Cross-reactivity of the CEDIA buprenorphine assay in drugs-of-abuse screening: influence of dose and metabolites of opioids

Figures & data

Table 1 Results from urine samples (n=15) after ingestion of two tablets of a prescription drug containing 400 mg paracetamol +30 mg codeine phosphate by a healthy drug-naïve male volunteer

Timea	CEDIA (ng/mL)	HPLC-MS/MS (ng/mL)
	Buprenorphine	C6G	Codeine	Norcodeine	Morphine	M3G	M6G
05.00	6.9	38,467	2,550	490	150	6,090	1,510
07.00	2.4	11,079	390	110	30	1,500	290
09.00	2.9	17,308	490	180	30	2,320	480
10.00	1.3	5,183	150	60	0	620	130
14.00	1.4	7,275	290	100	50	2,220	460
15.00	0.5	4,665	240	70	50	2,850	490
22.25	0.2	3,371	130	60	50	2,070	330
27.00	0	775	30	0	30	1,060	250
30.00	0.1	0	0	0	0	610	120
34.25	0	0	0	0	20	860	160
39.00	0	0	0	0	0	800	160
46.25	0	0	0	0	10	550	110
50.34	0.1	0	0	0	0	300	0
51.55	0.2	0	0	0	0	0	0
55.00	0	0	0	0	0	0	0

Note:

a Time is given in hours and minutes after drug ingestion.

Abbreviations: CEDIA, cloned enzyme donor immunoassay; HPLC-MS/MS, high-performance liquid chromatography–tandem mass spectrometry; C6G, codeine-6-glucuronide; M3G, morphine-3-glucuronide; M6G, morphine-6-glucuronide.

Table 2 Frequency and share of positive results for drugs-of-abuse in patient urine samples (n=2,272), as determined by screening with HPLC-MS/MS

Substance	Positive results
	n	%
6-Monoacetylmorphine	34	1.5
Amphetamine	258	11.4
Benzodiazepines	959	42.2
Buprenorphine	711	31.3
Codeine	56	2.5
Methadone	664	29.2
Methamphetamine	320	14.1
Morphine	123	5.4

Abbreviation: HPLC-MS/MS, high-performance liquid chromatography–tandem mass spectrometry.

Table 3 Results from CEDIA and HPLC-MS/MS in 25 urine samples with false-positive CEDIA buprenorphine

CEDIA (ng/mL)	HPLC-MS/MS (ng/mL)
Buprenorphine	C6G	Codeine	Norcodeine	Morphine	M3G	M6G
18.20	91,294	13,561	4,166	200	3,138	646
18.10	17,165	1,018	788	26	784	185
16.70	0	0	0	0	0	0
15.00	41,938	5,149	1,121	57	2,538	600
11.20	9,415	2,335	1,070	37,665	139,823	48,915
10.60	11,602	3,832	970	27,621	178,586	68,758
9.60	11,174	1,766	143	36,253	120,903	46,146
8.60	0	0	0	0	0	0
7.60	4,279	1,467	51	29,390	125,056	48,454
7.30	4,336	1,168	94	23,826	127,825	42,778
7.10	37,801	2,096	1,067	29	2,123	415
7.10	12,172	2,604	1,104	14,096	148,129	46,608
6.90	29,861	3,682	705	314	9,137	1,661
6.90	0	0	0	0	0	0
6.80	24,820	1,736	297	19,118	117,673	40,609
6.80	0	0	0	0	0	0
6.30	2,948	1,078	86	12,441	97,368	20,997
6.30	0	0	0	0	0	0
6.20	30,479	5,868	962	86	1,154	231
6.00	0	0	0	0	0	0
5.90	8,464	1,527	611	9,245	105,213	31,656
5.80	27,626	1,676	1,273	57	3,092	461
5.40	19,020	1,078	514	17	3,461	600
5.20	6,419	1,197	254	8,389	108,444	23,073
5.00	0	0	0	0	0	0

Abbreviations: CEDIA, cloned enzyme donor immunoassay; HPLC-MS/MS, high-performance liquid chromatography–tandem mass spectrometry; C6G, codeine-6-glucuronide; M3G, morphine-3-glucuronide; M6G, morphine-6-glucuronide.

Figure 1 PPV and NPV with different cutoff concentrations of CEDIA for buprenorphine in patient urine samples (n=2,272).

Abbreviations: PPV, positive predictive value; NPV, negative predictive value; CEDIA, cloned enzyme donor immunoassay.

Table S1 Analytical performance of HPLC-MS/MS for relevant analytes

Component	Calibration rangea	Limit of detectiona	Imprecisionb (%)
6-Monoacetylmorphine	33–3,275	6.0	10.5
7-Aminoclonazepam	29–5,737	2.33	6.7
7-Aminoflunitrazepam	28–5,690	2.25	6.8
7-Aminonitrazepam	25–5,031	0.7	15.3
Alprazolam	31–6,178	0.9	5.1
Amphetamine	27–2,708	4.4	11.7
Benzoylecgonine	29–2,894	0.3	5.8
Cocaine	6–608	0.2	12.0
Codeine	30–2,997	6.9	16.3
Codeine-6-glucuronide	95–9,511	5.0	15.3
Desmethyldiazepam	54–10,832	3.5	5.1
Diazepam	3–570	1.9	10.4
EDDP	28–2,775	0.8	29.1
Ephedrine	33–3,320	5.5	13.6
Ethylmorphine	31–3,137	7.7	13.4
Hydrocodone	60–5,994	10.9	17.6
MDA	36–3,589	3.3	9.8
MDMA	39–3,873	1.0	10.7
Methadone	31–3,102	1.4	32.0
Methamphetamine	30–2,992	5.8	8.2
Morphine	29–2,855	9.0	17.1
Morphine-3-glucuronide	231–23,074	8.1	6.7
Morphine-6-glucuronide	139–13,851	15.0	8.5
N-desmethylzopiclone	38–7,513	2.6	7.5
Norcodeine	29–2,880	8.8	20.2
Oxazepam	57–11,470	4.4	7.5
Oxycodone	63–6,313	7.2	10.1
Temazepam	60–12,069	2.6	4.8
Zopiclone-N-oxide	41–8,122	1.5	11.3

Notes:

a Concentrations are given in ng/mL;

b total analytical imprecision is calculated based on performance at lowest calibrator plus 25%.

Abbreviations: HPLC-MS/MS, high-performance liquid chromatography–tandem mass spectrometry; EDDP, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine; MDA, 3,4-methylenedioxy-amphetamine; MDMA, 3,4-methylenedioxy-methamphetamine.