1. Abstract: One of the most popular herbal agents in the US market is Saw Palmetto (Sabal, Serenoa repens), which is derived from the American dwarf palm tree. Among its important usages include the treatment for prostrate complaints, bladder irritation, urination problems in BPH stages I and II. However the significant limiting factor to our understanding of the use and effectiveness of this phytotherapy is the lack of standardization of the products and unknown active chemical constituents. It has become essential to standardize these products,so that the variations between products and within different batches of the same manufacturer can be assessed. In this study, various finished commercial Saw palmetto preparations purchased from pharmacies or well reputed health stores in the USA were analyzed for their content based upon the concentrations of four free fatty acids (oleic, myristic, linoleic and lauric acid) after solvent extraction with heptane followed by derivatization with phenacyl bromide/triethylamine in acetone by an HPLC method on a C 18 column. The compounds were eluted using Acetonitrile/Water and the eluents were monitored at a wavelength of 242 nm. The content of these four free fatty acids varied from 0.3 – 31.3 % lauric acid, 0.2- 13 % for linoleic acids; 0.06- 7.78 % for myristic acid and 0.7 – 71.99 % for oleic acid. It was interesting to observe that the concentration of lauric and oleic acids were more in the soft gel formulations as compared to tablet or capsule. The variability in concentration of these marker compounds, suggest a need for standardization of saw palmetto products for maximum benefits to our population. Introduction: The lipidosterolic extract of Saw Palmetto plant or Serenoa repens [Batr] has been used for many years to treat symptomatic Benign Prostatic Hyperplasia [BPH] throughout Europe [1,2]. It’s use in the United States is increasing rapidly [1]. Chemical investigation of Serenoa repens revealed the presence of numerous free fatty acids, fatty acid esters and glycerides, phytosterols, carbohydrates, and aromatic acids. The n-hexane (lipidosterolic) extracts have been reported to contain 35,30,11 and 5 % of oleic, lauric, myristic and linoleic acids respectively as free fatty acids, and less than 2 % of phytosterol, aliphatic alcohols and polypyrenic compounds [2,3]. The lipidosterolic extract is commonly blended with excipients to form powders with or without standardization. Saw palmetto preparations are standardized to 20 –25% fatty acids. It is difficult to quantitate fatty acids directly with UV or fluorescence detection due to lack of chromophores in the structure. HPLC methods of fatty acids after derivatization, have been reported in the literature. Various derivatization techniques have also been investigated [4]. The increasing demand for Saw palmetto has led to the production of various commercial products in the market. However, it is difficult to find products that are uniform in content. The purpose of this pilot study was to develop a simple HPLC method for standardization of Saw Palmetto products. Conclusion: Our data show that US Saw palmetto products contain lauric acid, linoleic acid, myristic acid and oleic acid in various concentrations. The variability in the concentration of these marker compounds, suggests that standardization may be necessary for quality assurance. References [1]http//www.nutritional-supplement-info.com/saw palmetto html. [2] Bombardelli, E. and Morazzoni, P. (1997) Serenoa repens (Bartram) J.K. Small. Fitoterapia LXVIII:99 - 113. [3] Raynaaud, J.P., Cousse, H. and Martin, P.M. (2002) Inhibition of type 1 and type 2 5-alpha-reductase activity by free fatty acids, active ingredients of Permixon. J. Steroid Biochem. Mol. Biol. 82: 233 – 239. [4] Mehta, A., Oeser, A.M., Carlson, MG. (1998) Rapid quantitation of free fatty acids in human plasma by high-performance liquid chromatography. J. Chromatography B 719: 9 - 23. VARIATIONS IN SAW PALMETTO FORMULATIONS IN THE US MARKET N. M. Enwerem 1 *, K. Kumar 1, F. Ayorinde 2, K. R. Scott 1 and A.K. Wutoh 1 1 Department of Pharmaceutical Sciences and 2 Department of Chemistry, Howard University, College of