Effects of Chronic Administration of Indomie Noodles on the Activity of Alkaline Phosphatase of Rat Small Intestine

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL SCIENCES

Sanni et al. Effects of chronic administration of indomie noodles on the activity of alkaline phosphatase of rat small intestine. Journal of pharmaceutical and biomedical sciences (J Pharm Biomed Sci.) 2013 September; 34(34): 1682-1687.

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ISSN NO- 2230 – 7885 CODEN JPBSCT NLM Title: J Pharm Biomed Sci.

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Original article Effects of chronic administration of indomie noodles on the activity of alkaline phosphatase of rat small intestine

1SANNI, Momoh, *1EJEMBI, Daniel, 2EMMANUEL, T. Friday, 1ABBAH, Okpachi C. & 1Okwutachi Sunday O

Affiliation:- further divided into equal number of males and 1Department of biochemistry, Kogi State University, females. Group 1 was fed with the noodle cooked Anyigba, Nigeria. and spiced, group 2 was fed with raw noodle 2Department of medical biochemistry, Kogi State spiced, and group 3 was fed with a mixture of the University, Anyigba, Nigeria. cooked and raw noodles and spiced with the seasoning. Groups 4-6 were fed in a repeated Author’s contributions- All authors contributed fashion as 1, 2 & 3 with the exclusion of the equally to this paper. seasoning spice. Group 7 was fed with normal rat feed. Water was provided ad libitum. Animals *Correspondence to:- were fed for thirty days and were sacrificed on EJEMBI Daniel. the thirty first day after commencement of Department of Biochemistry, Kogi State University, administration of test feed (indomie) and the Anyigba, Nigeria. organs of interest was collected and kept for [email protected] analysis. The results obtained were statistically treated (p<0.05) at 95% confidence level using Abstract: F-test with a positive association of consumption The effects of chronic administration of indomie of indomie noodles with damage to small instant noodles on the activity of alkaline intestinal tissues was observed. phosphatase of rat small intestine were investigated. Forty eight (48) albino rats (Rattus novergicus) of average weight 210g were divided Key words: Indomie instant noodles; Alkaline th into 7 groups containing 6 rats except the 7 phosphatase. group which 12 rats as control; each group was

Article citation:- Sanni et al. Effects of chronic administration of indomie noodles on the activity of alkaline phosphatase of rat small intestine. Journal of pharmaceutical and biomedical sciences (J Pharm Biomed Sci.) 2013 September; 34(34):1682-1687.Available at http: //www.jpbms.info

INTRODUCTION The noodles are quick and easy to make, and can ndomie is a brand of instant noodles by be eaten as a snack or as part of a main meal. Their Indofood, the world's largest instant noodle versatility means that people can carry them to I manufacturer, located in Indonesia. Indofoods work or on trips, and cook them simply by adding is one of the world’s largest manufacturers of dried hot water from a kettle. Indofood is one of the instant noodles based in Indonesia, the world’s largest pre-packaged food companies in indonesia, largest Muslim country with a population of about and was founded in 1972 as Panganjaya 140 million people (Indofood, 2010)12. Indomie Intikusuma before changing to its current name in (the name derived from the Indonesian MiGoreng, 1990 by Sudono Salim under the Salims group21. which means fried noodle) is an instant noodle sold dried in packets or cups. The company’s INGREDIENTS products sell throughout Indonesia, Malaysia, Noodles: wheat flour, vegetable oil, iodized salt, Australia, Nigeria, the United States, and 25 other sodium polyphosphate, sodium carbonate, countries – and not just to expatriate Indonesians potassium carbonate, guargum, tartrazine (CI (Indofood, 2010)12. 19140), antioxidant (TBHQ).

