, Volume 19, Issue 4, pp 227–233 | Cite as

A toxicological investigation of a celery seed extract having anti-inflammatory activity

  • M. C. Powanda
  • K. D. Rainsford
Research Article


Background and aims

An extract of the seed from celery (Apium graviolens) (CSE), and fractions thereof, have been found to possess anti-inflammatory activity, gastro-protective activity, and anti-Helicobacter pylori activity. In view of the potential for employing these extracts for therapeutic use, toxicological investigations were undertaken with an alcoholic extract (A-CSE) which has previously been shown to have the above pharmacological activities.


A 28-day toxicity study was performed in rats according to Good Laboratory Practice (GLP) conditions. Eighteen adult male and 18 adult female rats were randomly assigned to 3 treatment groups of 6 rats/sex/group and were dosed orally with A-CSE of 0, 150 or 5,000 mg/kg per day. Daily observations of vital signs and body weights were recorded and ophthalmological investigations were performed. At autopsy, the principal organs were weighed and sections collected for histological analysis. Serum and urine samples were collected at termination for routine clinical chemistry. Under non-GLP conditions alpha-2-μ-globulin immunohistochemistry was performed on kidney tissues and hepatic cytochrome P450 protein was determined, as well as, the enzymatic activities of the principal isoforms.


All animals survived treatments with no visible or behavioral signs of toxicity being observed during the study. There were no statistically significant differences in body weight gains, body weight gains per day or cumulative absolute body weight gains, for either sex, in any treatment groups when compared with controls. Slightly increased liver weight and liver to body and brain weight ratios were observed in female rats and in liver to body weight ratios in male rats given high dose A-CSE which was a test article effect, but the absence of any microscopic correlates for the liver weight increases suggests that these were not toxicologically significant. Treatment related macroscopic changes were not observed at necropsy and microscopic findings were limited to minimal increases in gastric eosinophils in several male and female rats in the 5,000 mg/kg per day treatment groups. Minimal focal degeneration of renal tubules was observed sporadically in both sexes assigned to all treatment groups including control and was consistent with early spontaneous nephropathy of laboratory rats and thus was not considered to represent a pathologic change associated with the test article. Increased serum globulin and phosphorus levels were observed in male rats given 5,000 mg/kg per day A-CSE and decreased serum triglycerides levels in female animals given 150 or 5,000 mg/kg per day A-CSE. The increase in serum globulin and phosphorus in male animals was small in magnitude and not considered toxicologically significant. The mechanism for the decrease in serum triglycerides in female rats was not apparent. Changes in urinalysis parameters were limited to small decreases in urine pH in female animals in the 150 and 5,000 mg/kg per day groups and were not deemed toxicologically significant. Alpha-2-μ-globulin immunohistochemistry was performed on kidney tissues from all animals and found to be within normal physiologic limits. Minor corneal mineralization occurred in some animals from all treatment groups. Cataracts were observed in one in the control and one in an animal that had 5,000 mg/kg per day but since the cataracts occurred in the metabolically inactive region of the lens, these were not considered indicative of test article related lesions. There were no changes in total hepatic microsomal protein or in total cytochrome P450 protein. Although male rats appeared to have to higher levels of total microsomal protein than female rats, there appeared to be no treatment effect in either male or female animals. As regards the activity of the various isoforms tested (CYP2B1/2, CYP1A1/2, CYP3A1/2), with the large range of activities detected for each P450 isoform, no clear change in activity or protein were observed, however, these data were not statistically analyzed.


These results suggest that there are no toxicologically significant sub-chronic effects of oral A-CSE in rats. The no adverse effect level for systemic toxicity would appear to be 5,000 mg/kg per day.


Apium graveolens Celery seed Toxicology Cytochrome P450 Anti-inflammatory activity 



We thank Drs Mary Beth Bauer, Steven Meller, and Amy L Roe and Kara E Woeller of The Proctor and Gamble Company Cincinnati, OH, USA) for their generous support and encouragement of this study. Our thanks also to Mr Vern Murdock and Mr Paul Sweeney of Beagle International Pty Ltd (Nerang, QLD, Australia) for generous donations of A-CSE used in these studies, and Professor Michael W Whitehouse (Griffiths University, QLD, Australia) for valuable advice.


