Effects of Aqueous and Ethanolic Extracts of Cyperusesculentus (Tiger Nut) on Bio-Markers of Hepatoxicity, Oxidative Stress and Lipidemic Indices.
Mfem CC1Archibong AN1* Oyama SE2Okon UE1Njoku AN1LeleiSA3
1 Physiology Department, Faculty of Basic Medical Sciences, University of Calabar,
2Department of Family Medicine, University of Alberta, Canada
3Physiology Department, Niger Delta University, Wilberforce Island, Amassoma. TX
Archibong A N1*; [email protected] +2348063074679
ABSTRACT
Cyperusesculentus(Tiger nuts) a perennial plant has been reported to be of great therapeutic value, consequently this study seeks to investigate the effect of aqueous and ethanolic extract of tiger nut on biomarkers of hepatoxicity, oxidative stress and lipidemic indices in albino Wistar rats. Twenty one (21) male albino Wistar rats weighing 180 – 240g were randomly assigned into three groups of seven rats each. Group 1 served as control, while Groups 2 and 3 were orally administered 400 mg/kg bw aqueous and ethanolic extract of tiger nut respectively for six weeks, after which biomarkers of hepatoxicity, oxidative stress and lipid profile were determined in the sera.It was observed thatTc, LDL and VLDL level in the Tiger nut extract treated groups were significantly reduced (p<0.001 and p<0.05) when compared with that of the control group, but this decrease was more pronounce in the aqueous extract treated group. Conversely the HDL in the extract treated groups were significantly increased (p<0.01) when compared with that of control group respectively, but this increase was more pronounce in the aqueous extract treated group. Also CAT, SOD and GPx activities in the aqueous extract treated groups were significantly increased (p<0.001) while, MDA, ALT, ALP and AST activities were significantly decreased (p<0.001) when compared with that of the control group. In conclusion,Consumption of tiger nut extracts (especially aqueous extract) possess anti-atherogenic, antioxidant and hepato-protective effects therefore its consumption in decoction of alcohol should be discouraged.
Keywords:
Tigernut, Hepatoprotective, Oxidative stress, anti-atherogenic, Extract
INTRODUCTION
Cyperusesulentus(Tiger nut), a grass-like plant is a cosmopolitan perennial crop commonlyfound in seasonally flooded wetlands [7]. It is native to the Old World and is a lesser knownvegetable that produces sweet nut-like tubers known as “earth almonds” [38]. It is also knownby other names such as earth nut, yellow nut sedge, chufa (Spanish), rush nut and ediblegalingale [25].
In Nigeria, it is called „ofio‟ by the Igbos, “Aya” by the Hausas and „imumu‟by the Yorubas[30].Despite its name, tiger nut is not a real nut. It is actually a tuber [38]. C.
esculentushad beenreported to be a „health‟ food with rich nutritional composition and disease preventingproperties [38].It serves as a daily ingredient of the diet for many people in North Africa as well as Spain [29].It is also consumed as a snack due to its rich milky taste [4] and rich sugar content [32][14].
Itis rich in vitamins (B1, C & E) and minerals such as calcium, magnesium and iron [3][14].Bioactive phytochemicals and nutrients in tiger nut include salicyclic acid, alkaloids,terpenoids, saponins, steroids, phosphorus and potassium that have a widerange of health promoting properties [31].
Tiger nut is considered to have adequate properties to fight respiratory infections and somestomach illnesses. It is considered as an effective remedy for diarrhea and is a preventivemeasure for cyst, prostate, hernia and rectum deformation (REF). It also prevents endometriosisor fibrosis as well as blockage of the tip of the fallopian tube [7]. Aqueous extract of tiger nutcould be used as a possible fertility booster and to attenuate sperm toxicity and as a powerfulaphrodisiac [14]. It is used in the management of diabetes mellitus [42] debility indigestion,dyspepsia and colitis [32][1]. Tiger nut has been shown to be hepatoprotective because itsconsumption was able to bring about a lowering in the biomarkers of liver injury (ALT, ASTand ALP) it also lowers oxidative stress by boosting antioxidant enzymes [31].
Many peopleconsume this plant extract as a decoction of alcohol claiming it to be of therapeutic value. Thisresearch is aimed at investigating the comparative effect of aqueous and ethanolic extract oftiger nut on biomarkers of hepatoxicity, oxidative stress and lipidemic indices.
MATERIALS AND METHODS Experimental animals
Twenty one (21) male albino Wistar rats weighing between 180- 250g were used for this study.
