Sampling Small Volumes of Saliva for Determination of the Stress Hormone α-Amylase: A Comparative Methodological Study

DOI: 10.4236/jbbs.2011.13026   PDF   HTML     5,166 Downloads   11,129 Views   Citations


Two sampling devices that allow saliva collection through absorption to a cotton roll (Salivette®-method) or to small cotton pellets (VectaSpinTM Micro [VSM]-method) were studied. Any loss of salivary alpha-amylase (sAA) activity in relation to the saliva volume absorbed and harvested by centrifugation was examined. A pooled saliva sample prepared from stimulated whole saliva (collected by drooling) of 30 subjects was used. Three different saliva volumes (2.9 ml, 1.5 ml, and 0.8 ml) were tested on cotton rolls and two (0.03 ml, and 0.015 ml) on cotton pellets. The sample sAA activity was determined from the hydrolysis of 2-chloro-4-nitrophenyl-α-D-maltotrioside. In comparison with the original drooling sample, no sAA loss was observed in 1.5 ml samples tested with Salivette, while a significant decrease of activity was recorded with smaller volumes. VSM collected samples showed a non-volume dependent decrease of sAA activity of about 25%. Salivette requires large saliva volumes to allow an accurate sAA estimation. With cases of limited saliva access, VSM may be a suitable sampling device.

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A. Arhakis, V. Karagiannis and S. Kalfas, "Sampling Small Volumes of Saliva for Determination of the Stress Hormone α-Amylase: A Comparative Methodological Study," Journal of Behavioral and Brain Science, Vol. 1 No. 3, 2011, pp. 194-198. doi: 10.4236/jbbs.2011.13026.

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The authors declare no conflicts of interest.


