MSACL 2018 US Abstract

Topic: Small Molecules / Tox

Analysis of 44 Drugs and their Metabolites in Umbilical Cord by LC-MS/MS

Vandana Megaraj (Presenter)
Cincinnati Childrens Hospital

Bio: Currently, I am working as a Research Scientist in the Clinical Mass Spectrometry Core, Pathology division in Cincinnati Children’s Hospital Medical Center (CCHMC) in Cincinnati, Ohio. I have over 6 years’ experience in utilizing various mass spectrometry instruments to perform research and clinical drug testing. I have been working at CCHMC since 2013 where I have expanded my research efforts into biomedical and clinical applications. My Overall goal is to maintain and oversee day to day analysis of 46 pain panel drugs, troubleshoot and maintain LCMS instruments for high volume production, method development and validation of drug analysis in various matrices, writing and review SOPs, support research for disease management in newborns. I obtained my PhD degree in Toxicology at University of Kentucky followed by a postdoctoral training at Newyork State department of health, Albany,NY

Authorship: Vandana Megaraj (1), Christina Huth (1), Scott Wexelblatt (2), Kenneth D.R. Setchell (1,3)
(1) Division of Pathology & Laboratory Medicine, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH (2) Division of Neonatology and Pulmonary Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH (3) Department of Pediatrics, University of Cincinnati College of medicine, Cincinnati, OH

Short Abstract

Detection of in utero drug exposure in pregnant women at the time of delivery is critical to implementing effective treatment of the newborn for withdrawal symptoms of neonatal abstinence syndrome. Umbilical cord tissue offers an alternative to urine and meconium for the detection of drugs of abuse during pregnancy. A validated LC-MS/MS method for the simultaneous detection of 44 drugs and metabolites in umbilical cord was developed using a novel sample preparation followed by hydrolysis of the conjugates and solid phase extraction. The method was rapid, making it suitable for routine clinical use and had high sensitivity, accuracy and precision.

Long Abstract


Expedient detection of in utero drug exposure is critical to effectively treat newborns for symptoms of withdrawal in Neonatal Abstinence Syndrome (NAS). NAS occurs as a result of in utero exposure to opioids and various other drugs, and is characterized by symptoms such as tremors or seizures, fever, gastrointestinal issues, failure to thrive, and respiratory problems. Meconium is the most frequently used matrix in conjunction with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for confirmative testing for prenatal drug exposure. We evaluated the utilization of umbilical cord tissue as an alternative matrix for neonatal drug testing. Umbilical cord have shown comparable results to meconium and holds several advantages, some of which are immediate and universal availability, tissue belongs to the neonate and does not require the mother’s permission to test and does not reflect drugs administered to neonates after birth.

The aim of this study was to develop and validate the robust method for analysis of 44 pain panel drugs and their metabolites in umbilical cord with the goal of rapid (approximately 24 hr) turnaround time for use in evaluating exposure of neonatal to drug of abuse.


Umbilical cord specimens were obtained at the time of delivery and analyzed for 10 drug classes: methamphetamines, opioids, cocaine, benzodiazepines, barbiturates, buprenorphine, methadone, muscle relaxants, nicotine metabolite and phencyclidine. Approximately 10 cm segment of umbilical cord tissues were received from hospitals. The cord segments were stripped of blood and rinsed with saline and an amount equivalent to approximately 1gm tissue was taken and stored in sterile tubes at -20°C. Drug-free cord samples were spiked with known amounts of 44 drugs standards and the internal standards to establish a calibration curve and quality control samples. Beta-glucuronidase enzyme was added to the tissue and homogenized using a bead homogenizer. Hydrolysis was performed by incubating at 60°C for 90 minutes. The samples were then extracted by SPE method using Strata X-C mixed mode columns and analyzed for the 10 drug classes using an LC-MS/MS method previously developed for urine analysis.


A fully validated LC-MS/MS method for simultaneous quantification of 44 drugs and metabolites in umbilical cord tissue was developed. The method had excellent limits of quantification (ranging from 0.1- 8 ng/mL) for different drug classes and linearity over a wide dynamic range (0.1– 1250 ng/mL). The calculated inter-assay accuracies (% bias) was (<13.6%) for all concentrations with an overall inter-batch precision lower than 13.8 % for the cord samples used as Quality Controls. Similar values were obtained for the intra-assay accuracy (<12%) and precision (<14%) for all QC samples across 10 different drug classes comprising 44 drugs. No detectable carryover was observed for all 44 drugs.

Conclusions & Discussion

Rapid detection of fetal exposure to illicit drugs is of considerable clinical value to determining the treatment of newborns with NAS. This highly sensitive and specific LC/MS/MS method for analysis of drug classes in umbilical cord tissue was suitable for routine clinical testing of fetal exposure to drug of abuse during pregnancy. The use of umbilical cord tissue has the advantage over meconium analysis in that it enables a faster turnaround time for resulting the findings to the clinician because waiting for meconium to be passed can take up to several days, or in some instances is passed in utero making collection impossible, while umbilical cord tissue is always available for drug testing. This assay represents one of the many improvements over previous methods for determining fetal drug exposure.

References & Acknowledgements:

Financial Disclosure

SalaryyesCincinnati Childrens Hospital
Board Memberno

IP Royalty: no

Planning to mention or discuss specific products or technology of the company(ies) listed above: