micro- and macroelemental composition and safety evaluation of the nutraceutical moringa oleifera...

7212019 Micro- and Macroelemental Composition and Safety Evaluation of the Nutraceutical Moringa oleifera Leaves

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Research ArticleMicro- and Macroelemental Composition and Safety Evaluationof the Nutraceutical Moringa oleifera Leaves

I J Asiedu-Gyekye1 S Frimpong-Manso2 C Awortwe3 D A Antwi4 and A K Nyarko1

983089 Department of Pharmacology and oxicology College of Health Sciences University of Ghana School of PharmacyPO Box LG 983092983091 Legon Ghana

983090 Department of Pharmaceutical Chemistry College of Health Sciences University of Ghana School of PharmacyPO Box LG 983092983091 Legon Ghana

983091 Division of Clinical Pharmacology Faculty of Health Sciences University of Stellenbosch PO Box 983089983097983088983094983091 Cape own South Africa983092 Department of Physiology College of Health Sciences University of Ghana Medical School PO Box 983092983090983091983094 Korle-Bu Ghana

Correspondence should be addressed to I J Asiedu-Gyekye asiedugyekyeyahoocouk

Received 983094 February 983090983088983089983092 Revised 983090983093 June 983090983088983089983092 Accepted 983090983093 June 983090983088983089983092 Published 983090983090 July 983090983088983089983092

Academic Editor Margaret James

Copyright copy 983090983088983089983092 I J Asiedu-Gyekye et al Tis is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Moringa oleifera is a multipurpose plant used in Ghana and most parts o Arica Its high mineral protein and vitamins contenthasenabled its use as a nutraceutical and panacea or various diseases Tis study aimed at measuring the micro- and macroelementscontent o dried Moringa oleifera leaves using energy dispersive X-ray 1047298uorescence spectroscopic (EDXRF) and assessing itstoxicological effect in rats Acute toxicity (983093983088983088983088 mgkg) and a subacute toxicity studies o the lea (983092983088 mgkg to 983089983088983088983088 mgkg) extractwere conducted in rats Blood samples were assessed or biochemical and haematological parameters Results showed signi1047297cantlevels o thirty-1047297ve (983091983093) elements (983089983092 macroelements and 983090983089 microelements) in M oleifera extract Tere were no observed overtadverse reactions in the acute and subacute studies Although there were observed elevations in liver enzymes AL and ALP( lt 0001) and lower creatinine levels in the extract treated groups no adverse histopathological 1047297ndings were ound Moringaoleifera dried lea extract may thereore be reasonably sae or consumption However the consumption o Moringa oleifera leavesshould not exceed a maximum o 983095983088 grams per day to prevent cumulative toxicity o these essential elements over long periods

1 Introduction

Moringa oleifera Lam ound in most parts o Ghana belongs

to the monogenetic amily Moringaceae (order Brassicales)It is a plant that has multipurpose nonmedicinal andmedicinal uses Its nonmedical uses include use o the seedsin wastewater treatment due to their coagulant properties[983089 983090] Its medicinal uses stem rom the act that the entireplant has high protein vitamins mineral and carbohydratecontent It is thus o high nutritional value or both humansand livestock Moringa leaves are rich in minerals such asiron potassium and calcium as well as vitamins essentialamino acidsand a numbero glycosides [983091 983092] Te seeds havehigh content (983092983090) o edible oil that also has medicinal uses

Moringa is used or the management o various ailmentsas a galactogogue in mothers o preterm inants [983093 983094] It is

also used to manage heart diseases and eye problems as wellas in1047298ammations and dyspepsia [983095 983096]

Pharmacological studies have shown that the extracts o

the plant have antioxidant [983097ndash983089983089] anticarcinogenic [983089983090] anti-in1047298ammatory antispasmodic and antidiuretic [983089983091] proper-ties Others include antiulcer antibacterial and antiungalproperties [983089983092] Recent studies indicate that it also hasantinociceptive [983095] as well as wound healing ability [983096]Studies on the root bark have shown it to have analgesicalexeteric and antihelminthic properties It has also beenreported to alter blood lipid pro1047297les [983089983093] oxicity studies haveshown that aqueous extract o moringa lea extract has nosigni1047297cant adverse effects in rats rabbits [983089983093ndash983089983096] or poultry [983089983096] However there are signi1047297cant differences in the saety and composition o various moringa species rom differentlocations [983089983097]

Hindawi Publishing CorporationJournal of Toxicology Volume 2014 Article ID 786979 13 pageshttpdxdoiorg1011552014786979



983090 Journal o oxicology

In Ghana and most parts o West Arica powdered leaveso M oleifera are marketed under different brand names andconsumed daily as a nutraceutical prophylaxis or cure or

various conditions Te prevailing socio-culturaleconomicconditions and consequent difficulty in accessing healthcareservices especially by rural populations as well as the general

perception that plant medicines are efficacious and reerom side effects [983089983097 983090983088] will increase the requent andwidespread spread use o moringa lea powder In view o the reported differences in moringa lea products which may impact adversely on itssaety the micro-and macroelementalcontent o dried leaves o Moringa oleifera were measured Inaddition the lea extract was investigated or its toxicologicaleffects in Sprague-Dawley rats

2 Materials and Methods

983090983089 Preparation of Extracts of Moringa oleifera Leaves forAnal- ysis In order to mimic the traditional method o extraction983090983096 kg o sample were blended with boiled distilled water

Te mixture covered with water was lef to stand overnightin a water bath maintained at 983094983088∘C and the watery portion(inusion) was 1047297ltered off using 983088983092983093 1038389m millipore celluloseester 1047297lters and reeze dried to obtain M oleifera extract(MOE) Portions were homogenized and labelled as sampleA or elemental analysis

983090983090 Preparation of Moringa oleifera Leaves for AnalysisLeaves o the Moringa oleifera species were collected romAccra separated rom other plant parts washed and driedin the shade or 1047297ve days at room temperature ground into1047297ne powder sieved (983090983089983090 1038389m mesh size) to obtain very 1047297nesamples and kept in separate containers Tis was labelled

sample B

983090983091 Pelleting of Samples for Analysis Te loosepowdery nature o samples (plant extracts A and lea powder B)required that they were pelleted beore analysis Beore pel-leting 983092 g o each sample was weighed and 983088983097 g o the binderFluxana (H Elektronic BM-983088983088983088983090-983089 (Licowax C micropowderPM-Hoechstwax)) was added and homogenized or 983091 min-utes Te mixture was pressed at 983090983088 t or 983090 minutes into pelletso 983091983090 mm in diameter or the subsequent XRF measurementsTree separate pellets were prepared rom each o samples Aand B

983090983092 Energy Dispersive X-Ray (ED XRF) MeasurementsEnergy dispersive X-ray (ED XRF) was used or simultaneousanalysis and measurement o the elemental content o thesamples Te procedure which used three-axial geometryreduced background noise due to radiation polarization Temonochromatic radiations emitted rom the X-ray tube wereapplied to excite the atoms o the sample

983090983093 Experimental Animals and Housing Conditions MaleSprague-Dawley rats (983089983093983088 gndash983089983096983088 g body weight) were pur-chased rom the Centre or Scienti1047297c Research into PlantMedicine Mampong Te rats were housed in plastic cageswith stainless steel tops in the animal care acility o theUniversity o Ghana Medical School and kept under standard

983089983090 h light and 983089983090 h dark schedule where room temperaturehumidity and ventilation werecontrolled during the acclima-tization period o seven (983095) days

983090983094 Acute and Subacute oxicity Studies Te reconstitutedpowdered MOE mixturewas preparedusingdistilled water as

the vehicle and administered as a single dose (983093983088983088983088 mgkg)by oral gavage or the acute toxicity studies Afer theadministration animals were observed every hour or the1047297rst 983094 hours then daily or the next 983089983091 days

In the subacute studies 1047297ve groups (983089ndash983093) o maleSprague-Dawley Rats (eight weeks old seven animalsgroup) o meanweight 983089983093983088 g were prepared Animals in each o the ourgroups were administered MOE extract over a dose range o 983088 mgkg to 983089983088983088983088kg by oral gavage daily or 983089983092 days Animalsin the 1047297fh group received the vehicle (distilled water)andserved as controls

