Subject(s)
Busulfan , Polyethylene Glycols , Tandem Mass Spectrometry , Tandem Mass Spectrometry/methods , Humans , Busulfan/pharmacokinetics , Busulfan/blood , Chromatography, High Pressure Liquid/methods , Drug Monitoring/methods , Immunosuppressive Agents/blood , Immunosuppressive Agents/pharmacokineticsABSTRACT
BACKGROUND: Busulfan is widely used in conditioning regimens to prepare patients for hematopoietic stem cell transplantation. Therapeutic drug monitoring (TDM) is critical due to large inter- and intra-individual variability in busulfan pharmacokinetics, and the risk of adverse consequences of toxicity including hepatic veno-occlusive disease. Busulfan is most commonly measured by liquid chromatography-mass spectrometry (LC-MS/MS), which is not as widely available in clinical laboratories as automated routine clinical chemistry analyzers. The objective was to perform analytical verification of a busulfan immunoassay on the Abbott Alinity c platform. METHODS: The MyCare Oncology busulfan immunoassay was configured as a third-party reagent on the Abbott Alinity c. Imprecision, linearity, sample carryover, and onboard stability of reagent studies were evaluated. The performance of the busulfan immunoassay using the Abbott Alinity c was compared to the Beckman Coulter AU480 using sodium heparinized plasma, as well as to LC-MS/MS using lithium heparinized plasma. RESULTS: The imprecision goal of 8% was met, and linearity within the analytical measurement range of 240 to 1700â ng/mL was verified. Sample carryover was negligible, and the reagents were stable onboard for at least 84 days. The busulfan immunoassay correlated well with LC-MS/MS (slope = 0.949, y-intercept = -7.8â ng/mL, r2 = 0.9935) and the Beckman Coulter AU480 (slope = 1.090, y-intercept = -34.5â ng/mL, r2 = 0.9988). CONCLUSIONS: This study demonstrated successful analytical verification of a busulfan third-party immunoassay on the Abbott Alinity c platform. The ability to perform TDM of busulfan on a routine clinical chemistry analyzer will positively impact turnaround times to improve patient outcomes.
Subject(s)
Busulfan , Drug Monitoring , Busulfan/blood , Busulfan/pharmacokinetics , Humans , Immunoassay/methods , Drug Monitoring/methods , Drug Monitoring/instrumentation , Tandem Mass Spectrometry , Chromatography, Liquid/methods , Hematopoietic Stem Cell Transplantation/adverse effects , Reproducibility of ResultsABSTRACT
Therapeutic drug monitoring (TDM) of busulfan (BU) is currently performed by plasma sampling in patients undergoing hematopoietic stem cell transplantation (HSCT). Saliva samples are considered a noninvasive TDM matrix. Currently, no salivary population pharmacokinetics (PopPKs) model for BU available. This study aimed to develop a PopPK model that can describe the relationship between plasma and saliva kinetics in patients receiving intravenous BU. The performance of the model in predicting the area under the concentration-time curve at steady state (AUCss ) based on saliva samples is evaluated. Sixty-six patients with HSCT were recruited and administered 0.8 mg/kg BU intravenously. A PopPK model for saliva and plasma was developed using the nonlinear mixed effects model. Bayesian maximum a posteriori (MAP) optimization was used to estimate the model's predictive performance. Plasma and saliva PKs were adequately described with a one-compartment model and a scaled central compartment. Body surface area correlated positively with both clearance and apparent volume of distribution (Vd), whereas alkaline phosphatase correlated negatively with Vd. Simulations demonstrated that the percentage root mean squared prediction error and lower and upper limits of agreements reduced to 10.02% and -16.96% to 22.86% based on five saliva samples. Saliva can be used as an alternative matrix to plasma in TDM of BU. The AUCss can be predicted from saliva concentration by Bayesian MAP optimization, which can be used to design personalized dosing for BU.
