Skip to content
2000
image of Role of Chitosan Nanoparticles loaded with Pomegranate Peels Extract on High Fat Diet-Induced Obesity in Rats

Abstract

Introduction

Obesity is a chronic disease that is widespread throughout the world. Pomegranate is a fruit that has proven to provide beneficial health effects due to its high nutritional content and bioactive antioxidants. The current study aims to investigate the effect of chitosan nanoparticles loaded with pomegranate peel extract (PPE-CSNPs) on high-fat diet (HFD) induced obesity in male rats.

Methods

Obesity was induced by feeding a high-fat diet for 4 weeks. Thirty rats were assigned into five main groups (6 rats/group) as follows: control (dist. water, orally), HFD control (dist. water, orally), HFD + pomegranate peel extract (PPE) (500 mg/kg, orally), HFD + chitosan-NP (60 mg/kg, orally), and HFD + PPE-CSNPs (60 mg/kg, orally).

Results

The treatment of PPE-CSNPs decreased body weight, glucose, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, creatinine, urea, total cholesterol, triglycerides, low density lipoprotein-cholesterol, malondialdehyde, and nitric oxide, while high density lipoprotein-cholesterol, glutathione reduced and catalase levels increased. The histological examination revealed a significant enhancement in the structures of the liver and kidneys.

Discussion

The antioxidant and hypolipidemic activities of PPE-CS surpassed those of PPE or CS individually, confirming the synergistic effect in obesity.

Conclusion

The combination of pomegranate extract with chitosan in the form of nanoparticles enhances their hypolipidemic efficacy in the treatment of obese rats. PPE-CSNPs are promising nanonatural products that could be used in obesity management.

© 2025 Bentham Science Publishers
Loading

Article metrics loading...

/content/journals/ccb/10.2174/0122127968402312250903083111
2025-09-16
2026-02-28

  • From This Site

    /content/journals/ccb/10.2174/0122127968402312250903083111
    dcterms_title,dcterms_subject,pub_keyword
    -contentType:Contributor -contentType:Concept -contentType:Institution
    10
    5
Loading full text...

Full text loading...

