Lipids were extracted from the sperm samples using a modified version of the Bligh and Dyer’s method as previously described [31]. Briefly, 750 µl of ice-cold chloroform: methanol (1:2, v/v) with 10 % deionized H2O was added to inactivate the samples. Then samples were homogenized on an automated bead ruptor (Omni, USA) and further centrifuged at 1500 rpm and 4 °C for 1 h. Next, 350 µl of deionized H2O and 250 µl of chloroform were added to generate separate phase. After transferring the lower organic phase into another tube, 500 µl of chloroform was added for a second extraction. The two extractions were pooled and dried using SpeedVac (Genevac, UK). The dried samples were stored at − 80 °C for mass spectrometric analysis.

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