measuring the changes of anion concentrations of saliva
DESCRIPTION
By Derek Menefee and Nathan Tompkins . Measuring the Changes of Anion Concentrations of Saliva. What is Saliva?. Saliva is produced in response to stimulation (usually taste) as well as movements of the jaw muscles, joints, and pressure from chewing. - PowerPoint PPT PresentationTRANSCRIPT
MEASURING THE CHANGES OF ANION CONCENTRATIONS OF SALIVA
By Derek Menefee and Nathan Tompkins
What is Saliva? Saliva is produced in response to stimulation
(usually taste) as well as movements of the jaw muscles, joints, and pressure from chewing.
It is produced mainly by three glands: the parotid, submandibular, and the sublingual.
Saliva controls the environment that the teeth are in, as well as protects against bacteria, and helps break down certain foods by using enzymes.
Saliva is made up mostly of water, electrolytes, mucus and enzymes.
What is Saliva Cont.
Contains 2 main enzymes: Amylase (starch breakdown) and maltase (maltose breakdown)
Saliva also contains bicarbonate, which acts as a pH buffer by neutralizing acid production and controlling plaque pH (Hogg, 2011).
What is being tested and Why? 1.) Human Salivary Anionic Analysis Using Ion Chromatography Z.F.
Chen, B.W. Darvell and V.W. Leung - 10 subjects were tested and analyzed for pH of saliva after they have been fasting overnight. - Saliva samples looked for concentrations of chloride, phosphate, nitrate, sulfate, thiocyanate, and acetate were analyzed. Acetate ion was the most abundant ion in saliva samples with nitrate as the second most abundant.
2.) Effectiveness In Learning Complex Problem Solving and Salivary Ion Indices of Psychological Stress and Activation P.Richter, J.W. Hinton, S. Reinhold-33 students were tested for samples of saliva fasting and a rest, and after being stressed after playing a challenging computer game for 2-3 hours. -Studies showed that a change in ion concentration (increase in potassium ion and decrease in sodium ion) in saliva occurred after psychologically stressful activity (Hinton, 1998).
Our Experiment/Hypothesis
In this experiment, involving the use of Ion Chromatography, is testing the concentrations of ions in saliva during different stimuli and analyzing the changes (if any) that take place.
The hypothesis is that the different samples that may induce enzyme reactions, stimulation, and changes in pH change may also result in changes in ion concentrations in saliva after the sample is consumed.
Reference Instrument: Varian Inductively Coupled Plasma Used to identify element intensities between saliva
after fasting and after ingestion of sample.
Materials and Methods Use of the Dionex ICS-90 instrument to run the
samples of saliva Parameters:
-Pressure: at 890 Psi-Flow Rate: 1 mL/min-Pressure Gauge: 9
4 different experiments are run on the IC to determine ion concentration of saliva after different stimuli
Samples Used:1.) Monster Energy Drink2.) Protein/starch meal3.) Vegetable/fruit meal4.) Coffee (no sugar or milk added)
Materials and Methods Cont. A sample of saliva is taken from the experimenter, who
has been fasting for 10-12 hours prior to consuming food or drink. This is done so the saliva may be used as a control to compare with the sample analysis.
Experimenter must rinse with DI water before each sample is taken as to not induce any false readings from the sample itself, but only to obtain the salivary reaction to the sample.
2 different samples are taken for each experiment (one from each experimenter).
A pH sample is also be taken before and after consuming to check for any differences.
The same procedure is also taken for the analysis of the ICP instrument.
