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Aim: Human 14C radiotracer studies provide information-rich data sets that enable informed decision making in clinical drug development. These studies are supported by liquid scintillation counting after conventional-sized 14C doses (50–200 μCi) or complex accelerator mass spectrometry (AMS) after microtracer-sized doses (∼0.1–1 μCi). Mid-infrared laser-based ‘cavity ring-down spectroscopy’ (CRDS) is an emerging platform for the sensitive quantitation of 14C tracers. Results & methodology: We compared the total 14C concentrations in plasma and urine samples from a microtracer study using both CRDS and AMS technology. The data were evaluated using statistical and pharmacokinetic modeling. Conclusion: The CRDS method closely reproduced the AMS method for total 14C concentrations. With optimization of the automated sample interface and further testing, it promises to be an accessible, robust system for pivotal microtracer investigations

Pharmacokinetic studies in the neonatal population are often limited by the small volume of blood that can be collected. The high sensitivity of (14) C-accelerator mass spectrometry (AMS) enables pharmacokinetic studies to be conducted with greatly reduced sample volumes. We demonstrated the utility of AMS in infants by studying the plasma pharmacokinetic behavior of nanogram doses of (14) C-ursodiol administered as a non-perturbing microdose or as a microtracer with therapeutic doses of non-labeled ursodiol in infants. Five non-cholestatic infants were administered 3 consecutive oral microdoses of (14) C-ursodiol: 8 ng (1.0 nCi), 26 ng (3.3 nCi), and 80 ng (10 nCi) 48 hours apart. Three additional infants with cholestasis were administered a single 80 ng (10.0 nCi) oral dose of (14) C-ursodiol together with a therapeutic dose of 40 mg/kg of non-labeled ursodiol. A pharmacokinetic model describing ursodiol concentrations was developed using nonlinear mixed-effects modeling. The pharmacokinetics of ursodiol in this pilot study were best described by a two-compartment model with first-order elimination. This study demonstrates the feasibility and utility of microdose and microtrace methodology in pediatric research.

14C-radiolabeled (radiocarbon) drug studies are central to defining the disposition of therapeutics in clinical development. Concerns over radiation, however, have dissuaded investigators from conducting these studies as often as their utility may merit. Accelerator mass spectrometry (AMS), originally designed for carbon dating and geochronology, has changed the outlook for in-human radiolabeled testing. The high sensitivity of AMS affords human clinical testing with vastly reduced radiative (microtracing) and chemical exposures (microdosing). Early iterations of AMS were unsuitable for routine biomedical use due to the instruments’ large size and associated per sample costs. The situation is changing with advances in the core and peripheral instrumentation. We review the important milestones in applied AMS research and recent advances in the core technology platform. We also look ahead to an entirely new class of 14C detection systems that use lasers to measure carbon dioxide in small gas cells., Lay Abstract: Radiocarbon (14C) is produced continuously in the upper atmosphere and incorporated into biological systems through photosynthesis. Incorporation of 14C ceases at death, hence by measuring natural levels of radiocarbon in deceased organic materials the age of the objects (e.g. vegetation, wood and historical artifact, among others) can be determined. Early carbon dating determinations (starting in the late 1940s) relied on monitoring the small levels of radioactivity decay energy emerging from the sample. In the late 1970s nuclear physicists in collaboration with archeologists developed a far more sensitive and efficient measuring technique known as accelerator mass spectrometry (AMS), which can identify individual carbon atoms. The technology was extended to biomedical research in the early 1990s. Here it had an immediate impact on in vivo human research across diverse areas such as carcinogenesis and human nutrition. The same carbon dating technology now supports human 14C-tracer metabolism studies conducted as part of early drug development. The high sensitivity of AMS enables scientists to explore new drug candidates safely in any human subpopulation at very low chemical (microdosing) and radioactive doses (microtracing). A ‘big physics’ tool originally developed for dating archeological artifacts is now benefiting modern drug development in unanticipated ways. The full benefits of the sensitive measurement technology were impeded by the scale and operational complexity of the early instruments, but this situation has changed significantly over the last decade after the emergence of purpose-built biomedical instruments. The evolving relationship between radiocarbon, AMS, and modern drug development is discussed in this review.

Accelerator Mass Spectrometry is an established technology whose essentiality extends beyond simply a better detector for radiolabeled molecules. Attomole sensitivity reduces radioisotope exposures in clinical subjects to the point that no population need be excluded from clinical study. Insights in human physiochemistry are enabled by the quantitative recovery of simplified AMS processes that provide biological concentrations of all labeled metabolites and total compound related material at non-saturating levels. In this paper, we review some of the exploratory applications of AMS (14)C in toxicological, nutritional, and pharmacological research. This body of research addresses the human physiochemistry of important compounds in their own right, but also serves as examples of the analytical methods and clinical practices that are available for studying low dose physiochemistry of candidate therapeutic compounds, helping to broaden the knowledge base of AMS application in pharmaceutical research.