Pharmacy, 2300 4 th Street, NW, Washington DC, 20059-0001 Aim: To determine the variability in a range of Saw palmetto products in the United States. To do this, we identified and measured the concentration of four marker compounds [Lauric acid, Linoleic acid, Myristic acid and Oleic acid] by HPLC with UV detection. Results: A typical HPLC separation of fatty acids, Lauric acid, Linoleic acid, Myristic acid and Linoleic acid is shown for acid standards (Fig 2) and in a representative sample (Fig 4). The on-line UV spectra are shown in Figs 3 and 5 respectively.The retention times for lauric acid, linoleic acid, myristic acid and oleic acid were 8.4,12.1,13.4,and 16.6 min, respectively. The four marker compounds (Lauric acid, Linoleic acid, Myristic acid and Oleic acid) showed linearity over the range 62.5 – 1000 µM and the accuracy and precision data are shown in tables 2 and 3. Experimental Standards and Controls  8 mM of Lauric acid, Myristic acid, Linoleic acid and Oleic acid were prepared in heptane.  1.0 ml of each solution was transferred to a sample tube, vortexed and labeled as mixed working standard.  1:1 serial dilution of this mixture were prepared in heptane to give calibration standards [1000, 500, 250,125, 62.5, 31.25, 15.6 and 7.8  M] and quality control standards [62.5 – 1000  M].  The calibration and quality control standards, were derivatized with phenacyl bromide, triethylamine and acetic acid, using modified method of Adosh et al [4] as shown in Fig. 1.  The derivatized solution was evaporated to dryness under an atmosphere of nitrogen using an integrated Speed vac system for 30 minutes.  The residue was reconstituted in acetonitrile/water mixture [83/17] and analyzed by HPLC.  Sample: Saw palmetto commercial dietary supplement products were purchased from local pharmacies, herbal and grocery shops in the District of Columbia and Maryland, USA. To protect the manufacturers’ identity, the samples are labeled A to J.  Sample Preparation  Five dosage units of each product to be tested were triturated to form a uniform mix.  50 mg of the sample was weighed into a sample tube & extracted with 15 mL of heptane for 1 hr.  The mixture was centrifuged at 800 x g for 10 minutes and filtered.  1 ml of the resulting extract was transferred to a reaction vial, derivatized with phenacyl bromide (100  L), triethylamine (100  L) and acetic acid (150  L) and analyzed by HPLC.  Chromatography: HPLC measurements were made using a Waters 2695 Alliance HPLC system, equipped with a 996 photodiode array detector. Separation of the analytes was performed on a 5  m VYDAC, C 18 column (250 cm x 4.6 mm id) at a flow rate of 1.0 ml/min using a gradient. The eluents were monitored at 242 nm at ambient temperature. The column temperature was maintained at 25  5º C.The chromatographic separation was accomplished by a gradient solvent system (Table 1)  Intraday and interday assay precision and accuracy are shown in Tables 2 and 3. Variability was below 7 % over the concentration range ( 62.5 – 1000  M) with accuracy between 99-100% for Lauric acid, 93-103% for Linoleic acid, 89-104%for Myristic acid and 93-103%for Oleic acid.  There was a significant product-to-product variability in the amount of these fatty acids (Table 4).Total lauric acid varied 92 fold (0.3-31.3 % by weight), Linoleic acid varied 65 fold ( 0.2 –13 % by weight), Myristic acid varied by 128 fold ( 0.06-7.78 % by weight) and oleic acid varied by 100 fold (0.7 – 71.99 % by weight). The concentration of these acids were more in gel preparations than in powder formulations (capsules and tablet). Fig 2&3: Chromatogram of a mixture of standards & their on-line spectra Fig 4&5: Chromatogram of a represent-ative sample & it’s on- line spectra Fig. 1 Time (mins) Flow% A% B 0.01.08020 8.01.08515 16.01.09010 20.01.01000 22.01.01000 23.01.08010 Table 1: Gradient table; (A = acetonitrile, B = water) Table 2: Intraday accuracy & precision (n=5) Table 3: Interday accuracy & precision (n=4) Table 4: Concentration of marker compounds in commercial Saw palmetto products Acknowledgement: The funding for this project was provided by Center for Excellence (COE), Howard University, HRSA. Fig 2Fig 3 Fig 5 Fig 4