Sampling Small Volumes of Saliva for Determination of the Stress Hormone α-Amylase: A Comparative Methodological Study
- 1
- 2
- 3
Abstract
Two sampling devices that allow saliva collection through absorption to a cotton roll (Salivette<sup>®</sup>-method) or to small cotton pellets (VectaSpin<sup>TM</sup> 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.
- 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
- 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.
- 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
- 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
- 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
- D. B. Ferguson, “Current Diagnostic Uses of Saliva,” Journal of Dental Research, Vol. 66, No. 2, 1987, pp. 420-424. doi:10.1177/00220345870660020601
- 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
- I. D. Mandel, “A Contemporary View of Salivary Research,” Critical Reviews in Oral Biology & Medicine, Vol. 4, No. 3-4, 1993, pp. 599-604.
- 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.
- 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
- 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
- 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