For the subacute studies afer the983089983092 days o MOE admin-istration o MOE each animal was observed every hour orsix hours daily or the next 983091 days and subsequently every day or 983089983088 days Animals were ed ad libitum with standard chow diet (AIN-983097983091G ormulation obtained rom GAFCO-Ghana)

983090983094983089 Observed Clinical Signs of oxidromes Body weightswere observed daily beore and afer administration Inaddition animals were observed daily or clinical signs o excitability twitching salivation morbidity miosis mydria-sis rising ur sluggish movement draping tremors and soorth

983090983095 Laboratory Examinations

983090983095983089 Blood Samples Under chloroorm anaesthesia blood

was obtained via cardiac puncture and the animals euth-anized by exsanguinations Samples o the blood collectedwere aliquoted into EDA-983090K tubes and plain tubes respec-tively Te EDA blood was immediately analysed or haema-tological parameters using the SYSMEX Haematology Auto-analyser [Kobe Japan] while serum prepared rom blood inthe plain tubes was used or biochemical examinations

983090983096 Necropsy and Histopathological Studies Gross patholog-ical investigations were conducted on each animal ollowingexsanguination Te heart kidney liver and gastrointesti-nal tract were prepared or histopathological examinationsAll experimental procedures and assays were conducted inaccordance with the international guidelines or evaluatingthe saety o herbal medicines [983090983089ndash983090983091]

983090983097 Statistical Analysis Results are presented as means plusmnSEM and analysis or statistical differences was done usingone-way ANOVA ollowed by Bonferroni post-hoc test

values less than 983088983088983093 were considered statistically signi1047297cant

3 Results

Te ED-XRF analyses led to the detection o a total o thirty-1047297ve (983091983093) elements comprising eleven (983089983089) major elementsand twenty-our (983090983092) minor elements Te major elementsdetected included Na Mg Al Si P S Cl K Ca Mn and Fe



Journal o oxicology 983091

983137983138983148983141 983089 Concentrations o major oxides per total mass o powderedlea in milligrams

Major oxides Level in sample (mg)

Na983090O 983091983095983093

MgO 983094983088983094

Al983090O

983091 983089983091983088

SiO2

983091983088983088

P983090O983093 983091983095983088

SO3

983089983090983090983096

Cl 983089983095983094

K983090O 983095983096983092

CaO 983089983089983096983088

MnO 983088983092

Fe983090O983091 983091983088

983137983138983148983141 983090 Concentrations o minor elements per total mass o powdered lea in milligrams

Minor elements Level in sample (mg)V 983090983092983088983088

Cr 983088983093983095983096

Co 983088983088983089983090

Ni 983088983088983089983092983096

Cu 983088983088983091983089983096

Zn 983088983089983089983093983094

Ga 983088983088983088983091983090

As 983088983088983088983090983090

Rb 983088983088983095983093983096

Y 983088983091983093983092983090

Zr 983088983088983088983090983092

Nb 983088983088983089983089983094

Mo 983088983088983088983093983094Sn 983088983088983093983095983094

Cs 983088983088983090983092983090

Ba 983088983096983097983088983088

La 983088983088983092983097983088

Ce 983088983088983094983097983092

H 983088983088983089983093983092

a 983088983088983089983089983092

Pb 983088983088983088983092983092

T 983088983088983088983091983090

and other heavy metals ables 983089 and 983090 show details o the

elements detected and their concentrations

983091983089 Clinical Symptoms In the acute toxicity studies as wellas the subacute studies at all the dose levels groomingrepetitive circling with arched-back posture were observed inall the rats except those in the control group Rats given highdoses o MOE (983089983088983088983088mgkg and 983093983088983088983088 mgkg) showed moreexcitability twitching and salivation Necropsy revealed noabnormalities

983091983090 Haematology Tese results o haematological investiga-tions are presented in able 983091 (acute toxicity) and able 983092(subacute toxicity studies) In the acute toxicity studies

983137983138983148983141 983091 Haematological analysis o a 983093983088983088983088 mgkg single doseadministration o MOE in male SDRs Values are expressed as meanplusmn SEM (1103925 = 7) Values o lt 005 were considered as statistically signi1047297cant lowast lt 005 lowastlowast lt 001 and lowastlowastlowast lt 00001 when controlwas compared with MOE

Parameters Groups

Control males 983093983088983088983088mgkg malesWBC (983089983088983091 1038389L) 48 plusmn 032 732 plusmn 004lowastlowastlowast

RBC (983089983088983094 1038389L) 83 plusmn 040 884 plusmn 004HGB (gdL) 151 plusmn 059 1482 plusmn 004HC () 536 plusmn 231 52 plusmn 032MCV (L) 65 plusmn 038 582 plusmn 007lowastlowastlowast

MCH (pg) 183 plusmn 016 1664 plusmn 004MCHC (gdL) 2817 plusmn 016 286 plusmn 004PL (983089983088983091 1038389L) 3003 plusmn 4444 1128 plusmn 037lowastlowast

LYM 8433 plusmn 139 8314 plusmn 005LYM number 41 plusmn 028 61 plusmn 004

WBC increased signi1047297cantly (by 983093983090983093) MCV on the otherhand decreased by 983089983088 Similarly platelets levels droppedsigni1047297cantly (983094983090983093) ollowing treatment with MOE

In the subacute studies WBC increased by 983095983091983095983093 and983094983095 ( lt 00001) at dose levels o 983092983088 mgkg and 983096983088 mgkgrespectively MCV dropped signi1047297cantly at the 983090983088983088 mgkgand 983089983088983088983088 mgkg dose levels Similarly platelets levels rosesigni1047297cantly over baseline level at the 983096983088 mgkg dose levelHowever at 983089983088983088983088 mgkg they dropped signi1047297cantly

983091983091 Biochemical Analysis Figure 983089 shows results obtainedrom the biochemical parameters used as markers or renalunction Te 1047297gure shows that blood urea levels were ele-

vated at dose levels o 983096983088 mgkg and 983089983088983088983088 mgkg Creatininelevels however signi1047297cantly decreased at all dose levelscompared to the controls

Figures 983090 and 983091 show results or the biochemical testor liver unction and lipid pro1047297les respectively Terewere increases in the levels o liver enzymes during MOEadministration in a non-dose dependent manner AS levelshowever reduced at all dose levelsexcept the 983096983088 mgkg groupTere were decreases in the lipid parameters during MOEadministration For the lipid pro1047297les MOE administrationgenerally led to a decrease in total cholesterol and triglyc-erides as well as LDL and to some extent HDL cholesterol

983091983092 Histopathology Photomicrographs o tissues prepared

rom control and animals treated with MOE at differentdose levels are presented in Figures 983093ndash983096 No observablecardiomyopathy was noted in the heart (Figure 983093) Figure 983094showed no observable ulceration Te epithelial cells o the stomach are intact Figure 983095 also showed no observablehistological lesions in the glomerulus and the tubules Terewere no observable histological lesions in the sinusoids andcentral vein o the liver (Figure 983096)

4 Discussion

Normally herbal preparations are considered relatively saeand devoid o numerous adverse effects probably because




983137983138983148983141 983092 Haematological Analysis o a 983089983092-day administration o MOE in male SDRs Values are expressed as mean plusmn SEM (1103925 = 7) Values o lt 005 were considered as statistically signi1047297cant lowast lt 005 lowastlowast lt 001 and lowastlowastlowast lt 00001 when control was compared with MOE

Parameters Groups

Control males 983092983088 mgkg males 983096983088mgkg males 983090983088983088mgkg males 983089983088983088983088 mgkg males value

WBC (983089983088983091 1038389L) 48 plusmn 032 834 plusmn 002lowastlowastlowast 72 plusmn 003lowastlowastlowast 42 plusmn 032 46 plusmn 014 00001

RBC (983089983088983094

1038389L)

83 plusmn 040 606 plusmn 008

lowastlowastlowast

655 plusmn 049

lowast

771 plusmn 004 764 plusmn 019 00001HGB (gdL) 151 plusmn 059 107 plusmn 004 1375 plusmn 008 1374 plusmn 004 128 plusmn 066 0934

HC () 536 plusmn 231 39 plusmn 049 412 plusmn 325 4622 plusmn 075 468 plusmn 171 00451