Subject(s)
Busulfan , Drug Monitoring , Hematopoietic Stem Cell Transplantation , Saliva , Humans , Bayes Theorem , Busulfan/administration & dosage , Busulfan/analysis , Busulfan/blood , Busulfan/pharmacokinetics , Drug Monitoring/methods , East Asian People , Prospective Studies , Saliva/chemistry , Computer SimulationABSTRACT
OBJECTIVES: To apply therapeutic drug monitoring and dose-individualization of intravenous Busulfan to paediatric patients and evaluate the impact of syringe-pump induced Busulfan infusion lag-time after in vitro estimation. METHODS: 76 children and adolescents were administered 2 h intravenous Busulfan infusion every 6 h (16 doses). Busulfan plasma levels, withdrawn by an optimized sampling scheme and measured by a validated HPLC-PDA method, were used to estimate basic PK parameters, AUC, Cmax, kel, t1/2, applying Non-Compartmental Analysis. In vivo infusion lag-time was simulated in vitro and used to evaluate its impact on AUC estimation. KEY FINDINGS: Mean (%CV) Busulfan AUC, Cmax, clearance and t1/2 for pediatric population were found 962.3 µm × min (33.1), 0.95 mg/L (41.4), 0.27 L/h/kg (33.3), 2.2 h (27.8), respectively. TDM applied to 76 children revealed 6 (7.9%) being above and 25 (32.9%) below therapeutic-range (AUC: 900-1350 µm × min). After dose correction, all patients were measured below toxic levels (AUC < 1500 µm × min), no patient below 900 µm × min. Incorporation of infusion lag-time revealed lower AUCs with 17.1% more patients and 23.1% more younger patients, with body weight <16 kg, being below the therapeutic-range. CONCLUSIONS: TDM, applied successfully to 76 children, confirmed the need for Busulfan dose-individualization in paediatric patients. Infusion lag-time was proved clinically significant for younger, low body-weight patients and those close to the lower therapeutic-range limit.
Subject(s)
Bone Marrow Transplantation , Busulfan/administration & dosage , Drug Monitoring , Infusions, Intravenous/methods , Administration, Intravenous , Adolescent , Adult , Age Factors , Area Under Curve , Body Weight , Busulfan/blood , Busulfan/pharmacokinetics , Child , Child, Preschool , Dose-Response Relationship, Drug , Female , Humans , Immunosuppressive Agents/administration & dosage , Immunosuppressive Agents/blood , Immunosuppressive Agents/pharmacokinetics , Infant , Male , Pediatrics , Young AdultABSTRACT
BACKGROUND: Personalizing busulfan doses to target a narrow plasma exposure has improved the efficacy and lowered the toxicity of busulfan-based conditioning regimens used in hematopoietic cell transplant. Regional regulations guide interlaboratory proficiency testing for busulfan concentration quantification and monitoring. To date, there have been no comparisons of the busulfan pharmacokinetic modeling and dose recommendation protocols used in these laboratories. Here, in collaboration with the Dutch Association for Quality Assessment in Therapeutic Drug Monitoring and Clinical Toxicology, a novel interlaboratory proficiency program for the quantitation in plasma, pharmacokinetic modeling, and dosing of busulfan was designed. The methods and results of the first 2 rounds of this proficiency testing are described herein. METHODS: A novel method was developed to stabilize busulfan in N,N-dimethylacetamide, which allowed shipping of the proficiency samples without dry ice. In each round, participating laboratories reported their results for 2 proficiency samples (one low and one high busulfan concentrations) and a theoretical case assessing their pharmacokinetic modeling and dose recommendations. All participants were blinded to the answers; descriptive statistics were used to evaluate their overall performance. The guidelines suggested that answers within ±15% for busulfan concentrations and ±10% for busulfan plasma exposure and dose recommendation were to be considered accurate. RESULTS: Of the 4 proficiency samples evaluated, between 67% and 85% of the busulfan quantitation results were accurate (ie, within 85%-115% of the reference value). The majority (88% round #1; 71% round #2) of the dose recommendation answers were correct. CONCLUSIONS: A proficiency testing program by which laboratories are alerted to inaccuracies in their quantitation, pharmacokinetic modeling, and dose recommendations for busulfan in hematopoietic cell transplant recipients was developed. These rounds of proficiency testing suggests that additional educational efforts and proficiency rounds are needed to ensure appropriate busulfan dosing.