[1] D. J. Adam, A. A. Milne, S. M. Evans, J. E. Roulston, A. J. Lee, C. V. Ruckley and A. W. Bradbury, “Serum Amylase Isoenzymes in Patients Undergoing Operation for Ruptured and Non-Ruptured Abdominal Aortic Aneurysm,” Journal of Vascular Surgery, Vol. 30, No. 2, 1999, pp. 229-235. doi:10.1016/S0741-5214(99)70132-1
[2] P. M. Martinez, A. R. Torres, R. Ortiz de Lejarazu, A. Montoya, J. F. Martin and J. M. Eiros, “Human Immunodeficiency Virus Antibody Testing by Enzyme-Linked Fluorescent and Western Blot Assays Using Serum, Gingival-Crevicular Transudate, and Urine Samples,” Journal of Clinical Microbiology, Vol. 37, No. 4, 1999, pp. 1100-1106.
[3] E. Filaire and G. Lac, “Dehydroepiandrosterone (DHEA) Rather than Testosterone Shows Saliva Androgen Responses to Exercise in Elite Female Handball Players,” International Journal of Sports Medicine, Vol. 21, No. 1, 2000, pp. 17-20. doi:10.1055/s-2000-8851
[4] S. P. Humphrey and R. T. Williamson, “A Review of Saliva: Normal Composition, Flow, and Function,” Journal of Prosthetic Dentistry, Vol. 85, No. 2, 2001, pp. 162-169. doi:10.1067/mpr.2001.113778
[5] C. F. Streckfus and L. R. Bigler, “Saliva as a Diagnostic Fluid,” Oral Diseases, Vol. 8, No. 2, 2002, pp. 69-76. doi:10.1034/j.1601-0825.2002.1o834.x
[6] D. B. Ferguson, “Current Diagnostic Uses of Saliva,” Journal of Dental Research, Vol. 66, No. 2, 1987, pp. 420-424. doi:10.1177/00220345870660020601
[7] D. Malamud, “Saliva as a Diagnostic Fluid,” British Medical Journal, Vol. 305, No. 6847, 1992, pp. 207-208. doi:10.1136/bmj.305.6847.207
[8] I. D. Mandel, “A Contemporary View of Salivary Research,” Critical Reviews in Oral Biology & Medicine, Vol. 4, No. 3-4, 1993, pp. 599-604.
[9] H. C. Slavkin, “Toward Molecularly Based Diagnostics for the Oral Cavity,” Journal of the American Dental Association, Vol. 129, No. 8, 1998, pp. 1138-1143.
[10] M. Navazesh, “Methods for Collecting Saliva,” Annals of the New York Academy of Sciences, Vol. 20, No. 694, 1993, pp. 72-77. doi:10.1111/j.1749-6632.1993.tb18343.x
[11] N. Rohleder and U. M. Nater, “Determinants of Salivary Alpha-Amylase in Humans and Methodological Considerations,” Psychoneuroendocrinology, Vol. 34, No. 4, 2009, pp. 469-485. doi:10.1016/j.psyneuen.2008.12.004
[12] E. A. Shirtcliff, D. A. Granger, E. Schwartz and M. J. Curran, “Use of Salivary Biomarkers in Biobehavioral Research: Cotton-Based Sample Collection Methods Can Interfere with Salivary Immunoassay Results,” Psychoneuroendocrinology, Vol. 26, No. 2, 2001, pp. 165-173. doi:10.1016/S0306-4530(00)00042-1
[13] A. G. Harmon, L. C. Hibel, O. Rumyantseva and D. A. Granger, “Measuring Salivary Cortisol in Studies of Child Development: Watch out-What Goes In May Not Come out of Saliva Collection Devices,” Developmental Psychobiology, Vol. 49, No. 5 2007, pp. 495-500. doi:10.1002/dev.20231
[14] J. A. DeCaro, “Methodological Considerations in the Use of Salivary a-Amylase as a Stress Marker in Field Research,” American Journal of Human Biology, Vol. 20, No. 5, 2008, pp. 617-619. doi:10.1002/ajhb.20795
[15] J. A. Bosch, H. S. Brand, T. J. Ligtenberg, B. Bermond, J. Hoogstraten and A. V. Nieuw Amerongen, “Psychological Stress as a Determinant of Protein Levels,” Psychosomatic Medicine, Vol. 58, No. 4, 1996, pp. 374-382.
[16] U. M. Nater, R. La Marca, L. Florin, A. Moses, W. Langhans, M. M. Koller and U. Ehlert, “Stress-Induced Changes in Human Salivary Alpha-Amylase Activity-Associations with Adrenergic Activity,” Psychoneuroendocrinology, Vol. 31, No. 1, 2006, pp. 49-58. doi:10.1016/j.psyneuen.2005.05.010
[17] D. A. Granger, K. T. Kivlighan, M. el-Sheikh, E. B. Gordis and L. R. Stroud, “Salivary Alpha-Amylase in Biobehavioral Research: Recent Developments and Applications,” Annals of the New York Academy of Sciences, Vol. 1098, 2007, pp. 122-144. doi:10.1196/annals.1384.008
[18] M. Neu, M. Goldstein, D. Gao and M. L. Laudenslager, “Salivary Cortisol in Preterm Infants: Validation of a Simple Method for Collecting Saliva for Cortisol Determination,” Early Human Development, Vol. 83, No. 1, 2007, pp. 47-54. doi:10.1016/j.earlhumdev.2006.04.003
[19] S. L. Udupa, A. R. Prabhakar and S. Tandon, “Alpha-Amylase Inhibitors in Foodstuffs,” Food Chemistry, Vol. 34, No. 2, 1989, pp. 95-101. doi:10.1016/0308-8146(89)90077-0
[20] K. T. Kivlighan and D. A. Granger, “Salivary Alpha-Amylase Response to Competition: Relation to Gender, Previous Experience, and Attitudes,” Psychoneuroendocrinology, Vol. 31, No. 6, 2006, pp. 703-714. doi:10.1016/j.psyneuen.2006.01.007
[21] R. Nagler, S. Lischinsky, E. Diamond, N. Drigues, I. Klein and A. Z. Reznick, “Effect of Cigarette Smoke on Salivary Proteins and Enzyme Activities,” Archives of Biochemistry and Biophysics, Vol. 379, No. 2, 2000, pp. 229-236. doi:10.1006/abbi.2000.1877
[22] E. S. Winn-Deen, H. David, G. Sigler and R. Chavez, “Development of a Direct Assay for Alpha-Amylase,” Clinical Chemistry, Vol. 34, No. 10, 1988, pp. 2005-2008.
[23] A. Y. Foo and R. Bais, “Amylase Measurement with 2-Chloro-4-Nitrophenyl Maltotrioside as Substrate,” Cli- nica Chimica Acta, Vol. 272, No. 2, 1998, pp. 137-147. doi:10.1016/S0009-8981(98)00009-6
[24] H. Ben-Aryeh, M. Fisher, R. Szargel and D. Laufer, “Composition of Whole Unstimulated Saliva of Healthy Children: Changes with Age,” Archives of Oral Biology, Vol. 35, No. 11, 1990, pp. 929-931. doi:10.1016/0003-9969(90)90075-L
[25] M. Kilian and B. Nyvad, “Ability to Bind Salivary Alpha-Amylase Discriminates Certain Viridans Group Streptococcal Species,” Journal of Clinical Microbiology, Vol. 28, No. 11, 1990, pp. 2576-2577.

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