MCV (L) 65 plusmn 038 6514 plusmn 022 6278 plusmn 073lowast 6127 plusmn 016lowastlowastlowast 611 plusmn 066lowastlowastlowast 00001

MCH (pg) 183 plusmn 016 1866 plusmn 002 185 plusmn 009 1774 plusmn 007 1673 plusmn 061 09705

MCHC (gdL) 2817 plusmn 016 2836 plusmn 002 2925 plusmn 017 286 plusmn 024 2732 plusmn 093 09846

PL (983089983088983091 1038389L) 3003 plusmn 4444 377 plusmn 2087 5772 plusmn 4578lowastlowastlowast 3112 plusmn 396 137 plusmn 1309lowastlowast 00001

LYM 8433 plusmn 139 758 plusmn 004 6002 plusmn 632lowast 8242 plusmn 059 8442 plusmn 097 00098

LYM number 41 plusmn 028 634 plusmn 005 89 plusmn 098 336 plusmn 024 378 plusmn 004 09535

Extract concentration (mgmL)

0

2

4

6

8

10

Control

U r e a ( m m o l L )

40mgmL

80mgmL

200mgmL

1000mgmL

(a)

0

20

40

60

80

100

Control

40mgmL

80mgmL

200mgmL

1000mgmL


lowastlowast

lowast

C r e a t i n i n e ( m o l L )

(b)

F983145983143983157983154983141 983089 Renal unction test during a 983089983092-day administration o MOE in male SDRs Values are expressed as means plusmn SEM (1103925 = 7) Values o lt 005 were considered as statistically signi1047297cant lowast

lt 005 lowastlowast lt 001 and lowastlowastlowast

lt 00001 when control was compared with MOE

they are considered to be ldquonaturalrdquo Tis study attemptedto analyse the elemental content o Moringa oleifera and to

ascertain i a 983089983092-day dosing could have any observable adverseeffects Tis is in view o the widespread use o moringalea powder as a ood supplement and treatment or variousdisease conditions

X-ray 1047298uorescence (XRF) is a ast accurate and non-destructive analytical technique used or the elemental andchemical analysis o powdered solid and liquid samplesAnalyses o the samples produced a total o 983091983093 elements(983089983092 macroelements and 983090983089 microelements) in M oleiferaTe concentrations o macroelements in the powdered lea samples are shown in Figure 983092 and included S Ca K MgNa P Si Cl Al Fe and Mn Te minor elements in thedecreasing order were V Ba Cr Y Ba Zn Rb Ce La

Cu Cs Sn H Co Ni Nb a U Mo Pb Bi Ga T Asand Zr All concentrations were within the recommended

daily allowance (RDA) limits RDA the average daily dietary intake level is expected to be sufficient to meet the nutrientrequirementso all healthy individuals [983090983092ndash983090983094] Tese resultsthereore would suggest that consumption o moringa lea powder can provide users with some o the essential mineralsthat the human body requires or optimum unction

Blood parameter analysis is relevant to risk evaluationas the haematological system has a higher predictive valueor toxicity in humans (983097983089) [983090983095] Moringa oleifera extractadministration was accompanied by a reduction in thehaematopoietic system (ables 983091 and 983092) at a dosage o 983092983088 mgkg which is evidenced by the levels o RBC HCHGB WBC and lymphocytes Te slightly reduced HGB





0

10

20

30

40

50

lowast

A l b u m i n ( g L )

(a)


0

20

40

60

80

G l o b u l i n ( g L )

lowastlowastlowast

(b)

0

20

40

60

80

T o t a l p r o t e i n ( g L )


(c)

00

05

10

15

20

25


I n d i r

b i l i r u b i n ( m o l L )

(d)

0

1

2

3

4

5


Control

40mgmL

80mgmL

200mgmL

1000mgmL

D i r


(e)

0

2

4

6


Control

40mgmL

80mgmL

200mgmL

1000mgmL

T

o t a l b i l i r u b i n ( m o l L )

()

F983145983143983157983154983141 983090 Continued




0

50

100

150

200


A L T ( m o l L

)

lowastlowast

(g)

0

50

100

150

200

A L P ( m o l L )


lowastlowastlowast

(h)

0

100

200

300

400

500

lowastlowast

A S T ( m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(i)

F983145983143983157983154983141 983090 Liver unction tests o a 983089983092-day administration o MOE in male SDRs Values are expressed as means plusmn SEM (1103925 = 7) Values o lt 005 were considered as statistically signi1047297cant lowast



HC RBC and total protein in the rats suggest potentialintererence with the haematopoietic system that wouldrequire urther investigations Te high RBC could also bedue to hypoxic conditions that might have resulted due tothe high doses o MOE administered Te absence o a majorsigni1047297cant change in haematological parameters is consistentwith observations by Awodele et al [983089983094]

Te statistically signi1047297cant increase in lymphocytes atMOE doses o 983092983088ndash983096983088 mgkg and the 983093983088983088983088 mgkg BW in theanimals might suggest the presence o inectionstress or apotential immune boosting effect o MOE at the stated doselevels Since the animals were speci1047297c pathogen-ree andwere

kept in a barrier system the observed effects are likely tobe due to potential immune boosting effects o the extract[983089983091 983089983092 983090983096] Tis needs urther investigation in a subchronicor chronic study

AS AL and ALP are major markers o liver unctionoxic injury to the liver leads to elevation in levels o allthese liver enzymes Tus the observed rise in AL and ALPollowing administration o MOE (ables 983093 and 983094) wouldsuggest a potential adverse effect o MOE on the liver SinceAL is localized primarily in the cytosol o hepatocytes itis a more sensitive marker o hepatocellular damage thanAS and ALP [983090983095 983090983096] Withinlimits these can provide




0

2

4

6

8

C h o l e s t e r o l ( m m o l L )


lowastlowastlowast

(a)

00

02

04

06

08

10

T r i g

( m m o l L )


lowastlowastlowast

(b)

0

1

2

3

4

5

L D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(c)

00

05

10

15

H D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(d)

F983145983143983157983154983141 983091 Lipid pro1047297le o a 983089983092-day administration o MOE in male SDRs Values are expressed as means plusmn SEM (1103925 = 7) Values o lt 005were considered as statistically signi1047297cant lowast



a quantitative assessment o the degree o damage sustainedby the liver Histopathological examinations however didnot reveal anyhistological lesionsin the sinusoids or central

vein (Figure 983096)Te copper component in the extract (able 983090) which is

a component o a number o enzymes that are involved inreducing molecular oxygen metabolizing substances suchas histamine serotonin epinephrine norepinephrine anddopamine could pose a threat during abnormal consumptiono the extract Copper de1047297ciency although rare results inhypochromic anaemia [983090983094] possible side-effects may includeliver damage and Wilsonrsquos syndrome

Direct indirect and total bilirubin re1047298ects the liverrsquosability to take up process and secrete bilirubin into bile andcan also be considered as a true test o liver unction [983090983095 983090983096]Tese were all high at the different doses especially at the983093983088983088983088 mgkg single oral high dose compared to the controls(ables 983093 and 983094) However total protein globulin andalbumin remained relatively unchanged or slightly increasedin all the groups in comparison with the controls Tesewere not entirely dose dependent and signi1047297cant but requireurther monitoring in subchronic studies

It must also be noted that gross pathological examinationo the treated animals did not reveal any abnormalities such




0

20

40

60

80

100

120

140

Average of sample T and K (mg)


S O 3

C a O

K 2 O

M g O

N a 2 O

P 2 O 5

S i O 2

C I

A I 2 O 3

F e 2 O 3

M n O

F983145983143983157983154983141 983092 Chart showing the decreasing order o concentration o elements in the leaves o Moringa oleifera

10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()

F983145983143983157983154983141 983093 Photomicrographs o the heart rom male Sprague-Dawley rats No observable cardiomyopathy Keys (a) control (b) 983093983088983088983088mgkgbwt (c) 983089983088983088983088 mgkg bwt (d) 983090983088983088 mgkg bwt (e) 983096983088 mgkg bwt and () 983092983088 mgkg bwt




10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()