These animals were obtained from the animal house of the Department of Physiology,University of Calabar, Nigeria. The animals were handled in accordance with standardprocedures. Prior to the commencement of the experiments, the animals were acclimatized forone week and given free access to rodent chow (Vital Feeds Nigeria, Limited) and water adlibitum [16]. Ethical approval was obtained from the Faculty of Basic Medical SciencesEthics Committee. The experiment was carried out in compliance with the National Instituteof Health Guide for care and use of Laboratory animals (NIH Publication No 823 revised 1978).
Preparation of C. esculentusextracts
Fresh tuber of Tiger nut was obtain from Gbogobiri market in Calabar, Cross River State, and was identified by the Chief Hebarium, Department of Botany University of Calabar.
Voucherspecimen deposited in the Departments herbarium number 2639 was kept for futurereference. The tiger nut was washed to remove debris including other physical contaminants dried at room temperature and ground to powder form. The dried samples were pulverized topowder form using grinding machine.1400 g of the dried sample was soaked in
7000 ml of distilled water for 24 h. The mixture wasthen filtered with a white cotton (satin) material, followed with filter paper (Whatmann No.1)into beakers and placed in an oven. The filtrate was evaporated to dryness using a rotaryevaporator at 40ºC. The extract was then collected into a sample bottle and stored in arefrigerator.
Similarly, the tiger-nut tuber was finely homogenized and 1400g was dissolved in 7000 ml of60% ethanol (v/v) and left to stand for 24 h at 40C. It was then filtered with a muslin cloth,followed with filter paper No. 1 to obtain the ethanol soluble extract. The filtrate was thenevaporated to dryness using a rotary evaporator at 40ºC. An oily crude extract was obtained.
The extract was stored in a beaker at 4ºC until use.
Toxicity Study on Tiger nut
Acute toxicity test was done according to standard procedure by Lorke[46] and as used by Archibong et al[47]. Thirty six mice were used for the study. They were randomly selected and assigned to six batches containing six animals each. They were allowed a week for adaptation.
Each batch received doses of extract intraperitoneally (1.64-104.48 mg/kg). Only the control group received normal saline intraperitoneally. They were all returned to their home cages and allowed free access to food and drinking water. The mortality in each group was assessed 24 hours after administration of the extract. The percentage mortalities were converted to probits and plotted against the log10 of the dose of the extract [46].
Experimental design
Twenty one (21) male albino wister rats weighing 180 – 240g were randomly assigned intothree (3) groups of seven (7) rats each.
Group 1 (Control) - received normal rat feed + drinking water
Group 2 (Aqueous Extract) – received same as group 1 + Aqueous extract (400mg/kg bodyweight once daily).
Group 3 (Ethanolic Extract) - received same as group 1 + Ethanolic extract (400 mg/kg bwonce daily).
The dose of 400mg/kg bw was arrived at following the estimation of Lethal toxicity(LD50) value for bothextracts. The administration was done orally and the experiment lasted for a period of sixweeks.
Collection of blood samples and analysis of different parameters
The animals were anaesthetized using chloroform and blood samples were collected via cardiacpuncture. The samples were emptied into plain sample bottles and allowed to clot for 2h.
It was thencentrifuged at 3000rpm for 10 minutes using centrifuge (model). The serum was collected intoclean test tubes and then used for the estimation of various parameters.
Lipid profile assessment
Measurement of total cholesterol
Total cholesterol was measured according to the method of Hassarajani, et al [16].
Measurement of triglyceride
Triglyceride was measured according to standard procedure as described by Hassarajani, etal [16].
Measurement of high density lipoprotein cholesterol
The High Density Lipoprotein was measured according to standard procedure [16].
Measurement of low and very low density lipoprotein
Low density and Very low density lipoprotein concentrations were calculated based onFriedwaldsformular[15].
VLDLC = Triglyceride 2.22
LDLC = Total Cholesterol – (HDLC+ VLDLC) Measurement of antioxidant enzymes
The different antioxidants enzymes were analyzed according to standard procedures.
Glutathione Peroxidase (GPx) activity was determined using the Rice Evans method [34].
The concentration of the product of the reaction between malondialdehyde (MDA)(a proxy for lipid peroxidation) and thiobarbituric acid - thiobarbituric acid reactive substances(TBARS) was spectrophotometrically measured [27].
Catalase (CAT) activity in the serum was determined as described by Aebi[2]. The change in the absorbance was monitored spectrophotometrically at 240nm over a 5min period.