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Seasoning powder: iodized salt, monosodium in foods and drinks in the European Union (EU) glutamate (621), hydrolyzed vegetable protein, soy over a specific period (FSA Board, powder, pepper, garlic powder, chicken flavor, 2008)9.Tartrazine has a noticeable effect on the Chili powder. behavior of young mice; Indomie noodles contain Tartrazine, a synthetic  Tartrazine inflamed the stomach lining lemon yellow azo dye primarily used as food (increased the number of lymphocytes and colouring agent (Food standards agency website, eosinophils of rats) when given in the diet for a 2011). It has a batch category code of E102 C.I. prolonged time14. 19140, FD&C Yellow 5, acid yellow 23, food yellow  Tartrazine was found to adversely affect 4 and trisodium-1-(4-sulfonatophenyl)-4-(4- and alter biochemical markers in vital organs e.g. sulfonatophenylazo)-5-(pyrazolone-3carboxylate). liver, kidney, small intestines etc of rat not only at It is water soluble and has a maximum absorbance higher doses but also at low doses4. in aqueous solution at 427±2nm13. Tartrazine is a commonly used colorant all over the world, mainly Organ studied (Small intestine): for yellow, but can also be used with brilliant blue In rat, the small intestine is about six times the (FD&C Blue; E133) or Green (E142) to produce length of the body from snout to anus16. This various green shades. enormous length is necessary to hold the food long Tartrazine was believed to cause most of the enough for thorough digestion as well as providing allergic and intolerance reaction experienced the large surface area needed for absorption. In amongst all the azo dyes, particularly in asthmatic humans, it is a long tube where major events of patients and those with aspirin intolerance (UK digestion and absorption occur19. food Guide, 2007)20. Symptoms of tartrazine The walls of the small intestine are thin but contain sensitivity can occur from either ingestion or bands of circular and longitudinal muscles which cutaneous exposure to substances containing produce peristaltic and ant peristaltic movement tartrazine. Symptoms could either be mild to responsible for rocking the content to and fro, srevere3.A variety of immunological responses mixing it, and gradually passing it towards the have been attributed to tartrazine ingestion, large intestine16. including anxiety, migraine3, clinical depression, blurred vision, itching, general weakness, feeling of Enzyme studied (Alkaline phosphatase): suffocation, purple skin patches and sleep Alkaline phosphatase (ALP, ALKP) (EC3.1.3.1) is a disturbance15. People who are exposed to the dye hydrolase enzyme responsible for removing experience symptoms of tartrazine sensitivity even phosphate groups from many types of molecules, at extremely small dose, some for periods up to 72 including nucleotides, proteins, and alkaloids. The hours after exposure6. In children, asthma attack process of removing the phosphate group is called and hives have been claimed, as well as supposed dephosphorylation. As the name suggests, alkaline link to thyroid tumors, chromosomal damage and phosphatase are most effective in an alkaline hyper reactivity7. environment. It is sometimes used synonymously as basic phosphatase18. Possible health effects of tartrazine: The British food standards agency on September Distribution and multiple forms of alkaline 6th, 2007, revised, advice on certain artificial food phosphatase: additives, including tartrazine (BBC news, 2007)5. In humans, alkaline phosphatase is present in all Professor Jim Stevenson of Southampton tissues throughout the entire body, but is University, the author of the report said, “This is an particularly concentrated in liver, bile duct, small important investigating area of research”. The intestine, kidney, bone, and the placenta (Harada et result suggests that consumption of certain al., 1986)10. Humans and most other mammals mixture of artificial food color and sodium contain the following alkaline phosphatase benzoate preservatives are associated with isozymes increase in hyperactive behavior in children. a. ALPI – intestinal The food standards agency (FSA) in April 10th, b. ALPL – tissue non-specific (liver/bone/kidney) 2008, called for voluntary removal of the colors c. ALPP – placental (Regan isozyme) (but not sodium benzoate). In addition, it recommended that the colors should be phased out