F983145983143983157983154983141 983094 Photomicrographs o the stomach rom male Sprague-Dawley rats No observable ulceration Te epithelial cells are intact Keys(a) control (b) 983093983088983088983088 mgkg bwt (c) 983089983088983088983088 mgkg bwt (d) 983090983088983088 mgkg bwt (e) 983096983088 mgkg bwt and () 983092983088 mgkg bwt

as presence o lesions or changes in colour o internal organsand relative organ weights as compared to the controls

Te kidneys are concerned with the elimination o drugsrom the body and are likely to be affected during such toxi-city studies at high doses In the treated male rats creatininelevelswerereduced compared to the controls ( lt 005)whileurea levels were inconsistent among the groups Tis suggeststhat MOE did not adversely affect the integrity o the renalsystem [983090983097] Also histopathological examinations did notreveal any observablehistologicallesions in theglomeruli andthe tubules (Figure 983095) However the reduction in creatininelevels may be deemed to be positive effect o MOE on therenal system which could be exploited therapeutically Tese1047297ndings are consistent with those reported by Isitua andIbeh983090983088983089983091 [983089983095]

Ingestion o chemicals substances including those o plant origin in excess and especially over long periods may

adversely affect major organs like the heart kidney liverand even the gastrointestinal system Tus the evaluation o histopathological changes in organs remains a cornerstonein assessing the saety o medicines and other substances[983091983088 983091983089] Histopathological examinations o the heart andstomach did not reveal any observable cardiomyopathies(Figure 983093) and ulcerations o the epithelial cells in the malerats (Figure 983094) Tese 1047297ndings are consistent with otherstudies [983091983090] It might be expected that at high doses o MOE may result in accumulation o MOE-derived iron Tismineral though an important component o haemoglobinand other proteins and enzymes may cause gastrointestinaldistress hemochromatosis and so orth [983090983090 983090983091] Te absence




10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()

F983145983143983157983154983141 983095 Photomicrographs o the kidney rom male Sprague-Dawley rats Tere were no observable histological lesions in the glomerulusand the tubules Keys (a) control (b) 983093983088983088983088 mgkg bwt (c) 983089983088983088983088 mgkg bwt (d) 983090983088983088 mgkg bwt (e) 983096983088 mgkg bwt and () 983092983088 mgkg bwt

o these effects may be explained by the act that the variouselements were within normal limits [983089983094]

It is important to note that the effects o chemicals

produced in laboratory animals when properly conductedprovide a useul indication o saety in humans [983091983091] Tusrom the above results it would be expedient to conductsubchronic or chronic toxicity studies with regard to thehematopoietic renal hepatic and reproductive changesbecause MOE is used as a ood supplement and is usedover a long period o time by consumers Moreover thecontamination o moringa leaves by heavy metals like lead(Pb) and arsenic (As) may pose a threat to humans becausethey are not biodegradable With reerence to the RDAlimits and other studies conducted regarding the presence o heavy metals in the leaves o MOE [983091983092 983091983093] it is imperativethat prevention o any possible cumulative toxicity o some

o the elements is most desirable during prolonged use o MOE Tere is thus the need or toxicological studies to beconducted on moringa rom different locations in order to

give an indication o their comparative saetyIt must however be stated that because plants may

absorb elements rom the soil and environment some o which may be toxic to humans plant nutrition climateand soil conditions and locations could also determine theelemental contents in the leaves [983089983097 983091983092 983091983093]

5 Conclusion

Tis study has provided some evidence that Moringa oleiferathat was collected in Accra Ghana in West Arica is reason-ably sae or consumption taking into account the elemental




10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()

F983145983143983157983154983141 983096 Photomicrographs o the liver rom male Sprague-Dawley rats Tere were no observable histological lesions in the sinusoids andcentral vein Keys (a) control (b) 983093983088983088983088 mgkg bwt (c) 983089983088983088983088 mgkg bwt (d) 983090983088983088 mgkg bwt (e) 983096983088 mgkg bwt and () 983092983088 mgkg bwt

composition when administered to rats Te acute studies

showed that the median lethal dose (LD50) could be greaterthan 983093983088983088983088 mgkg as all the animals survived at his doselevel It is also important to monitor the concentration o itselemental composition as a possible reason o serious healthalterations

6 Recommendation

Based on the levels o these minerals and the permissibleamount in the human body it is recommended that theconsumption o Moringa oleifera leaves be limited to amaximum o 983095983088 grams per day in order to prevent excessiveconsumption and subsequent accumulation o some o these

essential elements At 983095983088 grams per day most o these

elements in the leaves could be ound in high quantitiesapproaching the RDA limit

Conflict of Interests

Te authors declare that there is no con1047298ict o interestsregarding the publication o this paper

Acknowledgment

Te authorsrsquo appreciation goes to Dr Daniel Boamah and hisstaff at the geochemistry department o the Ghana geological




983137983138983148983141 983093

Parameters Groups

Control males 983093983088983088983088 mgkg males

Urea 983093983091 plusmn 983088983089983096 983093983096983091 plusmn 983088983096

Creatinine 983095983092983095 plusmn 983091983088983095 2714 plusmn 087lowastlowastlowast

protein 983093983097983097983094 plusmn 983089983092983092 983094983095983093 plusmn 983090983097983088

Albumin 983091983093 plusmn 983088983091983089 983091983094983088983096 plusmn 983088983097983097

Globulin 983090983092983094983094 plusmn 983089983092983094 983091983089983089983094 plusmn 983090983091

Dir Bil 983090983088983090 plusmn 983088983089983094 542 plusmn 100lowastlowastlowast

Indirct Bil 983089983093 plusmn 983088983090983097 983090983095983090 plusmn 983088983093983097

Bil 983091983092983092 plusmn 983088983090983097 616 plusmn 114lowast

AL 983093983090983090983096 plusmn 983088983090983097 983089983089983091983093 plusmn 983095983089983097

AS 983089983097983088983089 plusmn 983092983097983094 983090983090983092983092 plusmn 983093983091983096983097

ALP 983089983093983089983090 plusmn 983088983089983097983094 2769 plusmn 0045lowastlowastlowast

Cholesterol 983093983094983097 plusmn 983088983090 418 plusmn 001lowastlowastlowast

rig 983088983096983095 plusmn 983088983088983089 04 plusmn 0003lowastlowastlowast

LDL 983092983088983090 plusmn 983088983088983092 311 plusmn 0004lowastlowastlowast

HDL 983089983091983088 plusmn 983088983088983089 983088983096983096 plusmn 983088983088983089

Tese are statistical signi1047297cance lowast lt 005 lowastlowast

lt 001 lowastlowastlowast lt 00001

983137983138983148983141 983094

Parameters Groups

Control males 983089983088983088983088 mgkg males 983090983088983088mgkg males 983096983088 mgkg males 983092983088mgkg males value

Urea 983093983091 plusmn 983088983089983096 983094983089983091 plusmn 983088983093983090 983093983088983089 plusmn 983088983090983095 983096983091983092 plusmn 983088983090983095 983093983093983097 plusmn 983088983092983097 983088983090983089983092983091

Creatinine 983095983092983095 plusmn 983091983088983095 983091983097983094983094 plusmn 983089983091983090983091lowast 983093983090983092 plusmn 983092983088983091 983094983089983088983092 plusmn 983091983096983092 983091983095983093983090 plusmn 983095983090983095lowastlowast 983088983088983088983088983093

protein 983093983097983097983094 plusmn 983089983092983092 983094983089983093983090 plusmn 983090983094983096 983094983089983096983090 plusmn 983089983091983096 983094983088983097983090 plusmn 983089983090983094 983094983093983096983092 plusmn 983089983097983096 983088983088983097983089983091

Albumin 983091983093 plusmn 983088983091983089 983092983088983093983094 plusmn 983090983090983092lowast 983091983094983097983090 plusmn 983088983095983093 983091983090983089983090 plusmn 983088983094983094 983091983095983095 plusmn 983089983090983091 983088983088983088983089983094

Globulin 983090983092983094983094 plusmn 983089983092983094 983090983091983094983092 plusmn 983090983094983088 983090983093983089 plusmn 983089983095983089 983090983096983096983088 plusmn 983088983097983089 983093983089983095983092 plusmn 983094983096983091lowastlowastlowast 983088983088983088983088983089

