ABSTRACT
OBJECTIVE To study active components and antitussive effect of aboveground part of Pinellia ternata （non- medicinal stems and leaves）， and compare them with the underground part of P. ternata （medicinal underground tubers）， providing scientific basis for the comprehensive utilization and product development of P. ternata. METHODS TLC， GC， HPLC and UPLC- MS/MS were used for qualitative and quantitative analysis of amino acids， volatile oil， total flavonoids and succinic acid from the aboveground and underground parts of P. ternata. The antitussive effects of the aboveground and underground parts of P. ternata were compared and studied through cough inducing experiment with concentrated ammonia water. RESULTS Results of TLC showed that at the corresponding positions on the chromatograms of the reference substances of P. ternata， and arginine， alanine， valine， leucine and rutin control， the aboveground and underground parts of P. ternata showed spots of the same color. Results of GC showed that the similarity among characteristic chromatograms of volatile oil from aboveground and underground parts of P. ternata was 0.767； results of HPLC and UPLC-MS/MS showed that compared with underground parts of P. ternata， the contents of succinic acid， quercetin， kaempferol and isorhamnetin increased by 0.15%， 0.15%， 0.09% and 0.03%， and aspartate content decreased by 2.5 mg/g. Pharmacodynamics results showed that compared with model control group， the cough incubation period of rats was prolonged significantly in administration groups （P＜0.05）， and the cough frequency within 3 min was significantly decreased （P＜0.05）； there was no statistical significance in the cough frequency within 3 min among administration groups （P＞0.05）. CONCLUSIONS The composition of amino acids， volatile oils and total flavonoids in aboveground part of P. ternata are similar to underground part of P. ternata， while the content of aspartic acid is lower than that in underground part. The aboveground part of P. ternata can prolong the cough incubation period of rats and reduce the number of coughs， which has a certain antitussive effect， but the effect is slightly weaker than that of the underground part.
ABSTRACT
Objective:To compare the immunogenity of the AST artificial antigen with two different carrier proteins, bovine serum albumin(BSA) and keyhole limpet hemocyanin(KLH).Methods: The hapten Astragaloside Ⅳ(AST) was linked to carrier protein BSA or KLH as immunogen,and linked ovalbumin(OVA) as coating antigen by the sodium periodate method.Then the 6-8 weeks BALB/c mice were immunized with AST-BSA or AST-KLH.The titer and specificity of the antiserum were detected by indirect enzyme-linked immunosorbent assay ( iELISA ) and indirect competitive enzyme-linked immunosorbent assay ( icELISA ) respectively.Results:The antiserum of the mice immunized with AST-KLH had better specificity than which immunized with AST-BSA.The IC50 of AST-KLH antiserum was about 5 μg/ml in the detection by icELISA.The high titer of both antiserums was about 1∶51 200 after immunizing mouse.Conclusion:The conjugate of AST-KLH was testified to have a better immunogenity in comparison with AST-BSA,this investigation would be the foundation of the further research of AST immunoassay.