Effect Of Sulfur Fumigation And Heat Treatment Desulfurization On The Quality Of Codonopsis Pilosula

Codonopsis Radix（CR）is the root of Codonopsis spilosula（Franch.）Nan-nf.of Campanulaceae.It is a commonly used tonic Chinese medicine in clinic.As CR is susceptible to mildew,sulfur fumigation is often used in processing and storaging this herb.At present,Chinese Pharmacopoeia（2015 edition）stipulates that the sulfur dioxide residue of CR should not be higher than 400 mg·kg^-1.In order to make the sulfur dioxide residue of sulfur-fumigated CR meet the limit requirements of Chinese Pharmacopoeia（2015 edition）,there is a phenomenon that CR is heat-treated to desulfurize in the market.As sulfur fumigation can increase harmful sulfur dioxide residues,and may cause transformation of active ingredients,thus affect the quality of medicinal materials,in this paper,the effects of sulfur fumigation and heat-treating desulfurization on the quality of CR are systematically studied as follows,which can provide scientific data for the feasibility evaluation of sulfur fumigation and heat treating desulfurization of CR,and also provide research ideas for the quality control of sulfur fumigated CR.Chapter1 Effects of sulfur fumigation and heat-treating desulfurization on sulfur fumigation derivatives and sulfur dioxide residues in CRUsing UPLC-QTOF-MS/MS based metabolome and multivariate statistical analysis method,the characteristic sulfur-containing derivatives,atractylolide II sulfur-containing derivatives and atractylolide III sulfur-containing derivatives were rapidly discovered in sulfur fumigated CR,indicating that sulfur fumigation affects the chemical composition of CR and produces sulfur-fumigated sulfur-containing derivatives.The characteristic ions m/z 327.0902 and m/z295.0999 of sulfur derivatives were used as screening ions for sulfur fumigation screening of commercial CR.The effects of sulfur fumigation and heat-treating desulfurization on contents of sulfur fumigation derivatives（Atractylenolide II sulfur derivatives and Atractylenolide III sulfur derivatives）,derivative precursors（Atractylenolide II and Atractylenolide III）and sulfur dioxide residues in CR were investigated by UPLC-QTOFMS/MS semi-quantitative method.1)Correlation between the sulfur dioxide residues in fumigated CR and the time course of sulfur fumigation.With the prolongation of sulfur fumigation,the content of sulfur fumigation derivatives（sulfur derivatives of Atractylenolide II and Atractylenolide III）first increases and then tends to be stable,while the content of Atractylenolide II and Atractylenolide III changes correspondingly with sulfur fumigation derivatives,and the content decreases first and then tends to be stable.There is no correlation between sulfur dioxide residues and sulfur derivatives.Sulfur fumigation increased the sulfur dioxide residue of CR and transformed its chemical composition,indicating that there was no correlation between the sulfur dioxide residue and the internal quality of sulfur fumigated CR.2)Correlation between sulfur dioxide residues in sulfur fumigated CR and the time of heat-treating desulfurization.The contents of sulfur fumigation derivatives（Atractylenolide II sulfur derivative and Atractylenolide III sulfur derivative）and derivative precursors（Atractylenolide II and Atractylenolide III）did not significantly change with the time of heat-treating desulfurization,indicating that their properties are stable.There is no correlation between sulfur dioxide residues and the sulfur fumigation derivatives.Heat-treating desulfurization could reduce the sulfur dioxide residues of sulfur fumigated CR,but could not eliminate the change of the internal quality of sulfur fumigated CR.Chapter2 Effects of sulfur fumigation and heat-treating desulfurization on non-saccharide small molecular components in CRUPLC-QTOF-MS/MS method was established to characterize the chemical profile of sulfur fumigated and heat-treating CR samples.A total of 60 compounds were identified,including alkynes and polyacetylenes,phenylpropanoids,alkaloids,flavonoids,terpenoids,amino acids,acids and nucleosides in CR.An HPLC method was developed for simultaneous determination of 13 compounds belonging to 7 types in