Dir bil 983090983088983090 plusmn 983088983089983094 983092983089983090 plusmn 983088983090983094 983090983097983090 plusmn 983088983091983095 983090983094983096 plusmn 983088983090983096 983090983097983096 plusmn 983088983088983093 983088983088983088983088983092

Indirdt bil 983089983093 plusmn 983088983090983097 983088983097983090 plusmn 983088983088983095 983089983097 plusmn 983088983091983093 983089983092983094 plusmn 983088983088983094 983089983095983092 plusmn 983088983088983090 983088983088983089983090983088 bil 983091983092983092 plusmn 983088983091 983092983097 plusmn 983088983091983088 983092983094983092 plusmn 983088983090983089 983091983097983090 plusmn 983088983088983095 983092983091983092 plusmn 983088983090983090 983088983088983089983096983096

AL 983093983090983090983096 plusmn 983088983090983097 983089983088983097983096 plusmn 983097983088983094 983097983091983097983090 plusmn 983090983097983091 983089983091983089983091 plusmn 983090983093983090983097lowastlowast 983095983095983097983096 plusmn 983089983094983094983095 983088983088983088983092983097

AS 983089983097983088983089 plusmn 983092983097983094 983091983097983095983093 plusmn 983088983088983092 983091983094983092983093 plusmn 983090983090983090983089 983091983091983095983091 plusmn 983089983089983090983092 983090983093983091983091 plusmn 983095983096983097983089 983088983089983097983090983096

ALP 983089983093983089983090 plusmn 983088983089983097983094 983089983088983094983088 plusmn 983088983088983092983097lowastlowastlowast 983089983088983091983089 plusmn 983088983088983092lowastlowastlowast 983089983094983093983089 plusmn 983088983088983091983095lowastlowastlowast 983097983093983096983092 plusmn 983088983088983093983089lowastlowastlowast 983088983088983088983088983089

Cholesterol 983093983094983097 plusmn 983088983090 983091983096983094 plusmn 983088983088983090lowastlowastlowast 983093983090983096 plusmn 983088983088983092lowastlowastlowast 983092983091983097 plusmn 983088983088983088983093lowastlowastlowast 983092983090983091 plusmn 983088983089983088lowastlowastlowast 983088983088983088983088983089

rig 983088983096983095 plusmn 983088983088983089 983088983092 plusmn 983088983088983089lowastlowastlowast 983088983092 plusmn 983088983088983089lowastlowastlowast 983088983094983097 plusmn 983088983088983089lowastlowastlowast 983088983091983096 plusmn 983088983088983089lowastlowastlowast 983088983088983088983088983089

LDL 983092983088983090 plusmn 983088983088983092 983090983092983097 plusmn 983088983088983089 983091983095983096 plusmn 983088983088983089 983091983089983097 plusmn 983088983088983088983092 983088983096 plusmn 983088983088983088983093 983088983088983088983088983089

HDL 983089983091983088 plusmn 983088983088983089 983089983090983088 plusmn 983088983088983089 983089983090983090 plusmn 983088983088983089 983088983096983096 plusmn 983088983088983089 983088983097983096 plusmn 983088983088983089 983088983088983088983088983089



survey department or the assistance and usage o their

EDXRF equipment

References

[983089] P F Raguindin L F Dans and J F King ldquo Moringa oleifera as aGalactagoguerdquo Breastfeeding Medicine vol 983097 pp 983091983090983091ndash983091983090983092 983090983088983089983092

[983090] M K iti E Harijono and S W Endang ldquoEffect lactagoguemoringa leaves ( Moringa oleifera Lam) powder in rats whiteemale wistarrdquo Journal of Basic and Applied Scienti1047297c Research vol 983091 no 983092 pp 983092983091983088ndash983092983091983092 983090983088983089983091

[983091] JW Fahey ldquoA review o the medical evidence or itsnutritionaltherapeutic and prophylactic properties Part 983089rdquo rees for Life Journal vol 983089 pp 983093ndash983089983093 983090983088983088983093

[983092] S WadhwaM S Panwar N SainiS S Rawat andS Singhal ldquoA

review on commercial traditional uses phytoconstituents andpharmacological activity o Moringa oleiferardquo Global Journal of raditional Medicinal Systems vol 983090 no 983089 pp 983089ndash983089983091 983090983088983089983091

[983093] rees or Lie(FL) Moringa Book 983090983088983088983093 httpwwwtreeorlieorg

[983094] A H Mollik ldquoPlants rom Sundarbans to the diet o lactatingmothers during puerperium o Barguna district o BangladeshrdquoPediatric Nephrology vol 983090983093 article 983089983097983088983092 abstract 983075983090983097983096 983090983088983089983088

[983095] M R Sulaiman Z A Zakaria A S Bujarimin M N SomchitD A Isra and S Moin ldquoEvaluation o moringa oleiera aque-ous extract or antinociceptive and anti-in1047298ammatory activitiesin animal modelsrdquo Pharmaceutical Biology vol 983092983094 no 983089983090 pp983096983091983096ndash983096983092983093 983090983088983088983096




[983096] B S Rathi S L Bodhankar and A M Baheti ldquoEvaluationo aqueous leaves extract o Moringa oleiera Linn or woundhealing in albino ratsrdquo Indian Journal of Experimental Biology vol 983092983092 no 983089983089 pp 983096983097983096ndash983097983088983089 983090983088983088983094

[983097] S Luqman S Srivastava R Kumar A K Maurya and DChanda ldquoExperimental assessment o Moringa oleifera lea andruit or its antistress antioxidant and scavenging potential

using in vitro and in vivo assaysrdquo Evidence-Based Complemen-tary and Alternative Medicine vol 983090983088983089983090 Article ID 983093983089983097983088983096983092 983089983090pages 983090983088983089983090

[983089983088] S Luqman and R Kumar ldquoAttenuation o hydroxyl radicalormation by extracted constituent o moringa oleiera lamrdquoCurrent Chemical Biology vol 983093 no 983091 pp 983090983089983091ndash983090983089983096 983090983088983089983089

[983089983089] A R Verma M Vijayakumar C S Mathela and C V Rao ldquoIn vitro and in vivo antioxidant properties o different ractions o Moringa oleiera leavesrdquo Food and Chemical oxicology vol 983092983095no 983097 pp 983090983089983097983094ndash983090983090983088983089 983090983088983088983097

[983089983090] I L Jung ldquoSoluble extract rom Moringa oleifera leaves with anew anticancer activityrdquo PLoS ONE vol 983097 no 983092 983090983088983089983092

[983089983091] D K Dubey J Dora A Kumar and R K Gulsan ldquoAmultipurpose treemdashMoringa oleierardquo International Journal of Pharmaceutical and Chemical Sciences vol 983093 no 983090 pp 983089983088983090ndash983089983088983093983090983088983089983092

[983089983092] J Mehta A Shukla V Bukhariya and R Charde ldquoTe magicremedy o Moringa oleiera an overviewrdquo International Journal of Biomedical and Advance Research vol 983090 no 983093 pp 983090983089983093ndash983090983090983095983090983088983089983089

[983089983093] A A Adedapo O M Mogbojuri and B O Emikpe ldquoSaety evaluations o the aqueous extract o the leaves o Moringaoleifera in ratsrdquo Journal of Medicinal Plants Research vol 983091 no983096 pp 983093983096983094ndash983093983097983089 983090983088983088983097

[983089983094] O Awodele I A Oreagba S Odoma J A eixeira Da Silvaand V O Osunkalu ldquooxicological evaluation o the aqueouslea extract o Moringa oleiera Lam (Moringaceae)rdquo Journal of

Ethnopharmacology vol 983089983091983097 no 983090 pp 983091983091983088ndash983091983091983094 983090983088983089983090[983089983095] C C Isitua andI N Ibeh ldquooxicological assessment o aqueousextract o Moringa oleiera and Caulis bambusae leaves inrabbitsrdquo Journal of Clinical oxicology supplement 983089983090 article983088983088983091 983090983088983089983091

[983089983096] J O Ashong and D L Brown ldquoSaety and efficacy o Moringaoleifera powder or growing poultryrdquo Journal of Animal Science vol 983096983097 E-supplement 983089 p 983096983092 983090983088983089983089