References

  1. Fahmy S.R. Zaki N.I. Eid S.Z. Mohamed A.S. Hassanein S.S. Effectiveness of echinochrome on hfd-induced hyperlipidemia in rats. Nat. Prod. Bioprospect. 2019 9 5 337 344 10.1007/s13659‑019‑00221‑4 31628663
    [Google Scholar]
  2. World Health Organization 2021 Available from: https:// www.who.int/news-room/fact-sheets/detail/obesity-and-overweight
  3. Umbro I. Baratta F. Angelico F. Del Ben M. Nonalcoholic fatty liver disease and the kidney: A review. Biomedicines 2021 9 10 1370 10.3390/biomedicines9101370 34680486
    [Google Scholar]
  4. Belfort-DeAguiar R. Seo D. Food cues and obesity: Overpowering hormones and energy balance regulation. Curr. Obes. Rep. 2018 7 2 122 129 10.1007/s13679‑018‑0303‑1 29619632
    [Google Scholar]
  5. Maharjan B.R. McLennan S.V. Yee C. Twigg S.M. Williams P.F. The effect of a sustained high-fat diet on the metabolism of white and brown adipose tissue and its impact on insulin resistance: A selected time point cross-sectional study. Int. J. Mol. Sci. 2021 22 24 13639 10.3390/ijms222413639 34948432
    [Google Scholar]
  6. Obesity and overweight 2024 Available from: https:// www.who.int/news-room/fact-sheets/detail/obesity-and-overweight
  7. Karri S. Sharma S. Hatware K. Patil K. Natural anti-obesity agents and their therapeutic role in management of obesity: A future trend perspective. Biomed. Pharmacother. 2019 110 224 238 10.1016/j.biopha.2018.11.076
    [Google Scholar]
  8. Reddy Mooli R.G. Mukhi D. Ramakrishnan S.K. Oxidative stress and redox signaling in the pathophysiology of liver diseases. Compr. Physiol. 2022 12 2 3167 3192 10.1002/cphy.c200021 3557896910074426
    [Google Scholar]
  9. Abdelmawgood I.A. Mohamed A.S. Mahana N.A. Abdel Wahab A.H.A. Badr A.M. Abdelkader A.E. Chrysin-loaded PLGA nanoparticle attenuates ferroptosis in lipopolysaccharide-induced indirect acute lung injury by upregulating Nrf2-dependent antioxidant responses. Respir. Physiol. Neurobiol. 2025 336 104451 10.1016/j.resp.2025.104451 40379234
    [Google Scholar]
  10. Coulter A.A. Rebello C.J. Greenway F.L. Centrally acting agents for obesity: Past, present, and future. Drugs 2018 78 11 1113 1132 10.1007/s40265‑018‑0946‑y 30014268
    [Google Scholar]
  11. Tak Y.J. Lee S.Y. Long-Term efficacy and safety of anti-obesity treatment: Where do we stand? Curr. Obes. Rep. 2021 10 1 14 30 10.1007/s13679‑020‑00422‑w 33410104
    [Google Scholar]
  12. Mohamed A.S. Ibrahim W.M. Zaki N.I. Ali S.B. Soliman A.M. Effectiveness of Coelatura aegyptiaca extract combination with atorvastatin on experimentally induced hyperlipidemia in rats. Evid. Based Complement. Alternat. Med. 2019 2019 9726137 10.1155/2019/9726137 307135806332942
    [Google Scholar]
  13. Mohamed R.G. Mohamed A.S. Dakrory A.I. Abdelaziz M.H. Protective impacts of quercetin and echinochrome on female fertility and pregnancy outcomes in polycystic ovary syndrome in rats. Birth Defects Res. 2025 117 5 e2487 10.1002/bdr2.2487 40396197
    [Google Scholar]
  14. Moustafa M.A. Mohamed A.S. Dakrory A.I. Abdelaziz M.H. Lepidium sativum extract alleviates reproductive and developmental toxicity in polycystic ovary syndrome induced by letrozole and high-fat diet in rats. Reprod. Sci. 2025 32 4 1338 1361 10.1007/s43032‑025‑01820‑y 40048056
    [Google Scholar]
  15. Gaber R. Mohamed A.S. Baiomy A.A.A. Ovothiol-A exhibited protective and therapeutic effect against hyperthyroidism in rats. Curr. Top. Med. Chem. 2025 25 10.2174/0115680266349656250318020702 40207760
    [Google Scholar]
  16. Zuorro A. Moreno-Sader K.A. González-Delgad Á.D. Inherent safety analysis and sustainability evaluation of chitosan production from shrimp exoskeleton in Colombia. Water 2021 13 4 553 10.3390/w13040553
