Neuropathic pain is defined as "pain initiated or caused by a primary lesion or dysfunction in the nervous system". In contrast to physiological pain that warns of noxious stimuli likely to result in tissue damage, neuropathic pain serves no protective function. Examples of neuropathic pain states include postherpetic neuralgia (shingles) and phantom limb / stump pain. This pain state also exists in the orofacial region, with the possibility of several variants including atypical odontalgia and burning mouth syndrome. There is a paucity of information on the prevalence of neuropathic pain in the orofacial region. One study assessed patients following endodontic treatment and found that approximately 3 to 6percent of patients reported persistent pain. Patients predisposed to the condition atypical odontalgia (phantom tooth pain) include those suffering from recurrent cluster or migraine headaches. Biochemical and neurobiological processes leading to a neuropathic pain state are complex and involve peripheral sensitisation, and neuronal plasticity of the central and peripheral nervous systems. Subsequent associated pathophysiology includes regional muscle spasm, sympathetic hyperfunction, and centralisation of pain. The relevant clinical features of neuropathic pain are: (i) precipitating factors such as trauma or disease (infection), (ii) pain that is frequently described as having burning, paroxysmal, and lancinating or sharp qualities, and (iii) physical examination may indicate hyperalgesia, allodynia and sympathetic hyperfunction. The typical patient complains of persistent, severe pain, yet there are no clearly identifiable clinical or radiographic abnormalities. Often, due to the chronicity of the problem, afflicted patients exhibit significant distress and are poor pain historians, thus complicating the clinician's task of obtaining a detailed and relevant clinical and psychosocial history. An appropriate analgetic blockade test for intraoral sites of neuropathic pain is mucosal application of topical anaesthetics. Other, more specific, tests include placebo controlled lignocaine infusions for assessing neuropathic pain, and placebo controlled phentolamine infusions for sympathetically maintained pain. The treatment and management of neuropathic pain is multidisciplinary. Medication rationalisation utilises first-line antineuropathic drugs including tricyclic antidepressants, and possibly an anticonvulsant. Topical applications of capsaicin to the gingivae and oral mucosa are a simple and effective treatment. Neuropathic pain responds poorly to opioid medication. Psychological assessment is often crucial in developing strategies for pain management. Psychological variables include distress, depression, expectations of treatment, motivation to improve, and background environmental factors. To enable a greater understanding of neuropathic pain, thereby leading to improved treatments, high-performance liquid chromatography-mass spectrometry is one analytical technique that has the potential to contribute to our knowledge base. This technique allows drugs and endogenous substances to be assayed from one sample in a relatively short time. The technique can identify, confirm, and measure the concentrations of multiple analytes from a single sample.

For pain research, the orofacial region is unique in a number of ways. The region has complex local anatomy, including substantial sensory innervation from neural pathways, and muscles of facial expression that convey important information concerning pain intensity and associated psychological traits. Although chronic orofacial pain conditions appear prevalent, useful documentation on pain intensity ratings using well established instruments is sparse. In particular, two conditions, atypical facial pain and atypical odontalgia, are poorly understood in aetiology so that definitive treatment modalities are severely limited. The region's local biofluid, saliva, has been used to diagnose various local and systemic disease states, and to quantitate drug concentrations. However, recent studies indicate that saliva also contains some of the same peptides, e.g. bradykinin, that are involved in pain mechanisms. It may be that pharmacological-pharmacokinetic studies of these peptides could shed more information on thesignificance of their presence in saliva. This thesis consists of four major sections. Section 1 comprises of three clinical studies investigating orofacial pain. Section 2 deals with clinical laboratory studies of saliva. Section 3 is concerned with the development of chromatographic methods to assay bradykinin and its pharmacokinetics in saliva. Section 4 uses chromatography for the identification of novel salivary peptides. This thesis, then, presents clinical studies of orofacial pain and pharmacological investigations of saliva as the local biofluid.

