ABSTRACT A new, non-pathogenic bioaugmentation product was formulated specifically for underground use in South African mines, using local bacterial isolates. This was designed for the remediation of various hydrocarbons via biochemical breakdown by sub-surface microorganisms. The active microorganisms were isolated from hydrocarbon-polluted areas of a gold mine. Many commercially available bioaugmentation products are already in existence however, all, to our knowledge, have been developed and tested primarily for use in the northern hemisphere. None have been formulated and tested in Africa. Our series of bacterial isolates are the first to be isolated from mine soils for hydrocarbon biodegradation purposes. Such isolates have further, not previously been tested on sub-surface contamination. The safety associated with the use of such a product in a closed mine-environment is of paramount importance. Initial batch-flask experiments were conducted using a readily-available commercial bioremediation product. This was tested on simple surfactant molecules and compared to the biodegradation observed under standard waste water treatment plant conditions. The bioremediation product increased biodegradation by 6% on average. Bacteria in the product were identified by 16S rDNA gene sequence analysis and found to be homologous to potentially pathogenic Bacillus cereus, known especially to effect immunocompromised individuals, this was of particular concern in the closed mine system. South African isolates were sourced from various hydrocarbon-polluted sources, with six bacteria ultimately being selected from deep sub-surface mine soil and water samples. The ability of these isolates to biodegrade waterborne monograde engine oil was assessed via GC-FID. The isolate showing average percentage growth increase, homologous to Pseudomonas pseudoalcaligenes, was found to degrade the motor oil by 98%. The new isolates were, on average, 16% more efficient at biodegrading petroleum hydrocarbons than the commercial bioremediation product isolates. Formulation of these isolates into the first commercially-available South African developed and tested bioaugmentation product will prove a successful conclusion to this study.

The international harbors and fishing ports in Taiwan are nearly not installed the wastewater or oil treatment facilities, which would cause the ocean pollution by the ballast and bilge water from the boats. The purpose of this research is to study the feasibility of treatment of diesel fuel and the polycyclic aromatic hydrocarbons (PAHs) in diesel by aerobic biofiltreation in salty environment. According to the experimental results, the mineral oil and PAHs in the diesel fuel present high good removal efficiencies. Both of the contaminants were decomposed fast at the first 8 hours, and then slow down. The emulsified diesel fuel were found able to be reduced 83% within 8 hours, and the concentrations of phenanthrene would found decreased from 1.5 to 0.1 mg/L. After 8 hours, we observed that the PAHs with high number of rings kept a mild diminish trend, while the PAHs with less rings exhibited arrearage and irregular indicated by some peaks appearing in the chromatography diagram of HPLC, which were not found in the diagram in the initial of test run. In conclusion, the aerobic biofiltreation systems were found able to degrade n-alkanes and PAHs in diesel fuel effectively. Thus we suggested to install the systems at the greasiness accepting facilities in the harbor. However, we still have to keep studying the influence of the interrelationship between the contaminants in diesel fuel and the environment factors.

The Abraham solvation parameter model is a good approach for analyzing and predicting biological activities and partitioning coefficients. The general solvation equation has been used to predict the solute property (SP) behavior of drug compounds between biological barriers. Gas chromatography (GC) retention time can be used to predict molecular descriptors, such as E, S, A, B & L for existing and newly developed drug compounds. In this research, six columns of different stationary phases were used to predict the Abraham molecular descriptors more accurately. The six stationary phases used were 5% phenylmethyl polysiloxane, 6% cyanopropylphenyl 94% dimethylpolysiloxane, 5% diphenyl 95% dimethylpolysiloxane, 100% dimethylpolysiloxane, polyethylene glycol and 35% diphenyl 65% dimethylpolysiloxane. Retention times (RT) of 75 compounds have been measured and logarithm of experimental average retention time Ln(RTexp) are calculated. The Abraham solvation model is then applied to predict the process coefficients of these compounds using the literature values of the molecular descriptors (Acree Compilation descriptors). Six correlation equations are built up as a training set for each of the six columns. The six equations are then used to predict the molecular descriptors of the illegal drugs as a test set. This work shows the ability to extract molecular information from a new compound by utilizing commonly used GC columns available with the desired stationary phases. One can simply run the new compound in GC using these columns to get the retention time. Plugging in the retention time into the developed equations for each of the column will predict the molecular descriptors for the test compound and will give some information about the properties of the compound.

The aim of the diploma thesis was the aroma active compounds assessment and sensory profile determination of various types of sea buckthorn (Hippopha rhamnoides L.). The theoretical part provides the detailed description of this plant, comprising the occurence, taxonomy, chemical composition and use. Experimental part summarizes the data obtained: identification and quantification of aroma compounds using the SPME – GC –FID and GC-MS method and evaluation of complete sensory profile. Various cultivars of sea buckthorn harvested in 2009-2012 were analysed. In total 23 various types of sea buckthorn were measured, 23 alcohols, 10 aldehydes, 11 esters, 14 ketones and 5 acids were found.