CR,including polyacetylenes Lobetyolin;flavonoids baicalin and rutin;terpenoids atractylenolide II,atractylenolide III;phenylpropanoid glycosides syringin;anthraquinone emodin;amino acid tryptophan;Furan 5-hydroxymethyl furfural.1)The total amount of the analytes did not significantly change after sulfur fumigation;the contents of emodin,tryptophan,lobetyolin,baicalin,rutin,atractylenolide II and atractylenolide III decreased after sulfur fumigation,and the contents of baicalin and rutin were negatively correlated with the time course of sulfur fumigation;the contents of 5-hydroxymethylfural increased significantly,but there was correlation the duration of sulfur fumigation.2)Heat-treating desulfurization had no significant effect on the total amount of components in sulfur fumigated CR;after heat-treating desulfurization,the content of 5-hydroxymethyl furfural increased significantly and positively correlated with the heat-treating duration;the contents of emodin,tryptophan,baicalin,rutin,syringin,atractylenolide II decreased,and the contents of tryptophan,rutin and baicalin were negatively correlated with the heat-treating duration.An UPLC-QTOF-MS/MS semi-quantitative method was developed to investigate the content change of 60 compounds in CR after sulfur fumigation and heat-treating.The results showed that the content of 31 compounds decreased after sulfur fumigation,mainly including phenylpropanoid glycosides,flavonoid glycosides and nitrogen-containing organic compounds.The content of other 18 compounds increased,of which 4 were sulfur-containing derivatives.After heat-treating,the contents of 7 compounds decreased,including phenylpropanoid glycosides,flavonoid glycosides and nitrogen-containing organic compounds.The contents of other 7 compounds increased,mainly including 5-hydroxymethyl furfural.Chapter3 Effects of sulfur fumigation and heat-treating on carbohydrates in CRThe content of total polysaccharides in CR before/after sulfur fumigation and heat-treating was compared by phenol-sulfuric acid method.The results showed that the content of total polysaccharides in CR increased significantly after sulfur fumigation,and decreased with the prolongation of heat-treating desulfurization time.The molecular weight range of polysaccharides from CR before/after sulfur fumigation and heat-treating was characterized by HPGPC-ELSD.It was found that the molecular weight range of polysaccharides from CR was 6.1×10⁵-3.1×10⁶ Da;and the molecular weight range of polysaccharides from 1.4×10⁶-3.1×10⁶ Da after sulfur fumigation and heat-treatment;The polysaccharides with molecular weight of 3800 Da-9700 Da increased significantly after sulfur fumigation for 10 h,and the polysaccharides with molecular weight of 3800 Da-9700 Da decreased significantly after heat-treatment.The changes of monosaccharide composition and content of CR polysaccharide before and after sulfur fumigation and heat-treating were compared by PMP derivatization method.The results showed that the polysaccharide of CR mainly consisted of Man,GalA,Rha,Glc,Gal and Ara.The monosaccharide composition of CR polysaccharide remained unchanged after sulfur fumigation and heat-treatment,but the composition quantitative results showed that the total monosaccharide content of CR polysaccharides decreased after sulfur fumigation,and the contents of Glc and Rha decreased significantly.After heat-treatment for 32 hours,the total monosaccharide content of polysaccharides of sulfur fumigated CR and the contents of Glc and Rha decreased significantly.The content changes of free monosaccharides and oligosaccharides in CR before/after sulfur fumigation and heat-treating were compared with pre-column derivatization HPLC-ELSD and PPM HPLC-UV.Free monosaccharides in CR are mainly Fru,Glc,Man and GalA,while oligosaccharides are mainly sucrose.After sulfur fumigation,the content of free monosaccharides in CR increased significantly,and the content of sucrose decreased.After heat-treating,the content of free monosaccharides decreased.In summary,both sulfur fumigation and heat-treating desulfurization have obvious effects on the internal quality of CR.Therefore,sulfur fumigation and heat-treating desulfurization are not suitable for the processing of CR.