| Summary: | Changes in the concentration of some constituents in women's saliva during the menstrual cycle were studied. Saliva was used because it is easier to collect than other body fluids and is continuously available for analysis. Glucose, the enzyme IT-Acetyl- Jý-D-glucosaminidase (NAG) and Calcium which are saliva constituents and belong to three different chemical groups were selected for the study. Several analytical techniques were investigated. The fluorometric assay procedure was found to 'be the best because of its specificity and sensitivity for the estimation of these constituents* Besides the fluorometric method a spectrophotometric method was used in the NAG determination and an atomic absorption method in the calcium estimation. Glucose was estimated by an enzymatic method, This is based on the reaction of glucose with the enzymes glucose oxidase and peroxidase to yield hydrogen peroxidey which in turn oxidises a non-fluorescent substrate, p-hydroxyphenylacetic acid, to a highly fluorescent producte The saliva samples in this determination had to be centrifuged at high speedl heated in a boiling water bath, centrifuged again and then treated with a mixture of cation and anion resins to remove the substances that inhibited the enzyme system. In the determination of the NAG activity the saliva samples were diluted with citric aoid/phosphate bufferg and then centrifuged at high speed. The assay was based on the enzymic hydrolysis of the nonfluorescent substrate j 4, -I. Iethyl-umbelliferyl-p-D-glucosaminide , to the highly fluorescent 4-Mothyl-umbelliferone. Calcium was estimated by a fluorometric procedure based upon the measurement of the fluorescence produced by the complex formed between calcein blue and calcium, at pH 9- 13- From the results obtained from the analysis of saliva samples of several women it was found that glucose showed a significant increase in its level around the exected time of ovulation. This was found in seven cycles out of ten. Similar results were found with the enzyme NAG* No significant change in the"Calcium levels was observed at arq particular time of the cycle, The levels of the glucose, the activity of the enzyme NAG and the concentration of the calcium were found to change daily, and to differ from one subject to another and in the sane subject from cycle to cycle, The increase observed it salivary glucose levels and the enzyme NAG activity could be monitored to predict the time of ovulation.
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