Subject(s)
Busulfan , Hematopoietic Stem Cell Transplantation , Busulfan/blood , Busulfan/pharmacokinetics , Humans , Laboratory Proficiency Testing , Quality Control , Transplantation ConditioningABSTRACT
Busulfan (Bu) is commonly used in myeloablative conditioning regimens for children undergoing hematopoietic stem cell transplantation. The standard target area under the concentration-time curve (AUC) of Bu is approximately 900-1500 µM min. In previous studies using five fixed doses (0.8-1.2 mg/kg) for Bu without dose adjustment, 75% patients achieved the target AUC. The aim of this pilot study was to determine the percentage of target AUC for intravenous (IV) Bu in Thai children. IV Bu was administered every 6 h over 16 doses. Blood samples were collected for pharmacokinetic (PK) analysis after the first, ninth, and thirteenth doses of Bu. Seven patients (2-14 years; median 6 years) were diagnosed with thalassemia (n = 4), acute myeloid leukemia (n = 2), and pure red cell aplasia. Three, two, and two patients received Bu at 1.1, 1.2, and 0.8 mg/kg, respectively. The AUC of Bu varied from 292-1714 µM min (median = 804). Nine (42.86%), eleven (52.38%), and one (4.76%) AUC values were within, below, and above the target, respectively. The median (range) Bu clearance was 5.93 (1.91-14.65) mL/min/kg. In this study, 42.86% AUC value achieved the target, which was lower than that in previous studies. Therapeutic drug monitoring (TDM) of Bu should be considered in Thai children receiving five fixed doses of IV Bu, and dose adjustment should be performed as necessary. Further PK studies for Bu with a larger sample size are warranted for confirming the necessity of TDM in every step dose of Bu.(Trial registration numbers; TCTR20190528003).
Subject(s)
Busulfan/therapeutic use , Hematopoietic Stem Cell Transplantation , Myeloablative Agonists/therapeutic use , Administration, Intravenous , Adolescent , Busulfan/administration & dosage , Busulfan/blood , Child , Child, Preschool , Drug Monitoring , Female , Humans , Male , Myeloablative Agonists/administration & dosage , Myeloablative Agonists/blood , Pilot Projects , Thailand , Transplantation ConditioningABSTRACT
BACKGROUND AND OBJECTIVES: Busulfan (Bu) is an old drug, but is still well recommended as an alkylating agent during conditioning therapy, before hematopoietic stem cell transplantation. Although its dose administration is standardized and based on patient weight, therapeutic drug monitoring is required in order to maintain its exposure [as area under the concentration-time curve (AUC) from 0 to infinity AUC0-∞] within a narrow therapeutic range and, if necessary, to adjust the dose with as short a lead time as possible. The aim of the study is to evaluate the agreement (as calculated AUC) between a gold standard analytical method and a new one that is faster and easier. METHODS: We analyzed 221 plasma samples from 37 children (0.25-16 years; 4-62.5 kg) and 11 adults (21-59 years; 45-80 kg), corresponding to 52 AUC values (ng h/mL). The drug exposure was calculated, simultaneously, by two validated analytical methods. The reference method was a high-performance liquid chromatography (HPLC) assay combined with an ultraviolet detector (UV). The test method had a triple quadrupole mass spectrometer (MS) as detector; the clean-up procedures of the samples were different and faster. RESULTS: The agreement between the two methods (reference and test) was evaluated in terms of Bu exposure differences based on Lin's concordance correlation coefficient (CCC) and represented by the Bland-Altman plot. The CCC between the AUC of the two methods was excellent (0.868; 95% CI: 0.802-0.935). The precision of the measures (expressed by Pearson's italic "r") was 0.872, and the accuracy (accounted by the bias correction factor) was 0.996. CONCLUSIONS: We can conclude that the HPLC-MS/MS assay represents a very good alternative to the reference.
