Hplc

Analysis and Separation of Organic irates in White wine Using HighPerformanceLiquidChromatography Atis,ArnelsonArwinG. andGaitos,GeraldM. InstituteofChemistry,UniversityofthePhilippines,Diliman,QuezonCity Performed declination9,2011 SubmittedDecember15,2011 RESULTSANDDISCUSSION solutions containing increasing amounts of the ingrained loony toons. These solutions were contained in flasks numbered 5, 6, 7, 8, and 9. (Refer to the Appendix to probe components of all(prenominal) flask). delays 2, 3, 4, and 5 showthepeakareasandtheequationofthe normalisation lift obtained for each(prenominal) thorough venomous in the chromatogramsobtainedforflasks5to9. Table2. efflorescence playing areasofTartaricAcidinFlasks5,6,7,8, and9 Flask retention vizorArea assiduousness time(min) (g/L) 5 2. 863 491364 0. 5 6 2. 905 918187 1 7 3. 027 5949164 5 8 3. 044 8363261 8 9 3. 065 9475446 10 CalibrationCurve comparison y=984371. 3561x+216064. 755 R2=0. 9912 Table3. tipAreasofMalicAcidinFlasks 5,6,7,and 8 Flask Retention PeakArea density time(min) (g/L) 5 3. 02 159742 0. 5 6 3. 760 1114051 1 7 3. 756 4892727 5 8 3. 795 6883424 10 CalibrationCurveEquation y=688260. 8175x+475029. 6892 R2=0. 9766 Table 4. Peak Areas of lactic Acid in Flasks 5, 6, 7, 8, and9 Flask Retention PeakArea Concentration time(min) (g/L) 5 3. 533 575620 0. 1 6 3. 598 1248888 1 7 3. 621 3762922 2. 5 The retention times of different organic acids commonly found in white wine were obtained using broad(prenominal) performance liquid chromatography.The peak areas of the acids were wherefore obtained through chromatogramsandplottedagainstthe preoccupations of the hold(a) solutions of the organic acids to construct a calibration curve. Finally, the calibration curves obtained were used to finalise the concentration of the organic acids in samples of white wine. Standard solutions of various organic acids commonly found in white wine (tartaric, malic, lactic, citric) were assigned to be contained in fl asks 1, 2, 3, and 4, respectively, and were run through the chromatograph.Themobilephaseusedfortheanalytes was H3PO4 at pH 3, to prevent dissociation of the organic acids. The resulting chromatograms of each standard were then analyzed to obtain the retention times of the organic acids. Table 1 shows the experimental retention times of the different organic acids. Table 1. observational Retention snips of Organic Acids OrganicAcid RetentionTime(min) Tartaric 3. 088 Malic 3. 812 Lactic 3. 620 Citric 3. 68 Since there were no expel peaks from the chromatograms obtained for the tartaric, malic, and lactic acid samples, the retention time at which the peak eyeshade is greatest was obtained as the experimentalretentiontimeoftheorganicacids. The experimental retention times were then used to identify the peak areas alike to each organicacidanalyzedintheresultingchromatogramsof 8 3. 597 5048614 3 9 3. 619 6519896 5 CalibrationCurveEquation y=1300341. 246x+414396. 3089 R2=0. 9815 Ta ble 5.Peak Areas of Citric Acid in Flasks 5, 6, 7, 8, and9 Flask Retention PeakArea Concentration time(min) (g/L) 5 3. 140 361921 0. 05 6 3. 208 907232 0. 1 7 3. 306 4736427 0. 5 8 3. 325 7508481 0. 75 9 3. 347 9612454 1 CalibrationCurveEquation y=9836731. 501x+96328. 12036 R2=0. 9994 The equations of the calibration curves were then used to calculate the concentration of each organic acid in the white wine samples. The peak area corresponding to each organic acid was first obtained, and substituted in the calibration curve equation.The concentrationofeachacidisshowninTable6. Table 6. Experimental Concentration of Various OrganicAcidsinWhiteWineSamples. Organic Retention PeakArea Concentration Acid Time(min) (g/L) Tartaric 2. 966 2675651 2. 499 Malic 3. 752 3576021 4. 506 Lactic 3. 393 4709737 3. 303 Citric 3. 144 794394 0. 091 Theresultsindicatethatmalicacidisthemajor(ip) componentofwhitewine. Butinreality,thisisnotthe case. Themajorcomponentofwhitewineisfoundtobe tartaricacid. T he chromatograms (See Appendix) of flasks 1 to 9, and of the sample are not well resolved.This discrepancy may be caused by several factors. These factors include poor solution preparation, contaminationofthe declarationorthesample,bubblesin the detector, impurities in the mobile phase, bleeding ofthecolumn,inadequate registrationofequilibriumin gradient operation, and carry? over from previous injection. repayabletothesefactors,itishighlyadvisedthatthe future researchers should cautiously execute each purpose of the experiment to eliminate the discrepancy and accordingly, they could attain better results.They could also pack use of theoretical retentiontimesoftheorganicacidstodetermineeach of them and which could further help the future researchers to analyze the wine sample more efficiently. REFERENCE Meyer,VeronikaR. PracticalHighPerformanceLiquid Chromatography. 2nded. 1993. England trick Wiley&SonsLtd. APPENDIX DataSheets Concentrationoftartaricacidstandard50. 0g/L Conce ntrationofmalicacidstandard50. 0g/L Concentrationoflacticacidstandard25. 0g/L Concentrationofcitricacidstandard5. 0g/L CompositionofFlasks1? 9 Volumeofstandardstocksolutions(inmL) RetentionTimeMeasurement CalibrationCurve Flask 1 2 3 4 5 6 7 8 TartaricAcid 5. 00 0. 00 0. 00 0. 00 0. 25 0. 50 2. 50 4. 00 MalicAcid 0. 00 7. 50 0. 00 0. 00 0. 10 0. 50 2. 50 5. 00 LacticAcid 0. 00 0. 00 5. 00 0. 00 0. 10 1. 00 2. 50 3. 00 CitricAcid 0. 00 0. 00 0. 00 5. 00 0. 25 0. 50 2. 50 3. 75 9 5. 00 7. 50 5. 00 5. 00 *Chromatogramsofflasks1? 9andofthewhitewinesample bedbeseenintheremainingpagesafterthis.