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The Near Infrared Spectroscopy-Based Quality Control of Drug and the Detection of Nucleic Acid with Nano Fluorescent Probe

Author: YinJieHong
Tutor: XieHongPing
School: Suzhou University
Course: Medicinal Chemistry
Keywords: Near infrared spectroscopy Rhizoma Compound clorprenaline Quantum dots Magnetic nanoparticles Fluorescent probe Nucleic acid detection
CLC: TB383.1
Type: Master's thesis
Year: 2009
Downloads: 137
Quote: 4
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Abstract


Based on near infrared (NIR) spectroscopy, the paper has studied the quality control for the traditional Chinese medicine Rhizoma pardis and the quality evaluation for compound clorprenaline in the first section.The whole process of the separating and purifying steroidal saponin in Rhizoma by using AB-8 macroporous resin was monitored with the help of near infrared spectrometer, which carried out the process quality control in real time. Based on the discriminant variable of the NIR spectra, the four stages were determined in the whole separating process by the elution with 70% ethyl hydroxide. The steroidal saponin was mainly included in the first three stages, in which the separation efficiencys of the 24α-hydroxy-pennogenin, pennogenin and diosgenin were 96.22%, 92.90% and 92.34%, respectively. The paper believes that the method has the potential of separating steroidal saponin in Rhizoma according to class 1 new drug in traditional Chinese medicine.The NIRS also provides a new method for the online quality control during production process for pharmaceutical preparation. Based on the near infrared fiber diffuse reflectance spectroscopy, the analytical methods of the fast and simultaneous determinations for clorprenaline hydrochloride, bromhexine hydrochloride and decloxizine hydrochloride contained in the compound clorprenaline have been established by the partial least squares method. The multivariate calibration models for clorprenaline hydrochloride, bromhexine hydrochloride and decloxizine hydrochloride have the good correlations with the correlation coefficients of 0.997, 0.994 and 0.990, and the root mean square errors of 0.028, 0.145 and 0.250 for the calibration sample sets, and the strong prediction abilities with the root mean square errors of 0.055, 0.120 and 0.210 for the prediction sample sets, respectively. Because of the fast and simultaneous fiber analyses with no pre-treating, the established method could be used in the process quality control of compound clorprenaline.In the second section, the quantum dots (QDs, i.e. the fluorescent nanomaterials) and magnetic nanopaticles (MNP) have been synthesized, and their applications on the detection of nucleic acids have been studied.CdTe QDs of the different particle sizes with the ligand thioglycolic acid have been synthesized in water by using the materials CdCl2, Te power, NaBH4 and thioglycolic acid in the conditions of heating and refluxing. The influencing factors on the CdTe QDs synthesis have been investigated, and the high quality CdTe QDs with green, yellow, orange and red fluorescence have been obtained, whose fluorescence quantum yields were up to 40%. By using the synthesed CdTe QDs and the sulfur source thioacetamide, we have synthesed the core-shell CdTe/CdS QDs. Their fluorescence quantum yields were further improved (up to 70%), and their stability for light and oxygen was obviously enhanced. Moreover, the paper synthesized initially the core-shell CdTe/SiO2 composite particles with good dispersibility and homogeneity by using the method of reverse microemulsion.Fe3O4 magnetic nanopaticles have been synthesized by using co-precipitation method. They had good magnetic properties (i.e. their specific saturation magnetization up to 70.970emu/g, their remanence low to 3.493emu/g and coercive force low to 23.57Oe). Their particle diameters were about 15nm. They were the stable colloid water solution without lay separation during several months. Based on the rich hydroxyl groups on the surface of Fe3O4 magnetic nanopaticles, the amino-coated Fe3O4 magnetic nanopaticles have been obtained by using silanizing agent, and this kind of modification has been confirmed by infra-red spectrum.QDs-DNA probes (CdTe-P,CdTe/CdS-P1) and MNP-DNA probes (MNP-P2) have been obtained by using the EDC cross-linking method and the glutaraldehyde one, respectively. Morever, we applied these two kinds of probes to the quantitative detection and identification of nucleic acid. Based on the probe CdTe-P, we established the fluorescent resonance energy transfer system between the CdTe QDs with the emission wavelength 540nm and ethidium bromide for the quantitative determination of nucleic acid. This method had the liner range 25-188 nM, the relation coefficient 0.997 and the limit of quantification 25nM. The sandwich-type method for the nucleic acid qualitative detection has been established using CdTe/CdS-P1 as the detecting probe and MNP-P2 as the capture probe. This method could recognize the completely-matched oligonucleotide sequence and the single-base-mismatched one with the combined probes (i.e. CdTe/CdS-P1 and MNP-P2), which may be applied to disease detection with genetic mutation in clinical medicine.

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