no1-2 pp 133ndash149 1999

[57] T C Broyer C M Johnson and R E Paull ldquoSome aspects oflead in plant nutritionrdquo Plant and Soil vol 36 no 1 pp 301ndash3131972

[58] A de Pasquale E Paino R de Pasquale and M P GermanoldquoContamination by heavy metals in drugs from different com-mercial sourcesrdquo Pharmacological Research vol 27 no 1 pp 9ndash10 1993

[59] A A K Abou-Arab M S Kawther M E El Tantawy R IBadeaa and N Khayria ldquoQuantity estimation of some con-taminants in commonly used medicinal plants in the Egyptianmarketrdquo Food Chemistry vol 67 no 4 pp 357ndash363 1999

[60] I A Khan J Allgood L A Walker E A Abourashed DSchlenk and W H Benson ldquoDetermination of heavy metalsand pesticides in ginseng productsrdquo Journal of AOAC Interna-tional vol 84 no 3 pp 936ndash939 2001

[61] E D Caldas and L L Machado ldquoCadmium mercury and leadinmedicinal herbs in BrazilrdquoFood andChemical Toxicology vol42 no 4 pp 599ndash603 2004

[62] I J Asiedu-Gyekye S Frimpong-Manso C Awortwe D AAntwi and A K Nyarko ldquoMicro- and macroelemental compo-sition and safety evaluation of the nutraceuticalMoringa oleiferaleaves rdquo Journal of Toxicology vol 2014 Article ID 786979 13pages 2014

[63] A Siddiq F Anwar M Manzoor and A Fatima ldquoAntioxidantactivity of different solvent extracts of Moringa oleifera leavesunder accelerated storage of sunflower oilrdquo Asian Journal ofPlant Sciences vol 4 no 6 pp 630ndash635 2005

[64] S Sreelatha and P R Padma ldquoAntioxidant activity and totalphenolic content of Moringa oleifera leaves in two stages ofmaturityrdquo Plant Foods for Human Nutrition vol 64 no 4 pp303ndash311 2009

[65] O K Chun S J Chung and W O Song ldquoEstimated dietaryflavonoid intake and major food sources of US adultsrdquo TheJournal of Nutrition vol 137 no 5 pp 1244ndash1252 2007

[66] L Dauchet P Amouyel S Hercberg and J Dallongeville ldquoFruitand vegetable consumption and risk of coronary heart diseasea meta-analysis of cohort studiesrdquoThe Journal of Nutrition vol136 no 10 pp 2588ndash2593 2006

[67] J A Nascimento K L G V Araujo P S Epaminondas et alldquoEthanolic extracts ofMoringa oleifera Lamrdquo Journal ofThermalAnalysis and Calorimetry vol 114 no 2 pp 833ndash838 2013

[68] J P Coppin Y Xu H Chen et al ldquoDetermination of flavonoidsby LCMS and anti-inflammatory activity in Moringa oleiferardquoJournal of Functional Foods vol 5 no 4 pp 1892ndash1899 2013

[69] P G Matshediso E Cukrowska and L Chimuka ldquoDevelop-ment of pressurised hot water extraction (PHWE) for essentialcompounds from Moringa oleifera leaf extractsrdquo Food Chem-istry vol 172 pp 423ndash427 2015

[70] B Sultana and F Anwar ldquoFlavonols (kaempeferol quercetinmyricetin) contents of selected fruits vegetables and medicinalplantsrdquo Food Chemistry vol 108 no 3 pp 879ndash884 2008

[71] T Marrufo S Encarnacao O M D Silva et al ldquoChemicalcharacterization and determination of antioxidant and antimi-crobial activities of the leaves of Moringa oleiferardquo RevistaInternacional de Gestao de Conflitos Ambientais vol 2 no 1 pp1ndash15 2013

[72] C Rodrıguez-Perez R Quirantes-Pine A Fernandez-Gutierrez and A Segura-Carretero ldquoOptimization ofextraction method to obtain a phenolic compounds-richextract fromMoringa oleifera Lam leavesrdquo Industrial Crops andProducts vol 66 pp 246ndash254 2015

