Analysis of Bacterial Vaginosis-Related Amines in Vaginal Fluid by Gas Chromatography and Mass Spectrometry

Abstract

The presence of various amines in vaginal fluid from women with malodorous vaginal discharge has been reported before. The investigations have used several techniques to identify the amines. However, an optimized quantification, together with a sensitive analysis method in connection with a diagnostic procedure for vaginal discharge, including the syndrome of bacterial vaginosis, as defined by the accepted "gold standard," has not been done before. We now report a sensitive gas chromatographic and mass spectrometric method for identifying the amines isobutylamine, phenethylamine, putrescine, cadaverine, and tyramine in vaginal fluid. We used weighted samples of vaginal fluid to obtain a correct quantification. In addition, a proper diagnosis was obtained using Bacterial vaginosis (BV) commonly occurs in women of childbearing age. Prevalences of 10 to 31% have been reported in various populations (7). Several investigations have been performed to identify one or several bacteria comprising the decisive pathogenic factor in the syndrome, but so far no specific bacteria have been implicated in causing BV. Rather, it seems that BV is accompanied by a shift in the normal lactobacillus flora to a mixed vaginal anaerobic flora including Gardnerella vaginalis, Bacteroides spp., and Mobiluncus spp. (for a review, see reference 18). The importance of BV with respect to women's health is emphasized by the association between BV and pelvic inflammatory diseases, adverse outcome of pregnancy, postpartum endometritis, and cuff cellulitis (8,12,14,15).Amsel et al.(1) proposed a set of practical diagnostic criteria for the clinical diagnosis of BV that is now often accepted as the "gold standard." Three of the four criteria, i.e., a vaginal pH of Ͼ4.5, the presence of an adherent white discharge, a fishy amine odor after addition of KOH, and clue cells, must be met for the diagnosis of BV. A scoring system based on microscopically detectable changes in vaginal fluid is now also commonly used; Gram-stained vaginal smears and wet-mount smears in particular are well documented for use in BV diagnostics (11, 13). Several other alternative methods have also been used to develop easy, inexpensive, and reproducible diagnostic methods such as the rapid nucleic acid hybridization test (5), proline aminopeptidase activity (20), and the amine test (17).The syndrome of BV as defined by Amsel's clinical criteria seems to be a well-defined polythetic concept of major importance to women's health. Consequently, it is important to describe the most important dimensions that can be of use in women's health care. One dimension is the increased numbers of anaerobic bacteria in the vagina that lead to the characteristic amine production and odor. Amine production is thus an important property of the concept of BV and is clearly related to scoring with Amsel's criteria or Nugent scores. Production of amines is a property of several anaerobic bacterial species occurring in the human vagina, and these amines are released into the vaginal fluid in BV ...

Genomic Comparisons of Lactobacillus crispatus and Lactobacillus iners Reveal Potential Ecological Drivers of Community Composition in the Vagina

“…The production of hydrogen peroxide is thought to be one mechanism by which Lactobacillus species can prevent anaerobes from colonizing the vagina (70). L. crispatus also has the capability to breakdown putrescine, a malodorous amino acid commonly found in vaginal secretions during episodes of bacterial vaginosis (71). We found the core genome of L. crispatus to contain an iron transport system that is absent in the core genome of L. iners.…”

Rifaximin Modulates the Vaginal Microbiome and Metabolome in Women Affected by Bacterial Vaginosis

“…Metabolomic data showed that the restoration of a Lactobacillus-dominated flora, which occurred following rifaximin treatment, was strongly associated with the increase of lactic acid concentration and the decrease of the pool of molecules associated with the metabolism of BV-related bacteria, such as acetic acid, succinate, short-chain fatty acids, and biogenic amines (6,(45)(46)(47)(48). Placebo treatment did not cause any significant variation in the metabolome in cases of spontaneous remission and BV persistence.…”

“…The spectrum was dominated in intensity and variance by the peaks ascribable to lactate, whose area ranged between 7% and 72% of the entire spectrum. Molecules associated with the growth of BV-related bacteria could be identified, such as short-chain fatty acids (acetate, propionate, butyrate, isobutyrate, and isovaleric acid) (45,46), organic acids (succinate, formiate, and fumarate) (47), and biogenic amines (putrescine, spermidine, and tyramine) (6,48). Peaks from amino acids and molecules from purine catabolism (hypoxanthine and xanthine) were also identified.…”

“…Table 3 lists the molecules whose concentrations significantly changed between V1 and V3 in women who were in remission at V3. Most of these molecules are known to be modulated by the balance between lactobacilli and BV-related bacteria, such as lactate, which increased with remission, and biogenic amines, shortchain fatty acids, acetate, and succinate, which decreased in parallel (6,(45)(46)(47)(48).…”

Immobilization of Antibodies and Enzymes on 3-Aminopropyltriethoxysilane-Functionalized Bioanalytical Platforms for Biosensors and Diagnostics

“…Putrescine is associated with cancer markers, 248,249 plant stress markers, 250 bacterial vaginosis, 251 and food spoilage. 252 The GCE fabricated with MWCNTs, Nafion, putrescine oxidase (POx), and APTES offers a fast response time of 10 s and a detection limit of 500 nM.…”