Demands for plant-based diets sufficient in all required amino acids are increasing due to health and environmental advantages. As legumes offer a complementing source of quality protein to the already heavily consumed cereals, methods for determining amino acid compositions in legumes are needed. The aim of this thesis was to establish a GC-FID method for analyzing amino acids in legumes and to apply the method to analyze six locally grown legumes: Black beans, brown beans, red kidney beans, white beans, yellow and grey peas. White bean flour was used as an inhouse control sample. Legumes were hydrolyzed with acid and alkaline treatments;hydrolysates were purified and derivatized using an amino acid analysis kit (EZ:faast, Phenomenex) and quantified with GC-FID. All legumes were rich in Asp + Asn, Glu + Gln, Lys and Leu while limited in Met, Cys and Trp. Quality control trials showed a generally good precision of the method (between day CVs were ≤ 10% for 12 of 16amino acids ) and good linearity (r2 were 0,99 or above for 16 of 19 calibration curves). Limits of detection and quantification ranged from 2-46 nmol/mL and 7-154nmol/mL, respectively, depending on the amino acid. No accuracy trial was performed. In conclusion, a rapid GC-FID method for amino acid analysis was established and amino acid compositions of locally grown legumes was tentatively determined. However, further quality control trials are required to validate the method. / Efterfrågan av växt-baserade dieter med komplett innehåll av aminosyror ökar på grund av både hälso- och miljömässiga fördelar. Då baljväxter erbjuder en kompletterande källa till protein av hög kvalitet till cerealier, som redan konsumeras i hög grad, krävs metoder för att analysera aminosyra innehåll i baljväxter. Målet med detta arbete var att etablera en GC-FID metod för att analysera aminosyror i baljväxter och att applicera metoden för att analysera aminosyror för att analysera sex lokala baljväxter: Svarta bönor, bruna bönor, röda kidney bönor, vita bönor, gula och gråa ärtor. Mjöl på vita bönor användes som in-house kontroll prov. Baljväxterna hydrolyserades med både syra och bas behandlingar, hydrolysaterna renades och derivatiserades med ett kit för aminosyraanalys (EZ:faast, Phenomenex) och kvantifierades med GC-FID. Alla baljväxter hade höga halter av Asp + Asn, Glu + Gln, Lys och Leu men låga halter av Met, Cys och Trp. Kvalitetskontroller visade på en generellt hög precision (CV mellan dagar ≤ 10% för 12 av 16 aminosyror) och god linjäritet (r2var 0,99 eller högre för 16 av 19 kalibreringskurvor). Detektions- och kvantifierings-gränser varierade mellan 2-46 nmol / mL respektive 7 – 154 nmol / mL beroende på aminosyran. Inga tester gjordes för att bestämma metodens riktighet.Slutsatsen drogs att en snabb GC-FID metod för aminosyra analys har framtagits och indikativa värden av lokala baljväxters aminosyra innehåll fastställts. Däremot krävs flera kvalitetskontroller för att validera metoden.

Syftet med detta examensarbete var att utveckla en GC-metod för analys av etylenglykoler i förorenade vattenprover, exempelvis industriavloppsvatten. Detta skulle åstadkommas med en GC-FID och MMI (Multimode Inlet), samt helst utan provupparbetning. Ett annat av målen var att kunna kvantifiera inom området 1 ppm till 100 ppm, och att metoden skulle vara tillräckligt stabil för att kunna ackreditera den.Ett flertal kolonner testades, där en HP-5 med dimensionerna 30 m*0,53 mm*0,88 μm från Agilent var den som gav bäst resultat. Även två olika liners provades, dels en med glasull och dels en 2 mm dimpled liner, där dimpled linern gav bäst resultat, och minst ”carry-over” mellan analyserna. Bland de parametrar som gav störst effekt utmärker sig trycket i injektorn, och tiderna som de olika flödena är igång. För fullständiga inställningar, se bilaga 2.En metod redo för ackreditering hann ej tas fram, det finns ett flertal parametrar som bör undersökas för en effektivare och stabilare analys. Men metoden som är framtagen är redo för omfattande repeterbarhetstest. / The purpose of this bachelor’s thesis was to develop a GC method for analysis of ethylene glycols in polluted water samples, for example industrial sewage water. This was meant to be accomplished with a GC FID and MMI (Multimode Inlet), preferably without sample preparation. One of the goals was to be able to quantify between 1 ppm and 100 ppm, and that the method was stable enough for accreditation.A number of columns were tested, from where a HP-5 with the dimensions 30 m*0,53 mm*0,88 μm from Agilent gave the best results. Also, two different liners were tested, one with glass wool and one 2 mm dimpled liner, where the dimpled liner gave the best results and the least carry-over between samples. Amongst the parameters which gave the biggest effect, the inlet pressure and the different flow times stands out. For a full set of settings, see Bilaga 2.A method ready for accreditation was not developed due to short of time. There are still a number of parameters which should be investigated for a more efficient and stabile analysis. But the method developed is ready for extensive repeatability tests.