[73] H Zhao J Dong J Lu et al ldquoEffects of extraction solventmixtures on antioxidant activity evaluation and their extractioncapacity and selectivity for free phenolic compounds in barley(Hordeum vulgare L)rdquo Journal of Agricultural and Food Chem-istry vol 54 no 19 pp 7277ndash7286 2006

[74] J Coppin A Study of the Nutritional and Medicinal Val-ues of Moringa oleifera Leaves from Sub-Saharan AfricaGhana Rwanda Senegal and Zambia Rutgers University NewBrunswick NJ USA 2008

[75] N Nakatani S-I Kayano H Kikuzaki K Sumino K Katagiriand T Mitani ldquoIdentification quantitative determination andantioxidative activities of chlorogenic acid isomers in prune(Prunus domestica L)rdquo Journal of Agricultural and Food Chem-istry vol 48 no 11 pp 5512ndash5516 2000

[76] A-S Cho S-M Jeon M-J Kim et al ldquoChlorogenic acidexhibits anti-obesity property and improves lipid metabolismin high-fat diet-induced-obese micerdquo Food and Chemical Toxi-cology vol 48 no 3 pp 937ndash943 2010

[77] X Zhang H Huang T Yang et al ldquoChlorogenic acid protectsmice against lipopolysaccharide-induced acute lung injuryrdquoInjury vol 41 no 7 pp 746ndash752 2010

[78] T Suzuki Y Honda and Y Mukasa ldquoEffects of UV-B radiationcold anddesiccation stress on rutin concentration and rutin glu-cosidase activity in tartary buckwheat (Fagopyrum tataricum)leavesrdquo Plant Science vol 168 no 5 pp 1303ndash1307 2005

[79] T Walle ldquoMethoxylated flavones a superior cancer chemopre-ventive flavonoid subclassrdquo Seminars in Cancer Biology vol 17no 5 pp 354ndash362 2007

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy


Carbohydrate Chemistry

International Journal of


Journal of

Chemistry


Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of


Chromatography Research International


Applied ChemistryJournal of



Theoretical ChemistryJournal of


Journal of

Spectroscopy

Analytical ChemistryInternational Journal of


Journal of


Quantum Chemistry


Organic Chemistry International

ElectrochemistryInternational Journal of



CatalystsJournal of


It is widely distributed throughout Africa Saudi ArabiaSoutheast Asia the Caribbean Islands and South AmericaEvery part of M oleifera has medicinal properties and iscommercially exploitable for the development of medicinaland industrial byproducts [3]

Traditionally the leaves fruits flowers and immaturepods of this tree are edible they are used as a highly nutritivevegetable in many countries particularly in India Pakistanthe Philippines Hawaii and some African nations [4ndash6] Indeveloping nations M oleifera is used as an alternative toimported food supplements to treat and combat malnutri-tion especially among infants and nursingmothers by virtueof its chemical constituents [7]

Several valuable reviews of the ethnobotanical usesof M oleifera are available [8ndash11] Moringa has beenfound to be a good source of polyphenols and antiox-idants [11] Phytochemicals such as vanillin omega fattyacids carotenoids ascorbates tocopherols beta-sitosterolmoringine kaempferol and quercetin have been reported inits flowers roots fruits and seeds The leaves in particularhave been found to contain phenolics and flavonoids [12 13]these compounds have various biological activities includingantioxidant anticarcinogenic immunomodulatory antidia-betic antiatherogenic and hepatoprotective functions andthe regulation of thyroid status [14ndash16] Moreover leavescontain trace elements that are essential to human health Forinstancemagnesium iron selenium and zinc play an impor-tant role in metabolism and interest in these elements isincreasing together with reports relating trace element statusand oxidative diseases [17 18] However a recent study hasshown that dried M oleifera leaves contain lead at very highvalues of 3520mgL [19] Therefore it is very important toidentify the mineral composition ofM oleifera leaves that arewidely consumed by humans and animals