Superoxide dismutase (SOD) activity in the serum was determined by assessing the inhibition of pyrogallol-auto oxidation [24]. Changes in the absorbance at 40nm were recorded at 1min interval for 5min. SOD activity was determined from standard curve of % inhibition of pyrogallol auto-oxidation with SOD activity
Measurement of liver enzymes
Measurement of Aspartate aminotransferase (AST) and Alanine aminotransferase (ALT) activities were done using enzymatic colorimetric methods as described by Reitman and Frankel [35]
Measurement of alkaline phosphatase (ALP)
ALP activity was measured using standard method [9] and as used by Archibonget al [6], Measurement of serum protein and bilirubin concentration
The determination of total bilirubin and total protein was done following standard procedures [43][44], respectively. Assay kits procured from reputable companies were used for all the determinations and assays, following the manufacturers‟ instructions.
Statistical analysis
Results obtained were presented as mean± standard error of mean. Data obtained were analyzed by one way analysis of variance (ANOVA) followed by post hoc student‟s Newman-keuls test using the SPSS computer program and p value less than 0.05 was considered statistically significant. The results were all presented in tabular form.
RESULTS Lethality study
Accordingly the acute toxicity study carried out on Tiger nut extracts revealed a safe margin, the LD50 value was about 919.21mg/kg (Aqueous extract) and 732.25mg/kg (Ethanolic extract) Lipid profile analysis
Total cholesterol (Tc) concentration.
The serum total cholesterol concentration of the aqueous and ethanolicextract treated groups are presented in Table 2. The Tc concentrations of the aqueous and ethanol extracts treated groups were significantly lower (p<0.01 and p<0.05) when compared with that of the control group.
However, there was no significant difference between the Tcconcentration of the aqueous extract group and that of the ethanolic extract group.
Triglyceride concentration (TG)
The serum triglyceride concentration of the aqueous and ethanolic extract treated groups are presented in Table 2. The TG concentration in the aqueous and ethanolicextract treated groups were significantly lower (p<0.001) when compared with that of the control group. Also the TG concentration in the ethanolic extract group was significantly lower (p<0.001) when compared with that of the aqueous extract treated group.
High Density Lipoprotein (HDL-c)
The serum HDL-c of the aqueous and ethanolic extract treated groups are presented in Table 2.
The HDL-c concentration in the aqueous and ethanolic extract treated groups were significantly higher (p<0.01 and p<0.05).when compared with that of the control group.
Low Density Lipoprotein (LDL-c )
The serum low density lipid concentration of the aqueous and ethanolic extract treated groups are presented in Table 2. The LDL-c concentration of the aqueous and ethanolic extract treated groups were significantly lower (p<0.01 and p<0.05) compared with that of the control group.
Also the LDL-c concentration in the aqueous extract treated group was lower when compared with that of the ethanolic extract treated group.
Very Low Density Lipoprotein (VLDL-c)
The serum very low density lipid concentration of the aqueous and ethanolic extract treated groupsare presented in Table 2. The VLDL-c concentration of the aqueousand ethanolic extract treated groups were significantly lower (p<0.01 and p<0.001 )when comparedwith that of control group. Also the VLDL-c concentration in the aqueous extract treated groupwas significantly lower (p<0.001 ) when compared with that of the ethanolic extract treatedgroup.
Table 1: Serum enzymes and Protein result of the different experimental groups (IU/L and mg/dl)
AST ALT ALP TP ALB GLB
Control 143.12±0.34 113.43±0.47 99.43±0.95 69.14±0.96 31.43±1.
04
38.00±0.62 Extract(Aqueou
s)
127.14±0.83*** 73.29±0.67*** 55.00±0.83*** 81.71±0.68*** 39.43±0.
95***
41.71±1.02**
Extract(Ethanol ic)
132.86±0.94***,c 79.14±0.95***,c 88.86±1.04***,c 74.57±0.48***,c 34.14±0.
34*,c
40.14±0.34* Values are expressed as mean + SEM, n = 7 *, **, *** = p< 0.05. 0.01, 0.001 vs control;
c = p<0.001 vs extract (aqueous)
Table 2: Lipid profile result of the different experimental groups (mmol/L)
Tc TG HDL LDL vLDL
Control 2.03±0.06 1.08±0.00 0.45±0.02 1.03±0.05 0.49±0.00 Extract(Aqueous) 1.79±0.03** 1.07±0.00 0.51±0.01** 0.85±0.02** 0.46±0.00**
Extract(Ethanolic) 1.86±0.05* 1.02±0.01***,c 0.47±0.01* 0.91±0.03* 0.48±0.00***,c Values are expressed as mean + SEM, n = 7 *, **, *** = p< 0.05. 0.01, 0.001 vs control;
c = p<0.001 vs extract (aqueous). Tc – Total cholesterol, TG – Triglyceride, DHL – High Density Lipoprotein, LDL – Low Density Lipoprotein, vLDL – Very Low Density Lipoprotein.