    [Google Scholar]
  17. de Souza A.F. Galindo H.M. de Lima M.A.B. Ribeaux D.R. Rodríguez D.M. Andrade R.F.S. Gusmão N.B. de Campos-Takaki G.M. Biotechnological strategies for chitosan production by mucoralean strains and dimorphism using renewable substrates. Int. J. Mol. Sci. 2020 21 12 4286 10.3390/ijms21124286 325602137352823
    [Google Scholar]
  18. Lovskaya D. Menshutina N. Mochalova M. Nosov A. Grebenyuk A. Chitosan-Based aerogel particles as highly effective local hemostatic agents. production process and in vivo evaluations. Polymers 2020 12 9 2055 10.3390/polym12092055 32927608
    [Google Scholar]
  19. Wawrzyniak N. Skrypnik K. Suliburska J. Dietary supplements in therapy to support weight reduction in obese patients. Acta Sci. Pol. Technol. Aliment. 2022 21 1 67 80 [PMID: 35174689
    [Google Scholar]
  20. Saad D.Y. Khalil H.A. Farouk A.E. Soliman A.M. Mohamed A.S. Therapeutic effect of chitosan-zinc oxide nanoparticles on acute kidney injury induced by gentamicin in wistar rats. Curr. Nanomed. 2023 13 3 217 227 10.2174/2468187313666230821153922
    [Google Scholar]
  21. Rostami E. Progresses in targeted drug delivery systems using chitosan nanoparticles in cancer therapy: A mini-review. J. Drug Deliv. Sci. Technol. 2020 58 101813 10.1016/j.jddst.2020.101813
    [Google Scholar]
  22. Shagdarova B. Konovalova M. Varlamov V. Svirshchevskaya E. Anti-Obesity effects of chitosan and its derivatives. Polymers 2023 15 19 3967 10.3390/polym15193967 37836016
    [Google Scholar]
  23. Sorrenti V. Randazzo C.L. Caggia C. Ballistreri G. Romeo F.V. Fabroni S. Timpanaro N. Raffaele M. Vanella L. Beneficial effects of pomegranate peel extract and probiotics on pre-adipocyte differentiation. Front. Microbiol. 2019 10 4 660 10.3389/fmicb.2019.00660 31001233
    [Google Scholar]
  24. Mo Y. Ma J. Gao W. Zhang L. Li J. Li J. Zang J. Pomegranate peel as a source of bioactive compounds: A mini review on their physiological functions. Front. Nutr. 2022 9 887113 10.3389/fnut.2022.887113 35757262
    [Google Scholar]
  25. Grabež M. Škrbić R. Stojiljković M.P. Rudić-Grujić V. Paunović M. Arsić A. Petrović S. Vučić V. Mirjanić-Azarić B. Šavikin K. Menković N. Janković T. Vasiljević, Nađa, Beneficial effects of pomegranate peel extract on plasma lipid profile, fatty acids levels and blood pressure in patients with diabetes mellitus type-2: A randomized, double-blind, placebo-controlled study. J. Funct. Foods 2019 64 103692
    [Google Scholar]
  26. Bahari H. Pourreza S. Goudarzi K. Mirmohammadali S.N. Asbaghi O. Kolbadi K.S.H. Naderian M. Hosseini A. The effects of pomegranate consumption on obesity indices in adults: A systematic review and meta‐analysis. Food Sci. Nutr. 2024 12 2 641 660 10.1002/fsn3.3739 38370057
    [Google Scholar]
  27. Zielińska A. Carreiró F. Oliveira A.M. Neves A. Pires B. Venkatesh D.N. Durazzo A. Lucarini M. Eder P. Silva A.M. Santini A. Souto E.B. Polymeric nanoparticles: Production, characterization, toxicology and ecotoxicology. Molecules 2020 25 16 3731 10.3390/molecules25163731 32824172
    [Google Scholar]
  28. Abdelmawgood I.A. Badr A.M. Abdelkader A.E. Mahana N.A. Mohamed A.S. Abdelfattah H.H. Chrysin-loaded poly (lactic-co-glycolic acid) nanoparticles alleviate sepsis-induced splenic injury by regulating myeloid-derived suppressor cells. Immunol. Res. 2025 73 1 80 10.1007/s12026‑025‑09634‑5 40358797
    [Google Scholar]
  29. Misra S.K. Mohn D. Brunner T.J. Stark W.J. Philip S.E. Roy I. Salih V. Knowles J.C. Boccaccini A.R. Comparison of nanoscale and microscale bioactive glass on the properties of P(3HB)/Bioglass® composites. Biomaterials 2008 29 12 1750 1761 10.1016/j.biomaterials.2007.12.040 18255139
