| High Performance Liquid Chromatography?HPLC?has the advantages of high pressure,high efficiency,high sensitivity,simple operation,and wide application.Now,it was widely used in chemical,medical,agronomy,commodity inspection and food analysis and other fields.The analytical principle of high performance liquid chromatography is to use the interaction between the component to be measured and the stationary phase to affect the retention time of different components of the column,so that the components flow out sequentially with under the action of the mobile phase to achieve separation detection.Before using HPLC to analyze the test item,proper pretreatment is required for the sample to be measured,and the use of impurity removal,enrichment or extraction can better achieve the purpose of analysis,thereby improving the detection sensitivity and accuracy.In recent years,a variety of sample pretreatment technologies have emerged in an endless stream,and environmental protection,rapid,and convenient pre-treatment methods have been pursued by the majority of research workers.This paper is divided into two parts:introduction and research report.The first part introduces the development and application of high-performance liquid chromatography,traditional Chinese medicines,Chinese patent medicines,and sample preparation techniques.The second part of the research reported the extraction and separation conditions and content determination of flavonoids and organic acids from two flavors of traditional Chinese medicine Rosa laevigata,pomegranate peel and the throat tablets of Chinese patent medicine.1.Simultaneous Determination of 6 Active Compounds in Rosa laevigata Michx by HPLC Combined with Ultrasonic-assisted Extraction-Gradient WavelengthTo establish a methed for separation and simultaneous determination of gallic acid,catechin,rutin,Quercetin,Kaempferol and apigenin in Rosa laevigata Michx by ultrasonic extraction combined with HPLC.The 6 compounds in Rosa laevigata Michx were separated on InertSustain C18?150 mm×4.6 mm,5?m?.Mobile phase consisted of methanol and 0.1%acetic acid?pH=3.0?with gradient elution at a flow rate of 1.0ml/min.The UV detection gradient wavelength was set at 280 nm?05.00 min?and350 nm?5.0020.00 min?.The column temperature was 35?and injection volume was 20?L.The separation of 6 active ingredients was completely achieved under the condition.Calibration curves of the six components showed good linear relationship?R>0.999 1,n=6?.The average recoveries were within 96.12%110.39%,RSD were between 1.06%and 3.14%.The limit of detection?S/N=3?was 0.027-0.75?g·L-1.The results showed that there were gallic acid 450.1?g·g-1,catechin 2324.6?g·g-1,rutin182.4?g·g-1,quercetin 20.8?g·g-1,kaempferol 8.2?g·g-1,apigenin 12.1?g·g-1.This method is simple,rapid and accurate.It is also a theoretical basis for the quantitative detection and quality control of Rosa laevigata Michx.2.Simultaneous Determination of 7 Organic Acids and Flavonoid Compounds in Pomegranate Rind by Ionic liquid Combined with Ultrasonic-assisted Extraction-HPLCSingle factor and orthogonal array designs were applied to optimize the Organic Acids and Flavonoid Compounds,and simultaneous separation and determination of gallic acid,catechin,chlorogenic acid,epicatechin,caffeic acid,quercetin,kaempferol in Pomegranate Rind was established by the ultrasonic extraction combined with HPLC method with 1-Butyl-3-methylimidazolium hexafluorophosphate as an extractant.Methods:The 7 compounds in Pomegranate Rind were separated on InertSustain C18?150 mm×4.6 mm,5?m?.Mobile phase consisted of methanol and 0.2%phosphoric acid?pH=3.25?with gradient elution at a flow rate of 0.7 ml·min-1.The UV detection wavelength was set at 280 nm.The column temperature was kept at 35?and injection volume was 20?L.The separation of 7 active ingredients was completely achieved under the condition.Calibration curves of the 7 components showed good linear relationship?R>0.9992,n=6?.The average recoveries were within91.52%105.41%?RSD%<2.19%?.The limits of detection?LODs?were in the range of0.086-0.503?g·L-1.The optimal extraction conditions were established as follows:detection wavelength 280 nm,ionic liquid concentration 0.8 mol·L-1,solid-liquid ratio1:80(g·mL-1),ultrasonic time 20 min,ultrasonic power 160 W.3.Simultaneous Determination of Eight Organic Acids and Flavonoids in Four Kinds of Throat Tablets by HPLCSingle-factor tests were conducted to optimize the determination of organic acids and flavonoids in the four Chinese patent medicines of Soothing throat tablets,Jian Min Pharynx tablets,Clear throat tablets,and Pharyngitis tablets,and Ultrasound-assisted extraction-HPLC method was established to determine the 8 active ingredients of gallic acid,protocatechuic acid,chlorogenic acid,caffeic acid,ferulic acid,quercetin,kaempferol,apigenin.Methods:The 8 compounds were separated on Inert Sustain C18?150 mm×4.6 mm,5?m?.Mobile phase consisted of methanol and 0.2%Acetic acid?pH=3.15?with gradient elution at a flow rate of 0.7 ml·min-1.The UV detection wavelength was maintained at 254 nm.The column temperature was kept at 30?and20?L per injection.The 8 active components was separated perfectly under this condition.Calibration curves of the eight ingredients showed expected linear relationship?r>0.9994,n=8?.The average recoveries were between 89.62%and 108.43%?RSD%<3.08%?.The experimental results appered that the contents of the 8 active ingredients in Jian Min Pharynx tablets are the highest among the 4 types of Throat treated tablets.This method established a simple and definite experimental condition to simultaneously determine the content of active ingredients of various substances,which was provided for the quality management of throat pills. |
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