Four US communities with high prevalence of MAMP abuse participated in the IDEAL study: Los Angeles, California; Des Moines, Iowa; Tulsa, Oklahoma; and Honolulu, Hawaii. The study was approved by Institutional Review Boards at each hospital, the University of Maryland, Warren Alpert Medical School of Brown University, and National Institute on Drug Abuse (NIDA); a NIDA Certificate of Confidentiality was obtained to facilitate honest recall of drug use history. Procedures and protocols were standardized across locations.6 (link) Maternal and infant exclusion criteria were fully described by Arria et al.6 (link) Among the most frequent reasons for ineligibility were the inability to speak English, maternal age < 18 years, multiple births and opiate use during pregnancy.
After providing informed consent, mothers completed a Substance Use Inventory to recall the amount and frequency of alcohol, tobacco, and drug (cannabis, hashish, MAMP, ecstasy, AMP, benzodiazepine/tranquilizers, barbiturates/sedatives, and cocaine/crack) consumption during each trimester and the three months prior to becoming pregnant. Quantity descriptions varied by drug, and frequency was classified by the number of days per week drugs were consumed: every day, almost every day, 3–4 times per week, 1–2 times per week, 2–3 times per month, once a month, or 1–2 times in 3 months. Route of administration, insufflation, ingestion, smoking and/or intravenous use, were recorded.
Meconium was collected from diapers until the appearance of milk stool. Specimens remained refrigerated until transported overnight (2-day for Hawaii) to the United States Drug Testing Laboratories (Des Plaines, IL) for analysis. Meconium (0.5 g) was homogenized in methanol and centrifuged. Supernatants were buffered and extracted using mixed mode solid phase extraction columns (ZSDAU020, United Chemical Technologies, Bristol, PA). Specimens were screened within 24 hours of receipt with Syva enzyme multiplied immunoassay technique (EMIT) II Plus (Dade Behring, Cupertino, CA) for cannabinoids (cutoff: 40 ng/g), cocaine metabolite (75 ng/g), opiates (150 ng/g) and amphetamines (500 ng/g) on an Olympus AU640 analyzer (Center Valley, PA). According to the manufacturer, the following compounds substantially cross-reacted (>5%) with the amphetamines immunoassay: d,l-MAMP (71%), benzphetamine (71%), phenmetrazine (70%), phentermine (55%), d,l-AMP (48%), l-MAMP (38%), mephentermine (33%), 3,4-methylenedioxyamphetamine (MDA) (29%), l-AMP (13%), fenfluramine (12.5%), 4-chloramphetamine (11%), tranylcypromine (8%), 3,4-methylenedioxyethylamphetamine (7%), norpseudoephedrine (7%), and MDMA (5%).7 If positive, separate aliquots were extracted and confirmed by GCMS within 48 h at the following cutoffs: 2 ng/g 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THCCOOH); 5 ng/g cocaine, cocaethylene, benzoylecgonine, and/or m-hydroxybenzoylecgonine; 5 ng/g codeine and morphine; and 5 ng/g AMP, MAMP, MDMA, and MDA.8 The tobacco biomarker cotinine was screened by enzyme-linked immunosorbent assay (ELISA, International Diagnostics, St. Joseph, MI) with a 10 ng/g cutoff. A more specific chromatographic method for cotinine in meconium was not available for confirmation. Maternal opiate use was exclusionary and cocaine use without concurrent MAMP use also was exclusionary; thus, meconium results for these drug classes are not presented, as data are not representative.
SPSS version 16.0 for Windows (Chicago, IL) and Microsoft Excel were employed for data analysis and statistical evaluation. Mann-Whitney tests evaluated analyte concentration and metabolite ratio differences between groups; p-values <0.05 were considered statistically significant.
After providing informed consent, mothers completed a Substance Use Inventory to recall the amount and frequency of alcohol, tobacco, and drug (cannabis, hashish, MAMP, ecstasy, AMP, benzodiazepine/tranquilizers, barbiturates/sedatives, and cocaine/crack) consumption during each trimester and the three months prior to becoming pregnant. Quantity descriptions varied by drug, and frequency was classified by the number of days per week drugs were consumed: every day, almost every day, 3–4 times per week, 1–2 times per week, 2–3 times per month, once a month, or 1–2 times in 3 months. Route of administration, insufflation, ingestion, smoking and/or intravenous use, were recorded.
Meconium was collected from diapers until the appearance of milk stool. Specimens remained refrigerated until transported overnight (2-day for Hawaii) to the United States Drug Testing Laboratories (Des Plaines, IL) for analysis. Meconium (0.5 g) was homogenized in methanol and centrifuged. Supernatants were buffered and extracted using mixed mode solid phase extraction columns (ZSDAU020, United Chemical Technologies, Bristol, PA). Specimens were screened within 24 hours of receipt with Syva enzyme multiplied immunoassay technique (EMIT) II Plus (Dade Behring, Cupertino, CA) for cannabinoids (cutoff: 40 ng/g), cocaine metabolite (75 ng/g), opiates (150 ng/g) and amphetamines (500 ng/g) on an Olympus AU640 analyzer (Center Valley, PA). According to the manufacturer, the following compounds substantially cross-reacted (>5%) with the amphetamines immunoassay: d,l-MAMP (71%), benzphetamine (71%), phenmetrazine (70%), phentermine (55%), d,l-AMP (48%), l-MAMP (38%), mephentermine (33%), 3,4-methylenedioxyamphetamine (MDA) (29%), l-AMP (13%), fenfluramine (12.5%), 4-chloramphetamine (11%), tranylcypromine (8%), 3,4-methylenedioxyethylamphetamine (7%), norpseudoephedrine (7%), and MDMA (5%).7 If positive, separate aliquots were extracted and confirmed by GCMS within 48 h at the following cutoffs: 2 ng/g 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THCCOOH); 5 ng/g cocaine, cocaethylene, benzoylecgonine, and/or m-hydroxybenzoylecgonine; 5 ng/g codeine and morphine; and 5 ng/g AMP, MAMP, MDMA, and MDA.8 The tobacco biomarker cotinine was screened by enzyme-linked immunosorbent assay (ELISA, International Diagnostics, St. Joseph, MI) with a 10 ng/g cutoff. A more specific chromatographic method for cotinine in meconium was not available for confirmation. Maternal opiate use was exclusionary and cocaine use without concurrent MAMP use also was exclusionary; thus, meconium results for these drug classes are not presented, as data are not representative.
SPSS version 16.0 for Windows (Chicago, IL) and Microsoft Excel were employed for data analysis and statistical evaluation. Mann-Whitney tests evaluated analyte concentration and metabolite ratio differences between groups; p-values <0.05 were considered statistically significant.
