Assessment of urine sample quality by the simultaneous measurement of urinary gamma-glutamyltransferase and lactate dehydrogenase enzyme activities: possible application to unravel cheating in drugs of abuse testing

Objectives: Evaluation of the simultaneous measurement of urinary gamma-glutamyltransferase (gamma GT) and lactate dehydrogenase (LDH) to discriminate fresh from previously frozen specimens in urine drug monitoring. Methods: Two widely available photometric tests (Siemens Healthineers Atellica) were used to determine the range of urinary gamma GT and LDH excretion and to study the decay in urinary enzyme activity under various storage conditions (room temperature, 4-8 degrees C, -18 degrees C, -80 degrees C). From these data, cut-off values were established and evaluated in split (fresh/frozen) specimens. Results: Both assays allow robust, reliable, and simultaneous determination of urinary gamma GT and LDH. In healthy subjects, the 95% reference intervals for enzyme activity in native urine were gamma GT: 24.4-100.4 U/g Crea (creatinine) and LDH: 2.5-45.8 U/g Crea. Frozen storage for at least 7 days at -18 degrees C resulted in a loss of activity to less than 50% in both enzymes. Cut-offs for frozen samples were gamma GT <= 33.2 U/g Crea and LDH <= 8.4 U/g Crea. When applied to 100 sample pairs (fresh/frozen), 86.5% (173/200) of the measurements were conclusive and the combination of concordant enzyme measurements (low gamma GT/low LDH or high gamma GT/high LDH) was able to predict the mode of storage with a sensitivity of 96.3% and a specificity of 96.7%. Conclusions: The additional measurements of urinary gamma GT and LDH can be used to detect previously frozen urine specimens. A simple protocol is proposed to provide additional information on sample quality when deceit is suspected. The procedure can be easily integrated into the standard workflow of urinary drug monitoring. Conclusions: The additional measurements of urinary gamma GT and LDH can be used to detect previously frozen urine specimens. A simple protocol is proposed to provide additional information on sample quality when deceit is suspected. The procedure can be easily integrated into the standard workflow of urinary drug monitoring.