[983089983097] K Freer Exposing the Lies and Deception Behind Organic Moringa oleifera Leaf Powder Not All Moringa Oleifera Products Are Created Equal You Might Be Shocked at the ruth Kate Freer Yahoo Contributor Network 983090983088983089983088

[983090983088] D Jaiswal P K Rai S Mehta et al ldquoRole o Moringa oleierain regulation o diabetes-induced oxidative stressrdquo Asian Paci1047297c

Journal of ropical Medicine vol 983094 no 983094 pp 983092983090983094ndash983092983091983090 983090983088983089983091[983090983089] World Health Organization Quality ControlMethods for Medic-

inal Plant Materials WHO Offset Publication WHO GenevaSwitzerland 983089983097983097983096

[983090983090] WHO Guidelines for the Assessment of Herbal MedicinesWHORM983097983089983092 WHO Geneva Switzerland 983089983097983097983089

[983090983091] Organization or Economic Cooperation and Development(OECD) Guidelines OECD Guidelines for esting of Chemicals Acute Oral oxicity-Fixed Dose Procedure 983092983090983088 983090983088983088983089

[983090983092] R E Lopez de Ruiz R A Olsina and A N Masi ldquoDifferentanalytical methodologies or the preconcentration and deter-mination o trace chromium by XRF in medicinal herbs witheffects on metabolismrdquo X-Ray Spectrometry vol 983091983089 no 983090 pp983089983093983088ndash983089983093983091 983090983088983088983090

[983090983093] M J Anjos R Lopes E F O Jesus S M Simabuco and RCesareo ldquoQuantitative determination o metals in radish usingx-ray 1047298uorescence spectrometryrdquo X-Ray Spectrometry vol 983091983089no 983090 pp 983089983090983088ndash983089983090983091 983090983088983088983090

[983090983094] C Vazquez N Barbara and S Lopez ldquoXRF analysis o micronutrients in endive grown on soils with sewage sludgerdquo X-Ray Spectrometry vol 983091983090 no 983089 pp 983093983095ndash983093983097 983090983088983088983091

[983090983095] aconic echnical Library Hematological Clinical Chemistry values Sprague- Dawley Rats 983090983088983088983091

[983090983096] C K Janeway ldquoImmunobiologyrdquo in Te Immune System inHealth and Disease E Lawrence Ed pp 983092983094983089ndash983092983094983091 GarlandScience New York NY USA 983094th edition 983090983088983088983093

[983090983097] W Arneson and J Brickell ldquoAssessment o liver unctionrdquo inClinicalChemistry A Laboratory Perspective pp 983090983091983091ndash983090983094983094DavisDavis Company Philadelphia Pa USA 983089st edition 983090983088983088983095

[983091983088] S K Ramaiah ldquoA toxicologist guide to the diagnostic interpre-tation o hepatic biochemical parametersrdquo Food and Chemical oxicology vol 983092983093 no 983097 pp 983089983093983093983089ndash983089983093983093983095 983090983088983088983095

[983091983089] P Greaves Histopathology of Preclinical oxicity Studies Inter- pretation and Relevance in Drug Safety Evaluation Academic

Press New York NY USA 983091rd edition 983090983088983088983095[983091983090] P I Zvinorova L Lekhanya K Erlwanger and E Chivandi

ldquoDietary effects o Moringa oleifera lea powder on growthgastrointestinal morphometry and blood and liver metabolitesin Sprague-Dawley ratsrdquo Journal of Animal Physiology and Animal Nutrition 983090983088983089983092

[983091983091] H Olson G Betton D Robinsdon et al ldquoConcordance o tox-icity o pharmaceuticals in humans and in animalsrdquo Regulatory oxicology and Pharmacology vol 983091983090 pp 983093983094ndash983094983095 983090983088983088983088

[983091983092] C Limmatvapirat S Limmatvapirat J Charoenteeraboon and Phaechamud ldquoInductively coupled plasma mass spectro-metric determination o heavy metals in Moringa oleieraLam leavesrdquo Research Journal of Pharmaceutical Biological and Chemical Sciences vol 983092 no 983089 pp 983089983094983089ndash983089983094983096 983090983088983089983091

[983091983093] K Annan R Dickson I Amponsah and I Nooni ldquoTe heavy metal contents o some selected medicinal plants sampled romdifferent geographical locationsrdquo Pharmacognosy Research vol983093 no 983090 pp 983089983088983091ndash983089983088983096 983090983088983089983091



Submit your manuscripts at

httpwwwhindawicom




In Ghana and most parts o West Arica powdered leaveso M oleifera are marketed under different brand names andconsumed daily as a nutraceutical prophylaxis or cure or

various conditions Te prevailing socio-culturaleconomicconditions and consequent difficulty in accessing healthcareservices especially by rural populations as well as the general

perception that plant medicines are efficacious and reerom side effects [983089983097 983090983088] will increase the requent andwidespread spread use o moringa lea powder In view o the reported differences in moringa lea products which may impact adversely on itssaety the micro-and macroelementalcontent o dried leaves o Moringa oleifera were measured Inaddition the lea extract was investigated or its toxicologicaleffects in Sprague-Dawley rats

2 Materials and Methods

983090983089 Preparation of Extracts of Moringa oleifera Leaves forAnal- ysis In order to mimic the traditional method o extraction983090983096 kg o sample were blended with boiled distilled water

Te mixture covered with water was lef to stand overnightin a water bath maintained at 983094983088∘C and the watery portion(inusion) was 1047297ltered off using 983088983092983093 1038389m millipore celluloseester 1047297lters and reeze dried to obtain M oleifera extract(MOE) Portions were homogenized and labelled as sampleA or elemental analysis

983090983090 Preparation of Moringa oleifera Leaves for AnalysisLeaves o the Moringa oleifera species were collected romAccra separated rom other plant parts washed and driedin the shade or 1047297ve days at room temperature ground into1047297ne powder sieved (983090983089983090 1038389m mesh size) to obtain very 1047297nesamples and kept in separate containers Tis was labelled

sample B

983090983091 Pelleting of Samples for Analysis Te loosepowdery nature o samples (plant extracts A and lea powder B)required that they were pelleted beore analysis Beore pel-leting 983092 g o each sample was weighed and 983088983097 g o the binderFluxana (H Elektronic BM-983088983088983088983090-983089 (Licowax C micropowderPM-Hoechstwax)) was added and homogenized or 983091 min-utes Te mixture was pressed at 983090983088 t or 983090 minutes into pelletso 983091983090 mm in diameter or the subsequent XRF measurementsTree separate pellets were prepared rom each o samples Aand B

983090983092 Energy Dispersive X-Ray (ED XRF) MeasurementsEnergy dispersive X-ray (ED XRF) was used or simultaneousanalysis and measurement o the elemental content o thesamples Te procedure which used three-axial geometryreduced background noise due to radiation polarization Temonochromatic radiations emitted rom the X-ray tube wereapplied to excite the atoms o the sample

983090983093 Experimental Animals and Housing Conditions MaleSprague-Dawley rats (983089983093983088 gndash983089983096983088 g body weight) were pur-chased rom the Centre or Scienti1047297c Research into PlantMedicine Mampong Te rats were housed in plastic cageswith stainless steel tops in the animal care acility o theUniversity o Ghana Medical School and kept under standard

983089983090 h light and 983089983090 h dark schedule where room temperaturehumidity and ventilation werecontrolled during the acclima-tization period o seven (983095) days

983090983094 Acute and Subacute oxicity Studies Te reconstitutedpowdered MOE mixturewas preparedusingdistilled water as

the vehicle and administered as a single dose (983093983088983088983088 mgkg)by oral gavage or the acute toxicity studies Afer theadministration animals were observed every hour or the1047297rst 983094 hours then daily or the next 983089983091 days

In the subacute studies 1047297ve groups (983089ndash983093) o maleSprague-Dawley Rats (eight weeks old seven animalsgroup) o meanweight 983089983093983088 g were prepared Animals in each o the ourgroups were administered MOE extract over a dose range o 983088 mgkg to 983089983088983088983088kg by oral gavage daily or 983089983092 days Animalsin the 1047297fh group received the vehicle (distilled water)andserved as controls

