| High Performance Liquid Chromatography (HPLC), a kind of new separation and analysis technique, which is important in the modern society. It has many merits, such as the good separation efficiency, efficient and high sensitivity in detection spirit, fast analysis speed, wide applicable range and convenient operation, etc. The high pressure infusion system of HPLC puts moving phase into the chromatographic column, after all ingredients are separated in the column, then enter the detector for testing, at last, it would realize the analysis of the sample. At present, the method has been widely used in food analysis, environment monitoring, bioengineering, pharmaceutical industry and other fields of science.China is the birthplace of Chinese medicinal herb, the effective components of Chinese medicinal herb are the material basis for prevention and treatment of the disease, how much the content of Chinese traditional medicine is has become an important symbol to assess its quality. In recent years, the continuous improvement of living standards has made people’s require of "return to nature" becomes increasingly intensity. The international and domestic academics have come up with many new methods and ideas in studying the natural ingredient of the Chinese traditional medicine. The modern analysis technics has also been widely used in Chinese traditional medicine research. Among them, HPLC is one of the most important means in analysis the Chinese traditional medicine quality.Sample pretreatment is an important step before chromatographic analysis. Because Chinese traditional medicine is a kind of typical, complicated and unknown sample, the physieochemical property of each ingredient is different, the discrepancy of each content is also very big. So, whether the sample pretreatment technique is advance or not, is the key point of chromatographic analysis in success or failure. This paper consists of two parts, the introduction and the research report. The introduction part gives an outline of the characteristic and develop condition of HPLC, the research progress of phenolic compound and sample pretreatment technique. The research report part has set up the separation and determining of gallic acid, catechin. epicatechin in Xiaoer xiesuting particles; quercetin. kaempferol, galangin in galangal; the extract and determine of resveratrol in polygonum cuspidatum.1、Micelle-mediated extraction for the determination of resveratrol in polygonum cuspidatum by high performance liquid chromatography (HPLC)A simple and green method was developed for the determination of resveratrol in polygonum cuspidatum, nonionic surfactant Triton X-100was used to extract resveratrol from polygonum cuspidatum prior to their determination by high performance liquid chromatography (HPLC). The samples were analyzed on a Phenomenex C18column with methanol-0.25%acetic acid glacial as a mobile phase with isocratic elution. The flow rate was0.8mL-min-1, and UV detection wavelength was set at306nm, the results showed that there was a good linear correlation between the concentration and the peak area of resveratrol in the range of0.04-200μg/mL(r=0.9998), the limit of detection (LOD) was9.46ng/mL (S/N=3).5%sodium chloride was added to the solution to facilitate the phase separation and increase the extraction recovery. The spiked recovery was94.47%.2、Ultrasonic extraction for the determination of Quercetin, Kaempferol and Galangin in Alpinia officinarum Hance by high performance liquid chromatography (HPLC)The non-ionic surfactant-based ultrasonic extraction coupled with high performance liquid chromatography was developed for the simultaneous separation and determination of quercetin, kaempferol and galangin from Alpinia officinarum Hance. Methods:Three compounds from the Triton X-100extract have been isolated by using a Phenomenex C18column with a gradient elution. the mobile phase was acetonitrile-0.25%acetic acid. The flow rate was1.0mL/min, and UV detection wavelength was set at360nm. The condition for the extraction of three compounds was optimized by the orthogonal test as:ultrasonic extraction by100times of8%Triton X-100solution for40min at40℃. Results:The calibration curves for quercetin, kaempferol and galangin were linear between0.0076-76μg/mL,0.0064-64μg/mL,0.0152-152μg/mL (r=0.9999). respectively. The limit of detection (LOD) was7.32 ng/mL,1.92ng/mL, and1.13ng/mL (S/N=3) respectively. Conclusion:This analytical method was proved to be simple, quick, sensitive, low cost, environment-friendly and accurate for the quantity determination of flavonoids from Alpinia of Bcinarum Hance. It provides a new and scientific means for assaying flavonoids in the Alpinia officinarum Hance.3、Micelle-mediated extraction for the determination of polyphenols in Xiaoer Xiesuting Pellets by high performance liquid chromatography (HPLC)A method based on micelle-mediated extraction was developed for the simultaneous separation and determination of gallic acid, catechin, and epicatechin from Xiaoer Xiesuting Pellets by reverse-phase high performance liquid chromatography (RP-HPLC). Methods:Three compounds (gallic acid, catechin and epicatechin) from the Triton X-100extract have been isolated by using a Phenomenex Cis column with a gradient elution, the mobile phase was acetonitrile-0.2%acetic acid. The flow rate was0.8mL/min, and UV detection wavelength was set at280nm. Chromatographic condition was investigated, and to optimize the extraction and preconcentration process. Results:The calibration curves for gallic acid, catechin and epicatechin were linear between0.0104-223μg/mL,0.0152-108μg/mL,0.0184-132μg/mL (r=0.9999), respectively. The limit of detection (LOD) was0.087ng/mL,0.43ng/mL, and0.64ng/mL, respectively. Under optimum conditions:5%of Triton X-100surfactant (w/v), solid/liquid ratio of1:100(g/mL), ultrasonic-assisted extraction at30℃for30min, the extraction yield reached the highest value.5%sodium chloride was added to the solution to facilitate the phase separation and increase the extraction recovery. The average recoveries were91.85～98.48%. Conclusion:The method is simple, rapid, sensitive, low cost and environment-friendly, and suitable for use in the quality control of Xiaoer Xiesuting Pellets. |
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