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High price and difficult to get of reference substance have become obstacles to HPLC assay of ethnic medicine. A new method based on quantitative reference herb (QRH) was proposed. Specific chromatograms in fruits of Capsicum frutescens were employed to determine peak positions, and HPLC quantitative reference herb was prepared from fruits of C. frutescens. The content of capsaicin and dihydrocapsaicin in the quantitative control herb was determined by HPLC. Eleven batches of fruits of C. frutescens were analyzed with quantitative reference herb and reference substance respectively. The results showed no difference. The present method is feasible for quality control of ethnic medicines and quantitative reference herb is suitable to replace reference substances in assay.
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Objective To investigate the neuroprotective effect and its mechanism of therapeutic hypothermia induced by dihydrocapsaicin (DHC) on cerebral ischemia reperfusion injury in mice.Methods Twenty adult wild type (WT) mice were randomly divided into WT group and WT + DHC group,ten mice in each group.Twenty transient receptor potential receptor 1 (TRPV1) knockout mice were randomly divided into TRPV1 KO group and TRPV1 KO + DHC group,ten mice in each group.The model of focal cerebral ischemia reperfusion injury was established in all mice.The mice in the WT group and TRPV1 KO group were subcutaneously injected with physiological saline 1.25 mg · kg-1 · h-1 after reperfusion.The mice in the WT + DHC group and TRPV1 KO + DHC group were subcutaneously injected with DHC 1.25 mg · kg-1 · h-1 after reperfusion.All the mice were moved into the cage at 22 degrees centigrade after 90 minutes of reperfusion.The core body temperature during the reperfusion period was recorded in every 5 minutes to 24 hours after reperfusion.The neurobehavioral score was performed before anesthesia and 24 hours after anesthesia.After the neurobehavioral test,the brain tissue sections of mice were stained with 2,3,5-triphenyltetrazolium chloride;and the infarction rate of the brain tissue was calculated.Results There was no significant difference in the core body temperature among the four groups at 0,30 and 60 minutes after reperfusion (P ＞ 0.05).There was no significant difference in the core body temperature between 90 minutes and 0,30 and 60 minutes after reperfusion (P ＞ 0.05);but the core body temperature at 2,3,4,6,12 and 24 hours after reperfusion was significantly lower than that at 0,30,60 and 90 minutes after reperfusion in WT group,TRPV1 KO group and TRPV1 KO + DHC group(P ＜ 0.05).There was no significant difference in the core body temperature among the WT group,TRPV1 KO group and TRPV1 KO + DHC group at 2,3,4,6,12 and 24 hours after reperfusion (P ＞ 0.05).The core body temperature at 90 minutes and 2,3,4,6,12,24 hours after reperfusion was significantly lower than that at 0,30 and 60 minutes after reperfusion in WT + DHC group(P ＜ 0.05);and the core body temperature in WT + DHC group was significantly lower than that in WT group,TRPV1 KO group and TRPV1 KO + DHC group at the same time point(P ＜ 0.05).There was no significant difference in the total neurobehavioral score among the four groups before anesthesia (P ＞ 0.05).The total neurobehavioral score at 24 hours after reperfusion was significantly lower than that before anesthesia in the four groups (P ＜ 0.05).The total neurobehavioral score in WT + DHC group was significantly higher than that in WT group,TRPV1 KO group and TRPV1 KO + DHC group at 24 hours after reperfusion(P ＜0.05).There was no significant difference in the total neurobehavioral score among WT group,TRPV1 KO group and TRPV1 KO + DHC group at 24 hours after reperfusion (P ＞ 0.05).There was no significant difference in the score of spontaneous activity,climbing test,body proprioception and response to vibrissae touch among the four groups at 24 hours after reperfusion (P ＞ 0.05).The score of symmetry test of limbs movement and forepaw stretching test in WT + DHC group was significantly higher than that in WT group,TRPV1 KO group and TRPV1 KO + DHC group at 24 hours after reperfusion(P ＜ 0.05).There was no significant difference in the score of symmetry test of limbs movement and forepaw stretching test among the WT group,TRPV1 KO group and TRPV1 KO + DHC group at 24 hours after reperfusion (P ＞ 0.05).The infarction rate of brain tissue in WT + DHC group was significantly lower than that in the other three groups at 24 hours after reperfusion (P ＜ 0.05);but there was no significant difference in the infarction rate of brain tissue among the WT group,TRPV1 KO group and TRPV1 KO + DHC group (P ＞ 0.05).Conclusion Subcutaneous injection of DHC after focal cerebral ischemia and repeffusion of mice can induce therapeutic hypothermia by activating TRPV1 receptor,and reduce brain tissue damage and improve neurological function.