    [Google Scholar]
  30. Balaji E.V. Selvan A.T. Nanopharmacology: A novel approach in therapeutics. Asian J. Res. Pharm. Sci 2019 9 1 09 16 10.5958/2231‑5659.2019.00003.1
    [Google Scholar]
  31. Fath-All A.A. Atia T. Mohamed A.S. Khalil N.M. Abdelaziz T.D. Mahmoud N.A. Elagali A.M. Sakr H.I. Abd El-Ghany M.N. Efficacy of yeast-mediated SeNPs on gastric ulcer healing and gut microbiota dysbiosis in male albino rats. Tissue Cell 2025 96 102953 10.1016/j.tice.2025.102953 40334393
    [Google Scholar]
  32. Rahnemoon P. Sarabi-Jamab M. Bostan A. Mansouri E. Nano-encapsulation of pomegranate (Punica granatum L.) peel extract and evaluation of its antimicrobial properties on coated chicken meat. Food Biosci. 2021 43 101331 10.1016/j.fbio.2021.101331
    [Google Scholar]
  33. Kotb M.A. Abdelmawgood I.A. WalyEldeen, A.A.; Abdelaziz, M.H.; Khalaf, M.L.; Mohamed, A.S.; Dakrory, A.E.; Ibrahim, S.A. Chrysin-loaded PLGA nanoparticles alleviate the implantation of endometriotic lesions via attenuation of peritoneal inflammation and downregulating NF-κB activation-driven expression of angiogenic factors. J. Drug Deliv. Sci. Technol. 2025 109 106984 10.1016/j.jddst.2025.106984
    [Google Scholar]
  34. Soltanzadeh M. Peighambardoust S.H. Ghanbarzadeh B. Mohammadi M. Lorenzo J.M. Chitosan nanoparticles as a promising nanomaterial for encapsulation of pomegranate (punica granatum l.) peel extract as a natural source of antioxidants. Nanomaterials 2021 11 6 1439 10.3390/nano11061439 34072520
    [Google Scholar]
  35. Kumar N. Pratibha; Trajkovska Petkoska, A.; Khojah, E.; Sami, R.; Al-Mushhin, A.A.M. Chitosan edible films enhanced with pomegranate peel extract: Study on physical, biological, thermal, and barrier properties. Polymers 2021 14 12 3305
    [Google Scholar]
  36. Wang Z. Pan Z. Ma H. Atungulu G.G. Extract of phenolics from pomegranate peels. Open Food Sci. J. 2011 5 1 17 25 10.2174/1874256401105010017
    [Google Scholar]
  37. Piazzini V. Landucci E. D’Ambrosio M. Tiozzo Fasiolo L. Cinci L. Colombo G. Pellegrini-Giampietro D.E. Bilia A.R. Luceri C. Bergonzi M.C. Chitosan coated human serum albumin nanoparticles: A promising strategy for nose-to-brain drug delivery. Int. J. Biol. Macromol. 2019 129 267 280 10.1016/j.ijbiomac.2019.02.005 30726749
    [Google Scholar]
  38. Chinedu E. Arome D. Ameh F. A new method for determining acute toxicity in animal models. Toxicol. Int. 2013 20 3 224 226 10.4103/0971‑6580.121674 24403732
    [Google Scholar]
  39. Reed M.J. Meszaros K. Entes L.J. Claypool M.D. Pinkett J.G. Gadbois T.M. Reaven G.M. A new rat model of type 2 diabetes: The fat-fed, streptozotocin-treated rat. Metabolism 2000 49 11 1390 1394 10.1053/meta.2000.17721 11092499
    [Google Scholar]
  40. Bergfeld W.F. Belsito D.V. Hill R.A. Klaassen C.D. Liebler D.C. James G. Shank R.C. Safety assessment of camellia sinensis-derived ingredients as used in cosmetics. Int. J. Toxicol. 2019 38 3 Suppl. 48S 70S 10.1177/1091581819889914 31840549
    [Google Scholar]
  41. Bancrof J.D. Stevens A. Theory and Practice of Histological Techniques. New York, NY, USA Churchill Levingstone 1996
    [Google Scholar]
  42. Al-Muammar M.N. Khan F. Obesity: The preventive role of the pomegranate (Punica granatum). Nutrition 2012 28 6 595 604 10.1016/j.nut.2011.11.013 22342388
    [Google Scholar]
  43. Cheung R. Ng T. Wong J. Chan W. Chitosan: An update on potential biomedical and pharmaceutical applications. Mar. Drugs 2015 13 8 5156 5186 10.3390/md13085156 26287217
    [Google Scholar]
  44. Keys A. Fidanza F. Karvonen M.J. Kimura N. Taylor H.L. Indices of relative weight and obesity. Int. J. Epidemiol. 2014 43 3 655 665 10.1093/ije/dyu058 24691951
