Study On Correlation Impurities Profile Of Coenzyme Q₁₀ And Its Capsules

Coenzyme Q₁₀(CoQ₁₀)also known as ubiquinone,is a fat-soluble ubiquinone compound.Natural CoQ₁₀ is present in animal and plant tissues,particularly mammalian mitochondria-rich biofilms.Physiologically,it mainly participates in the supply of energy and antioxidation of the tricarboxylic acid cycle as a proton transmitter.It is mainly used for the adjuvant treatment of various chronic diseases and cancers in clinical practice as well as is widely used in dietary supplements.In 2016,Waigaoqiao Laboratory of Shanghai Food and Drug Control undertook the national drug evaluation sampling test for CoQ₁₀ solid preparations.It was found that CoQ₁₀ capsules might have problems in quality standards,prescription design and preparation process.Several batches of the 202 batches of CoQ₁₀0 capsules did not meet the requirements of the Pharmacopoeia of The People's Republic of China?ChP?.However,due to the short time for exploratory research,there was no further systematic study on the impurity profile of the variety.Therefore,on the basis of this paper,the research was carried out with reference to the relevant requirements of ICH?Q3A,Q3B?.The main research work included the following aspects:?1?Examined the overall situation of CoQ₁₀and its capsules.Referring to the related substances inspection method of CoQ₁₀ in ChP,a methodological study was conducted on it.The related impurities of CoQ₁₀ and its capsules were well separated and detected.As a result,five impurities?impurity 1 to impurity 5?containing more than 0.1%in the liquid chromatogram of CoQ₁₀ and its capsules were present,and the impurity 1 was characterized by coenzyme Q₉,the other four were unknown impurities.?2?The impurities preparation method of CoQ₁₀ was established,and three impurities with purity greater than 97.0%were obtained by separation and purification.CoQ₁₀ was destroyed by alkali and high temperature,then the content of impurity 2 and impurity 4 was increased,and the enrichment solutions of impurity 2and impurity 4 can be achieved.The solutions obtained were pre-separated by silica gel column chromatography and purified by preparative HPLC.The effluent was handled by collected,concentrated,dried,and then separately prepared to obtain impurity 2 and impurity 4,respectively.The impuritiy 4 were separated and enriched by normal phase high performance liquid chromatography to prepare two isomer impurities?impuritiy 4-1 and impurity 4-2?.?3?Structure confirmation of CoQ₁₀ related impurities.UPLC-QTOF/MS,NMR,IR and other methods were combined with foreign pharmacopoeia and literature reports to confirm the structure of impurity 2,impurity 3,impurity 4-1,impurity 4-2 as well as impurity 5,respectively.Among them,impurity2 was confirmed to be the impurity E contained in the British Pharmacopoeia.Impurity 3 was 5-demethoxy CoQ₁₀.Impurity 4-1 and impurity 4-2 were confirmed as ethoxy monosubstituted CoQ₁₀,which were not reported before.Impurity 5 was coenzyme Q₁₁,a homologous impurity of CoQ₁₀.?4?Source analysis of CoQ₁₀0 impurities and structural speculation of potential impurities.The CoQ₁₀ and its capsules were investigated for the generation of impurities under the stability test conditions and then the source of the impurities was analyzed.The test results showed that impurity 1,impurity 3 and impurity 5 were from the synthesis process.Impurity 2,impurity 4-1 and impurity 4-2 were impurities which were easily introduced during the synthesis process or storage.For CoQ₁₀ capsules in the accelerated test of 6 months,two potential impurities except impuritiy 1 to impuritiy 5 were found,and the impurity structures were estimated by UPLC-QTOF/MS.In addition,a CoQ₁₀ material compatibility test was conducted to investigate the path of impurity generation.The results showed that CoQ₁₀0 was incompatible with the auxiliary calcium sulfate in the CoQ₁₀ capsules.?5?A method for the determination of residual solvent by gas chromatography was established.The residual solvent in CoQ₁₀ and its capsules was investigated.The methodological investigation showed that the established method for determination of residual solvent was specific and sensitive.The method was applied to determine the residual solvent content in CoQ₁₀ and its capsules.It was found that some enterprises had capsules with ethanol residue.The research results of this thesis can provide reference for the production of CoQ₁₀ and optimization of the formulation process,lay a foundation for the improvement of the quality standard of CoQ₁₀ and its capsules.