Assessment of Blood Contamination in Biological Fluids Using MALDI-TOF MS

Abstract

Biological fluid sample collection often includes the risk of blood contamination that may alter the proteomic profile of biological fluid. In proteomics studies, exclusion of contaminated samples is usually based on visual inspection and counting of red blood cells in the sample; analysis of specific blood derived proteins is less used. To fill the gap, we developed a fast and sensitive method for ascertainment of blood contamination in crude biological fluids, based on specific blood-derived protein, hemoglobin detection by MALDI-TOF MS. The MALDI-TOF MS based method allows detection of trace hemoglobin with the detection limit of 0.12 nM. UV-spectrometry, which was used as reference method, was found to be less sensitive. The main advantages of the presented method are that it is fast, effective, sensitive, requires very small sample amount and can be applied for detection of blood contamination in various biological fluids collected for proteomics studies. Method applicability was tested on human cerebrospinal and follicular fluid, which proteomes generally do not contain hemoglobin, however, which possess high risk for blood contamination. Present method successfully detected the blood contamination in 12 % of cerebrospinal fluid and 24 % of follicular fluid samples. High percentage of contaminated samples accentuates the need for initial inspection of proteomic samples to avoid incorrect results from blood proteome overlap.

Estradiol and Progesterone Levels are Related to Redox Status in the Follicular Fluid During In Vitro Fertilization

“…One of the strengths of our study was the use of FF samples with very low levels of blood contamination. Certainly, the use of equipment with increased sensitivity in the detection of blood molecules would allow more accurate results [ 74 ]. However, such methods use expensive equipment and require well-trained professionals; therefore, these methods are not fast or inexpensive.…”