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Objective:To develop an HPLC method for the determination of capsaicine, dihydrocapsaicin, imperatorin and isoim-peratorin in Guanjie Jietong plasters. Methods:A CAPCELL PAK C18 column was used as the chromatographic column, and the flow rate was 0. 8 ml·min-1. The mobile phase A consisted of methanol-acetonitrile(1∶1), the mobile phase B was of 1% phosphoric acid. The detection wavelength was 280 nm and 254 nm, respectively. Results:There was a good linear relationship between the con-centrations and the peak areas within the range of 0. 298-5. 956μg(r=0. 9998) for capsaicine, 0. 152-3. 044μg(r=0. 999 6) for di-hydrocapsaicin, 0. 018-0. 352 μg(r=0. 999 5) for imperatorin and 0. 010-0. 204 μg(r=0. 999 3) for isoimperatorin. The average re-covery was 97. 8%(RSD=1. 02%), 97. 0%(RSD=0. 76%), 96. 6%(RSD=0. 65%) and 98. 1%(RSD=1. 35%), respectively. Conclusion:The method is convenient, accurate, sensitive and repeatable, which can be used in the determination of capsaicine, di-hydrocapsaicin, imperatorin and isoimperatorin in Guanjie Jietong plasters.
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Introducción. En el desarrollo de nuevos procesos e ingredientes para la industria alimentaria, las técnicas de extracción permiten obtener principios activos a partir de productos naturales, aptos para el consumo humano y fuertemente relacionados con la tendencia hacia una alimentación más saludable y libre de contaminantes. Las oleorresinas de especies vegetales permiten el manejo simultáneo de componentes pigmentantes y aromatizantes como en el caso de los pimientos, para los cuales la extracción con solventes orgánicos es una práctica común que debe refinarse de tal modo que se empleen sustancias de extracción seguras y aprobadas para el consumo humano. Objetivo. Comparar la composición de la oleorresina de dos variedades de ají picante (tabasco y habanero) extraída con dos solventes diferentes (hexano y acetato de etilo). Materiales y métodos. Se tomó material seco de dos variedades de ají picante (tabasco y habanero) y se sometieron a extracción con dos solventes orgánicos diferentes (hexano y acetato de etilo). Las oleorresinas resultantes fueron concentradas para calcular el rendimiento en extracción y luego fueron analizadas en un HPLC para determinar su contenido en capsaicina y dihidrocapsaicina. Resultados. Se encontró que el acetato de etilo es un mejor solvente que el hexano, pues presenta un porcentaje general de extracción del 11,23% frente a 7,14%. En la caracterización por HPLC se encontraron concentraciones aceptables de los principios pungentes (capsaicina y dihidrocapsaicina) las cuales corresponden con los valores teóricos en los cuales el ají habanero presenta entre 150.000 y 325.000 Unidades Scoville y tabasco presenta entre 50.000 y 100.000 Unidades Scoville. Conclusión. En un proceso de extracción con solventes sobre ají picante, se obtiene un mejor rendimiento en extracción empleando acetato de etilo como solvente y una mayor concentración de capsaicina y dihidrocapsaicina empleando ají habanero.
Introduction. In the development of new processes and ingredients for food industry, extraction techniques allow us to obtain active principles from natural products, good for human consuming and strongly related to the tendency towards a healthier and free of contaminants feeding. Oleoresins from vegetables allow us to work with dye and aromatic components (as in the case of peppers, for which, extraction with organic solvents is a common practice that must be refined in such a way that only safe and apt for human consumption substances are used). Objective. To compare the composition of oleoresin from two kinds of chili pepper (Tabasco and habanero) extracted with two different solvents (hexane and ethyl acetate). Materials and methods. Dry material from both kinds of chili pepper were taken and submitted to extraction with two different organic solvents (hexane and ethyl acetate). The oleoresins that resulted from this procedure were concentrated to calculate their performance in extraction and then analyzed in a HPLC to determine their contents of capsaicin and dihydrocapsaicin. Results. We found that ethyl acetate is a better solvent than hexane; it shows a general percentage of extraction of 11.23% compared to a 7.14%. In the characterization with the HPLC we found acceptable concentrations of the pungent principles (capsaicine and dihydrocapsaicin), corresponding to the theoretical values in which habanero pepper represents a value between 150000 and 325000 Scoville units and the Tabasco, between 50000 and 10000 Scoville units. Conclusion. In an extraction process for chili peppers, a better extraction performance is obtained by using ethyl acetate as solvent and a better concentration of capsaicin and dihydrocapsaicin by using habanero chili pepper.
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Objective To establish an HPLC method to determine the two principal constituent-capsaicin and dihydrocapsaicin in general alkaloids of Capsaicin.Method The analysis was performed on C18 column(4.6 mm?250 mm,5 ?m),with methanol-water(70:30) as the mobile phase.The flow rate was 1.0 mL/min and the detection wavelength was set at 280 nm.Result The linear relationships of capsaicin and dihydrocapsaicin were within the range of 1.955～7.820 ?g(r=0.999 9) and 2.709～7.224 ?g(r= 0.995 5),respectively.The average recoveries were 101.57% and 108.87%,respectively.Conclusion The method is convenient,the result is accurate and reliable,and can be used to determine the content of two principal constituent in general alkaloids of Capsaicin.