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Inhibition and activation: of feeding and were sacrificed by decapitation. The All mammalian ALP isoenzymes except placental organ of interest (small intestine) were removed (ALPP) are inhibited by homoarginine and and put in a 0.25M ice cold sucrose solution. similarly all except the intestinal and placental ones are blocked by levamisole. Heating for 2 Preparation of tissue homogenate: hours at 65°C inactivates most isoenzymes except The tissue was weighed and homogenized using an placental isoforms 11. automated homogenizing machine in a 0.25M Alkaline phosphatase is a metalloenzyme, Mg2+and sucrose solution this was kept in a clean dry Co2+ ions appear to be essential for its activation1. sample bottle and kept in the freezer before being used for enzyme activity determination2. MATERIALS AND METHODS Determination of alkaline phosphatase activity Enzyme substrate and reagents: (Schmidt and schmidt, 1963)17: Alkaline phosphatase enzyme (RANDOX KIT) This is a colorimetric method of enzyme assay consists of diethanolamine buffer, magnesium according to the recommendations of the Deutsche. chloride and paranitrophenylphosphate as Gesellschaft, Furklinischechemie. Alkaline substrate. phosphatase (ALP) activity was activated by Mg2+ with a resultant yellow colour solution. Test animals: Fourty eight (48) albino rats of average weight 210g were obtained from Faculty of Agriculture Principle: farm Kogi State University Anyigba, Nigeria. P-nitrophenylphosphate + H2O → Phosphate + P- nitrophenol. Animal groupings: Forty eight (48) rats (Rattus novergicus) were Procedure (Schmidt and schmidt, 1963)17: randomly grouped into seven (7) groups. Groups 1- The reagent composition is 1mmol/L of 6 being the test groups consists of 6 rats each, each diethanolamine buffer and 0.5mm/L of magnesium group was further sub-grouped by sex into 3 males chloride (MgCl2) while the substrate is 10mmol/L and 3 females respectively. Group 7 is the control P-nitrophenylphosphate. The buffer in 60ml was group and was fed with normal rat feed; the reconstituted with the substrate at a rate of 3ml of control group had twelve rats of which six were buffer in 1 bottle of substrate to produce 60ml; males and six females. The experimental design is 0.01ml of sample was pipetted into the test tube as follows; and 0.5ml of the reagent was added and mixed, dry a. Group 1 was fed with indomie instant noodles air was used as reagent blank. The reading was cooked and spiced with the seasoning. taken using spectrophotometer. The initial b. Group 2 was fed with raw indomie instant absorbance was taken and a timer was started noodle spiced with the seasoning. simultaneously, the reading was taken c. Group 3 was fed with a mixture of both cooked consecutively after 1, 2 and 3 minutes. and raw indomie instant noodle spiced with seasoning. RESULTS d. Group 4 was fed with indomie instant noodles When indomie was administered, the following cooked without seasoning. was observed. This is represented in the graph e. Group 5 was fed with raw indomie instant below; From figure 1 above, the blue charts noodle without seasoning. represents the values for ALP activity in male rats f. Group 6 was fed with a mixture of both cooked while the green charts represent the values of ALP and raw indomie instant noodle without activity in female rats. In males, taking control as seasoning. the reference group; in group1, the activity of ALP Water was provided ad libitum. The rats were was observed to have decreased by 85.53%, group examined within a thirty (30) days scheme in 2, decreased by 93.25%, group3,decreased by order to ascertain levels of chronic toxicity. 94.85%, group4, decreased by 70.74%, group 5, decreased by 88.10% and group6, decreased by Animal sacrifice: 83.28%. ALL THE experimental rats were starved for 24 hours prior to the day of sacrifice after the 30th day

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Figure1. Graph of alkaline phosphatase activity

In females, taking control as the reference group; the cells which consequently results in the release group 1, decreased by 75.77%, group 2, decreased of this enzyme into extracellular fluid, thereby by 88.67%, group3, decreased by 89.87%, group 4, decreasing the enzyme activity in the small decreased by 87.67%, group 5, decreased by intestine. Furthermore, rats of group 1-3 showed a 17.62%, and group6, decreased by 75.33%. greater decrease in activity which may possibly be The results for the control groups 858.36U/I for due to the combined action of agents from the male and 626.52U/I for female indicate a high seasoning powder, the noodles inherent degree of enzyme activity in the small intestinal constituents and additives. Tartrazine was found to tissues of these rats. However, consumption of adversely affect and alter biochemical markers in indomie noodles significantly reduces the activity vital organs e.g. liver, kidney, small intestines etc of of ALP in the small intestine. rat not only at higher doses but also at low doses4.