The separating characteristics of high performance liquid chromatography (HPLC) columns, measured in terms of the height equivalent of a theoretical plate (HETP) and skewness of the eluted peak, are investigated using computational fluid dynamics (CFD). Gradually expanding and contracting sections are introduced at the inlet and outlet, respectively, in columns with and without frits and their performance was compared with that of the conventional columns without expanding and contracting regions. / Singapore-MIT Alliance (SMA)

Many mixtures important to research consist of hundreds or even thousands of individual components of interest. These types of mixtures are far too complex to separate by a single chromatographic dimension in any reasonable amount of time. However, if a multidimensional approach is used, where a complex mixture is separated by an initial dimension, simpler fractions of that separation are collected and each of those fractions are analyzed individually, highly complex mixtures can be resolved in relatively short amounts of time. This dissertation serves as a guide to multidimensional chromatography, in particular, two-dimension liquid chromatography. There are many aspects of multidimensional separations that have been investigated to show its aspects, drawbacks and potential ability to separate highly complex mixtures. Measurements for the performance of multidimensional chromatography, the effects of the first and subsequent dimensions and the approaches to pairing dimensions are shown with experimental examples. Fundamental and practical features of multidimensional chromatography are explained as well as theoretical discussions on current and future multidimensional chromatography performance. Experimentally, very high peak capacities were obtained (ca. 7000) and an algorithm to predict how to best optimize a two-dimensional separation based on the time used and performance was created for designing experiments.

Liquid chromatography coupled to mass spectrometry (LC‐MS) has due to its superiorresolving capabilities become one of the most common analytical instruments fordetermining the constituents in an unknown sample. Each type of sample requires a specificset‐up of the instrument parameters, a procedure referred to as method development.During the requisite experiments, a huge amount of data is acquired which often need to bescrutinised in several different ways. This thesis elucidates data processing methods forhandling this type of data in an automated fashion.The properties of different commonly used digital filters were compared for LC‐MS datade‐noising, of which one was later selected as an essential data processing step during adeveloped peak detection step. Reconstructed data was further discriminated into clusterswith equal retention times into components by an adopted method. This enabled anunsupervised and accurate comparison and matching routine by which components fromthe same sample could be tracked during different chromatographic conditions.The results show that the characteristics of the noise have an impact on the performanceof the tested digital filters. Peak detection with the proposed method was robust to thetested noise and baseline variations but functioned optimally when the analytical peaks hada frequency band different from the uninformative parts of the signal. The algorithm couldeasily be tuned to handle adjacent peaks with lower resolution. It was possible to assignpeaks into components without typical rotational and intensity ambiguities associated tocommon curve resolution methods, which are an alternative approach. The underlyingfunctions for matching components between different experiments yielded satisfactoryresults. The methods have been tested on various experimental data with a high successrate. / De analysinstrument som används för att ta reda på vad ett prov innehåller(och till vilken mängd) måste vanligtvis ställas in för det specifika fallet, för attfungera optimalt. Det finns ofta en mängd olika variabler att undersöka som harmer eller mindre inverkan på resultatet och när provet är okänt kan man oftast inteförutspå de optimala inställningarna i förtid.En vätskekromatograf med en masspektrometer som detektor är ett sådantinstrument som är utvecklat för att separera och identifiera organiska ämnen lösta ivätska. Med detta mycket potenta system kan man ofta med rätt inställningar delaupp de ingående ämnena i provet var för sig och samtidigt erhålla mått som kanrelateras till dess massa och mängd. Detta system används flitigt av analytiskalaboratorer inom bl.a. läkemedelsindustrin för att undersöka stabilitet och renhethos potentiella läkemedel. För att optimera instrumentet för det okända provetkrävs dock att en hel del försök utförs där inställningarna varieras. Syftet är attmed en mindre mängd designade försök bygga en modell som klarar av att peka åtvilket håll de optimala inställningarna finns. Data som genereras från instrumentetför denna typ av applikation är i matrisform då instrumentet scannar och spararintensiteten av ett intervall av massor varje tidpunkt en mätning sker. Om enanalyt når detektorn vid aktuell tidpunkt återges det som en eller flera överlagdanormalfördelade toppar som ett specifikt mönster på en annars oregelbundenbakgrundssignal. Förutom att alla topparna i det färdiga datasetet helst ska varavälseparerade och ha den rätta formen, så ska tiden analysen pågår vara så kortsom möjlig. Det är ändå inte ovanligt att ett färdigt dataset består av tiotalsmiljoner uppmätta intensiteter och att det kan krävas runt 10 försök med olikabetingelser för att åstadkomma ett godtagbart resultat.Dataseten kan dock till mycket stor del innehålla brus och andra störandesignaler vilket gör de extra krångligt att tolka och utvärdera. Eftersom man ävenofta får att komponenterna byter plats i ett dataset när betingelserna ändras kan enmanuell utvärdering ta mycket lång tid.Syftet med denna avhandling har varit att hitta metoder som kan vara till nyttaför den som snabbt och automatiskt behöver jämföra dataset analyserade medolika kromatografiska betingelser, men med samma prov. Det slutgiltiga målet harfrämst varit att identifiera hur olika komponenter i provet har rört sig mellan deolika dataseten, men de steg som ingår kan även nyttjas till andra applikationer.