Table 3: Antioxidants enzymes and Malondialdehyde results in the different experimental groups (uIu/L)
CAT MDA SOD GPx
Control 21.41±0.36 1.28±0.00 1.09±0.03 44.43±0.48 Extract(Aqueous) 26.44±0.56*** 0.38±0.01*** 1.66±0.05*** 48.57±0.48***
Extract(Ethanolic) 12.77±0.08***,c 0.67±0.01***,c 0.56±0.02***,c 27.29±0.64***,c Values are expressed as mean + SEM, n = 7 *** = p< 0.001 vs control; c = p<0.001 vs extract (aqueous). CAT – Catalase, MDA – malondialdehyde, SOD – Superoxide dismutase, GPx – Glutathione peroxidase.
Table 4: Bilirubin results in the different experimental groups (mmol/L)
TB CB UB
Control
Extract(Aqueous) Extract(Ethanolic)
25.31±0.26 15.27±0.21***
21.11±0.43***,c
18.06±0.50 7.77±0.10***
11.30±0.39***,c
7.26±0.56 7.50±0.20 9.81±0.13***,c
Values are expressed as mean + SEM, n = 7 *** = p< 0.001 vs control; c = p<0.001 vs extract (aqueous). TB – Total bilirubin, CB – Conjugated bilirubin, UB – Unconjugated bilirubin.
Antioxidant Enzymes and Malondialdihyde Superoxide dismutase (SOD)
The Superoxide dismutase activities of the aqueous and ethanolicextract treated groupsare presented in Table 3. The SOD activity in the aqueous extract treated group was significantly higher (p<0.001), while that of ethanolic extract treated group wassignificantly lower (p<0.001) when compared with that of the control group. Also the SOD activity in the aqueous extract treated group was significantly higher (p<0.001) whencompared with that of the ethanolic extract treated group.
Glutathione Peroxidase (GPx)
The Glutathione peroxidase activities of the aqueous and ethanolic extract treated groups are presented in Table 3. The GPxactivity in the aqueous extract treated group was significantly higher (p<0.001), while that of the ethanolic extract treated group wassignificantly lower (p<0.001) when compared with that of the control group. Also the GPx activity in the aqueous extract treated group was significantly higher (p<0.001) whencompared with that of the ethanolic extract treated group.
Catalase (CAT)
The catalase activities of the aqueous and ethanolic extract treated groups are presented in Table 3. The CAT activity in the aqueous extract treated group wassignificantly higher (p<0.001), while that of the ethanolic extract treated group wassignificantly lower (p<0.001) when compared with that of the control group. Also the CAT activity in the aqueous extract treated group was significantly higher (p<0.001) whencompared with that of the ethanolic extract treated group.
Malondialdehyde (MDA)
The malondialdehydeactivities of the aqueous and ethanolicextract treated groups are presented in Table 3. The MDA activity in the aqueous and ethanolicextract treated groups were significantly lower (p<0.001) when compared with that of thecontrol group. Also the MDA activity in the aqueous extract treated group wassignificantly lower (p<0.001) when compared with that of the ethanolic extract treated group.
Liver enzymes
Aspartate aminotransferase (AST)
The aspartate aminotransferase activities of the aqueous and ethanolic extract treated groups are presented in table 1. The ASTactivity of the aqueous andethanolic extract treated groups were significantly lower (p<0.001) when compared with thatof the control. Also the AST activity in the aqueous extract treated group was significantly lower (p<0.001) when compared with that of the ethanolic extract treated group.
Alanine aminotransferase (ALT)
The alanine aminotransferase activities of the aqueous and ethanolic extract treated groupsare presented in Table 1. The ALT activity in the aqueous andethanolic extract treated groups were significantly lower (p<0.001) when compared with thatof the control group. Also the ALT activity in the aqueous extract treated group wassignificantly lower (p<0.001) when compared with that of the ethanolic extract treated group.
Alkaline phosphatase (ALP)
The alkaline phosphatase activities of the aqueous and ethanolicextract treated groupsare presented in Table 1. The ALP activity in the aqueous and ethanolicextract treated groups were significantly lower (p<0.001) when compared with that of thecontrol group. Also the ALP activity in the aqueous extract treated group wassignificantly lower (p<0.001) when compared with that of the ethanolic extract treated group
Serum protein Total protein (TP)
The total protein concentration of the aqueous and ethanolic extract treated groups is presented in table 1. The TP concentration in the aqueous and ethanolic extract treatedgroups were significantly lower (p<0.001) when compared with that of the control group. Also the TP concentration in the aqueous extract treated group was significantly higher (p<0.001) when compared with that of the ethanolic extract treated group.