Subject(s)
Alkylating Agents/administration & dosage , Alkylating Agents/blood , Busulfan/administration & dosage , Busulfan/blood , Drug Monitoring/standards , Adolescent , Adult , Child , Child, Preschool , Chromatography, High Pressure Liquid/methods , Chromatography, High Pressure Liquid/standards , Cohort Studies , Drug Monitoring/methods , Female , Humans , Infant , Infusions, Intravenous , Male , Mass Spectrometry/methods , Mass Spectrometry/standards , Middle Aged , Young AdultABSTRACT
Busulfan (Bu) is an alkylating agent commonly used in preparative regimens for hematologic malignant and non-malignant patients undergoing hematopoietic stem cell transplantation (HSCT). The objective of the present study was to develop an UPLC-MS/MS method for quantification of Bu in human plasma. A total of 55 patients with hematologic malignancies (n = 34) and non- malignancies (n = 21) received myeloablative Bu therapy prior to HSCT. A tandem mass spectrometric method was developed and validated to quantify Bu levels in these patients. The method was fully validated over the concentration range of 25-2000 ng/mL (r > 0.99). The assay method demonstrated good precision and accuracy. Stability studies indicated that the drug was stable in various conditions. Incurred sample reanalysis findings were within acceptable ranges (<15% of the nominal concentration). Based on the 1st dose AUC results, one third of hematologic malignant patients and half of non-malignant patients needed dose adjustment. However, in subsequent doses (5th, 9th, and 13th), 77%, 82% and 82%, respectively, of hematologic malignant patients and 71%, 67% and 86%, respectively, of non-malignant patients achieved the target range of Bu AUC. The suitability of the developed method for routine TDM of Bu in HSCT patients was demonstrated. The study suggests that the pharmacokinetic profile of Bu varies in both groups.
Subject(s)
Antineoplastic Agents, Alkylating/blood , Busulfan/blood , Chromatography, High Pressure Liquid/methods , Tandem Mass Spectrometry/methods , Adolescent , Adult , Antineoplastic Agents, Alkylating/administration & dosage , Antineoplastic Agents, Alkylating/therapeutic use , Busulfan/administration & dosage , Busulfan/therapeutic use , Child , Child, Preschool , Dose-Response Relationship, Drug , Drug Monitoring/methods , Female , Hematologic Neoplasms/drug therapy , Humans , Male , Middle Aged , Young AdultABSTRACT
A rapid and sensitive method for the quantitative detection of busulfan (BU) in children's hemolytic samples by HPLC-tandem mass spectrometry (MS/MS) was established. In this study, the sample preparation procedure involved a one-step protein precipitation with acetonitrile (ACN) solution, and the HPLC-MS/MS method used Hypersil GOLD C18 . The mobile phase consisted of 10 mM ammonium acetate solution (containing 0.1% formic acid) and ACN with a flow rate of 0.4 mL/min. Multiple reaction monitoring modes were used for quantitative analysis and the ion pairs of BU and BU-d8 were m/z 263.9 â 150.9 and 272.0 â 159.0, respectively. BU had a good linearity in the range of 0.01-10 µg mL-1 . The intra- and inter-day relative error was between -7.21% and 8.26%, and the coefficient of variation was less than 12.64%. The average extraction recovery rate in plasma samples was 99.76% ± 6.53%, and the matrix in normal plasma and hemolyzed plasma had no significant effect on the detection results. Normal and hemolytic samples could maintain good stability at 4, 25 and -40°C. As a result, this method is particularly suitable for determining BU in hemolytic samples from children with hematopoietic stem cell transplantation (HSCT), and this study provides the methodological basis for further research on the pharmacokinetics of BU in children with HSCT.
