Influence of light and time on bilirubin degradation in CSF spectrophotometry for subarachnoid haemorrhage

Investigation of the influence of light on bilirubin degradation over time in cerebrospinal fluid (CSF) specimens from patients with computed tomography (CT) positive subarachnoid haemorrhage (SAH).

Twenty-nine CSF samples were analysed from 23 patients with CT-positive SAH. Samples were divided into two cohorts - one half being stored in the dark, the other in normal room light conditions. Samples were assayed by spectrophotometry 0, 1, 2, 6, 19, 24 and 48 h post-storage. Net bilirubin absorbance (NBA) was measured in absorbance units (AU) by determining the optical density at 476 nm.

In 27 of 28 (96%) samples stored in light conditions and 21 of 29 (72%) samples stored in dark, a decline in the amount of bilirubin detected in CSF over time was observed. The rate of bilirubin degradation over 24 h for CSF samples stored in the light ranged from 0.0001 to 0.0048 AU/h (mean 0.0020 AU/h), and was significantly greater (p < 0.01) than the rate of degradation in the dark, which ranged from 0 to 0.0024 AU/h (mean 0.0005 AU/h).

CSF bilirubin samples are susceptible to the same photodegradation as shown in serum bilirubin samples. The rate of degradation is not predictable or consistent. This photodegradation may potentially produce false-negative results in the diagnosis of SAH. If spectrophotometry cannot be carried out immediately, all such CSF samples should at least be centrifuged immediately, the supernatant protected from light as soon as possible and the collection and analysis times provided in the report.