The next generation Accelerator Mass Spectrometer system specifically designed to address the needs of the growing pharmaceutical science market has passed validation testing. The system dubbed BioMICADAS is based on a previously developed compact carbon dating instrument, the MICADAS. Like its predecessor, it has an overall footprint of only 2.5 × 3 m2 and uses a 200 kV high voltage platform for tandem based ion acceleration. The ion source can accommodate samples as graphite or gaseous CO2. It is equipped with two independently operating vacuum locks, allowing continuous measurement sequence and providing a capacity of ∼20,000 samples per annum. A barcoded cathode tracking system allows data capture into Laboratory Information Management System (LIMS) for Good Laboratory Practices (GLP) regulated work. It can be housed in research laboratories alongside other complementary bioanalytical equipment and operated by general laboratory staff as the system is designed to be robust and user-friendly. The system has undergone rigorous validation over the range from 0.1 to 100 Modern Carbon, including accuracy, linearity, robustness, and precision experiments over the course of 7 months. It has been shipped and installed at the site of our collaborative partner, Vitalea Science in Davis, California. The installation process took ∼2 weeks from boxes to beam. The feasibility of the system to determine the absolute specific activity of biogenic samples was also shown by using the method of isotopic dilution.

Accelerator mass spectrometers have an energy acceleration and charge exchange between mass definition stages to destroy molecular isobars and allow single ion counting of long-lived isotopes such as 14C (t½=5370 years.). ‘Low’ voltage accelerations to 200 kV allow laboratory-sized accelerator mass spectrometers instruments for bioanalytical quantitation of 14C to 2–3% precision and accuracy in isolated biochemical fractions. After demonstrating this accuracy and precision for our new accelerator mass spectrometer, we discuss the critical aspects of maintaining quantitative accuracy from the defined biological fraction to the accelerator mass spectrometry quantitation. These aspects include sufficient sample mass for routine rapid sample preparation, isotope dilution to assure this mass, isolation of the carbon from other sample combustion gasses and use of high-efficiency biochemical separations. This review seeks to address a bioanalytical audience, who should know that high accuracy data of physiochemical processes within living human subjects are available, as long as a 14C quantitation can be made indicative of the physiochemistry of interest.

Folate is a structurally diverse family of molecules that occur in low concentrations in biological tissues and fluids and therefore present complex challenges to the analytical chemist. As a result, techniques that quantify folates in biological specimens have emerged as a high scientific priority. In this manuscript, isotopic methods using traditional mass spectrometry for the measurement of whole blood total folate and 14 C-accelerator mass spectrometry (AMS) for human kinetic studies are presented. Additionally, recent developments with the intrinsic labeling of spinach folate from a [ 14 C]- para -aminobenzoic is discussed.

Vegetables and fruits provide an array of microchemicals in the form of vitamins and secondary metabolites (phytochemicals) that may lower the risk of chronic disease. Tracing these phytochemicals at physiologic concentrations has been hindered by a lack of quantitative sensitivity for chemically equivalent tracers that could be used safely in healthy people. Accelerator mass spectrometry is a relatively new technique that provides the necessary sensitivity (in attomoles) and measurement precision (

The effect of vitamin A supplements on metabolic behavior of an oral tracer dose of [14C]beta-carotene was investigated in a longitudinal test-retest design in two adults. For the test, each subject ingested 1 nmol of [14C]beta-carotene (100 nCi) in an emulsified olive oil-banana drink. Total urine and stool were collected for up to 30 days; concentration-time patterns of [14C]beta-carotene, [14C]retinyl esters, and [14C]retinol were determined for 46 days. On Day 53, the subjects were placed on a daily vitamin A supplement (10000 IU/day), and a second dose of [14C]beta-carotene (retest) was given on Day 74. All 14C determinations were made using accelerator mass spectrometry. In both subjects, the vitamin A supplementation was associated with three main effects: 1). increased apparent absorption: test versus retest values rose from 57% to 74% (Subject 1) and from 52% to 75% (Subject 2); 2). an approximately 10-fold reduction in urinary excretion; and 3). a lower ratio of labeled retinyl ester/beta-carotene concentrations in the absorptive phase. The molar vitamin A value of the dose for the test was 0.62 mol (Subject 1) and 0.54 mol (Subject 2) vitamin A to 1 mol beta-carotene. Respective values for the retest were 0.85 and 0.74. These results show that while less cleavage of beta-carotene occurred due to vitamin A supplementation, higher absorption resulted in larger molar vitamin A values.

Long-term physiologic tracing of nutrients, toxins, and drugs in healthy subjects is not possible using traditional decay counting of radioisotopes or stable isotope mass spectrometry due to radiation exposure and limited sensitivity, respectively. A physiologic dose of C-14-labeled folic acid (35 mu g, 100 nCi) was ingested by a healthy adult male and followed for 202 days in plasma, erythrocytes, urine, and feces using accelerator mass spectrometry. All samples and generated wastes were classified nonradioactive and the subject received a lifetime-integrated radiological effective dose of only II mu Sv. Radiolabeled folate appeared in plasma 10 min after ingestion but did not appear in erythrocytes until 5 days later. Approximately 0.4% of the erythrocytes were intrinsically labeled with an average of 130 C-14 atoms during erythropoiesis from the pulse of plasma [C-14]folate. An appropriate radiocarbon-labeled precursor can intrinsically label DNA or a specific protein during synthesis and obtain limits of quantitation several orders of magnitude below that of stable isotope methods. (C) 1999 Academic Press.