DATA
Anion Ret. time Peak Height ppmInjection Peak 0.46 0.315 N/A
Chloride 4.86 0.527 N/ANitrate 6.65 0.059 0.002
N/A 7.34 11.265 N/AN/A 8.42 1.34 N/A
Phosphate 8.91 228.652 N/AN/A 12.06 0.475 N/A
Derek Fasting Day 1 Sample
Nate Fasting Day 1 Sample
Anion Ret. time Peak Height ppmInjection Peak 0.49 .413 N/A
N/A N/A N/A N/AN/A N/A N/A N/A
Nitrite 7.5 12.859 N/APhosphate 9.08 242.354 N/A
N/A N/A N/A N/A
Nate Monster SampleAnion Ret. time Peak Height ppm
Injection Peak 0.33 0.915 N/AFluoride 3.17 0.046 0.009Chloride 3.93 0.035 N/A
N/A N/A N/A N/AN/A N/A N/A N/A
Phosphate 8.85 67.398 N/AN/A N/A N/A N/A
Derek Monster Sample
Anion Ret. time Peak Height ppmInjection Peak 0.49 0.268 N/A
N/A N/A N/A N/ANitrite 7.33 20.031 N/A
Phosphate 9.02 465.608 N/AN/A N/A N/A N/AN/A N/A N/A N/A
Anion Ret. time Peak Height ppmInjection Peak 0.37 0.365 N/A
N/A N/A N/A N/AN/A N/A N/A N/A
Nitrate N/A N/A N/APhosphate 8.97 350.996 N/A
N/A N/A N/A N/AN/A N/A N/A N/A
Derek Fasting Day 2
Anion Retention Time (min) Peak Hight (µS) Amount ppmInjection Peak 4.83 0 N/AFluoride 6.53 .40 N/AChloride N/A N/A N/ANitrite 7.30 18.442 N/ABromide N/A N/A N/ANitrate N/A N/A N/APhosphate 8.57 454.834 41.268Sulfate 10.28 324.391 N/A
Derek Protein Saliva Sample
Nate Fasting Day 3 Sample
Anion Retention Time (min) Peak Hight (µS) Amount ppmInjection Peak 4.84 0 N/AFluoride N/A N/A N/AChloride N/A N/A N/ANitrite N/A 18.442 30.238Bromide N/A N/A N/ANitrate N/A N/A N/APhosphate 8.65 355.265 30.238Sulfate 9.78 4.137 N/A
Nate Vegetable Sample
Anion Retention Time (min) Peak Hight (µS) Amount ppmInjection Peak 0.41 0 N/AFluoride N/A N/A N/AChloride N/A N/A N/ANitrite 4.88 18.442 N/ABromide N/A N/A N/ANitrate 7.21 N/A N/APhosphate 8.57 N/A 26.429Sulfate 10.28 324.391 N/A
Derek Day 3 Fasting Sample
Anion Retention Time (min) Peak Hight (µS) Amount ppmInjection Peak 1.83 0 N/AFluoride N/A N/A N/AChloride 4.84 .4 N/ANitrite N/A N/A N/ABromide N/A N/A N/ANitrate 7.32 18.821 N/APhosphate 8.73 470.635 42.092Sulfate 9.83 6.513 N/A
Derek Fruit Sample
Anion Retention Time (min) Peak Hight (µS) Amount ppmInjection Peak 0.41 0 N/AFluoride N/A N/A N/AChloride N/A N/A N/ANitrite 7.39 22.285 30.238Bromide N/A N/A N/ANitrate N/A N/A N/APhosphate 8.953 343.433 N/ASulfate N/A N/A N/A
Derek Day 4 Fasting Sample
Anion Retention Time (min) Peak Hight (µS) Amount ppmInjection Peak 2.83 0 N/AFluoride N/A .40 N/AChloride 4.39 6.674 .383Nitrite N/A 18.442 N/ABromide N/A N/A N/ANitrate N/A N/A N/APhosphate 5.20 244.846 N/ASulfate N/A N/A N/A
Derek Coffee Sample
Anion Retention Time (min) Peak Hight (µS) Amount ppmInjection Peak 0.56 0 N/AFluoride N/A N/A N/AChloride N/A N/A N/ANitrite N/A N/A N/ABromide N/A N/A N/ANitrate N/A N/A N/APhosphate 6.867 323.026 N/ASulfate N/A N/A N/A
ICP RESULTSNate Fasting Sample
Element IntensityAl 278.88As 1.4510B 142.37Br 1073.4C 138903Ca 19933Cl 2165.3Co 41.341Cu 152.12Fe 155.54H 454.17I 119.43K 1395923N 742.58Na 810814Ni 30.703O 4445252P 13783
Pb 100.92S 848.26
Zn 90.453
Element IntensityAl 383.52As 2.0548B 141.39Br 960.38C 252743Ca 56832Cl 1948.5Co 40.517Cu 159.73Fe 161.35H 194.54I 270.18K 2293814N 794.63Na 306923Ni 33.288O 4357401P 27941