Albumin (ALB)
The albumin concentration of the aqueous and ethanolic extract treated groups are presented in Table 1. The ALB concentration in the aqueous and ethanolic extract treatedgroups were significantly higher (p<0.001) when compared with that of the control group. Alsothe ALB concentration in the aqueous extract treated group was significantly higher (p<0.001)when compared with that of the ethanolic extract treated group.
Globulin (GLB)
The globulin concentration of the aqueous and ethanolic extract treated group are presented in Table 1. The GLB concentration in the aqueous and ethanolic extract treatedgroups were significantly higher (p<0.001) when compared with that of the control. Also theGLB concentration in the aqueous extract treated group was significantly higher (p<0.001).when compared with that of the ethanolic extract treated group.
Bilirubin
Total bilirubin:The total bilirubin concentration of the aqueous and ethanolic extract treated groups are presented in Table 4. The total bilirubin concentration in the aqueous and ethanolicextract treated groups were significantly lower (p<0.001) when compared with that of thecontrol. Also the total bilirubin concentration in the aqueous extract treated group wassignificantly lower (p<0.001) when compared with that of the ethanolic extract treated group.
Conjugated bilirubin:The conjugated bilirubin concentration of the aqueous and ethanolicextract treated groups are presented in Table 4. The Conjugated bilirubin concentration in the aqueousand ethanolic extract treated groups were significantly lower (p<0.001) when compared withthat of the control. Also the conjugated bilirubin concentration in the aqueous extract treatedgroup was significantly lower (p<0.001) when compared with that of the ethanolic extracttreated group.
Unconjugated bilirubin:The unconjugated bilirubin concentration of the aqueous and ethanolicextracttreated groups are presented in Table 4. The unconjugated bilirubin concentration in the controlgroups was significantly lower (p<0.001) when compared with that of the ethanolic extracttreated group but showed no significant difference when compared with that of the aqueousextract treated group. Also the unconjugated bilirubin concentration in the aqueous extracttreated group was significantly lower (p<0.001) when compared with that of the ethanolicextract treated group.
DISCUSSION AND CONCLUSION
Discussion
The result reveals significant decrease in the lipid profile in extract treated groups comparedwith that of control group. This decrease is very important because raised level of serum total cholesterol, triglyceride and low densitylipoprotein cholesterol are possible indication of coronary heart attack, risk of heart diseaseand stroke [45]. This result is of significance because accumulation of LDL-c predisposes oneto cardiovascular disease, but on the contrary accumulation of HDL-c is cardioprotective. A recent study had reported that tiger nut consumption reduced atherogenic indices inexperimental rat [17], this reduction is an indication of the cardioprotective effects of tiger nut.
Also the HDL-c concentration in the different extract treated groups were observed to increase significantly when compared with that of the control group respectively. These findings are in agreement with previous studies that reported a significant increase in serum HDL-cholesterol due to tiger nut consumption [41][17].
The cardiovascular protective effects observed in the tiger nut extracts group may be due tothe presence of monounsaturated fats in it which is cardioprotective[21][5]. Tiger nut areknown to contain omega 6 and omega 9 [13], which are known to reduce the level of LDLccholesterol and triglycerides whereas they induce increase in the rate of HDL-c cholesterollimiting the risk of cardiovascular diseases [8].Further observation also reveal that the level of HDL-c in the aqueous extract treatedgroup was significantly increased when compared with that of the ethanolic extract treatedgroup, while the reverse was the case in other parameters (TG, TC, LDL and vLDL). Maybe extraction with ethanol was able to reduce the potency of some of the important component oftiger nut.
The result also revealed a significant increase in CAT, SOD and GPx concentration in theextract treated group when compared with that of the control group. This result is consistentwith studies done by Olabiyiet al [28] that reported increased level of antioxidant enzymesfollowing tiger nut consumption. CAT, SOD and GPx are important antioxidant enzymes thataids in the scavenging
and degrading of free radicals. SOD perform important function bycatalyzing the conversion of superoxide radicals to O2 andH2O[39][37] while Catalasefurther decomposes the toxic H2O2 to O2 andH2O[11][22]. GPx is known to catalyzethe reduction of hydroperoxide by glutathione [20]and also protect againstoxidative stress. The observed rise in antioxidant enzymes in the present study might beattributed to the flavonoids and antioxidant vitamins C and E present in tiger nut which boostthe activities of the antioxidant enzymes [19][40]. Also tiger nut has been reported to containvitamin C and E that are potent scavenger of free radicals [19].