Subject(s)
Busulfan/blood , Chromatography, High Pressure Liquid/methods , Hematopoietic Stem Cell Transplantation , Tandem Mass Spectrometry/methods , Blood Specimen Collection , Busulfan/pharmacokinetics , Busulfan/therapeutic use , Child , Child, Preschool , Drug Monitoring/methods , Female , Hemolysis , Humans , Immunosuppressive Agents/blood , Immunosuppressive Agents/pharmacokinetics , Immunosuppressive Agents/therapeutic use , Linear Models , Male , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
Bone marrow transplantation is used to treat particular types of cancers such as lymphoma, leukemia, and multiple myeloma. Appropriate dosing of busulfan during the preparative phase is critical for a successful allograft; if blood concentrations get too high significant liver toxicity can occur, if blood concentrations are too low, then graft-versus-host disease (GVHD) can develop. Busulfan monitoring in blood allows hospitals with the opportunity to provide individualized medicine to patients and improve overall patient outcome. Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) is an important analytical method for quantification of busulfan in plasma in order to optimize the dose. © 2020 Wiley Periodicals LLC. Basic Protocol: Analysis of busulfan by liquid chromatography/mass spectrometry.
Subject(s)
Antineoplastic Agents, Alkylating/blood , Busulfan/blood , Chromatography, High Pressure Liquid , Drug Monitoring , Immunosuppressive Agents/blood , Spectrometry, Mass, Electrospray Ionization , Tandem Mass Spectrometry , Humans , Predictive Value of TestsABSTRACT
Determination of busulfan concentration in patients undergoing bone marrow transplantation is necessary in order to reduce toxic effects and/or graft rejection due to unadjusted dose exposure. A new extraction method namely dispersive micro solid phase extraction (DMSPE) based on mesoporous sorbent was used for cleaning-up the plasma samples. DMSPE coupling with liquid chromatography with tandem mass spectrometry (LC-MS/MS) was implemented for the determination of busulfan dosage in plasma samples. The linear range was found from 10 to 2000 ng/ml. The precision and accuracy were found better than 15% according to Food and drug Administration (FDA) guideline. This method was successfully used to determine the busulfan in patients administrated busulfan as part of the preparative regimen for bone marrow transplantation.
Subject(s)
Busulfan/isolation & purification , Immunosuppressive Agents/isolation & purification , Solid Phase Microextraction/methods , Adsorption , Busulfan/blood , Chromatography, High Pressure Liquid , Humans , Immunosuppressive Agents/blood , Plasma/chemistry , Tandem Mass SpectrometryABSTRACT
BACKGROUND: Therapeutic drug monitoring (TDM) for busulfan supports dose adjustment during conditioning for stem cell transplantation. The authors aimed to develop and validate limited sampling strategies (LSS) of 4-5 samples for a precise estimation of the area under concentration (AUC)-time curve of busulfan, in plasma as an alternative to an intensive sampling strategy (ISS) requiring 9-10 samples. METHODS: ISS TDM data from 297 patients (≤18 years of age) were used. AUCLSS was calculated using the trapezoidal rule and multiple linear regression (MLR). Unlike more complex modeling methods, MLR does not require sophisticated software or advanced training of personnel. MLR coefficients were estimated in the development subset containing randomly selected 50% of the records and were then used to calculate the AUCLSS of the remaining records (the validation subset). The agreement between dose adjustment recommendations (DAR) based on ISS and LSS, in the validation subset, was evaluated by a Bland-Altman analysis. A DAR deviating from an ISS-based reference by <15% was deemed acceptable. RESULTS: Twelve LSSs were acceptable. Sampling at 0, 120, 180, and 240 minutes after the start of the second infusion (LSS15) yielded the best performance, with DAR deviating from the reference by <10% for 95% of cases; the AUCLSS was determined as follows: AUCLSS = 74.7954 × C(0) + 81.8948 × C(120) + 38.1771 × C(180) + 138.1404 × C(240) + 54.1837. This LSS and LSS13 performed similarly well in an independent external validation. CONCLUSIONS: MLR-based estimates of AUCLSS provide DARs that deviate minimally from the reference. LSSs allow the reduction of patient discomfort, a â¼50% reduction of TDM-related workload for nursing staff and blood loss and a â¼25% reduction in laboratory workload. These benefits may encourage wider use of busulfan TDM, supporting safe and efficacious personalized dosing.