Pb 130.05S 888.14
Zn 97.298
Nate Monster
Element Intensity
Al 278.88
As 1.4510
B 142.37
Br 1073.4
C 138903
Ca 19933
Cl 2165.3
Co 41.341
Cu 152.12
Fe 155.54
H 454.17
I 119.43
K 1395923
N 742.58
Na 810814
Ni 30.703
O 4445252
P 13783
Pb 100.92
S 848.26
Zn 90.453
Derek Fasting
Element IntensityAl 377.27As 2.0997B 149.15Br 1578.4C 441089
Ca 45925Cl 2046.5Co 41.636Cu 156.63
Fe 153.91H 877.46I 238.73K 1927251N 817.16
Na 1839628Ni 32.872O 3829018P 24472
Pb 97.376S 2001.4
Zn 181.28
Derek Monster
Discussion of Results •In regards to the changes in ion concentration the only changes witnessed were due to the pH and protein samples. However the concentrations varied between the two different experimenters.
•In the first Nathan fasting sample a nitrate peak of .002 ppm appeared at retention time 6.65 minutes. When the Nathan Monster Sample was run the nitrate peak vanished and a fluoride peak of .009 ppm appeared at retention time 3.17 minutes.
•Derek’s phosphate group nearly doubled in height after drinking the Monster sample.
•Derek’s protein also showed an increase in peak height an a concentration of 41.268 pmm
• Average number peaks obtained by a sample was 8.3 (lowest number being 6, highest number being 11)
•ICP was used to look at the changes in element intensity. The ICP data for “fasting” to “Monster” matched up with the same pattern of changes in the “fasting” to “Monster” data for the IC.
•Standard “Fasting” pH for Derek : 6.5
•Standard “Fasting” pH for Nathan: 7.2
•pH of Monster being used: 3.5
•Monster Sample pH for Derek : 6.0
•Monster Sample pH for Nathan: 6.8
•All other sampling’s pHs only moved by .1 or by nothing at all.
Changes in pH while testing
Things that could be changed and Improved upon
•Use a good 7 anion standard.
• Have all the test subjects drink the same water while fasting. Most likely DI water.
•Consume the same exact meals
•Stress / sickness
•Time restraints, waiting on the IC to warm up. Having time to run more samples on the ICP. Only running 4 samples a day.
•Better Storage.
•More test subjects (to look at more samples, to see if there is more sample similarities.
•do more test with varying pH’s , and use more sensitive pH tests.
•Calculate the actual amount of time needed to produce a “fasting sample”. We could have run more than two samples a day, which would have saved on time.
•Further speculation of different ions inducting thiocyanate and acetate.
Future Research
Works Cited
Chen, Z.F., Darvell, B.W., Leung, V.W.H. Human saliva anionic analysis using ion chromatography. Archives of Oral Biology. (2004) Vol 49 (11). Pgs. 863-869.
Richter, P., Hinton, J.W., Reinhold, S. Effectiveness in learning complex problem solving and salivary ion indices in psychological stress and activation. International Journal of Psycology (1998). Vol 30 (3). Pgs. 329-337.
Hogg,Steve. Salivary Buffering, bicarbonate, and pH. The Oral Environment. (2011) http://www.ncl.ac.uk/dental/oralbiol/oralenv/tutorials/bicarbonate.htm
Kaufman, Eliaz., Lamster, I.B. Diagnostic Applications of Saliva-A Review. International and American Associations for Dental Research. (2002) Vol 13 (2). Pgs. 197-212.