Fluorescent Probes For Neurotransmitter Determination Of Fatty Acids And Its Application

For the powerful separation and analytical capacities, reversed-phase high performance liquid chromatographic method has been widely used in many fields of scientific research and application, such as biology, chemistry, toxicology, clinical medicine, food hygiene and environmental monitoring. It is also one of the fastest developing and the most widely used methods in analytical chemistry. However, not all compounds are accessible to be analyzed by chromatography due to the limitation of the compounds properties and the low content of detected components or the suitable detector of HPLC. Therefore, developing testing methods with high sensitivity has very important significance. At present, a wide range of derivatization regents have been used in HPLC, but both of them exist more or less shortcomings. Therefore, developing highly sensitive fluorescent regents to meet the actual needs of HPLC is very important. The dominant ideology of this paper is to design a new fluorescence derivatization reagent to enhance detection sensitivity and improve the shortcoming of existing reagents in actual analysis process. The main works of this paper involve designing and preparing new labeled reagents, determining spectral properties, developing chromatographic methods. Specific works are as follows:Chapter One:The developing progress and basic theory about high performance liquid chromatography and high performance liquid chromatography coupled with mass spectrometry were simply presented. The principles of fluorescence and chemical derivatization were introduced. The widely used fluorescence derivatization reagents were summarized.Chapter Two:Pre-column derivatization methods for the sensitive determination of neurotransmitter, respectively, using 2-[2-(7H-dibenzo[a,g]carbazole-ethoxy)]-ethyl chloroformate (DBCEC-C1) 2-[2-(7H-dibenzo[a,g]carbazole-ethoxy)]-ethyl chloroformate (DBCEC-C1) as labeling reagents followed by HPLC-MS analysis have been developed. The optimal derivatization and chromatographic separation conditions using DBCEC-C1 as labeling reagents were evaluated, respectively. The contents of neurotransmitter from real samples such as neurotransmitters in cerebrospinal fluid in rats were determined. Studies on derivatization conditions indicated that neurotransmitter could rapidly and smoothly react with DBCEC-C1 reagents under very mild conditions with maximal derivatization yields close to 100% with excellent linearity, good repeatability and higher sensitivity.Chapter Three:Pre-column derivatization methods for the sensitive determination of fatty acids, using DPPIPE and BAETS labeling reagents followed by HPLC-MS analysis have been developed. The optimal derivatization and chromatographic separation conditions using DPPIPE and BAETS as labeling reagents were elucidated. The contents of free fatty acids from real samples such as plasma, pollens were determined. Studies on derivatization conditions indicated that fatty acids could rapidly and smoothly react with the labeling reagents under very mild conditions, with maximal derivatization yields close to 100%, with excellent linearity, good repeatability and higher sensitiviy.