DISCUSSION The significant decrease in the activity of ALP in Most diseases results from defects in metabolic the small intestine of female rats of groups 1-6 routes of which hyperactivity, hypo activity and might plausibly be due to the inhibition of the inhibition of metabolic enzymes are largely enzymes activity by tartrazine or synergetic involved. These metabolic enzymes are inhibition by tartrazine, monosodium glutamate compartmentalized in different tissues. Their (MSG) and other components whose effects have elevations or decrease in amount serves as not been fully elucidated or as a result of damage biomarker for clinical diagnosis of some illness8. done to the cells which consequently results in the Ngaha and akanji have shown that biochemical release of this enzyme into extracellular fluid. parameters like enzyme assay can indicate cellular Furthermore, rats of group 1-3 showed a greater damage by conventional histological techniques2. decrease in activity which may possibly be due to The huge decrease in the activity of ALP in the the combined action of agents from the seasoning small intestine of male rats of all the test groups powder and the noodles inherent constituents and may be accrued to the inhibition of the enzyme additives. Tartrazine was found to adversely affect activity by tartrazine or synergetic inhibition by and alter biochemical markers in vital organs e.g. tartrazine, monosodium glutamate (MSG) and liver, kidney, small intestines of rat not only at other components whose effects have not been higher doses but also at low doses4. fully elucidated or as a result of damage done to

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CONCLUSION AND RECOMMENDATION animal. Table 1 suggest that indomie instant This research study showed that the consumption noodles be consumed but only on occasional basis of indomie instant noodles by any of the 6 especially during instances of need for a fast food, combinations experimented, significantly affects this is because it contains the necessary nutrient the activity of small intestinal Alkaline phosphatase for healthy living in moderate proportions. (p<0.05). However, the degree of effect varies, However, if taken in excess, it can result in serious depending on the nature of diet preparation, sex of medical complications. animal and possibly physiological state of the Table1. Alkaline phosphatase activity in rats of various groups GROUPS MALE (nm/min) I/U FEMALE (nm/min) I/U 1 124.20 151.80 2 57.96 69.00 3 44.16 63.48 4 251.16 77.28 5 102.12 516.12 6 143.52 154.56 CONTROL 858.36 626.52

Table 2. Analysis of variance (ANOVA) for alp activity in male albino rats Descriptive Male 95% Confidence Interval for

Std. Mean N Mean Deviation Std. Error Lower Bound Upper Bound Minimum Maximum Group1 1 1.2420E2 . . . . 124.20 124.20 Group2 1 57.9600 . . . . 57.96 57.96 Group3 1 44.1600 . . . . 44.16 44.16 Group4 1 2.5116E2 . . . . 251.16 251.16 Group5 1 1.0212E2 . . . . 102.12 102.12 Group6 1 1.4352E2 . . . . 143.52 143.52 Control 1 8.5836E2 . . . . 858.36 858.36 Total 7 2.2593E2 287.03305 1.08488E2 -39.5356 491.3870 44.16 858.36

Table 3. Analysis of variance (ANOVA) for alp activity in female albino rats Descriptive Female 95% Confidence Interval for

Mean N Mean Std. Deviation Std. Error Lower Bound Upper Bound Minimum Maximum Group1 1 1.5180E2 . . . . 151.80 151.80 Group2 1 69.0000 . . . . 69.00 69.00 Group3 1 63.4800 . . . . 63.48 63.48 Group4 1 77.2800 . . . . 77.28 77.28 Group5 1 5.1612E2 . . . . 516.12 516.12 Group6 1 1.5456E2 . . . . 154.56 154.56 Control 1 6.2652E2 . . . . 626.52 626.52 Total 7 2.3697E2 233.64248 88.30856 20.8825 453.0490 63.48 626.52

REFERENCES 1.Ahmed, Z., Abdul-Fadi, M A.M. and King, E.J., Properties of 3.Alvarez, C.E., Alcover, S.R., and Sainz M.T., Pharmaceutical alkaline phosphatase”.BiochemBiophys. Acta 1959; 36:228-23. preparations which contains tartrazine”.Allergol immunopathol 2.Akanji M. A. and Ngaha E. O. Effect of repeated administration (Madr) 1981; 9(1):45-54. of berenil on urinary excretion with corresponding tissue 4.Amin KA, Abdel Hameid H 2nd , Abd Elsttar AH.Effect of food pattern in rats. Pharmacol. Toxicol.1989; 64, 272-275. azo dyes tartrazine and carmoisine on biochemical parameters related to renal, hepatic function and oxidative stress

