METHOD FOR DETECTING OPIOIDS, OPIATES, CANNABINOIDS, OR BENZODIAZEPINES IN A SAMPLE WITH A B-GLUCURONIDASE ENZYME
The present invention discloses a highly efficient method for detecting opiods, opiates, cannabinoids, or benzodiazepines present in a sample, comprising the steps of adding to said sample an enzyme with β-glucuronidase activity originated from genus Brachyspira or any mutant derived thereof; incubating the sample with the enzyme; and detecting said opiods, opiates, cannabinoids, or benzodiazepines by means of a suitable technique.
1 . A method for detecting opiods, opiates, cannabinoids, or benzodiazepines in a sample, comprising the steps of:
a) providing the sample in which said opiods, opiates, cannabinoids, or benzodiazepines are to be detected;
b) adding to said sample an enzyme with β-glucuronidase activity originated from genus Brachyspira or any mutant derived thereof;
c) incubating the sample with the enzyme; and
d) detecting said opiods, opiates, cannabinoids, or benzodiazepines through any suitable technique.
2 . The method of claim 1 , wherein the sample is a biological sample.
3 . The method of claim 2 , wherein the biological sample is selected from the group consisting of saliva, whole blood, plasma, urine, hair, skin, teeth, soft tissues, meconium and vitreous humor.
4 . The method of claim 1 , wherein the enzyme with β-glucuronidase activity originates from species Brachyspira pilosicoli.
5 . The method of claim 4 , wherein the enzyme with β-glucuronidase activity is selected from the group consisting of:
a) an amino acid sequence comprising the amino acid sequence shown in SEQ.ID.NO. 1;
b) an amino acid sequence sharing at least an 80% of identity with the amino acid sequence defined in a).
6 . The method of claim 1 , wherein the step of incubating the sample with the enzyme is performed for a period between 1 and 30 minutes.
7 . The method of claim 1 , wherein the step of incubating the sample with the enzyme is performed at a temperature range between 20 and 60 degrees Celsius.
8 . The method of claim 1 , wherein the step of incubating the sample with the enzyme is performed at a range of pH between 4.0 and 9.0.
9 . The method of claim 1 , wherein the suitable technique for detecting opiods, opiates, cannabinoids, or benzodiazepines is selected from the group consisting of LC-MS, GC-MS, HPLC-MS, LC-HRMS, HPLC-HRMS, LC-MS-MS, GC-MS-MS and HPLC-MS-MS.