A new general method is developed for the optimization of HPLC ternary or pseudo-quaternary mobile phases which are represented by the trilinear coordinate system. This method can predict the global optimum of the mobile phase composition. The global optimum composition along each edge of the triangle and the corresponding selectivity factor of the worst-separated peak pair(s) are used in this method. This method is named the weighted pattern comparison optimization method (WPCO) and is applicable for both known and unknown samples. The WPCO method is simpler than those currently in use. The WPCO method was tested by using 68 literature data sets whose separation response surfaces are different. Results of the WPCO method agree with the results obtained by the minimum α plot method and by the grid search method, and do so with substantially fewer experimental measurements. Compared with the 5% (in eluent composition) step size grid-search procedure, the WPCO method using the same step size reduces the experimental work by 75%. For further reducing the experimental work, the original WPCO method is simplified. In an ordinary HPLC separation, the separation factor and resolution are approximately proportional to the logarithm of the selectivity factor. Based on this, the separation factor replaces the logarithm of selectivity factor in the original WPCO method. This further reduces the experimental work and avoids the error introduced in the measurement of the column dead volume. The simplified WPCO method has been tested in the normal-phase and reversed-phase chromatography separation cases. The simplified WPCO method has been tested by using 27 literature data sets whose separation response surfaces are different. Results of the simplified and original WPCO methods are nearly identical when the capacity factors of the solutes of the worst-separated peak pairs are greater than 5. When the capacity factors are less than 5, the simplified WPCO method is satisfactory in less complex, less critical applications. Two new metabolites of nefopam have been separated from greyhound urine. In the separation process, flash chromatography is used for cleaning up and preseparating the samples in a single step. Compared with other techniques, experimental work is reduced. The structure of one of the newly discovered metabolites is determined using MS and NMR. The most probable structure of the other metabolite is determined using MS. The main metabolic pathways at different doses in greyhounds are studied. / Graduation date: 1991

A reverse-phase high pressure liquid chromatographic method for the quantitative analysis of unchanged ascorbic acid in human urine is described. Selection of an appropriate mobile phase and discussion of some of the analysis problems are presented. Twenty-four hour ascorbate excretion profiles from two subjects were determined. Standard redox titration procedures were used to corroborate the chromatographic method. When compared to classical titration or colorimetric redox procedures, the new assay features straightforward sample preparation and improved sensitivity.Ball State UniversityMuncie, IN 47306

The production and distribution of counterfeit drugs is a serious worldwide health problem. One recent example is the appearance of fake artesunate antimalarial tablets in Southeast Asia. Due to the malevolent circumstances in which these fakes are produced, concern over the presence of toxic tablet ingredients is very much a legitimate health issue. Therefore, quantification of the amount of active ingredient present in tablets marketed as artesunate, a drug used for the treatment of the multidrug-resistant Plasmodium falciparum malaria in Southeast Asia required liquid chromatography coupled to mass spectrometry (LC-MS). This quantification allows the classification of the tablets as genuine, sub-therapeutic or fake and the validation of field results using colorimetric tests. During the LC-MS experiments, there were the observations that several of the samples contained a “wrong” active ingredient (AI). This was identified via accurate mass measurements, chromatographic retention time and in-source collision-induced dissociation (CID)as erythromycin, a common antibiotic. Using multivariate unsupervised clustering algorithms, the LC-MS data was utilized for “chemically fingerprinting” the fake tablet samples and investigating the similarities between them. The results of this initial survey show a correlation between sample origin, the different types of fake authentication holograms found in the packaging and sample composition.

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Thesis (Ph. D.)--University of Missouri-Columbia, 2001. / Typescript. Vita. Includes bibliographical references (leaves 114-126). Also available on the Internet.