Studies On The Improved Quality Control Of Earthworm Injection

Earthworm injection is a traditional Chinese medicine preparation,which is obtained by removing protein,lipid and other macromolecular substances from the decoction of Pheretima aspergillum which is extracted by water for many times and precipitated by alcohol,then sterilizing by hot pressing,adding various excipients and sterilizing again.It contains a variety of free amino acids,nucleotides and other chemical components.It has the effect of relieving asthma and cough,and can be used for cough and wheezing caused by bronchial asthma.There are many deficiencies in identification,inspection and content determination in the current standard(Standard No.WS-11340(ZD-1340)-2002),which can not reflect the quality of Earthworm injection effectively.To improve the quality control system of Earthworm injection and ensure the safety of medication,the thesis is related to improve the quality standard of Earthworm injection on the basis of the current standards,Technical Guidelines for Research on Quality Standards of New Traditional Chinese Medicine(Draft for comments),Guiding Principles for Validation of Analytical Methods,Guiding Principles for Formulation of Limit of Harmful Residues of Traditional Chinese Medicine and Technical Requirements for Fingerprint Study of Traditional Chinese Medicine Injection(Interim).The improved quality standard of Earthworm injection provides a theory foundation and methodology for the quality control of Earthworm injection.The thesis mainly includes the following contents and results:(1)A HPLC method was established to determine the contents of alanine,valine,lysine,isoleucine and leucine in Earthworm injection simultaneously;the HPLC fingerprints of free amino acids in 10 batches of Earthworm injection were established.The results of fingerprint similarity evaluation were over 0.90,proving that the preparation process had a good reproducibility,stability and feasibility.(2)A HPLC method was established for the determination of hypoxanthine,uridine,inosine,guanosine and phenol in Earthworm injection simultaneously,which was stable,reliable and reproducible.Both contents determination of active components(hypoxanthine,uridine,inosine and guanosine)and phenol detection were added to the improved quality standard.(3)Add the new inspection items of harmful heavy metals to the quality standard.The improved quality standard added the determination of the copper,arsenic,cadmium,mercury and lead contents in Earthworm injection,and a sensitive and accurate ICP-MS method was established.(4)Improve the identification method of Earthworm injection.The HPLC identification method of nucleosides(hypoxanthine,uridine,inosine and guanosine)and free amino acids(lysine,leucine,valine peaks)in Earthworm injection were used to replace the original TLC method.(5)An improved quality standard of Earthworm injection was drafted,which had a good scientificity,rationality and applicability and conformed to the requirements of relevant principles.The methods established in this thesis have good linear relationships in the determination range,high sensitivity,accuracy and precision.The new quality standard improves the quality control of Earthworm injection,and reflects the immanence of Earthworm injection,and provides a theory foundation and guarantee for its quality control and clinical safety.