CONFIDENTIAL ASIDEC 2018/CHM510 UNIVERSITI TEKNOLOGI MARA FINAL EXAMINATION COURSE : ANALYTICAL SEPARATION METHODS COURSE CODE CHMS510 EXAMINATION DECEMBER 2018 TIME 2HOURS ISTRUCTIONS TO CANDIDATES 1 This question paper consists of five (5) questions. 2 ‘Answer ALL questions in the Answer Booklet. Start each answer on a new page. 3. Do not bring any material into the examination room unless permission is given by the invigitator. 4. Please check to make sure that this examination pack consists of : the Question Paper i) _an Answer Booklet — provided by the Faculty 5, ‘Answer ALL questions in English DO NOT TURN THIS PAGE UNTIL YOU ARE TOLD TO DO SO This examination paper consists of & printed pages (© Hak Cipta Universiti Teknolog! MARA CONFIDENTIALCONFIDENTIAL 2 ASIDEC 2018/CHMS10 QUESTION 1 a) A mixture of fatty acids X, Y and Z was separated using gas. chromatography flame ionization detector (GC-FID) after derivatization using N,O- bis(trimethylsily!) trifluoroacetamide (BSTFA). Separation was achieved using. a 30 m x 0.25 mm x 0.32 um HP-SMS column with linear mobile phase velocity (11) of 60.0 cms”. The retention times for peak X, Y and Z were 111 s, 123 s and 178 s, respectively, i) Calculate the capacity factor (k’) based on compound X. (5 marks) Calculate the plate height of the column based on compound Y. The peak width of compound Y is 0.081 min. (4 marks) iii) Explain why the mixture of fatty acids need to be derivatized prior to chromatographic analysis. (3 marks) 5) Discuss the most significant term in the van Deemter equation that explains the advantage of using capillary column compared to packed column in gas chromatography. (3 marks) QUESTION 2 a) The analysis of four chlorinated pesticides was conducted using gas chromatography with electron capture detector (GC-ECD). The separation shown in Figure 1 was obtained using polydimethylsiloxane (PDMS) column with isothermal temperature at 150°C. The separation has resolution values of 0.9, 7.9 and 10.5. 1 mau 4 a 2 2 | 20 } | | 10 Lt x) enemas 1 2 3 min Figure 4 i) Define the term resolution in chromatography. (2 marks) (© Hak Cipta Universiti Teknologi MARA, CONFIDENTIALCONFIDENTIAL 3 ASIDEC 2018/CHM510 b) ii) Based on the given resolutions, comment on the efficiency of the separation. (4 marks) iii) With reasons, suggest a suitable temperature programming approach to improve the separation. (4marks) iv) _ Discuss the effect on the separation of compound 1 and 2 and compound 3 and 4, if'a 50% phenyl polydimethyisiloxane column was used, (6 marks) ‘Separation of six analytes was achieved using size exclusion chromatography (SEC) with the following conditions: polystyrene based column, 7.5 x 600 nm with molecular mass exclusion limit of 1 x 10°; Mobile phase: tetrahydrofuran. The first two analytes were eluted together immediately, while efficient separation was observed for the other four analytes. Comment on this observation. (4marks) QUESTION 3 a) b) ‘A mixture of alcohols was analysed on a 250 mm Cis column. The mobile phase consisted of a mixture of 30% ACN and 70% water. Explain how the retention time would be affected if: i) the composition of the mobile phase is changed to 70% ACN and 30% water (3 marks) ii) the Cg bonded phase contained unreacted silanol groups. (3 marks) ii) polyethylene glycol (PEG) column is used to replace Cis column. (3 marks) The retention in ion exchange chromatography is based on the attraction between solute ions and charged sites of the resin, Explain briefly what will happen to the retention time of analytes if the separation is carried out using a sulphonic acid resin with pH less than 4. (3 marks) Three water soluble vitamins; niacinamide (a neutral compound), riboflavin (a neutral compound), and thiamine (a cation) were separated using micellar electrokinetic capillary chromatography (MEKC) in 15 mM borate buffer (pH 8) with 50 mM sodium dodecyl sulfate (SDS). The migration times of niacinamide, riboflavin and thiamine were 7.8 min, 12.3 min and 13.9 min, respectively. (© Hak Cipta Universiti Teknologi MARA CONFIDENTIALCONFIDENTIAL 4 ASIDEC 2018/CHM510 i) Explain briefly the principle of separation of both neutral compounds (niacinamide and riboflavin). (4 marks) ii) The concentration of SDS compounds. Explain briefly. important in ensuring the separation of neutral (3 marks) QUESTION 4 a) The lectropherogram in Figure 2 shows a separation of five species (“A‘, B*, C°,"8D? and E>) using capillary electrophoresis (CE) with UV detector conducted at pH 8. lon nents" Figure 2 i) Discuss the factors that determine the rate of migration of these species. (3 marks) i) ‘Suggest the order of elution of the five species that corresponds to peak 1 to 5. Verify your answer. (4 marks) b) SFC is a hybrid of gas and liquid chromatography aimed at improving some of the shortcomings of each of the other techniques. i) Identify the compounds that are not conveniently handled by either gas or liquid chromatography but can easily determine by SFC. (4 marks) ii) State when should pressure programming applied in SFC. Discuss the effect of pressure programming in this separation. (4 marks) © Hak Cipta Universiti Teknologi MARA, CONFIDENTIAL,CONFIDENTIAL 5 ASIDEC 2018/CHM510 QUESTION 5 a) One of the extraction approach of solid phase microextraction (SPME) is by placing the ‘SPME fibre on the headspace of the aqueous sample. i) Give another extraction approach of SPME for the extraction of non-volatile sample. (1 mark) ii) Compare the principle of extraction in solid phase microextraction (SPME) with that of solid phase extraction (SPE). (4 marks) b) For each of the following, give a reason to the mentioned problem and suggest an approach to overcome the problem without changing the extraction method used. i) _ Non-polar contaminants from soil sample cannot be extracted using hexane with microwave extraction method (MAE), (3 marks) ji) Alkanes can be extracted from urban air particulates using supercritical fluid extraction (SFE) with COz at 75 bar (45°C), whereas the polyaromatic hydrocarbons (PAHs) remain unextracted. (3 marks) END OF QUESTION PAPER (© Hak Cipta Universiti Teknologi MARA CONFIDENTIAL,