| Ardisia japonica(Thunb)Blume is Ardisia and Myrsinaceae plant,also known as Japanese Ardisia Herb and Phylla-gathis fordii(Han ce)C.Chen,is commonly used Chinese medicinal materials.The whole plant and root can be used for medicine.In this experiment,the characters and microstructure of Ardisia japonica(Thunb)Blume were identified;the moisture,ash and extract in Ardisia japonica(Thunb)Blume were measured;the chemical components of the whole plant were extracted,isolated and identified.To establish methods to determine contents of 2-methyl-5-[8’(Z)-tridecylene]-resorcinol(Co mpound 2)in Ardisia japonica(Thunb)Blume by high performance liquid chromatography and to establish methods to determinate the total phenolic acid of Ardisia japonica(Thunb)Blume by spectropho tometry.Characterization and microscopic identification of Ardisia japonic a(Thunb)Blume: The whole plant of Ardisia japonica(Thunb)Blume was selected and appearance,shape and other characteristics were identified;the crosscut surface,leaf surface and powder of the stem of Ardisia japonica(Thunb)Blume were selected and observed under the microscope to identify the morphological characteristics of the tissue,cells and inclusions.The results show that the characteristics of Ardisia japonica(Thunb)Blum,the cross section of the stem,the surface of the leaves,the characteristics of tissues,cells and inclines in the powder are basically consistent with those of Ardisia japonica(T hunb)Blum in Chinese pharmacopoeia(2015 edition).The stem of A rdisia japonica(Thunb)Blume is cylindrical with loose roots.The surface of the stem is reddish brown with fine veins and nodes.The leaves are alternate,elliptic and pale green.The epidermal cell wall of cross section of the stem was thicker;the cork layer was more;the cortex was thicker and the thick horn cells were more closely arranged.Most of the xylem cells were completely lignified and the cathet ers were mostly arranged in a single row.The epidermal cells leaf surface view are mostly elliptic;stomata are mostly inequalities and a few are irregular.The whole grass powder is brown;the threaded catheter is more;the secretory cavity is mostly incomplete;contains brown secretion;the starch grain in the powder is less.The moisture,ash and extract of determination: According to the method of the fourth part of Chinese pharmacopoeia(2015 edition),the right amount of Ardisia japonica(Thunb)Blume was weighed,crushed and sifted,the samples were heated,dried,weighed and the moisture content was calculated;the right amount of Ardisia japonica(Thunb)Blume was weighed,crushed,sifted,and then added dei onized water,the Ardisia japonica(Thunb)Blume was cold-soaked and let stand for a period of time and the extract was filtered,heated and cooled to room temperature,and extract content was calculated;take the right amount of Ardisia japonica(Thunb)Blume,pulverize,burn,cool,weigh,calculate the total ash content.The results show that the moisture content of Ardisia japonica(Thunb)Blume is 7.20% and the extract content was 15.92 % and the total ash content was 6.29 %.The separation and identification of chemical constituents: The whole plant of Ardisia japonica(Thunb)Blume was crushed,soaked and extracted with anhydrous ethanol.The extract was decompressed and concentrated to make paste.The paste was separated by silicagel,D101 macroporous adsorption resin,C18 and other chromatographic methods.The results show that from the whole plant of Ardisia japonica(Thunb)Blum,four chemical components were obtained as 2-methyl-5-[10’(Z)-pentadecenyl]-resorcinol(1),2-methyl-5-[8’(Z)-tridecylene]-resorcinol(2),2-methyl-5-tridecyl-resorcinol(3),2-methyl-5-pentad ecyl-resorcinol(4)respectively.The determination of compound 2: The roots,stems and leaves of Ardisia japonica(Thunb)Blume were respectively crushed and screened,and the petroleum ether was reflux extracted by petroleum ether.The extract was filtered,steamed and concentrated,and the volume is fixed in methanol.High performance liquid chromatography was used to determine compound 2.The Chromatographic column was Wonda Sil C18 Superb column(250 mm×4.6 mm,5 μm),the mobile phase was methanol-water(75∶25),the flow rate was 1 m L/min,and the detection wavelength was 275 nm.The results show that the linear range of compound 2 was 0.054~0.870 mg/mL.The regression equation is Y = 17009c-317.67(r = 0.9997).Precision RSD was 0.48 %.Stability RSD was 0.84 %.The recovery was 102.59 %.The contents of compound 2 in roots,stems and leaves of dried Ardisia japonica(Thunb)Blum were 16.78 mg/g,17.39 mg/g and 19.56 mg/g,respectively.The determination of total phenolic acid: The roots,stems and leaves of Ardisia japonica(Thunb)Blume were crushed and screened respectively.Use anhydrous ethanol to perform ultrasonic extraction and filter the extraction solution.The reference substance was Gallic acid and the color reagent was 1% potassium ferricyanide-1% ferric chloride and the detection wavelength was 760 nm.The results show that the linear range of gallic acid was 0.16~0.96 g/m L.The regression equation is A = 0.8324C-0.0244(r= 0.9999).Precision RSD was 0.75%.Stability RSD was 0.73 %.The RSD was 0.51 %,0.45 % and 0.55 % respectively.The standard recovery is 99.1 %.The contents of total phenolic acid in roots,stems and leaves of dried Ardisia japonica(Thunb)Blum were 16.32 mg/g,21.13 mg/g and 8.28 mg/g,respectively.Four compounds,all of which are phloroglucinol,were isolated from Ardisia japonica(Thunb)Blum,among which compound 1 was a new compound,compound 2 was known,compound 3 and compound 4 were first discovered in Ardisia japonica(Thunb)Blum.A method for the determination of compound 2 by high performance liquid chromatography was established.The results showed that the content in roots and stems was lower than that in leaves.This method is simple,fast,stable and accurate.A spectrophotometric method for the determination of total phenolic acid was established.The results showed that the content in roots and stems was higher than that in leaves.This method is simple,fast and accurate. |
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