Hplc Analysis of Capsaicin
HPLC Analysis of Capsaicin
Eric Sorenson
Work Completed on 11/28/12
Quantitative analysis of Capsaicin was analyzed in three peppers that were expected to differ greatly in their capsaicin content and thus their relative spiciness. The idea was to have a nice range of capsaicin concentrations to clearly reflect the subjective pain one experiences from biting into a chili. Fresh Habanero, Jalapeno and Red Bell Pepper were purchased from the local Family Fresh grocery store to be analyzed via HPLC. This capsaicin project consisted of three stages to ensure proper and accurate analysis. The first stage was the preparation of the three samples for injection into the HPLC. One gram of fine powder of each pepper was obtained by first chopping up with a scissors into a mortar and freezing with liquid nitrogen and finally homogenizing with the pestle. Precisely as measured by the analytical balance, 1.014 g was obtained from the habanero, 1.015 g was obtained from the jalapeno and 1.0270 g was obtained from the red bell pepper. It was observed that the red bell pepper seemed to be significantly juicier from the other two peppers. This mass was put into test tubes and labeled A, B and C respectively. Using a graduated cylinder 10 mL of 60/40 water/ACN solution was added to each test tube which were vigorously shaken and placed in hot water bath <75˚C for no longer than 15 minutes. More shaking of the samples was done during the hot water bath. The three samples were allowed to cool to room temperature before filtering into auto sampler vials through a 0.45 micron syringe filter. The next stage in the capsaicin analysis was to properly set up the HPLC correctly and to prepare the capsaicin samples. The system suitability standard was prepared by diluting 10 µL of 1.0 mg/mL capsaicin standard with 990 µL of the water/ACN solution previously used into an auto sampler vial. This capsaicin calibration standard final concentration was 10 µg/mL. Two
Eric Sorenson
Work Completed on 11/28/12
Quantitative analysis of Capsaicin was analyzed in three peppers that were expected to differ greatly in their capsaicin content and thus their relative spiciness. The idea was to have a nice range of capsaicin concentrations to clearly reflect the subjective pain one experiences from biting into a chili. Fresh Habanero, Jalapeno and Red Bell Pepper were purchased from the local Family Fresh grocery store to be analyzed via HPLC. This capsaicin project consisted of three stages to ensure proper and accurate analysis. The first stage was the preparation of the three samples for injection into the HPLC. One gram of fine powder of each pepper was obtained by first chopping up with a scissors into a mortar and freezing with liquid nitrogen and finally homogenizing with the pestle. Precisely as measured by the analytical balance, 1.014 g was obtained from the habanero, 1.015 g was obtained from the jalapeno and 1.0270 g was obtained from the red bell pepper. It was observed that the red bell pepper seemed to be significantly juicier from the other two peppers. This mass was put into test tubes and labeled A, B and C respectively. Using a graduated cylinder 10 mL of 60/40 water/ACN solution was added to each test tube which were vigorously shaken and placed in hot water bath <75˚C for no longer than 15 minutes. More shaking of the samples was done during the hot water bath. The three samples were allowed to cool to room temperature before filtering into auto sampler vials through a 0.45 micron syringe filter. The next stage in the capsaicin analysis was to properly set up the HPLC correctly and to prepare the capsaicin samples. The system suitability standard was prepared by diluting 10 µL of 1.0 mg/mL capsaicin standard with 990 µL of the water/ACN solution previously used into an auto sampler vial. This capsaicin calibration standard final concentration was 10 µg/mL. Two