| Turnip and Highland barley are widely cultivated in plateau areas.Turnip has the functions of heat-clearing,detoxifying,nourishing and aerobic capacity improvement.It is reported to possess many biological activities,such as anti-hypoxia,anti-oxidation and hypoglycemia,and has been developed as various health foods.Highland barley is one of the main crops in plateau areas,of which seedling also has effects like anti-oxidation,anti-aging and immunity increasing.However,few studies have been done on the quality control standards of these two herbs.In this subject,components analysis and quality control standards of turnip and highland barley seedling(HBS)were studied.The overall research contents are summarized as follows:1.Literature reviewWe reviewed the components,analytical methods and pharmacological activities of turnip and HBS,in order to provide the research basis and theoretical support for the experiment.2.Component study by chromatography-mass spectrometry2.1 TurnipLC-MS/MS was applied in analysis.According to the obtained precise mass numbers and fragment information,some compounds were speculated as follows:in 0.9?1 min,components were mainly organic acids,hexose,hexobiose and trisaccharide;in 1?10 min,components were mainly amino acids and glycosides;in 11-21 min,components were mainly long-chain fatty acids,their derivatives and their glycosides.Through GC-MS,organic acids in turnip were analyzed.Based on NIST 2014,22 organic acids were identified.Monoacids account for 77.46%of the total peak area,diprotic acid account for 4.47%.There are 8 kinds organic acid normalized area more than 4%,including pentanoic acid,succinic acid,citric acid,hexadecanoic acid,linolenic acid,oleic acid,linoleic acid and hepta-2,4-dienoic acid.2.2 HBSAccording to the precise mass numbers and fragment information obtained through LC-HRMS/MS,compounds were speculated as follows:in 1?7 min,components were mainly organic acids,amino acids,and hexobiose;in 9?14 min,components were mainly flavonoids and their glycosides;in 15?26 min,components were mainly long-chain fatty acids and their derivatives.3.Quality detection method studyIn accordance with the terms of quality standard in Pharmacopoeia,water content,ash content and extract were determined on turnip and HBS,and TLC method for identification and content determination method were at the same time established.3.1 quality control of turnip(1)Conducted under relevant methods in Appendix of Chinese Pharmacopoeia,the water content of different batches of turnip was shown between 2.7%and 13.9%.The total ash content was between 4.0%and 6.9%,and the acid-insoluble ash content was between 0.1%and 0.2%.The extract content was between 49.0%and 57.7%.(2)A TLC method to indentify turnip was established,with silica gel G as thin layer plate,n-butyl alcohol,water and glacial acetic acid(8:1:1)as developing solvent,alcoholic solution of sulfuric acid(10%)as chromogenic reagent.The method is simple and specific.(3)Gas chromatography with precolumn derivatization was used to determine content of acetylpopionic acid,succinic acid,hexadecanoic acid,citric acid,linolenic acid,oleic acid,linoleic acid.The methodology study meets the requirement of content determination.The linear relationship was good in the concentration range of 0.02?3 g/L.The correlation coefficients of organic acids were greater than 0.9995,and the recovery was between 97.27%and 103.54%.The method exhibits a good reproducibility and operability.The results showed that different batches of turnip exhibited significant content differences on organic acids.(4)Sulfuric anthrone reaction was involved to establish content determination of total sugar and polysaccharide.The methodology study meets the requirement of content determination.In the range of 0.015-0.135 g/L,the correlation coefficient was 0.9996 and the recovery rate was 98.38%(RSD = 1.8%)with glucose as the control.The content of total sugar and polysaccharide in turnip were 31.6%?54.0%and 0.6%?0.9%respectively.(5)The fingerprint of turnip was established through HPLC.The fingerprint was built by software Similarity Evaluation System for Chromatographic Fingerprint of TCM published by GPC(Version 2004A).The Uridine was used as reference.The results of the methodological investigation meet the requirements of fingerprint establishment.The result shows that there were seven common fingerprint peaks in turnip with similarity larger than 0.9.However the graph need improve due to insufficient batches of samples.3.2 HBS quality control(1)Conducted under relevant methods in Appendix of Chinese Pharmacopoeia,the water content of different batches of HBS was 8.20%?8.7%.The extract content was between 36.7%and 43.1%.(2)A TLC method to indentify HBS was established,with silica gel G as thin layer plate,acetic ether,acetone and glacial acetic acid(7:1:0.01)as developing solvent,examined at 365nm wavelength.The method is simple and specific.(3)A UV-vis spectrophotometry method to determine the total flavonoids was established with 1%triethylamine solution as shift reagent.The methodology study meets the requirement of content determination.Taken apigenin as control sample,the linear relationship was good in the concentration range of 0.002-0.01 g/L,the correlation coefficient was 0.9990,and the recovery rate was 100.25%(RSD=1.8%).The method exhibits a good reproducibility and operability.The results showed that the total flavonoid content of HBS in different batches ranged froml.75%?1.93%.(4)HPLC method was also used to determine content of saponarin in HBS,using Thermo BDS HYPERSIL C18,with methanol-0.1%formic acid(30:70)as mobile phase,which met the requirements of content determination by methodological investigation.The linear relationship was good in the concentration range of 0.0496-0.496 g/L.The correlation coefficient was one,and the recovery rate was 102.15%(RSD=1.75%).The method is simple and operable.The result showed that the saponin content of HBS in different batches ranged from 0.8%to 1.6%.(5)The fingerprint of HBS was established through HPLC.The fingerprint was built by software Similarity Evaluation System for Chromatographic Fingerprint of TCM published by GPC(Version 2004A).The saponin was used as reference.The result showed that there were 13 common fingerprint peaks in HBS with similarity larger than 0.93(HBS from Rejie,Luobu,Tibet),but the similarity of HBS from Danzeng was smaller than 0.9.The graph still need improve due to insufficient batches of samples.4.Pharmacological activity4.1 bioactivity of turnipThe anti-acetylcholinesterase activity of turnip was conducted by ELLMAN in vitro.The results showed that different batches had different abilities.DPPH and hydroxyl radical-scavenging assay were used to investigate the antioxidant capacity of turnip.The results showed that when the concentration of turnip reached 25 g/L,the free radical-scavenging rate reached 50%.4.2 bioactivtiy of HBSDPPH and hydroxyl radical-scavenging assay were used to investigate the antioxidant capacity of HBS and compared with wheat seedling.The results showed that when the concentration of HBS reached 2 g/L,the scavenging rate of DPPH free radical reached 50%.Except HBS from Danzeng,when the concentration reached 2.5g/L,the free radical-scavenging rate reached 50%.At the same concentration,the scavenging ability of DPPH free radicals in wheat seedlings was higher than that in HBS,but the scavenging ability of hydroxyl free radicals was lower than that in barley seedlings. |
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