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Sanni et al. Available at jpbms.info biomarkers in young male rats.Food Chem Toxicol. 2010 12.Indofood (2010) “Our history”. Oct;48(10):2994-9. doi: 10.1016/j.fct.2010.07.039. Epub 2010 http://www.indofood.com/about_history.aspx Aug 3. Pubmed. 13.Jain, R., Bhargava, M., and Sharma, N. Electrochemical studies 5.BBC news in “parents warned of additives link”[updated on a pharmaceutical Azo dye: tartrazine.Industrial chemistry Thursday, 6, September, 2007. Assessed September 10, 2012]. research 2003; 42(2):243.doi:10.1021/ie 020228g. Available from; www.bbc.co.uk/2/hi/6979976.stm. 14.Mountinho, I.L., Bertges, L.C., and Assis, R.V. Prolonged use of 6.Dipalma J.R. Tartrazine sensitivity. Am Fam Physician. 1990 the food dye tartrazine and its effect on gastric mucosa of Nov;42(5):1347-50. Wistar rats. Brazilian journal of biology 2007;67 (1):141-5. doi 7.Donna M. Food additives and hyperactive behavior in 3 year- 10.1590/51519-69842007000100019PMID.17505761 old and 8/9-year-old children in the community: a randomized, 15.Rowe, K.S., and Rowe, K.J., (1994) “Synthetic food colouring double blinded, placebo-controlled trial” the lancet and behavior: a dose response effect in a double blind, placebo- 2007;370(9598):1560-1567.doi:10.1016/SOI 40- controlled, repeated-measures study”. The journal of pediatrics 673(07)61306-3. PMID 17825405. 1994;125(53t1):691-8. doi: 10.1016/S50022-3476(06) 80164- 8.Ejembi D and Sanni M. Studies on the consequences of the 2.PMID 7965420. administration of Aqueous extracts of Azadiracta Indica And 16.Rowett, H.G., (1974). In the rat as a small mammal”. 3rd Morinda Lucida on the Heamatological Parameters And The edition John Murry publishers Ltd. Great Britain. Activity of phosphatases In rats Cellular Tissues ” International 17.Schmidt E. & Schmidt F.W. Determination of GOT and GTP Journal Of Asian Social Science 2012; 2(8):1222-1230. enzyme. Enzymologia Biologica Et Clinica. 1963;3(1):1–5. 9.Food Standard Agency (2008) “board discussion colour 18.Tamás L, Huttová J, Mistrk I, and Kogan G. eﬀect of advice, available from carboxymethyl chitin-Glucan on the activity of some hydrolytic www.southampton.ac.uk/mediacentre/news/2008/apr/08_65. enzymes in maize plants".Chem.Pap.2002;56(5):326–329. shtml. 19.Tortora, G.J and Grabowiski, S.R (2003) “Principles of 10. Harada M, Udagawa N, Fukasawa K, Hiraoka BY, Mogi M. anatomy and physiology”.10th ed. John Wiley and sons Publisher Inorganic pyrophosphatase activity of purified bovine pulp Inc. Hobo Ken,N.J, Pp 231-9. alkaline phosphatase at physiological pH . J Dent Res. 1986 Feb; 20.UK food guide (2007) “E102 tartrazine, FD&C yellow no 65(2):125-7. 5”.www.ukfoodguide.net/e102.htm 11.Horiuchi,T., Horiuchi, S., and Mizuno, D., (1959). "A Possible 21. Witular, R.A., (2004) “Salim at driving seat at indofood.The Negative Feedback Phenomenon controlling Formation of Jakartapost. Retrieved 1 Feb.2010. Available from Alkaline Phosphomonoesterase in Escherichia coli". Nature 183 en.wikipedia.org/wiki/Sudono_Salim; June 26, 2004. (30 May 1959): 1529–1530.DOI:10.1038/1831529b0

Conflict of interest: - Author has not declared any conflict of interest.

Source of funding: - Not declared.

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