For the subacute studies afer the983089983092 days o MOE admin-istration o MOE each animal was observed every hour orsix hours daily or the next 983091 days and subsequently every day or 983089983088 days Animals were ed ad libitum with standard chow diet (AIN-983097983091G ormulation obtained rom GAFCO-Ghana)

983090983094983089 Observed Clinical Signs of oxidromes Body weightswere observed daily beore and afer administration Inaddition animals were observed daily or clinical signs o excitability twitching salivation morbidity miosis mydria-sis rising ur sluggish movement draping tremors and soorth

983090983095 Laboratory Examinations

983090983095983089 Blood Samples Under chloroorm anaesthesia blood

was obtained via cardiac puncture and the animals euth-anized by exsanguinations Samples o the blood collectedwere aliquoted into EDA-983090K tubes and plain tubes respec-tively Te EDA blood was immediately analysed or haema-tological parameters using the SYSMEX Haematology Auto-analyser [Kobe Japan] while serum prepared rom blood inthe plain tubes was used or biochemical examinations

983090983096 Necropsy and Histopathological Studies Gross patholog-ical investigations were conducted on each animal ollowingexsanguination Te heart kidney liver and gastrointesti-nal tract were prepared or histopathological examinationsAll experimental procedures and assays were conducted inaccordance with the international guidelines or evaluatingthe saety o herbal medicines [983090983089ndash983090983091]

983090983097 Statistical Analysis Results are presented as means plusmnSEM and analysis or statistical differences was done usingone-way ANOVA ollowed by Bonferroni post-hoc test

values less than 983088983088983093 were considered statistically signi1047297cant

3 Results

Te ED-XRF analyses led to the detection o a total o thirty-1047297ve (983091983093) elements comprising eleven (983089983089) major elementsand twenty-our (983090983092) minor elements Te major elementsdetected included Na Mg Al Si P S Cl K Ca Mn and Fe






Na983090O 983091983095983093

MgO 983094983088983094

Al983090O

983091 983089983091983088

SiO2

983091983088983088

P983090O983093 983091983095983088

SO3

983089983090983090983096

Cl 983089983095983094

K983090O 983095983096983092

CaO 983089983089983096983088

MnO 983088983092

Fe983090O983091 983091983088



Cr 983088983093983095983096

Co 983088983088983089983090

Ni 983088983088983089983092983096

Cu 983088983088983091983089983096

Zn 983088983089983089983093983094

Ga 983088983088983088983091983090

As 983088983088983088983090983090

Rb 983088983088983095983093983096

Y 983088983091983093983092983090

Zr 983088983088983088983090983092

Nb 983088983088983089983089983094

Mo 983088983088983088983093983094Sn 983088983088983093983095983094

Cs 983088983088983090983092983090

Ba 983088983096983097983088983088

La 983088983088983092983097983088

Ce 983088983088983094983097983092

H 983088983088983089983093983092

a 983088983088983089983089983092

Pb 983088983088983088983092983092

T 983088983088983088983091983090






Parameters Groups












4 Discussion






Parameters Groups



RBC (983089983088983094

1038389L)


lowastlowastlowast

655 plusmn 049

lowast










0

2

4

6

8

10

Control


40mgmL

80mgmL

200mgmL

1000mgmL

(a)

0

20

40

60

80

100

Control

40mgmL

80mgmL

200mgmL

1000mgmL


lowastlowast

lowast


(b)














0

10

20

30

40

50

lowast


(a)


0

20

40

60

80


lowastlowastlowast

(b)

0

20

40

60

80



(c)

00

05

10

15

20

25


I n d i r


(d)

0

1

2

3

4

5


Control

40mgmL

80mgmL

200mgmL

1000mgmL

D i r


(e)

0

2

4

6


Control

40mgmL

80mgmL

200mgmL

1000mgmL

T


()

F983145983143983157983154983141 983090 Continued




0

50

100

150

200


A L T ( m o l L

)

lowastlowast

(g)

0

50

100

150

200

A L P ( m o l L )


lowastlowastlowast

(h)

0

100

200

300

400

500

lowastlowast

A S T ( m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(i)











0

2

4

6

8



lowastlowastlowast

(a)

00

02

04

06

08

10

T r i g

( m m o l L )


lowastlowastlowast

(b)

0

1

2

3

4

5

L D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(c)

00

05

10

15

H D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(d)












0

20

40

60

80

100

120

140



S O 3

C a O

K 2 O

M g O

N a 2 O

P 2 O 5

S i O 2

C I

A I 2 O 3

F e 2 O 3

M n O


10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()









10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()








5 Conclusion





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()




6 Recommendation






Acknowledgment





983137983138983148983141 983093

Parameters Groups










AL 983093983090983090983096 plusmn 983088983090983097 983089983089983091983093 plusmn 983095983089983097

AS 983089983097983088983089 plusmn 983092983097983094 983090983090983092983092 plusmn 983093983091983096983097








983137983138983148983141 983094

Parameters Groups



















EDXRF equipment

References














































httpwwwhindawicom






Na983090O 983091983095983093

MgO 983094983088983094

Al983090O

983091 983089983091983088

SiO2

983091983088983088

P983090O983093 983091983095983088

SO3

983089983090983090983096

Cl 983089983095983094

K983090O 983095983096983092

CaO 983089983089983096983088

MnO 983088983092

Fe983090O983091 983091983088



Cr 983088983093983095983096

Co 983088983088983089983090

Ni 983088983088983089983092983096

Cu 983088983088983091983089983096

Zn 983088983089983089983093983094

Ga 983088983088983088983091983090

As 983088983088983088983090983090

Rb 983088983088983095983093983096

Y 983088983091983093983092983090

Zr 983088983088983088983090983092

Nb 983088983088983089983089983094

Mo 983088983088983088983093983094Sn 983088983088983093983095983094

Cs 983088983088983090983092983090

Ba 983088983096983097983088983088

La 983088983088983092983097983088

Ce 983088983088983094983097983092

H 983088983088983089983093983092

a 983088983088983089983089983092

Pb 983088983088983088983092983092

T 983088983088983088983091983090






Parameters Groups












4 Discussion






Parameters Groups



RBC (983089983088983094

1038389L)


lowastlowastlowast

655 plusmn 049

lowast










0

2

4

6

8

10

Control


40mgmL

80mgmL

200mgmL

1000mgmL

(a)

0

20

40

60

80

100

Control

40mgmL

80mgmL

200mgmL

1000mgmL


lowastlowast

lowast


(b)














0

10

20

30

40

50

lowast


(a)


0

20

40

60

80


lowastlowastlowast

(b)

0

20

40

60

80



(c)

00

05

10

15

20

25


I n d i r


(d)

0

1

2

3

4

5


Control

40mgmL

80mgmL

200mgmL

1000mgmL

D i r


(e)

0

2

4

6


Control

40mgmL

80mgmL

200mgmL

1000mgmL

T


()

F983145983143983157983154983141 983090 Continued




0

50

100

150

200


A L T ( m o l L

)

lowastlowast

(g)

0

50

100

150

200

A L P ( m o l L )


lowastlowastlowast

(h)

0

100

200

300

400

500

lowastlowast

A S T ( m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(i)











0

2

4

6

8



lowastlowastlowast

(a)

00

02

04

06

08

10

T r i g

( m m o l L )


lowastlowastlowast

(b)

0

1

2

3

4

5

L D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(c)

00

05

10

15

H D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(d)












0

20

40

60

80

100

120

140



S O 3

C a O

K 2 O

M g O

N a 2 O

P 2 O 5

S i O 2

C I

A I 2 O 3

F e 2 O 3

M n O


10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()









10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()








5 Conclusion





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()




6 Recommendation






Acknowledgment





983137983138983148983141 983093

Parameters Groups










AL 983093983090983090983096 plusmn 983088983090983097 983089983089983091983093 plusmn 983095983089983097

AS 983089983097983088983089 plusmn 983092983097983094 983090983090983092983092 plusmn 983093983091983096983097








983137983138983148983141 983094

Parameters Groups



















EDXRF equipment

References














































httpwwwhindawicom





Parameters Groups



RBC (983089983088983094

1038389L)


lowastlowastlowast

655 plusmn 049

lowast










0

2

4

6

8

10

Control


40mgmL

80mgmL

200mgmL

1000mgmL

(a)

