| Pipperine is the major component of pepper. It has many biological activities. But the extraction method of pipperine is not perfect and the method to investigate the content of piperine still can be improved. Solubility of pipperine in multiple solvent has not research. This article research extraction method, investigate method and solubility of pipperine.1. The content of piperine in the flower, young and ripen fruit of pepper were investigated by RP-HPLC with Diamonsil column (C18,5μm,150mm×4.6mm) at room temperature and 343 nm. The mobile phase was water-methanol (gradient elution) with flow rate of 1mL/min. The piperine was used as the standard. The result showed that the linear range was 3～150μg/mL,and correlation coefficient was 0.9994.The limit of detection was 1.2μg/mL. Retention time was 12.3min. Contents of piperine of young pepper was 4.153mg/mL and the ripen fruit of pepper was 27.2mg/mL.Their relative standard deviation were 1.9% and 0.95%,respectively. Flower of pepper contents none of piperine. The average recovery of green pepper was 97.06% while ripen pepper is 96.66%. The relative standard deviations of them were 0.95% and 1.21%, respectively. These result conclusion that the method is simple,accurate,and can be used for determining the content of pipperine in the flower, young and ripen fruit of pepper.2. In order to get a efficient method of extracting piperine from pepper fruits, the adsorption and elution effects of piperine by Resins were investigated. Piperine was soluted by 40% ethanol (EtOH) and 1.5M acetic acid (HAc) and adsorbed by resins. Then it was eluted by 70%/80% EtOH and 1.5M HAc. The piperine was detected by HPLC. After the experiments, we found the H103 adsorb most of piperine, but AB-8 has the best effect of adsorption and elution.3. The solution of piperine in multiple solvent including ethanol, acetic acid, water and HCl were investigated to extract more piperine from piper fruit. Piperine was determined by reverse phase high-performance liquid chromatography with Diamonsil column (C18,5μm,250 mm×4.6 mm) at 343 nm. Experiment data were simulated by Allometric model and the formula is Z=0.9+ 4.54×10-10×x5.675+1.8029×y2.12848+2.37×10-10×x5.675×y2.12848(Z:sample solution,mol/mL,x:the percentage of ethanol's volume, ml/100mL,y:the acetic acid in the authentic sample solution, g/100mL), the adj-R2=0.997, the comparative deviation less than 2%. These results are good in agreement with experimental data. It reveals that the model can meet the requirements of the selection and design in extracting piperine from piper fruit.4. The content of piperine and 6-gingerol in condiment were investigated by RP-HPLC with Diamonsil column (C18,5μm,250mm×4.6mm) at room temperature and 280 nm. The mobile phase was water-methanol (gradient elution) with flow rate of 1mL/min. The 6-gingerol oxime was used as the internal standard. There was no distinct difference between the external standard method and internal standard method. The result showed that the linear range of piperine was 3～150μg/mL, while 6-gingerol and 6-gingerol oxime were 20～640μg/mL. The correlation coefficient were 0.9995,0.9997 and 0.9995, respectively. This condiment contains 20.23 mg/g piperine and 4.33mg/g 6-gingerol. These result conclusion that the method has no need to make standard curve duplicatly and can detect piperine and 6-gingerol simultaneously.
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