    [Google Scholar]
  45. Manna P. Jain S.K. Obesity, oxidative stress, adipose tissue dysfunction, and the associated health risks: Causes and therapeutic strategies. Metab. Syndr. Relat. Disord. 2015 13 10 423 444 10.1089/met.2015.0095 26569333
    [Google Scholar]
  46. Jiang S.Z. Lu W. Zong X.F. Ruan H.Y. Liu Y. Obesity and hypertension. Exp. Ther. Med. 2016 12 4 2395 2399 10.3892/etm.2016.3667 27703502
    [Google Scholar]
  47. Sabi E.M. Bin Dahman L.S. Mohammed A.K. Sumaily K.M. Al-Daghri N.M. -2548G>A LEP polymorphism is positively associated with increased leptin and glucose levels in obese saudi patients irrespective of blood pressure status. Medicina 2022 58 3 346 10.3390/medicina58030346 35334523
    [Google Scholar]
  48. Zhao X. Yuan Z. Anthocyanins from Pomegranate (Punica granatum L.) and Their Role in Antioxidant Capacities in Vitro. Chem. Biodivers. 2021 18 10 e2100399 10.1002/cbdv.202100399 34388293
    [Google Scholar]
  49. Lavoro A. Falzone L. Gattuso G. Salemi R. Cultrera G. Leone G. Scandurra G. Candido S. Libra M. Pomegranate: A promising avenue against the most common chronic diseases and their associated risk factors.[Review] International Journal of Functional Nutrition 2021 2 2 6 10.3892/ijfn.2021.16
    [Google Scholar]
  50. Jurenka J.S. Therapeutic applications of pomegranate (Punica granatum L.): A review. Altern. Med. Rev. 2008 13 2 128 144 [PMID: 18590349
    [Google Scholar]
  51. Olvera-Sandoval C. Fabela-Illescas H.E. Fernández-Martínez E. Ortiz-Rodríguez M.A. Cariño-Cortés R. Ariza-Ortega J.A. Hernández-González J.C. Olivo D. Valadez-Vega C. Belefant-Miller H. Betanzos-Cabrera G. Potential mechanisms of the improvement of glucose homeostasis in type 2 diabetes by pomegranate juice. Antioxidants 2022 11 3 553 10.3390/antiox11030553 35326203
    [Google Scholar]
  52. Bai Y. Zheng J. Yuan X. Jiao S. Feng C. Du Y. Liu H. Zheng L. Chitosan oligosaccharides improve glucolipid metabolism disorder in liver by suppression of obesity-related inflammation and restoration of peroxisome proliferator-activated receptor gamma (PPARγ). Mar. Drugs 2018 16 11 455 10.3390/md16110455 30463189
    [Google Scholar]
  53. Ge J. Miao J.J. Sun X.Y. Yu J.Y. Huangkui capsule, an extract from Abelmoschus manihot (L.) medic, improves diabetic nephropathy via activating peroxisome proliferator–activated receptor (PPAR)-α/γ and attenuating endoplasmic reticulum stress in rats. J. Ethnopharmacol. 2016 189 238 249 10.1016/j.jep.2016.05.033 27224243
    [Google Scholar]
  54. Jalili V. Poorahmadi Z. Hasanpour Ardekanizadeh N. Gholamalizadeh M. Ajami M. Houshiarrad A. Hajipour A. Shafie F. Alizadeh A. Mokhtari Z. Shafaei H. Esmaeili M. Doaei S. The association between obesity with serum levels of liver enzymes, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and gamma‐glutamyl transferase in adult women. Endocrinol. Diabetes Metab. 2022 5 6 e367 10.1002/edm2.367 36039792
    [Google Scholar]
  55. Singh A. Sharma, Om Clinical biochemistry of hepatotoxicity. Journal of Clinical Toxicology 2011 4 Special Issue S4 S001 10.4172/2161‑0495.S4‑001
    [Google Scholar]
  56. Soliman A.M. Mohamed A.S. Abdel-Khalek A.A. Badran S.R. Impact of polyvinyl chloride nano-plastics on the biochemical status of Oreochromis niloticus under a predicted global warming scenario. Sci. Rep. 2025 15 1 3671 10.1038/s41598‑025‑87558‑8 39880882
    [Google Scholar]
  57. Bouasla A. Bouasla I. Boumendjel A. Abdennour C. El Feki A. Messarah M. Prophylactic effects of pomegranate (Punica granatum) juice on sodium fluoride induced oxidative damage in liver and erythrocytes of rats. Can. J. Physiol. Pharmacol. 2016 94 7 709 718 10.1139/cjpp‑2015‑0226 27124270