ROS degrade polyunsaturated lipids forming malondialdehyde which causes toxicstress in cells [33]. Malondialdehyde is a stable end product of lipid peroxidation and is abiological marker of oxidative stress [23]. That the level of MDA was reduced following thedifferent extract treatment shows that tiger nut extract has the ability of reducing lipidperoxidation therefore playing an important role in maintaining good health. MDAconcentration in the aqueous extract treated group was significantly decreased when comparedwith that of ethanolic extract treated group. This implies that the aqueous extract of tiger nuthad a higher antioxidant capacity than the ethanol extract.
The liver enzymes activities were significantly reduced in aqueous and ethanolic extracttreated groups when compared with that of the control group. This result is in agreement withthe study conducted by Onuohaet al [31]who reported that tiger nut consumption significantly preventedliver injury by reducing the levels of these liver function enzymes. This result is also corroborated by the total bilirubin concentration which was significantly reduced in the extracttreated group compared with that of the control. It is a common knowledge that elevation intotal bilirubin in serum is an indicator of liver or biliary tract disease [18] also increase in biliarysynthesis alongside biliary pressure (during intoxication) causes increase in serum ALPconcentration. It is therefore important that a healthy liver will secret less of bilirubin and ALP[12], and this was observed following treatment with tigernut extract. It was observed that thereductions in these liver enzymes and total bilirubin concentration in the aqueous extract treatedgroup exceeded those observed in the ethanolic extract treated group. Theseshows that consumption of aqueous extract of tiger nut is more hepatoprotective than ethanolic extract.
Levels of total protein, albumin and globulin makes up important part of blood plasma,which serves to maintain osmotic pressure and carry steroid hormones, lipids and fibrinogenwhich is essential for blood clotting [36]. Higher levels of these proteins are beneficial [10].Following treatment with the respective tiger nut extracts, the total protein, albumin andglobulin concentrations were observed to be significantly increased compared with that of thecontrol group. This suggests that both the aqueous extract treated group and ethanolic extracttreated group are hepato-protective and this is supported by recent article by Odeyepo&Odoje,[26]
though the aqueous extract seemed to be more potent than the ethanol extracts.
Limitations
This study was limited by few factors. It would have benefited from the determination of theactive ingredient in tiger nut, and induction of dyslipidaemia to test the hypolipidaemic effectof tiger nut. But unfortunately we were limited by resources yet subsequent studies may haveto take care of that.
Conclusion
In conclusion, consumtion of tiger nutextrat possess anti-atherogenic, antioxidant and hepato- protectiveeffects with the aqueous extract being more potent than the ethanolic extract, therefore its consumption in decoction of alcohol should be discouraged.
PLEASE NOTE THE FOLLOWING:
NO CONFLICT OF INTEREST BETWEEN AUTHORS
NO FUNDING SUPPORT FROM ANY INDIVIDUAL OR ORGANISATION
ETHICAL PERMISSION WAS OBTAINED FOR THIS RESEARCH
AUTHORS’ CONTRIBUTIONS: Mfem CC and ArchibongAN designed the research, and did the laboratory work. Oyama SE and NjokuANdid the literature review, Okon UE and Lelei SA did the writing and revision.
REFERENCES
1. Adejuyitan, J. A., Otunola, E.T., Akande, E. A., Bolarinwa, I. F., Oladokun, F. M. (2009). Some physiochemical properties of flour obtained from fermentation of tiger nut (Cyperusesculentus) Afri J Food Sci; 3: 51-55.
2. Aebi, H. (1984). Catalase in vitro Methods in Enzymology. Methods inEnzymology, 105, 121–6.
3. Al-Shaikh, M. N., Wahab, T. A., Kareem, S. A., Hamoudi, S. R. (2013). Protectiveeffect of chufa tubers (Cyperusesculentus) on induction of sperm abnormalities in mice treated with lead acelate. Int J Drug Res; 5: 387-392.
4. Amaal, A. M and Essraa, M. A. (2010). The effect of Cyperusesculentuson sperm function parameters in prepubertal mice as a model for human. J Baghdad Sci; 7: 389-393.
5. Arafat, S. M., Gaafar, A. M.,Nassef, S. L. (2009). Chufa tubers (CyperusesculentusL.): As a new source of food. World applied Sciences Journal; 7(2): 151-156.
6. Archibong, A. N., Akwari, A. A., Ofem, O. E., Bassey, I. O., Ukweni, S. U., Eno, A. E. (2017).
Alteration in biochemical indices following administration of seafood (Thaiscoronata) extract.
European Journal of Biological Research :7 (2) 131-138
7. Bamishaiye, E. I and Bamishaiye, O. M. (2011). Tiger nut: As a plant, its derivatives andbenefits.African Journal of Food, Agriculture, Nutrition and Development; 11 (5):5157-5170.
