Development of Analytical Methods Based on Near Infrared
Total Page：16
File Type：pdf, Size：1020Kb
ADVERTIMENT. Lʼaccés als continguts dʼaquesta tesi queda condicionat a lʼacceptació de les condicions dʼús establertes per la següent llicència Creative Commons: http://cat.creativecommons.org/?page_id=184 ADVERTENCIA. El acceso a los contenidos de esta tesis queda condicionado a la aceptación de las condiciones de uso establecidas por la siguiente licencia Creative Commons: http://es.creativecommons.org/blog/licencias/ WARNING. The access to the contents of this doctoral thesis it is limited to the acceptance of the use conditions set by the following Creative Commons license: https://creativecommons.org/licenses/?lang=en Development of analytical methods based on Near Infrared Spectroscopy for monitoring of pharmaceutical and biotechnological processes and control of new psychoactive substances Aira Yira Miró Vera Doctoral Thesis PhD Program in Chemistry Professor Manel Alcalà Bernàrdez, PhD, Director Department of Chemistry Faculty of Sciences 2019 Acknowlegments This research was funded by MINECO (Spain) through the project CTQ 2016-79696-P (AEI/FEDER, EU). I appreciate the academic support of Professor Manel Alcalà Bernàrdez. I am also grateful to Professor Francisco Valero and personnel of the Department of Chemical Engineering UAB, Professor Sergi Armenta and personnel of the Department of Analytical Chemistry University of Valencia for the kind provision of samples, instruments, as well as their valuable collaborative efforts. 2 ABSTRACT The Near Infrared Spectroscopy (NIRS) is an analytical technique based on the interaction of electromagnetic radiation in the wavelength range 780-2500 nm and matter. The NIR spectrum can be considered as a “fingerprint” of each chemical compound or mixture of them, which contains absorption bands that are the result of overtones and combinations of the fundamental vibrations observed in the Mid-Infrared region. Additionally, NIRS of solids is sensible to the scattering effect caused by physical characteristics of the samples, therefore provides simultaneous sensitivity to chemical and physical changes of solids. Because of NIR spectra show broad and overlapped bands makes it necessary the use of Chemometrics, which implies the application of statistical and mathematical methods to such spectral data, for achieving the maximal extraction and collection of useful information from it. The general objective of this doctoral thesis is developing analytical methods based on NIRS for monitoring pharmaceutical and biotechnological processes and for the control of illicit drugs. The following conditions have been considered i) extended active pharmaceutical ingredient (API) concentration ranges during granulation and tableting using the process spectrum (PS), ii) on-site identification of new psychoactive substances (NPS) during police seizing procedures with hand-held and bench-top instruments and iii) inline monitoring of the production of recombinant Lipase B from Candida antarctica in Pichia pastoris using Glycerol as carbon source. i) Extended API concentration ranges during granulation and tableting using the PS The PS is a methodology for preparing calibration sets by adding the changes due to the manufacturing process to NIR spectra of samples prepared at the laboratory, using an algebraic procedure. The PS has successfully included the process contributions during modelling in the central point of API concentration values of diverse formulations, however the properties of this methodology at extreme points of API concentration ranges have not been studied yet. For evaluating such properties, in this work the PS was applied to samples in the range of ± 30% of a nominal API value. Results have shown that the PS performance can be affected by API concentration changes in the studied range, and classical pre-treatments are not enough to overcome this condition. 3 ii) Comparison of the performance of bench-top and hand-held NIR instruments concerning the identification of NPS The NPS are 'legal highs' with molecular differences regarding the structures of illicit controlled drugs, whose emergence have expanded the current synthetic drugs market in a very important way. The feasibility of using portable NIRS instruments for the fast identification of NPS have been previously demonstrated, however, their performance has not been faced to the performance of bench-top instruments. Results presented in this thesis expose that, even when models developed using data from NIRS miniaturized instruments are limited in performance regarding those developed using data provided by bench-top instruments, classification models of NPS based on data from hand-held instruments can be useful to make real-time and on-site decisions that can be confirmed later using high performance analytical instrumentation. iii) Inline monitoring of the production of recombinant Lipase B from Candida antarctica in Pichia pastoris using glycerol as carbon source. The use of new constitutive promoters and recycled carbon sources in the recombinant production of industrial proteins, such as lipases, in the cell factory Pichia pastoris is advantageous for improving production yields and minimizing the cost of the culture medium. The capabilities of a NIR spectrometer with fiber optic coupling for immersion of a transflectance probe were employed for the inline monitoring of the cultivation mentioned in the headline. Quantitative models have been developed for Biomass, Total protein, Nitrogen and Activity, which have demonstrated better prediction capability during the feed batch stage than during the batch stage. Predictions of glycerol values has been probably affected by the formation of hydrogen bonds in the aqueos medium. 4 Table of content ABSTRACT .............................................................................................................................................. 3 OBJECTIVES ........................................................................................................................................... 7 1. Introduction ........................................................................................................................................... 8 1.1 An historical perspective ................................................................................................................. 8 1.2 Physical fundamentals................................................................................................................... 17 1.3 Acquisition modes ........................................................................................................................ 26 1.4 Instrumentation ............................................................................................................................. 30 1.5 Data analysis ................................................................................................................................. 36 1.6 General advantages and disadvantages of the technique .............................................................. 48 1.7 Applications of NIRS .................................................................................................................... 49 1.8 References ..................................................................................................................................... 66 2. Performance of the Process Spectrum calculated at extreme API concentration values in the inclusion of the process variability of pharmaceutical solids manufacturing into calibration sets of samples prepared at the laboratory ....................................................................................................................... 75 2.1 Introduction ................................................................................................................................... 75 2.2 Experimental ................................................................................................................................. 77 2.3 Results and discussion .................................................................................................................. 78 2.4 Conclusions ................................................................................................................................... 87 2.5 References ..................................................................................................................................... 87 3. Comparison of the performance of bench-top and hand-held Near infrared instruments concerning the identification of new psychoactive substances ....................................................................................... 89 3.1 Introduction ................................................................................................................................... 89 3.2 Experimental ................................................................................................................................. 91 3.3 Results and discussion .................................................................................................................. 97 3.4 Conclusions ................................................................................................................................. 108 5 3.5 References ................................................................................................................................... 108 4. Inline monitoring of recombinant production of Lipase B from Candida antarctica in Pichia pastoris using glycerol as carbon source .........................................................................................................................