P-042 Method validation for a multi-element panel in human seminal plasma by inductively coupled plasma tandem mass spectrometry (ICP-MS/MS)
Authors: Navratilova J , Myskova M , Kuta J , Franzova K , Ješeta M 
Published: June 2022

Abstract

Study question
Can we quantitatively, accurately, and simultaneously determine concentrations of essential and toxic elements in seminal fluid?

Summary answer
We developed and validated a multi-element panel to quantify essential and toxic elements in seminal plasma in one single assay.

What is known already
It is well known that certain elements are involved in normal spermatogenesis, sperm function and male fertility. Elements, such as zinc, copper, and calcium, are important for the maintenance of normal spermatogenesis, sperm maturation etc. On the other hand, even low-level exposure to elements, such as lead, cadmium, arsenic, mercury has been associated with adverse effects on male reproductive health.

Study design, size, duration
In this study we developed an analytical method based on inductively coupled plasma tandem mass spectrometry (ICP-MS/MS) to accurately quantify panel of several essential and toxic elements including vanadium, chromium, cobalt, manganese, nickel, copper, zinc, arsenic, selenium, strontium, molybdenum, cadmium, tin, antimony, mercury, lead in seminal plasma.

Participants/materials, setting, methods
Seminal plasma samples were analysed by inductively coupled plasma mass spectrometry (Agilent 8900 ICP-MS/MS, Agilent technologies) after 10x dilution of samples by solution containing deionized water, a non-ionic surfactant Triton X-100 (0.04 %), ammonia (1 %), butanol (2 %), EDTA (0.04 %) and internal standards for ICP-MS/MS (20 ng/ml of Scandium, Germanium, Indium, Lutetium and Bismuth).

Main results and the role of chance
We developed and validated multielement method for detection of essential and toxic elements in seminal fluid using ICP-MS/MS. The method validation was performed based on FDA guidelines. Validation criteria included limit of detection (LOD), limit of quantitation (LOQ), accuracy and precision. Due to the lack of the relevant certified reference material the validation criteria of the method were assessed in pool of spiked seminal samples. The accuracy of the ICP-MS/MS method was evaluated as a percent recovery of the amount of target analyte added into the sample. Recovery rates were within the range of 89 % and 110 % for all analytes. Additionally, certified reference materials Seronorm™ Trace Elements Serum L-1 and Seronorm™ Trace Elements Serum L-2 (Billingstad, Norway) were used for each batch of analysis. The LOD varied from 0.003 ng/mL for Cd to 0.03 μg/mL for Zn.

Limitations, reasons for caution
The limitation of the developed method is the analysis cost.

Wider implications of the findings
Laboratory testing for essential and toxic elements is important to diagnose metal toxicity and nutritional deficiency, particularly in occupationally exposed groups. This method developed to assess elemental deficiency or toxicity and to evaluate synergic effects between metals and chemicals present in the firefighting environment on spermatozoa.

Trial registration number
not applicable