Table 2.

Advantages and disadvantages of commonly used techniques to measure seminal oxidative stress. Reproduced with permission from Agarwal, A., Ther Adv Urol. 2016. (Table 1).

Assay	Advantages	Disadvantages
OS via chemiluminescence	Chemiluminescence is robust High sensitivity and specificity Luminol measures global ROS levels—both extracellular and intracellular (superoxide anion, hydrogen peroxide, hydroxyl radical)	Time-consuming method Requires large and expensive equipment Variables such as semen age, volume, repeated centrifugation, temperature control, and background luminescence may interfere with measurement
TAC	Rapid colorimetric method Measures total antioxidants in seminal plasma	Does not measure enzymatic antioxidants Length of the inhibition time is a critical aspect of the test Requires expensive microplate readers
ROS-TAC score	Better predictor compared with ROS and TAC alone	Requires statistical modeling Not a direct measure of ROS or TAC, rather a prediction of oxidative stress
MDA (TBARS adduct by colorimetry or fluoroscopy)	Measures lipid peroxidation Detects MDA-TBA adduct by colorimetry or fluoroscopy	Rigorous controls required Non-specific test providing post hoc measure only
ORP	Provides redox balance in real time Measures all known and unknown oxidants and antioxidants Less time-consuming and requires less expertise Can be measured in semen and seminal plasma, including frozen specimens	Affected by viscosity of the sample

MDA, malondialdehyde; ROS, reactive oxygen species; TAC, total antioxidant capacity; TBARS, thiobarbituric acid reactive substances.