In Mexico M oleifera is widely cultivated in differentzones of the country and is found inmore than ten states fromSonora to Oaxaca on the Pacific side Few studies have beenconducted on nutritional and phytochemical composition[20 21] however to date a detailed composition of the leavesof M oleifera that is native to Mexico has not been reportedyet In addition it is important to bear in mind that themineral and phenolics contents present in leaves depend onseveral factors such as geographical area where the plant iscultivated type of soil water and fertilizers industrializationprocess and storage conditions Taking these precedents intoconsideration the aim of this study was to evaluate thephytochemical constituents of methanolic extracts and traceelements and nutritional values present in M oleifera leavesgrown in the northeast and west regions of Mexico

2 Materials and Methods

21 Reagents and Standards The methanol and acetonitrileused in this study were of HPLC grade and were purchasedfrom JT Baker USA andCaledon Lab Canada HPLC-gradephenolic standards (gallic and chlorogenic acids rutin lute-olin quercetin apigenin and kaempferol) were of analyticalreagent grade and were purchased from Sigma-Aldrich (StLouis MO USA) Demineralized water was taken from a

Milli-Q Plus water purification system (Millipore BedfordUSA) The multielement standard mixture solutions con-tained Ba Bi Li Cu Co In Li Ni Pb and V the otherscontained Ba CaMg and Sr Analytical grade 65wv nitricacid (HNO

3) (JT Baker) was used in the decomposition

22 Samples Preparation M oleifera leaves were collectedfrom Mexican cultivars (Lombardia Michoacan locatedon 19∘0110158403010158401015840N and 102∘0510158403910158401015840W and San Pedro Coahuilalocated on 25∘4510158403510158401015840N and 102∘5810158405410158401015840WMexico April 2011)and a voucher of the specimen was deposited in the CIIDIR-IPN herbarium Durango Mexico for future references Theplants were identified and authenticated by M Sc AlbertoGonzalez Zamora Faculty of Biology UJEDThe leaves werecleaned and processed as described by previous researchers[22 23] A set of five samples from each geographical originwere randomly collected and analyzed All materials were air-dried and powdered and protected from light until furtheranalysis

23 Chemical Analysis Representative subsamples weredried in a forced drought oven at 105ndash110∘C to a constantweight for moisture determination Crude protein crudelipid crude fiber and ash were analyzed by triplicate accord-ing to AOAC [24] The total carbohydrates were determinedby the difference method [100 minus (proteins + fats + moisture+ ash in percentage)] [25 26] The energy values (kcal100 g)were determined by multiplying the values of carbohydrateslipids and proteins by factors of 4 9 and 4 respectively andtaking the sum expressed in kilocalories [27]

24 Extraction of Phenolic Compounds The extracts wereprepared using 23 g of dry-ground sample and 260mL of 80methanolic aqueous solution by successive maceration Themixtures were shaken in amagnetic grid at room temperaturefor 21 h They were then filtered through Whatman filterpaper number 1 The final extract was concentrated on arotator evaporator and the filtrate was placed in a deepfreezer for 24 h and lyophilized to obtain a powdered extractAll of the extractions were performed in triplicate and thesupernatants were kept at minus40∘C until further analysis

25 ICP-MS Analysis The instrumentation used for diges-tion consisted of a closed microwave oven (MARS-XpressCEM)Themultielemental analysis was performed by induc-tively coupled plasma mass spectrometry (ICP-MS) with areaction cell (XSeries 2 Thermo Scientific) Prior to analysisthe samples were homogenized in a mortar approximately05 g of the sample was accurately weighed in an acid-washedpolytetrafluoroethylene (PTFE) digestion tube and 5mL ofHNO

3(65wv) was added The tube was heated in a

microwave oven at a power setting of 50 and 150∘C for10min 70 and 220∘C for 20min and 10 and 100∘C for15min The digest was transferred into a 50mL acid-washedvolumetric flask which was filled with demineralized waterand stored in a polypropylene container Onewater blankwasrun with each batch of samples




