Herbal medicines?: a case study

Adulterated honey raises more than just questions!
Analytical Unit, St George’s - University of London, UK
Susannah Kenyon, Jennifer Button, Denise A. McKeown, Terry D. Lee and David W. Holt.
Introduction
Screening
Herbal remedies as prophylaxis and treatment of a A 50-year-old man was introduced to the herbal honey A reference standard was prepared by mixing a reference Screening indicated that both jars submitted by the variety of ailments have been sought after and used for by a friend. The patient reported good clinical efficacy to ‘blank’ honey (purchased from Sainsbury’s PLC) with patient and all sachets analysed, were positive for centuries. Herbal medicines and remedies for the his GP, who became suspicious about the contents of deionised water containing sildenafil (50mg/L), tadalafil sildenafil. Tadalafil and vardenafil were not detected treatment of impotence are widely available from Internet the product. Two jars of honey were sent to the (10mg/L) and vardenafil (16mg/L). The honey samples sites that promote them as “safe, herbal alternatives” Analytical Unit for toxicological analysis (one used by the and a blank were prepared by mixing with deionised Quantitation of the contents revealed a mean and “100% natural” products.1,2 In the United Kingdom water. Diluted standards, blanks and samples were concentration of 59mg of sildenafil per sachet (~20g there are 3 phosphodiesterase type 5 inhibitor drugs analysed using a Perkin Elmer Lambda 35 UV/VIS The manufacturer of the honey product was identified as of honey). Figure 3 shows the variation in content available on prescription to treat impotence: sildenafil, spectrometer (Wellesley Mass), scanning across a UV Etumax and a similar product was purchased directly within and between sachets analysed.
wavelength range 190-400nm. Diluted standards, blanks from the Etumax website10. Sachets (12 x 20g) of “Royal and samples were also extracted using pH7 buffer into Honey” were received in a presentation box at a cost of MTBE, evaporated under vacuum, reconstituted in 80% Herbal alternatives to impotence drugs are becoming $55 plus $10p&p. Overall packaging and description of methanol and injected onto the LC system. A Perkin increasingly popular as they appeal to individuals who the product were similar to those used by the patient Elmer Series 200 pump, autosampler and column oven are unable to take phosphodiesterase inhibitors. Herbal (figure 1). The listed ingredients in the purchased honey was used. Chromatography was achieved on an Alltech products for the treatment of erectile dysfunction have were Radix Eurycoma longifolia Extract (200mg), Radix Alltima C18 (150 x 2.1mm, 5µm) column maintained at been found to contain sildenafil and tadalafil before.5,6 Panax Ginseng powder (200mg), Bee Larva Powder 50°C. The mobile phase contained methanol:deionised A recent news article reported an Israeli beekeeper water (80:20 v/v), with ammonium formate added to claiming that his bees that feed on a local flower called achieve a final concentration of 2mM. Mass- ‘Zallouh’ on Mount Hermon on the borders of Israel, spectrometric detection was performed on a Sciex Syria and Lebanon, can produce honey that improves Pure reference standards of sildenafil (Viagra), tadalafil API2000 triple quadrupole equipped with a turbo-ion 1 1 2 2 3 3 1 1 2 2 3 3 1 1 2 2 3 3 1 1 2 2 3 3 1 1 2 2 3 3 1 1 2 2 3 3 (Cialis) and vardenafil (Levitra®) were generous gifts spray interface (Applied Biosystems) held at 400°C. “Lebanese Viagra” and its key ingredient is ferula from Pfizer, Lilly and Bayer, respectively. Two jars of Nitrogen was used as collision gas. Positive Honey sachet and aliquot number, with mean result
honey bought by the patient and 6 of 12 sachets ionisation was used, and the extracts were Figure 3: Sildenafil contents of the 6 sachets of
purchased by the Analytical Unit were analysed. analysed using both product ion scan and honey analysed (3 aliquots of each, injected in Aliquots of the jar honey (~5g), and the entire contents of multiple reaction monitoring (MRM) mode. The duplicate). Overall mean was 59.4mg of sildenafil each of the sachets (~20g) were screened by scanning precursor ions [M+H]+ for sildenafil, tadalafil (UV) and liquid and vardenafil were m/z 475, 390 and chromatography with tandem mass-spectrometric 489 respectively. Product ions used Conclusions
It is concerning to find another product, being sold as a herbal treatment for impotence, containing a Quantitation
prescription only medicine. It is also unusual being a food product, that may be consumed unsuspectingly. deionised water over a concentration range 10-500ng/mL. This is the second product sold by Etumax that has Honey samples (entire sachet contents ~20g) were been found to contain sildenafil5. Unlike the previous diluted 1 in 100 with deionised water. 3 separate aliquots product, it contained only sildenafil and no other Figure 1: (a) Jars of honey
from the initial dilutions were taken and diluted a further 1 phosphodiesterase type 5 inhibitor. Sildenafil was in 10,000 with deionised water. As a result the honey also found in quantities (59.4mg per sachet), that samples (diluted 1 in 1,000,000) were effectively aqueous would cause a notable pharmaceutical effect or solutions of sildenafil, therefore matching the calibrator interaction (typical dose 50-100mg per day3). Our matrix. Standards, blanks and samples were then findings further demonstrate the potential dangers for injected in duplicate directly onto the LC/MS/MS system. patients taking “natural” or “herbal” products that are Only the sachets of honey were quantified. not under the control of a regulatory agency. The case described here involves the use of honey References
claiming to be herbal and sold to improve fertility and R Thurairaja, B Barrass and R Persad. Internet websites selling herbal treatments for erectile dysfunction. Int J Impot Res. 6. Fleshner N, Harvey M, Adomat H, Wood C, Eberding A, Hersey K and Guns E. Evidence for contamination of herbal erectile dysfunction products with phosphodiesterase type 5 inhibitors. J Urol. 2005 Aug;174(2):636-41 Dennehy CE, Tsourounis C and Miller AE. Evaluation of herbal dietary supplements marketed on the internet for recreational 7. Matti Friedman. Guardian Sep 30, 2007. www.guardian.co.uk cause “distinguished sexual activity”. The product listed use. Ann Pharmacother. 2005 Oct;39(10):1634-9. 8. K. A. Hadidi, T. Aburjai and A. K. Battah. A comparative study of Ferula hermonis root extracts and sildenafil on British National Formulary (BNF), vol 47 2004 p405 copulatory behavious of male rats. Fitoterapia. 2003 Apr 74(3).
no prescription drugs among its ingredients. Smith KM and Romanelli F. Recreational use and misuse of phosphodiesterase 5 inhibitors. J Am Pharm Assoc (Wash 9. Ferula Hermonis: Discover the “Lebanese Viagra” that can Restore Your Libido and Sexual Potency found at Figure 2: LC/MS/MS chromatogram of 50ng/mL
DC). 2005 Jan-Feb;45(1):63-72; quiz 73-5.
www.thehealthierlife.co.uk/article/3691/ferula-hermonis-impotence.html. Accessed 4-Oct 2007.
Kenyon .S, Button .J, Perella .P, McKeown D.A and Holt .D.W. An Herbal Remedy: More Than Was Bargained For. Journal 10. Etumax website; wwwetumax.com accessed 16 Oct 2007.
sildenafil std, blank and honey sample (1 in 1,000,000). of Clinical Pharmacology. 2006 Nov46(11)1379-81

Source: http://www.forensic-toxicology.org/Honey.pdf