    [Google Scholar]
  58. Abou zaid, O.; Elsonbaty, S.; Moawad, F.; Abdelghaffar, M. Antioxidants and hepatoprotective effects of chitosan nanoparticles against hepatotoxicity induced in rats. Benha Vet. Med. J. 2019 36 1 252 261 10.21608/bvmj.2019.111628
    [Google Scholar]
  59. Tsuboi N. Okabayashi Y. Shimizu A. Yokoo T. The Renal Pathology of Obesity. Kidney Int. Rep. 2017 2 2 251 260 10.1016/j.ekir.2017.01.007 29142961
    [Google Scholar]
  60. Khan H.N. Pergulwar A. Siddiqui A.M. Shinde A.R. Estimation of serum urea, creatinine and uric acid in obese subjects. Int. J. Innov. Res. Med. Sci. 2017 2 8 2455 8737
    [Google Scholar]
  61. Mestry S.N. Gawali N.B. Pai S.A. Gursahani M.S. Dhodi J.B. Munshi R. Juvekar A.R. Punica granatum improves renal function in gentamicin-induced nephropathy in rats via attenuation of oxidative stress. J. Ayurveda Integr. Med. 2020 11 1 16 23 10.1016/j.jaim.2017.09.006 29555255
    [Google Scholar]
  62. Jing S.B. Li L. Ji D. Takiguchi Y. Yamaguchi T. Effect of chitosan on renal function in patients with chronic renal failure. J. Pharm. Pharmacol. 1997 49 7 721 723 10.1111/j.2042‑7158.1997.tb06099.x 9255718
    [Google Scholar]
  63. Loomba R. Friedman S.L. Shulman G.I. Mechanisms and disease consequences of nonalcoholic fatty liver disease. Cell 2021 184 10 2537 2564 10.1016/j.cell.2021.04.015 33989548
    [Google Scholar]
  64. Al-Moraie M.M.D. Arafat R.A. Al-Rasheedi A.A. Effect of pomegranate juice on lipid profile and antioxidant enzymes in hypercholesterolemic rats. Life Sci. J. 2013 10 3 2717 2728
    [Google Scholar]
  65. Chen R.Y. Liu S.H. Chiang M.T. Effects and mechanisms of chitosan and chitosan oligosaccharide on hepatic lipogenesis and lipid peroxidation, adipose lipolysis, and intestinal lipid absorption in rats with high-fat diet-induced obesity. Int. J. Mol. Sci. 2021 22 3 1 16
    [Google Scholar]
  66. Ahn S.I. Cho S. Choi N.J. Effectiveness of chitosan as a dietary supplement in lowering cholesterol in murine models: A meta-analysis. Mar. Drugs 2021 19 1 26 10.3390/md19010026 33435383
    [Google Scholar]
  67. Thomàs-Moyà E. Gómez-Pérez Y. Fiol M. Gianotti M. Lladó I. Proenza A.M. Gender related differences in paraoxonase 1 response to high-fat diet-induced oxidative stress. Obesity 2008 16 10 2232 2238 10.1038/oby.2008.340 18719679
    [Google Scholar]
  68. Sansbury B.E. Hill B.G. Regulation of obesity and insulin resistance by nitric oxide. Free Radic. Biol. Med. 2014 73 383 399 10.1016/j.freeradbiomed.2014.05.016 24878261
    [Google Scholar]
  69. Saghir F.S.A. Ibrahim F.S. Amom Z. Othman L. The role of oxidative stress and inflammation in prediabetes: A review. Malays J. Med. Health. Sci. 2023 19 4 326 331 10.47836/mjmhs.19.4.45
    [Google Scholar]
  70. Sutthasupha P. Lungkaphin A. The potential roles of chitosan oligosaccharide in prevention of kidney injury in obese and diabetic conditions. Food Funct. 2020 11 9 7371 7388 10.1039/D0FO00302F 32839793
    [Google Scholar]
/content/journals/ccb/10.2174/0122127968402312250903083111
Loading
Role of Chitosan Nanoparticles loaded with Pomegranate Peels Extract on High Fat Diet-Induced Obesity in Rats
Bentham Science Publishers ; https://doi.org/10.2174/0122127968402312250903083111
/content/journals/ccb/10.2174/0122127968402312250903083111
/content/journals/ccb/10.2174/0122127968402312250903083111
Loading

Data & Media loading...


  • Article Type:     Research Article
Keywords: Obesity ; oxidative stress ; high-fat diet ; nanoparticles ; natural product ; pomegranate

Most Cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error
Please enter a valid_number test