TOXICO-METABOLOMIC SIGNATURES OF BLOOD PLASMA AND SEMINAL PLASMA IN INFERTILE MALES USING LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY

Abstract

1.1. Background and Challenge

Infertility in males represents a worldwide substantial public health challenge, affecting millions of couples globally. Idiopathic and unexplained infertility remains the most frustrating subset of the lot as assigning a clear etiology becomes a challenge by routine diagnostic procedures.²² this decline in semen quality is justified, partially, by global scientific evidences suggesting the detrimental role of acute and chronic exposures of environmental as well as occupational xenobiotics such as Organophosphates (OP) and endocrine disrupting chemicals (EDCs) in reproductive health.³ which leads to a critical need for identification of non-invasive but quantifiable biomarkers that can bridge the gap between toxic exposures and their impairing effects on functional reproductive health.

 1.2. Methodology

This study follows on the toxico-metabolomic profiling conducted on matched blood plasma (BP) and seminal plasma (SP) samples collected from infertile patients exhibiting abnormal semen parameters (Oligo- Astheno- Teratozoospermia, OAT). Samples were prepared using high-throughput Toxtyper® liquid- liquid extraction (LLE) protocol and analysed using high resolution liquid chromatography- tandem mass spectrometry (LC-MS/MS) on a Bruker amazon speed ion trap instrument. The rapid and simultaneous detection of broad spectrum of small molecule’s, including basic, acidic and neutral compounds was ensured and encouraged by the fast polarity switching capabilities of the method. 

1.3. Key Results

The toxico-metabolomic analysis successfully identified a co-occurring molecular signature consisting of nine distinct xenobiotics across the study cohort. High-priority compounds detected included the potent EDCs Di(2-ethylhexyl) phthalate (DEHP) and Tetrabromobisphenol A (TBBPA), along with the OP insecticide Acephate. Comparative quantification revealed that eight of the nine identified toxins were present in both the systemic (BP) and reproductive (SP) compartments. Analysis of molecular abundance demonstrated a significant transfer rate of these lipophilic compounds into the seminal microenvironment, often reaching concentrations hypothesized to interfere with germ cell function.

 1.4. Conclusion

The toxico-metabolomic profile provides a novel and quantifiable etiology of previous idiopathic male infertility by confirming the direct exposure of germ sells to reproductive toxicants via seminal fluid. This finding validates the use of seminal plasma analysis as a critical, non-invasive diagnostic tool by offering molecular and targeted basis of clinical intervention for similar cases of environmental or occupational toxicity on reproductive functions.²¹