3 Results

















Referencecompound

Retentiontime (min)


2













4 Discussion




(min)10 20 30 40 50 60

(mAU

)

020406080

268

32

884

157

56

469

3148

261

519

22

560

5757

962

598

19 606

22

655

2665

736

C

LR

QK



10 20 30 40 50 60

020406080

100

264

7

585

6 670

3

149

00 390

03

424

7643

469

443

11

464

2147

291

480

5849

283

516

5849

663

533

7352

001

541

8153

672

626

08510

69

607

9760

506

553

6955

918

562

7757

805

583

58

545

66

(min)

(mAU

)

10 20 30 40 50 60

020406080

100120140

(min)

(mAU

)

146

70666

0

265

3 388

63

434

1544

284

464

2448

077

493

1649

682

625

7960

777

605

00

577

8658

301

558

8456

227

533

5954

162

536

95

510

7851

668

520

10

San Pedro

Lombardia

























5 Conclusion




Acknowledgments



References






























































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of




3 Results

















Referencecompound

Retentiontime (min)


2













4 Discussion




(min)10 20 30 40 50 60

(mAU

)

020406080

268

32

884

157

56

469

3148

261

519

22

560

5757

962

598

19 606

22

655

2665

736

C

LR

QK



10 20 30 40 50 60

020406080

100

264

7

585

6 670

3

149

00 390

03

424

7643

469

443

11

464

2147

291

480

5849

283

516

5849

663

533

7352

001

541

8153

672

626

08510

69

607

9760

506

553

6955

918

562

7757

805

583

58

545

66

(min)

(mAU

)

10 20 30 40 50 60

020406080

100120140

(min)

(mAU

)

146

70666

0

265

3 388

63

434

1544

284

464

2448

077

493

1649

682

625

7960

777

605

00

577

8658

301

558

8456

227

533

5954

162

536

95

510

7851

668

520

10

San Pedro

Lombardia

























5 Conclusion




Acknowledgments



References






























































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of






Referencecompound

Retentiontime (min)


2













4 Discussion




(min)10 20 30 40 50 60

(mAU

)

020406080

268

32

884

157

56

469

3148

261

519

22

560

5757

962

598

19 606

22

655

2665

736

C

LR

QK



10 20 30 40 50 60

020406080

100

264

7

585

6 670

3

149

00 390

03

424

7643

469

443

11

464

2147

291

480

5849

283

516

5849

663

533

7352

001

541

8153

672

626

08510

69

607

9760

506

553

6955

918

562

7757

805

583

58

545

66

(min)

(mAU

)

10 20 30 40 50 60

020406080

100120140

(min)

(mAU

)

146

70666

0

265

3 388

63

434

1544

284

464

2448

077

493

1649

682

625

7960

777

605

00

577

8658

301

558

8456

227

533

5954

162

536

95

510

7851

668

520

10

San Pedro

Lombardia

























5 Conclusion




Acknowledgments



References






























































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


(min)10 20 30 40 50 60

(mAU

)

020406080

268

32

884

157

56

469

3148

261

519

22

560

5757

962

598

19 606

22

655

2665

736

C

LR

QK



10 20 30 40 50 60

020406080

100

264

7

585

6 670

3

149

00 390

03

424

7643

469

443

11

464

2147

291

480

5849

283

516

5849

663

533

7352

001

541

8153

672

626

08510

69

607

9760

506

553

6955

918

562

7757

805

583

58

545

66

(min)

(mAU

)

10 20 30 40 50 60

020406080

100120140

(min)

(mAU

)

146

70666

0

265

3 388

63

434

1544

284

464

2448

077

493

1649

682

625

7960

777

605

00

577

8658

301

558

8456

227

533

5954

162

536

95

510

7851

668

520

10

San Pedro

Lombardia

























5 Conclusion




Acknowledgments



References






























































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

















5 Conclusion




Acknowledgments



References






























































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of















































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

Research Article Nutritional Content and Elemental and ...

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