I am performing malondialdehyde (MDA) measurements in Leydig cells based on the method of Heath and Packer (1968), with modifications adapted to our laboratory conditions. Could you suggest any alternative protocols or assays for the quantification of MDA in cellular samples?

The protocol is as follows: To each tube, I add 1500 µL of a TCA-TBA reagent (20% trichloroacetic acid and 0.5% thiobarbituric acid) and 150 µL of the cell suspension (1×10⁶ cells). For the blank tubes, I add 1500 µL of the same TCA-TBA reagent and 150 µL of TRIS-HCl buffer. All tubes are incubated at 100°C for 1 hour. After incubation, the tubes are cooled and centrifuged at 1000 g for 5 minutes. The absorbance of the supernatant is then measured at 532 nm using a spectrophotometer.

However, I am encountering two major issues: