High‐throughput quantification of drugs of abuse in biofluids via 96‐solid‐phase microextraction–transmission mode and direct analysis in real time mass spectrometry

Abstract

Rationale The workload of clinical laboratories has been steadily increasing over the last few years. High‐throughput (HT) sample processing allows scientists to spend more time undertaking matters of critical thinking rather than laborious sample processing. Herein we introduce a HT 96‐solid‐phase microextraction (SPME) transmission mode (TM) system coupled to direct analysis in real time (DART) mass spectrometry (MS). Methods Model compounds (opioids) were extracted from urine and plasma samples using a 96‐SPME‐TM device. A standard voltage and pressure (SVP) DART source was used for all experiments. Examination of SPME‐TM performance was done using high‐resolution mass spectrometry (HRMS) in full scan mode (100–500 m/z), whereas quantitation of opioids was performed using triple quadrupole MS in multiple reaction monitoring mode and by using a matrix‐matched internal standard correction method. Results Thirteen points (0.5 to 200 ng mL−1) were used to establish a calibration curve. Low limits of quantitation (LOQ) were obtained (0.5 to 25 ng mL−1) for matrices used. Acceptable accuracy (71.4–129.4%) and repeatability (1.1–24%) were obtained for validation levels tested (0.5, 30 and 90 ng mL−1). In less than 1.5 hours, 96 samples were extracted, desorbed and processed using the 96‐SPME‐TM system coupled to DART‐MS. Conclusions A rapid HT method for detection of opioids in urine and plasma samples was developed. This study demonstrated that ambient ionization mass spectrometry coupled to robust sample preparation methods such as SPME‐TM can rapidly and efficiently screen/quantify target analytes in a HT context.

Effect of reducing sample volume on the detection of drugs in urine by transmission mode direct analysis in real time mass spectrometry

“…6,7 TM-DART-MS has yet to be explored extensively as only a limited number of research papers have been published on this topic; 3,4 however, it has been used in some recent studies for rapid screening and quantification of pesticides in food 8 and drugs of abuse in biofluids. 9 While drugs can be detected in biological matrices without chromatographic or analyte separation, [8][9][10][11][12][13] the lack of separation presents challenges for detection and quantitation, particularly when working with samples containing low levels of drugs and relatively high concentrations of matrix molecules as is the case with urine, which contains polar molecules that are readily ionizable by DART. 14 According to Song et al, these urinary matrix molecules create a transient microenvironment (TME) that can shield the target analytes from direct ionization by DART and thus leave them undetectable.…”

Optimization of Coated Blade Spray for Rapid Screening and Quantitation of 105 Veterinary Drugs in Biological Tissue Samples

“…As such, the introduction of such matrices to the MS often requires multiple sample preparation steps and chromatographic separation to prevent interferences from matrix coextractives. Conventional sample preparation methods often include solvent extraction (SE) or protein precipitation (PP) followed by additional extract cleanup with either solid phase extraction (SPE) or dispersive SPE (d-SPE). ,− These steps can be very time-consuming and may not be suitable for the direct and high-throughput screening requirements of modern analysis of real samples. , An alternative approach to this lengthy workflow is to simplify sample preparation and eliminate chromatographic separation by directly introducing the sample to the MS. ,− Direct-to-MS analysis techniques offer the advantages of obtaining analytical results in the order of seconds and screening hundreds of samples in a short period of time . However, in case of tissue samples, minimal sample preparation can introduce an abundance of matrix interferences to the MS.…”

“…However, in case of tissue samples, minimal sample preparation can introduce an abundance of matrix interferences to the MS. Solid phase microextraction (SPME) has consistently demonstrated its ability to isolate and enrich a wide variety of analytes from biological samples with minimal coextraction of proteins, salts, and other matrix macromolecules. − SPME-based devices have also been successfully used to integrate sample preparation and direct sample introduction into the MS system in a single device. ,− One of these devices is the coated blade spray (CBS), which has been widely and successfully applied for rapid qualitative and quantitative analysis of biological fluids such as blood, plasma, and urine. ,− In addition to its direct-to-MS capability, CBS also offers high throughput capabilities and can be easily automated. ,, …”

“…5,13−15 These steps can be very time-consuming and may not be suitable for the direct and high-throughput screening requirements of modern analysis of real samples. 12,16 An alternative approach to this lengthy workflow is to simplify sample preparation and eliminate chromatographic separation by directly introducing the sample to the MS. 12,17−19 MS analysis techniques offer the advantages of obtaining analytical results in the order of seconds and screening hundreds of samples in a short period of time. 14 However, in case of tissue samples, minimal sample preparation can introduce an abundance of matrix interferences to the MS.…”

“…25,27−29 In addition to its direct-to-MS capability, CBS also offers high throughput capabilities and can be easily automated. 16,18,29 Several other direct-to-MS methods have been studied and demonstrated suitable for analysis of tissue samples, but none so far for multiresidue analysis of more than 100 analytes in both negative and positive mode in a single run. 8,12,15,30,31 CBS has not been investigated yet for analysis of tissue samples as, unlike mere direct-sample-to-MS technologies, there is need to remove any tissue debris potentially adhered to the device prior to instrumental analysis.…”