Study On The Separation Of Chemical Components Of Blumea Balsamifera Oil And Its Quality Standards

At present,Blumea balsamifera oil was only included in the"Quality Standards for Traditional Chinese Medicine and National Medicinal Materials of Guizhou Province"（2003 edition）.The monitoring content specified in this standard was too simple,and only the content of L-Borneol was monitored,which made the lack of a stable,reliable,controllable,effective and safe quality standard system for Blumea balsamifera oil.Since there was no comprehensive and unified quality standard on the market,the quality of Blumea balsamifera oil produced by different companies varies,and the quality was unstable,resulting in uneven product quality.Therefore,it is very necessary to study the active ingredients of Blumea balsamifera oil and its quality control standards.The main contents of this research are as follows:1.Study on the identification of chemical components in Blumea balsamifera oilQualitative inspection through chemical and foam tests confirmed that the Blumea balsamifera oil contains Flavonoids,Organic acids,and some Blumea balsamifera oil contains Anthraquinones,but no Alkaloids,total Saponins and total Polysaccharides.The content of total Flavonoids,Organic acids and total Anthraquinones were determined by UV in the range of 0.165～0.843,8.33～28.060,0～0.152 mg·g^-1.2.Study on separation of chemical components in Blumea balsamifera oil2.1 GC-MS chemical composition analysis of Blumea balsamifera oilGC-MS analysis technology was used to detect the chemical composition of Blumea balsamifera oil.The results showed that the kinds of 75 main components in Blumea balsamifera oil obtained by steam distillation were L-Borneol（20.18%）,trans-Caryophyllene（15.81%）,Camphor（10.44%）,Caryophyllene oxide（5.57%）,Xanthoxylin（5.31%）,thymine hydroquinone dimethyl ether（3.02%）,Cuminene（3.18%）,Linalool（2.09%）,1-Octen-3-ol（1.15%）.2.2 Separation of silica gel column chromatography of Blumea balsamifera oilSilica gel column chromatography was used to carry out gradient elution of petroleum ether（60-90℃）-ethyl acetate on Blumea balsamifera oil.GC-MS was used to identify Xanthoxylin（96.033%）,trans-Caryophyllene（34.446%）,Caryophyllene oxide（60.291%）,thymol hydroquinone dimethyl ether（49.306%）,L-borneol（81.982%）,,Methyl jasmonate（13.637%）7 components,of which Methyl jasmonate andα-Cedrol were isolated from Blumea balsamifera oil for the first time.2.3 Study on preparation and separation of Blumea balsamifera oil at medium pressureBlumea balsamifera oil was used for HPLC medium-pressure preparation and separation to obtain Xanthoxylin with a purity as high as 99.335%.3.Study on the improvement of the quality standard of Blumea balsamifera oil3.1 Physical and chemical propertiesAccording to the requirements of the 2015 edition of the Chinese Pharmacopoeia,the physical and chemical properties and inspection parameters of Blumea balsamifera oil were measured and studied.The results show that the specific rotation of Blumea balsamifera oil was-26.368°-28.773°,and the refractive index was 1.4812-1.4954,the relative density was 0.9458～0.9852.Blumea balsamifera oil was a yellow to brownish red,clear liquid,with a special aromatic odor,with partial volatility,some enamel oils can often precipitate and crystallize when cold,and then clarify after heating.Easily soluble in ethanol,chloroform,ether,almost insoluble in water.Inspection:Heavy metals must not exceed 10 parts per million;Adding 1m L of85%ethanol to 1m L of Blumea balsamifera oil,the solution should be clear.3.2 TLC identification.With silica gel G plate,ethyl acetate-cyclohexane-chloroform（1.5:10:3）as the best developing agent,which can effectively treat Xanthoxylin,β-Caryophyllene,Linalool,L-Borneol,andα-Humulene in Blumea balsamifera oil.3.3 Simultaneous determination of the contents of 7 components in Blumea balsamifera oil by HPLC-DAD.A method for simultaneous determination of seven active ingredients inβ-Caryophyllene,Caryophyllene oxide,Xanthoxylin,β-Pinene,Linalool,α-Pinene,andα-Humulene in Aina fragrant oil was established.Chromatographic conditions were Supersil-T C18 column（4.6×150 mm,5μm）,acetonitrile（A）-0.4%phosphoric acid（B）.Water gradient elution:0-5 min,28%A;5-50 min,30%A;50-55 min,28-48%A;55-135 min,48%A;135-145 min,48-70%A;145-155 min,70-80%A;155-165 min,80%A;165-180 min,80-28%A,column temperature 25℃,injection volume 10μL,flow rate 1.0 m L?min^-1,λ=202 nm.The contents of the 7 ingredients of 13 batches of Blumea balsamifera oil were 76.151-168.710,11.476-127.236,8.350-44.200,21.629-59.091,2.761-23.249,2.342-9.711,6.458-11.164 mg·g^-1.3.4 Study on HPLC fingerprint of Blumea balsamifera oilIn this experiment,13 batches of Blumea balsamifera oil medicinal materials from different sources were used as objects,and their fingerprints was established by high-performance liquid chromatography.The conditions were Supersil-T C18column（4.6×150 mm,5μm）,the mobile phase was acetonitrile-0.4%phosphoric acid water system,gradient elution,detection wavelength is 202 nm,injection volume was10μL,column temperature was 25℃and flow rate was 1.0 m L?min^-1.13 batches of Blumea balsamifera oil HPLC fingerprints were established through the fingerprint similarity evaluation system of Chinese medicinal materials（version 2.0）.A total of17 common peaks were calibrated,which can comprehensively reflect the common mode and internal quality of 13 batches of Blumea balsamifera oil.The similarity was calculated,and the similarity between the HPLC fingerprint of each test product and the fingerprint of the control characteristic fingerprint is above 0.93.3.5 Investigation of the stability of Blumea balsamifera oilThe results of influencing factors showed that when stored under high temperature conditions,the properties and quality of Blumea balsamifera oil had uncontrollable and irreversible significant changes within a short period of time.The appearance traits and active ingredients were relatively stable,and no significant changes have occurred.Therefore,it was recommended that Blumea balsamifera oil should be stored at a low temperature,protected from light,and sealed.The recommended storage time was 1 year.