Soil contamination from petroleum spills is a frequent environmental problem in the world. It is obvious that petroleum exploration has contributed immensely to the economic growth of Nigeria, but over the last few decades, the Niger Delta of Nigeria has suffered grave human health risk and ecosystem degradation resulting from oil spillages, petroleum products leakages and other involuntary effluent discharges from oil exploration activities. This research seeks to develop and optimize GC-FID methods for the analysis of Petroleum hydrocarbons. Crude oil spillage contamination of soil from the Niger Delta was investigated 3 months after a crude oil-pipeline spillage. 47 Soil samples (300-500g) were collected at several points in the South-South Niger Delta. Control samples were taken from four unaffected sites within the vicinity of spillage with similar soil characteristics. Samples were collected at depths of 0-15 cm, 15-30 cm and 30-60 cm. The soil samples were prepared for analysis using solvent extraction methods, passed through column of sodium sulphate and Florisil® to aid in column performance, remove moisture and gross impurities. Samples were analysed using gas chromatography with a flame ionisation detector. Penetration and migration of C10-C26 and C26-C34 hydrocarbons through the soil layers were assessed by cluster analysis to determine the spatial distribution, penetration and chemical similarity of these compounds over the contaminated area. This information is a useful guide for bioremediation purpose. It was found that total petroleum hydrocarbon concentrations varied from 9-289 mgkg-1 topsoil, 8-318 mgkg-1 subsoil and 7-163 mgkg-1 at the greatest depth measured.The results show elevated levels of total hydrocarbon contents when compared with the reference sites. Drastic steps should be taken to carefully monitor and remediate the environment. Bioremediation with plants and micro-organisms is endorsed.

This master´s thesis deals with optimization and validation of method for determination of free fatty acids. The theoretical part is focused on possibilities of extraction, extract separation, esterification, gas chromatography determination and brief description of the most important validation parameters. All with main emphasis on applying on free fatty acids. The experimental part describes the selection and procedure of optimizing individual steps of the method. Extraction of lipids was performed by solvent mixture (diethylether/petroleumether), based on the standard ČSN EN ISO 1735, for the extraction of free fatty acid solid phase extraction was tested. The acid catalyzed esterification based on ČSN EN ISO 5509 using methanolic solution BF3 was used for derivatization of free fatty acids and a gas chromatograph with a flame ionization detector was used for their determination. Subsequently, selected validation parameters of the method were verified: repeatability, reproducibility, linearity, limits of detection and quantitation. Sunflower oil and emmental type cheese, bought in common food market, were used as model samples for selection and optimatization of the method.

This thesis deals with the assessment of aroma active compounds in gooseberry and sensory analysis of it focusing on flavour. The theoretical part describes general characteristics, biological marks and properties of selected varieties, healing abilities, the active substances contained in gooseberry fruits and their use. The aim of the experimental part was to determine aroma-active substances contained in fruits of nineteen selected cultivars of gooseberry using SPME-GC-FID method. In total 45 volatile compounds were identified, among them 17 alcohols, 11 aldehydes, 6 ketones, 9 esters and 2 acids. For a better identification one sample (Captivátor variety) was also analysed using GC-MS with purge and trap extraction. Pentan-1-ol, hexanal, pentanal, ethylethanoate, ethylbutanoate, methylethanoate, methylbutanoate and ethanol were confirmed using mass spectra. Only several cultivars were selected for the assessment of overall sensory quality and palatability of fruits. Karmen and Remarka varieties were considered as the best samples by assessors.

This master's thesis deals with the characterization of elderberry fruit (Sambucus nigra L.) with emphasis on the aroma active compounds (AAC) and vitamin C (ascorbic acid (AA), dehydroascorbic acid (DHA)). The theoretical part gives a brief outlook on the chemical composition and attributes of Sambucus berries, its healing properties and the means of processing. The rest of the theoretical part thoroughly examines the AAC and vitamin C. The suitable assessment methods are also discussed. The experimental part of the work consists of two main tasks. The first one was to assess the aroma profile of Sambucus berries using the HS-SPME-GC-FID. 18 samples were examined (the wild elderberry and 17 cultivated varieties). In total, 24 different volatile aroma active compounds were identified and quantified, 12 of which belonged to alcohols, 5 to aldehydes, 4 to esters, 1 to acids and 2 “other” compounds (linalool and -damascenone). The chemical composition of the individual samples was not uniform; no single sample contained all of the identified compounds. The second task was to assess the total vitamin C content (the sum of DHA and AA) using the HPLC. For the purpose of reducing DHA to AA three reducing agents were examined (DTT, TCEP and L-cysteine) at two different pH levels (7 and 4). TCEP at pH 4 turned out to be most efficient and was subsequently applied on the samples of 5 selected elderberry varieties. The measured vitamin C content was within the range of 5,1 – 13,0 mg per 100 g of fruit.