8. Belewu, M.A and Abodunrin, A.O. (2008). Preparation of kunun from unexploited richfood source: Tigernut(Cyperusesculentus). Pakistan Journal of Nutrition, 7:109-111.
9. Bergemeyer, H. Uand Brent, E. (1974). LDU-UV assay with pyruvate and NADH.In: Method ofenzymatic analysis (Ed): H. U. Bergemeyer Academic press, NewYork, 574 – 579
10. Busher J. T. (1990). Serum albumin and globulin In: Walker H.K., Hall W. D., Hurst J.W.
Clinical methods: The history,physical and laboratory examinations (3rd ed.).Butterworts, Boston.
11. Chelikani, P., Fita, I., Loewen, P. (2004).Diversity of structures andproperties among catalases".
Cellular and Molecular Life Sciences, 61 (2), 192–208
12. Dash, D.,Yeligar, V., Nayak, S., Ghosh, T., Rajalingam, D., Sengupta, P. (2007). Evaluation of hepatoprotective and antioxidant activity of Ichnocarpusfrutescens(Linn.) R.Br. On aracetamol- induced hepatotoxicity in rats. Trop J PharmRes. ;6:755–65.
13. Dubois, V., Brenton, S., Linder, M., Fanni, J.,Parmentier, M. (2007). Fatty acidprofile of 80 vegetable oils with regard to their nutritional potential. European Journalof Lipid Science and Technology, 109, 720 -732.
14. Ekaluo. U. B., Ikpeme E. V., Etta S. E., Ekpo, P. B. (2015). Effect of aqueous extract oftiger nut (CyperusesculentusL.) on sperm parameters and testosterone level of malealbino rats. Asian Journal of Biotechnology; 7(1): 39-45.
15. Friedwald, W. T., Levy, R. I., Fredrickson, D. S. (1972). Estimation of the concentrationof low density lipoprotein cholesterol without the use of the preparative ultracentrifuge.Clin Chem;
18(6): 499- 502.
16. Hassarajani, S., Souzo, T., Mengi, S., Chattopadhaya Y. (2007). Efficacy studyof the bioactive fraction (F-3) of Acoruscalamus in hyperlipidemia. Indian Journalof harmacology,39, 196 – 200.
17. Ibitoye, O. B., Aliyu, N. O.,Ajiboye, T. O. (2018) Tiger nut based diet improves the lipidprofile and antioxidant status of male Wistar rats. Journal of food biochemistry 42 :5 –20.
18. Iyanagi, T., Emi, Y., Ikushiro, S. (1998) Biochemical and molecular aspects of geneticdisorders of bilirubin metabolism. BiochimBiophys Acta.1407:173–84.
19. Kummar, V., Abbas, K., Fausto, N., Robbsins, L.,Contran, R. (2004.).Pathology bases of disease. New York. WB Saunder.
20. Li, X. X., Zhao, X. Y., Huang, L. X., Hu, X. X., Liu, X. Y., Yang, J. J.,Zhang, P. J., Zhang, Y.
H. (2018) Effect of smoking and alcohol consumption on lipid profile in male adult inNorthwest rural China. Public Health 157 : 7 – 13.
21. Linssen, J. H., Kielman, G. M., Cozijnsen, J. L., Pilnik,S.W. (1998). Comparison of chufaand olive oils. Food Chemistry; 28(4): 279-285.
22. Liu, X. and Kokare, C. (2017). Microbial enzyme of use in industry In: Biotechnology ofmicrobial enzymes. Academic Press, Cambridge: pp 287-298.
23. Mao,C. C., Yuan, J. Q., Lv, Y. B., Gao, X. X., Yin, Z. X., Kraus, V.B., Luo, J. S., Chei, C.
L.,Matchar D. B. (2019). Association between superoxide dismutase, malondialdehyde andall cause mortality in older adult: BMC Geriatrics 19 (1) 104.
24. Marklund, S. L. (1985). Pyrogallol auto-oxidation. In Handbook of methods foroxygen radical research. Ed: Greenwald R. A, Boca Raton, F. L. CRC Press, pp43 – 247.
25. Oderinde, R. A. andTairu O. A. (1988). Evaluation of the properties of yellow nut sedge (Cyperusesculentus) tuber oil. Food Chem; 28: 233-237.
26. Odeyepo, T. A. andOboje, O. F. (2014). Hepato-protective activities of tiger nut (Cyperusesculentus) against hepato-toxicity induced by Carbon tetrachloride in rats.
Researchand reviews. Journal of Pharmacology and Toxicological studies; 2(4): 37-41.
27. Ohkawa, H., Ohishi, N., Yagi, K. (1979). Assay for lipid peroxides in animal tissues bythiobarbituric acid reaction. Anal Biochem; 95: 351-358.
