1. James Bergenti Jessica Melhorn Kimberly Duncan

2. http://www.youtube.com/watch?v=1tRq8ExAHzk

3. Paper Review “Determination of the Scoville Heat Value for Hot Sauces and Chilies: An HPLC Experiment James D. Batchelor, Bradley T. Jones Taken from the Journal of Chemical Education

4. Background Capsaicinoids are a group of compounds that give various peppers their characteristic taste. Capsaicin is comprised of seven capsacinoids, or closely related alkanoids. Upon contact with capsaicin, neurons in the nociceptors in the palate become excited, emitting a perception of pain, and release local inflammatory mediators. SHV = 16.1 million

5. Dihydrocapsaicin and Nordihydrocapsaicin Dihydrocapsaicin: accounts for 22% of total capsaicinoid mixture. The same pungency as capsaicin. SHV = 16.1 million Nordihydrocapsaicin: Accounts for 7% of the total capsaicinoid mixture. Half as pungent as capsaicin. SHV= 9.3 million

6. Scoville Heat Value First brought to light by in 1912 by Wilbur Scoville. The Scoville heat value is relative to the final concentration of capsaicin in a compound, expressed in grams of compound per gram of sample. The value is obtained by multiplying the literature established SHV of the pure compound by the final concentration.

7. Comparative Scoville Values

8. Purpose of Experiment To quantify and determine the Scoville Heat Value of various peppers through High Performance Liquid Chromatography. Three alkaloids were tested: Capsaicin, Dihydrocapsaicin, and Nordihydrocapsaicin. SHV was calculated from integrated absorbance data.

9. Materials Chemicals: Ethanol (95%) used as an extraction solvent Synthetic Capsaicin (97%) used to prepare working standards. Instruments: HPLC with a UV/Vis absorption detector Reference Instrument: UV/Vis A 50ppm sample was run through UV/Vis in order to find the wavelength of the capsaicin peak. Approx. 224 nm

10. Unknowns Tabasco Pepper Sauce- Red Pepper Homemade Hot Sauce – Jalapeno El Yucateco Hot Sauce – Habanero Dried Red Pepper

11. Procedure Preparation of samples: 1000 ppm stock was made by dissolving 25 mg in 25 mL of 95% ethanol. Serial Dilutions: (1000 ppm)(V1) = (50)(10) =.5 mL 1000 ppm stock (1000 ppm)(V1) = (40)(10) =.4 mL 1000 ppm stock (1000 ppm)(V1) = (30)(10) =.3 mL 1000 ppm stock (1000 ppm)(V1) = (20)(10) =.2 mL 1000 ppm stock (1000 ppm)(V1) = (10)(10) =.1 mL 1000 ppm stock (1000 ppm)(V1) = (5)(10) =.05 mL 1000 ppm stock (1000 ppm)(V1) = (1)(10) =.01 mL 1000 ppm stock

12. Preparation of Hot Sauce Ground Red Pepper: 2.9846g dissolved in 50mL of ethanol. Stirred continuously, held at a slow boil for 30 minutes. El Yucateco: 15.0218g dissolved in 50mL of ethanol. Stirred continuously, held at a slow boil for 30 minutes. Homeade: 14.9652g dissolved in 50mL of ethanol. Stirred continuously, held at a slow boil for 30 minutes. Tobasco – 14.9841g dissolved in 50mL of ethanol. Stirred continuously, held at a slow boil for 30 minutes.

13. Method Solutions were filtered via vacuum filtration, brought up to volume in a 100mL Erlenmeyer flask. Approximatley 10 uL was injected onto the column for each sample. All standards and samples were run three times.

16. Day 1 Concentration (ppm)Time (min)Area 502.68551844 402.56503746 302.55444197 203.39196666 103.37153268 53.44145662 13.37123038

17. Day 2 Concentration (ppm)Time (min)Area 501.15248147 401.17228091 301.18215945 201.18162185 101.18123636 51.18107843 11.1790089

18. Day 3 Concentration (ppm)Time (min)Area 501.19192246 401.19132122 301.18107133 201.1955208 101.1827337 51.2018407 11.188670

19. Sample Calculation

20. Habanero Literature Value: 100,000-300,00 SHV Calculated Value: 77,195 SHV

21. Cayenne Ground Red Pepper: Literature Value: 30,000-50,000 SHV Calculated Value: 65,912 SHV Tabasco Sauce Literature Value: 30,000-50,000 SHV Calculated Value: 71,635

22. Jalapeno Literature Value: 2,500-8,000 SHV Calculated Value: 5,121 SHV

23. Taste Test Scale of 1-10 1 = not hot. 10 = unbearable Jalapeno – 2 Tabasco – 6 Red Pepper – 5 Habanero - 8

24. Discussion Calculated values were not extremely accurate, however the pattern remained constant. Errors: Low baseline resolution Voltage range was not constant throughout experiment Peaks were truncated Heat of peppers could have varied based upon soil they were grown in and time of year they were harvested. Standards and calibration curves could have been made of the Dihydrocapsaicin and Nordihydrocapsaicin, however due to high cost and limited availability, this was not practical. Only 50mg of synthetic capsaicin was available, so only one stock solution was able to be made.

25. Future Experiments Stock solution would be remade each time standards were made and run. Dihydrocapsaicin and Nordihydrocapsaicin calibration curves could be made. Use of a wider variety of peppers.