| Objective The contents of 9 active ingredients in whole Mahuang Decoction and the split prescription compatibility decoction were detected to compare the traditional decoction and dispensing granule decoction.The sweating effect was observed and compared through the diaphoretic volume and the duct diameter of sweat gland at axillary region in mice,providing some scientific basis for the clinical application of granule.Methods ①Content assay of active ingredients in crude and dispensing granules by the pharmacopoeia method:A HPLC method was used for the contents of active ingredients in 4 medicinal herbs pieces consist of ephedra,bitter almond,cassia twig,licorice root,and the formula granules,according to the method of content determination in China Pharmacopoeia 2015 Edition.②Content assay of 9 active ingredients in formula granule decoction and traditional Mahuang Decoction by a HPLC-DAD method:Both the traditional decoction and granule decoction were set to 15 experimental groups according to compatibility of prescription.The active ingredients,ephedrine,pseudoephedrine,norephedrine,cinnamaldehyde,cinnamic alcohol,cinnamic acid,amygdalin,glycyrrhizic acid and liquiritin were detected by HPLC-DAD for each group.The differences between the traditional decoction and dispensing granule decoction were compared,and the influences on the content of each component by different compatibilities were observed.③Comparison experiment of sweating effect:The sweating effect among the experimental groups were observed and compared by measuring diaphoretic volume and duct diameter of sweat gland at axillary region in mice.The differences between the traditional decoction and dispensing granule decoction were observed.Results ①Results for the Pharmacopoeia method:The content of ephedrine hydrochloride in Herba Ephedrae was 6.21%,and 2.12%for pseudoephedrine hydrochloride.The content of cinnamic aldehyde in Radix Guizhi was 1.04%.The content of Amygdalin in Radix bitter almond was 15.99%.The content of glycyrrhizin in Radix Zhigancao was 1.05%,3.69%for glycyrrhizic acid.It’s 18.29%for ephedrine hydrochloride in dispensing granule,11.95%for pseudoephedrine hydrochloride.It’s 3.77%.for cinnamic aldehyde in Guizhi granules.It’s 38.59%for amygdalin in bitter almond granules.It’s 3.59%for glycyrrhizin in licorice dispensing granules,11.64%for glycyrrhizic acid.The 4 medicinal herbs for the experiment consist of ephedra,bitter almond,cassia twig and licorice were in line with the China Pharmacopoeia according to the above data.However,there were certain differences in the contents of active ingredients in formula granules of ephedra,bitter almond,cassia twig and licorice compared with medicinal herbs pieces.There was a loss in the content of active ingredients during the preparation of formula granules.②The contents of 8 active ingredients consist of ephedrine,pseudoephedrine,norephedrine,cinnamaldehyde,cinnamic alcohol,cinnamic acid,glycyrrhizic acid and liquiritinomponents in 15 compatibility groups were higher than the corresponding granule groups,while the contents of amygdalin in the compatibility groups were all lower than the granule groups.It was 8.74mg/g,4.47 mg/g,1.19 mg/g,10.24 mg/g,18.75 mg/g,1.46 mg/g,4.13 mg/g,23.48 mg/g,7.58 mg/g for ephedrine,pseudoephedrine,norephedrine,cinnamaldehyde,cinnamic alcohol,cinnamic acid,amygdalin,glycyrrhizic acid and liquiritin in Mahuang Decoction Pieces group,respectively.However,the content of the above 9 active ingredients in whole granule groups was only 4.71 mg/g,3.32 mg/g,0.76 mg/g,2.09 mg/g,5.86 mg/g,0.80 mg/g,6.32 mg/g,13.16 mg/g,5.25 mg/g,respectively.③Tthe contents of active ingredients in Mahuang Decoction group were all higher than these in single herb groups and other compatibility groups,whether it was a pieces group or a granule group.④The contents of active ingredients all showed some changes by the compatibility of Mahuang and Gancao,cassia twig and bitter almond,ephedra and bitter almond,ephedra and cinnamon,licorice and bitter almond,cassia twig and licorice.⑤There were different degrees for sweating effect in mice of 9 traditional decoction groups and 9 granule groups,by comparing the duct diameter of sweat gland at axillary region.The diameters were all increased compared with the control group,with a significant difference(P<0.01).⑥Whether the traditional decoction groups or the granule groups,the diaphoretic volume and the duct diameter of sweat gland in Mahuang Decoction group was the maximal,the sweating effect was stronger than the compatibility groups.The duct diameter of sweat gland in Mahuang Decoction group was 106.5 μm,while it was only 73.8 μm in formula granule group.It was better for the sweating effect in Mahuang Decoction group than in formula granule group(P<0.05).⑦The sweating effect in each compatibility group containing ephedra was larger than that off ephedra.The sweating effect of ephedra were better after adding to cinnamon.Conclusions ①The HPLC-DAD determination method established in the paper showed the high specificity and good precision,which could be used for the simultaneous determination of multicomponent in ephedra soup.Under the experimental chromatographic conditions,all components can achieve good separation with no interference by stray peaks.②The contents of 8 active ingredients consist of ephedrine and pseudoephedrine,norephedrine,cinnamaldehyde,cinnamic alcohol,cinnamic acid and glycyrrhizic acid and liquiritin in Mahuang Decoction group and its compatibility decoction groups were all higher than the corresponding granule groups,while the content of amygdalin in decoction groups was lower than that in granule groups.From the point of chemical content,traditional decoction groups are better than granule groups.The replace of traditional herbal medicine of Mahuang Decoction by dispensing granules is still arguably.③With the different combinations among ephedra,bitter almond,cassia twig and licorice,the corresponding active ingredient showed some changes.Among them,the contents of effective ingredients in Mahuang Decoction were higher than these in single herb groups and other compatibility groups.④Through the sweating test,we can see that all the experimental groups(decoction groups and granule groups)have different degrees of sweating.The inner diameters were increased compared with the blank control group.⑤The sweating effect in whole Ephedrae Decoction group was better than granule groups.The replace of traditional herbal medicine of Mahuang Decoction by dispensing granules is still arguably. |
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