28. Olabiyi, A. A., Oboh,G. G., Adefegha, S. A. (2016) Effect of dietary supplementation of tigernut and wallnt on sexual behaviour, hormonal level and antioxidants status in male rats.Journal of food Biochemistry 41(3)
29. Oladele, K. A., Osundahunsi, F.O., Adebowale, A.Y. (2009). Influence of processingtechniques on the nutrients and anti-nutrients of tiger nut (CyperusesculentusL.).World J Diary & Food sci; 2: 88-93.
30. Omode, A., Fatoki, A., Olagun, A.( 1995) Physiochemical properties. Some underexploitedand non conventional oil seed. Journal of Agricultural food chemistry. 11: 50– 53
31. Onuoha, N. O., Ogbusua, N.O., Okorie, A. N., Ejike, C.E. (2017). Tiger nut (CyperusesculentusL.) “milk” as a potent “nutri-drink” for the prevention of acetaminopheninduced hepatotoxicity in a murine model. J IntercultEthnopharmacol; 6(3):
290-295.
32. Pamplona-Roger, G.D. (2002). Encyclopedia of food and their healing power. Reviewand Herald Publishing Association, Maryland.
33. Pryor, W. A. and Stanley, J. P. (1975). Suggested mechanism for the production ofmalondialdehyde during the autoxidation of polyunsaturated fatty acids. Non- enzymicproduction of prostaglandin endoperoxides during autoxidation. J Org Chem; 40(24):
3615-3617.
34. Rice, E. C., Diplock, A. T., Symons, M. C. (1991). Techniques I free radicalresearch. Elsevier, Amsterdam London New York, Tokyo, 194 – 196
35. Rietman, S. and Frankel, S. (1957). A colorimetric method for aspartate and alanineamino transferases in serum. Am J Chem Path; 28:56-58.
36. Riris, I. (2017). MethoxyBergenin‟s effect on total protein, albumin and globulin levelsof wistar rats‟ blood. Der PharmaChemica; 9(13): 90-94.
37. Romao, S. (2015). Therapeutic value of oral supplementation with melonsuperoxide dismutase and wheat gliadin combination. Nutrition, 31(3), 430–6
38. Sanchez-Zapata, E., Fernández-Lòpez, J., Perez-Alvarez, J.(2012). Tiger nut(Cyperusesculentus) commercialization: Health Aspects, composition, properties andfood application. Comprehensive reviews in food science and food safety; 11(4):125
39. Vanaporn, M., Wand, M., Michell, L., SarkarTyson, M., Ireland, P., Goldman,S., Kewcharoenwong, C., Rinchai, D., Lertmemongkolchai, G.,Titball, R.(2011). Superoxide dismutase C is required for intracellular survival and virulence ofBurkholderia pseudomallei.
Microbiology, 157 (8), 2392–400
40. Wayah, S. and Shehu, S. (2013) Assessment of nutritional and anti-nutritionalcomponent of Tiger nut residue. International Journal of Scientific research 4(6) : 5 -9
41. Zommara, M. andImaizumi, K. (2017). Antiatherogenic effect of tiger nut tubers(CyperusesculentusL.) supplemented diet in apoliprotein E knockout mice. Journal ofSustainable Agricultural Sciences; 43(4): 197-204.
42. Borges, O. B., Goncalves, J.L., Carvalho, P. C., Silva, A.P. (2008). Nutritional quality of chestnut (Castanea sativa Mill.) cultivars from Portugal. Food Chem., 106: 976-984 43.Jendrassik, L. and Grof, P. (1938). Estimation of total serum bilirubin level byspectrophotometrically in serum and plasma. BiochemischeZeitschrift, 297: 81-89.44. Tietz, N.W., Pruden, E.L.,Siggaard-Andersen, O. (1994) Liver function. In TietzTextbook of Clinical Chemistry; Burtis, A.C., Ashwood, E.R., Eds.; Saunders: London,UK, 1994; pp.
1354–1374.
45. Drummond, K.E. and Brefere,L.M. (2013). Nutrition for Food Service and CulinaryProfessionals. 8th Edn., Wiley Global Education, USA., ISBN: 9781118476833, Pages:
656.
46. Lorke, D (1983) A new approach to practical acute toxicity testing. Arch Toxicol 54: 275– 87 47. Archibong, N. A.,Akwari, A. A., Ofem, E. O., Irene, O. B., Samuel, U. U.,Asim, E. E. (2015).
Effect of Egeriaradiata(Clam) Extract on Biochemical Parameters of Albino Wistar Rats.
Journal of Medical Sciences, 15 (2): 87-93.