0

20

40

60

80

100

Control

40mgmL

80mgmL

200mgmL

1000mgmL


lowastlowast

lowast


(b)














0

10

20

30

40

50

lowast


(a)


0

20

40

60

80


lowastlowastlowast

(b)

0

20

40

60

80



(c)

00

05

10

15

20

25


I n d i r


(d)

0

1

2

3

4

5


Control

40mgmL

80mgmL

200mgmL

1000mgmL

D i r


(e)

0

2

4

6


Control

40mgmL

80mgmL

200mgmL

1000mgmL

T


()

F983145983143983157983154983141 983090 Continued




0

50

100

150

200


A L T ( m o l L

)

lowastlowast

(g)

0

50

100

150

200

A L P ( m o l L )


lowastlowastlowast

(h)

0

100

200

300

400

500

lowastlowast

A S T ( m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(i)











0

2

4

6

8



lowastlowastlowast

(a)

00

02

04

06

08

10

T r i g

( m m o l L )


lowastlowastlowast

(b)

0

1

2

3

4

5

L D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(c)

00

05

10

15

H D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(d)












0

20

40

60

80

100

120

140



S O 3

C a O

K 2 O

M g O

N a 2 O

P 2 O 5

S i O 2

C I

A I 2 O 3

F e 2 O 3

M n O


10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()









10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()








5 Conclusion





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()




6 Recommendation






Acknowledgment





983137983138983148983141 983093

Parameters Groups










AL 983093983090983090983096 plusmn 983088983090983097 983089983089983091983093 plusmn 983095983089983097

AS 983089983097983088983089 plusmn 983092983097983094 983090983090983092983092 plusmn 983093983091983096983097








983137983138983148983141 983094

Parameters Groups



















EDXRF equipment

References














































httpwwwhindawicom





0

10

20

30

40

50

lowast


(a)


0

20

40

60

80


lowastlowastlowast

(b)

0

20

40

60

80



(c)

00

05

10

15

20

25


I n d i r


(d)

0

1

2

3

4

5


Control

40mgmL

80mgmL

200mgmL

1000mgmL

D i r


(e)

0

2

4

6


Control

40mgmL

80mgmL

200mgmL

1000mgmL

T


()

F983145983143983157983154983141 983090 Continued




0

50

100

150

200


A L T ( m o l L

)

lowastlowast

(g)

0

50

100

150

200

A L P ( m o l L )


lowastlowastlowast

(h)

0

100

200

300

400

500

lowastlowast

A S T ( m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(i)











0

2

4

6

8



lowastlowastlowast

(a)

00

02

04

06

08

10

T r i g

( m m o l L )


lowastlowastlowast

(b)

0

1

2

3

4

5

L D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(c)

00

05

10

15

H D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(d)












0

20

40

60

80

100

120

140



S O 3

C a O

K 2 O

M g O

N a 2 O

P 2 O 5

S i O 2

C I

A I 2 O 3

F e 2 O 3

M n O


10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()









10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()








5 Conclusion





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()




6 Recommendation






Acknowledgment





983137983138983148983141 983093

Parameters Groups










AL 983093983090983090983096 plusmn 983088983090983097 983089983089983091983093 plusmn 983095983089983097

AS 983089983097983088983089 plusmn 983092983097983094 983090983090983092983092 plusmn 983093983091983096983097








983137983138983148983141 983094

Parameters Groups



















EDXRF equipment

References














































httpwwwhindawicom




0

50

100

150

200


A L T ( m o l L

)

lowastlowast

(g)

0

50

100

150

200

A L P ( m o l L )


lowastlowastlowast

(h)

0

100

200

300

400

500

lowastlowast

A S T ( m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(i)











0

2

4

6

8



lowastlowastlowast

(a)

00

02

04

06

08

10

T r i g

( m m o l L )


lowastlowastlowast

(b)

0

1

2

3

4

5

L D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(c)

00

05

10

15

H D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(d)












0

20

40

60

80

100

120

140



S O 3

C a O

K 2 O

M g O

N a 2 O

P 2 O 5

S i O 2

C I

A I 2 O 3

F e 2 O 3

M n O


10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()









10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()








5 Conclusion





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()




6 Recommendation






Acknowledgment





983137983138983148983141 983093

Parameters Groups










AL 983093983090983090983096 plusmn 983088983090983097 983089983089983091983093 plusmn 983095983089983097

AS 983089983097983088983089 plusmn 983092983097983094 983090983090983092983092 plusmn 983093983091983096983097








983137983138983148983141 983094

Parameters Groups



















EDXRF equipment

References














































httpwwwhindawicom




0

2

4

6

8



lowastlowastlowast

(a)

00

02

04

06

08

10

T r i g

( m m o l L )


lowastlowastlowast

(b)

0

1

2

3

4

5

L D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(c)

00

05

10

15

H D L ( m m o l L )


Control

40mgmL

80mgmL

200mgmL

1000mgmL

(d)












0

20

40

60

80

100

120

140



S O 3

C a O

K 2 O

M g O

N a 2 O

P 2 O 5

S i O 2

C I

A I 2 O 3

F e 2 O 3

M n O


10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()









10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()








5 Conclusion





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()




6 Recommendation






Acknowledgment





983137983138983148983141 983093

Parameters Groups










AL 983093983090983090983096 plusmn 983088983090983097 983089983089983091983093 plusmn 983095983089983097

AS 983089983097983088983089 plusmn 983092983097983094 983090983090983092983092 plusmn 983093983091983096983097








983137983138983148983141 983094

Parameters Groups



















EDXRF equipment

References














































httpwwwhindawicom




0

20

40

60

80

100

120

140



S O 3

C a O

K 2 O

M g O

N a 2 O

P 2 O 5

S i O 2

C I

A I 2 O 3

F e 2 O 3

M n O


10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()









10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()








5 Conclusion





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()




6 Recommendation






Acknowledgment





983137983138983148983141 983093

Parameters Groups










AL 983093983090983090983096 plusmn 983088983090983097 983089983089983091983093 plusmn 983095983089983097

AS 983089983097983088983089 plusmn 983092983097983094 983090983090983092983092 plusmn 983093983091983096983097








983137983138983148983141 983094

Parameters Groups



















EDXRF equipment

References














































httpwwwhindawicom




10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()









10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()








5 Conclusion





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()




6 Recommendation






Acknowledgment





983137983138983148983141 983093

Parameters Groups










AL 983093983090983090983096 plusmn 983088983090983097 983089983089983091983093 plusmn 983095983089983097

AS 983089983097983088983089 plusmn 983092983097983094 983090983090983092983092 plusmn 983093983091983096983097








983137983138983148983141 983094

Parameters Groups



















EDXRF equipment

References














































httpwwwhindawicom




10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()








5 Conclusion





10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()




6 Recommendation






Acknowledgment





983137983138983148983141 983093

Parameters Groups










AL 983093983090983090983096 plusmn 983088983090983097 983089983089983091983093 plusmn 983095983089983097

AS 983089983097983088983089 plusmn 983092983097983094 983090983090983092983092 plusmn 983093983091983096983097








983137983138983148983141 983094

Parameters Groups



















EDXRF equipment

References














































httpwwwhindawicom




10x

(a)

10x

(b)

10x

(c)

10x

(d)

10x

(e)

10x

()




6 Recommendation






Acknowledgment





983137983138983148983141 983093

Parameters Groups










AL 983093983090983090983096 plusmn 983088983090983097 983089983089983091983093 plusmn 983095983089983097

AS 983089983097983088983089 plusmn 983092983097983094 983090983090983092983092 plusmn 983093983091983096983097








983137983138983148983141 983094

Parameters Groups



















EDXRF equipment

References














































httpwwwhindawicom




983137983138983148983141 983093

Parameters Groups










AL 983093983090983090983096 plusmn 983088983090983097 983089983089983091983093 plusmn 983095983089983097

AS 983089983097983088983089 plusmn 983092983097983094 983090983090983092983092 plusmn 983093983091983096983097








983137983138983148983141 983094

Parameters Groups



















EDXRF equipment

References














































httpwwwhindawicom






































httpwwwhindawicom

micro- and macroelemental composition and safety evaluation of the nutraceutical moringa oleifera...

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