Coumarin Compounds Detection Methods And Applied Research

Coumarins, a common drug, have various clinical drug effects. In medical fields, they have various medical effects, such as anticoagulant, antibacterial, sedation etc. And in Traditional Chinese Medicine, they are used as index compounds to reflect quality of.Material Chinese medicine, and they are regulated by Chinese Pharmacopoeia. Thus, Human take advantage of Coumarins'medical effect to overcome disease, protect health and improve the quality of living. However, recently many company exaggerate the medical effect of coumarins, public misunderstand the safety of coumarin and the illegal use of these compounds, therefore many adverse reactions are reported by medias. As a result, more and more publics pay attention to Coumarins. The problem government faced to is that how to control and inspect misuse and abuse coumarins and reduce adverse effect of coumarins.Because coumarins are widly used in the society, and there are many paths that people intake coumarins. So to restrict coumarins content in product is the key point to solve the problem. However, in China, Coumarins inspection and detection is not complete and detail, and there are few suitable methods to detect these compounds. Therefore, to establish these methods to detect Coumarins will benefit many industries and will help human leave far away "hidden killers"-Coumarin.First, although various detection methods were established before, lack a new method which determination scores of Coumarins in signal analysis. Because there are hundreds of samples must be tested and present methods only can determine 5 or 6 Coumarins in signal analysis, so hardly meet requirement of regulation and hardly fulfills daily test workload. Second, in sample test, matrix is far complicated than before, and it will affect determination and quantization Coumarins. Thus old methods will misreport the content of Couamrin in samples. Third every single method will have it limits in detection, so establish other detection methods will helpful to confirm results. Four, in recent years, market have more sample need to be tested, as a results, how to rapid, accurate, specify detect scores of Coumarins in complicated matrixes become a bottleneck to limit export and import industry. Then a new Ultra Performance Liquid Chromatography tandem Mass spectrapy (UPLC-MSMS) detection methods were establish by this research to apply in Coumarins determination fileds.This research analysis 24 common Coumarins by Mass spectrapy, then compare and contrast fragmentation mechanisms of Coumarins. Then based on this research, this research establish a UPLC-MSMS methods to simultaneous detect 22 Coumarins. Then take anvantage of this screen method determination Coumarins contend in Chinese medicine-Du Liang dropping pills. Furthermore, test this pill by High performance liquid chromatography (HPLC).This research establishes a UPLC-MSMS method for Coumarins detection, because Tradition Chinese medicine has complicated matrixes, complicated and variaous compounds, so it is hard to determine accurate. In this research, first compare and contrast the extract condition. This part contrast different ultrasonic extraction solvent and time. Then this part develops a Solid Phase Extraction (SPE) method. This results show a best purifies results and little matrix effect. And in this part determine contend of 5-methoxypsoraln,8-methoxypsoralen and imperatorin in Du Liang Dropping pill. And this is the first time determine 5-methoxypsoralen and 8-methoxypsoralen in Du Liang Dropping Pills. Thus this research pushes forward Du Liang Dropping pill development and improve quality standard of Du Liang Dropping pill. This determination method has advantage, such as rapid (less than 6min), sensitive, accurate, specify, stability. And this determination method is suitable for Du Liang Dropping pill test.This research also develop a HPLC-DAD methods simultaneous determine 5-methoxypsoralen, imperatorin, isoimperatorin and ferulic acid. This part also uses ultrasonic extraction. And choose extract solvent and extract time. And in 30min separation this methods have a best anlysis results. This method has advantage in simple and convenient, fast and sensitivity, accurate and specify. And this method can be used as a complementary method to UPLC-MSMS method. For the cheap and wide-spread instrument, this method will be popular in primary lab. And this method can not detect 8-methoxypsoralen will limit it using rang.Finally, this research establish a HPLC-DAD methods to simultaneous determine 17 Coumarins in cosmetics. For the Cosmetic industry always add flavours in products. Some illegal factory adds flavors contend furocoumarin; some compounds restricted by European Cosmetic Regulation, and cause threat to human health. Thus, in order to protect human health, this research develop a method to simultaneous determine 17 Coumarins in cosmetic. This methods also simple and convenient, fast and sensitivity, accurate and specify. The LOQs are 0.5ug/ml, linear range is 0.5～50 ug/ml, and it have good linear relationship. The RSD of intra-day and inter-day assays were 0.87%～2.05%and 0.81%～2.03%. The blank add recovery were 82.3%～102.9%, with RSD%(n=6) were 0.27～3.23%. This method satisfies the requirement of European cosmetic regulation, and this method can be used in daily cosmetic test.In conclusion, this research establishes 1 screen method and 2 determination methods of Coumarins. And successful use these methods in Traditional Chinese medicine and cosmetic filed. These methods improve Du Liang Dropping pill and cosmetic test standard, develop a Coumarin test platform. Based on this platform researcher can control Coumarins abuse and inspect Coumarins misuse. And because this research also establish 2 HPLC methods, it is much convenient for test center to choose the suitable method to determine Coumarins.