  1. Amacher DE (2010) The effects of cytochrome P450 induction by xenobiotics on endobiotic metabolism in pre-clinical safety studies. Toxicol Mech Methods 20(4):159–166PubMedCrossRefGoogle Scholar
  2. Bjeldanes LF, Kim I-S (1977) Phthalide components of celery essential oil. J Org Chem 42:2333–2335CrossRefGoogle Scholar
  3. Blumenthal M, Busse WR, Goldberg A, Gruenwald J, Hall T, Riggins CW, Rister RS (1998) The complete german commission E monographs. Therapeutic guide to herbal medicines. American Botanical Council/Integrative Medicine Communications, Boston/AustinGoogle Scholar
  4. Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  5. Burke MD, Thompson S, Halpert J, Haaparanta T, Mayer RT (1985) Ethoxy-, pentoxy-, and benzyloxyphenoxazone and homologues: a series of substrates to distinguish between different induced cytochromes P-450. Biochem Pharmacol 34:3337–3345PubMedCrossRefGoogle Scholar
  6. Butters DE, Whitehouse MW (2003) Treating inflammation: some (needless) difficulties for gaining acceptance of effective natural products and traditional medicines. Inflammopharmacology 11:97–110PubMedCrossRefGoogle Scholar
  7. Butters DE, Davis CKC, McGeary RP, Powanda MC, Rainsford KD, Whitehouse MW (1999) Extracts of celery seed for the prevention and treatment of pain, inflammation and gastrointestinal irritation. WO 00/40258. 3 November 1999. US Patent 6,352,728, 5 Mar 2002 and Continuation in part as US Patent 6,576,274, 10 June 2003Google Scholar
  8. CDER (2005) Guidance for industry: estimating the maximum safe starting dose in initial clinical trials for therapeutics in adult healthy volunteers.
  9. Craig WJ (1999) Health-promoting properties of common herbs. Am J Clin Nutr 70:491–499Google Scholar
  10. Flockhart DA (2009) Drug interactions: cytochrome P450 drug interaction table, version 5.0 released on January 12, 2009. Indiana University School of Medicine.
  11. Friedmen M, Henika PR, Mandrell RE (2002) Bactericidal activities of plant essential oils and some of their isolated constituents against Campylobacter jejuni, Escherichia coli, Listeria monocytogenes and Salmonella enterica. J Food Prot 65:1545–1560Google Scholar
  12. Kamal M, Adel MA, Ahmad D, Talal A (2009) Hypolipidemic effects of seed extract of celery (Apium graveolens) in rats. Pharmacogn Mag 1:301–305Google Scholar
  13. Kapoor LD (1990) CRC handbook of ayurvedic medicinal plants. CRC Press, Boca RatonGoogle Scholar
  14. Kitajima J, Ishikawa T, Satoh M (2003) Polar constituents of celery seed. Phytochemistry 64:1003–1011PubMedCrossRefGoogle Scholar
  15. Lewis DA, Tharib SM, Veitch GBA (1985) The anti-inflammatory activity of celery Apium graveolens L. (Fam. Umbelliferae). Int J Crude Drug Res 23:27–32Google Scholar
  16. Macleod AJ, Macleod G, Subramanian G (1988) Volatile aroma constituents of celery. Phytochemistry 27:373–375CrossRefGoogle Scholar
  17. Mdidea (2006) Products. Accessed 17th Nov 2006
  18. Mencherini T, Cau A, Bianco G, Della Loggia R, Aquino RP, Autore G (2007) An extract of Apium graveolens vardulce leaves: structure of the major constituent, apiin, and its anti-inflammatory properties. J Pharm Pharmacol 59:891–897PubMedCrossRefGoogle Scholar
  19. Michio T (2005) Anti-inflammatory agent and foods and drinks containing same compounds. US Patent 5916565Google Scholar
  20. Momin RA, Nair MG (2001) Mosquitocidal, nematicidal and antifungal compounds from Apium gravelolens L. seeds. J Agric Food Chem 49:142–145PubMedCrossRefGoogle Scholar
  21. Momin RA, Ramsewak RS, Nair MG (2000) Bioactive component and 1,3-di-[(cis)-9-octadecenoyl]-2-[(cis,cis)-9,12-octadecadienoyl]glycerol from Aprium graveolens L. seeds. J Agric Food Chem 48:3785–3788PubMedCrossRefGoogle Scholar
  22. Oiye SO, Muroki NM (2002) Use of spices in foods. J Food Technol Africa 7:39–44Google Scholar
  23. Raffa RB (2005) Analgesic patent applications: I. Rev Analg 8:71–84CrossRefGoogle Scholar
  24. Rainsford KD, Liu Z-P (2006) Anti-helicobacter activity of celery seed extract. US Patent, No. 6,352,728; 20060013906Google Scholar
  25. Riddle JM (2004) Kidney and urinary therapeutics in early medieval monastic medicine. J Nephrol 7:324–328Google Scholar
  26. Sonderfan AJ, Arlotto MP, Dutton DR, MCMillen SK, Parkinson A (1987) Regulation of testosterone hydroxylation by rat liver microsomal cytochrome P-450. Arch Biochem Biophys 255:27–41PubMedCrossRefGoogle Scholar
  27. Usher G (1974) A dictionary of plants used by man. Constable, LondonGoogle Scholar
  28. Wei A, Shibamoto T (2007) Antioxidant activities and volatile constituents of various essential oils. J Agric Food Chem 55:1734–1742Google Scholar
  29. Whitehouse MW, McGeary RP, Davis CKC (2000) Anti-inflammatory activity of celery seed extracts in rats with chronic inflammation. Inflammopharmacology 8:310–311Google Scholar
  30. Whitehouse MW, Butters DE, Clarke ML, Rainsford KD (2001) NSAID gastropathy: prevention by celery seed extracts in disease-stressed rats. Inflammopharmacology 9:201–209CrossRefGoogle Scholar
  31. Zheng G-q, Kenney PM, Zhang J, Lam LKT (1993) Chemoprevention of benzo[α]pyrene-induced forestomach cancer in mice by natural phthalides from celery seed oil. Nutr Cancer 19:77–86PubMedCrossRefGoogle Scholar
  32. Zhou Y, Taylor B, Smith TJ, Liu Z-P, Clench M, Davies NW, Rainsford KD (2009) A novel compound from celery seed with a bactericidal effect against Helicobacter pylori. J Pharm Pharmacol 61:1067–1077PubMedGoogle Scholar

Copyright information

© Springer Basel AG 2010

Authors and Affiliations

  1. 1.M/P Biomedical Consultants LLCMill ValleyUSA
  2. 2.Biomedical Research CentreSheffield Hallam UniversitySheffieldUK

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