Subject(s)
Busulfan/blood , Drug Monitoring/methods , Immunosuppressive Agents/blood , Adolescent , Age Factors , Area Under Curve , Body Surface Area , Body Weight , Busulfan/administration & dosage , Busulfan/pharmacokinetics , Child , Child, Preschool , Dose-Response Relationship, Drug , Female , Hematopoietic Stem Cell Transplantation/methods , Humans , Immunosuppressive Agents/administration & dosage , Immunosuppressive Agents/pharmacokinetics , Linear Models , Male , Sex FactorsABSTRACT
BACKGROUND: Busulfan (Bu) is one of the conditioning regimen components for pediatric hematopoietic stem cell transplantation. Bu therapeutic drug monitoring (TDM) is essential for a successful treatment outcome and toxicity evasion. Dried blood spot (DBS) sampling is a rapid and simple method for Bu TDM, compared with conventional plasma sampling. This study evaluated the feasibility of using the DBS method for Bu TDM. The hematocrit (Hct) and conditioning day were also examined for their impact on the DBS method's performance. METHODS: Venous blood collected from 6 healthy volunteers was diluted, using their plasma into 4 samples of varying Hct values. Each sample was spiked with Bu calibrators (300, 600, and 1400 ng/mL), prepared using DBS and dried plasma spot (DPS) sampling and analyzed using a validated liquid-chromatography tandem-mass spectrometry method. Clinical blood samples (n = 153) from pediatric patients (n = 15) treated with Bu (mainly from doses 1, 2, 5, and 9) were used to prepare paired volumetric DBS and DPS samples. A Bland-Altman plot and Deming regression were used to define the agreement between the paired DBS and DPS measurements. Passing-Bablok regression analyses investigated the effects of Hct and conditioning day on the linearity between both methods. RESULTS: In vitro analyses showed good agreement between DBS and DPS measurements, with a mean difference of -5.4% and a 95% confidence interval on the limits of agreement of -15.3% to 4.6%. Clinical samples showed good correlation (Pearson correlation coefficient = 0.96; slope = 1.00) between the DBS and DPS methods. The DBS method met the clinical acceptance limits for clinical samples, with a bias <±20%. Bland-Altman plots showed good agreement, with only 5.8% of paired measurements exceeding the limits of agreement (±1.96 SD), although within its 95% confidence interval. Hct observations ranged from 21.7% to 34.7% and did not affect Bu concentrations measured from DBS in either the in vitro or in vivo studies. CONCLUSIONS: These results show that DBS is a useful method for Bu TDM, provided samples are analyzed on the collection day. DBS sampling offers advantages over traditional plasma sampling in infants and younger children because only small volumes of blood are required.
Subject(s)
Antineoplastic Agents, Alkylating/blood , Busulfan/blood , Dried Blood Spot Testing , Drug Monitoring/methods , Child , Cohort Studies , HumansABSTRACT
Busulfan and melphalan are cytotoxic DNA alkylating agents that are used in many hematopoietic stem cell transplantation (HCT) conditioning regimens. We report the development of an assay using turbulent flow liquid chromatography (TFLC) and tandem mass spectrometry to simultaneously measure the concentration of busulfan (Bu) and melphalan (Mel) in human plasma. The method involves precipitating proteins in the plasma specimen with an organic solvent containing deuterated internal standards of both compounds. Following centrifugation, an aliquot of the supernatant was injected into the TFLC mass spectrometry system operated in the positive ion mode. The analytical measurement range for both compounds was 10-5000â¯ng/mL, and with validated dilutions the reportable range was extended to 25,000â¯ng/mL. Intra-day and inter-day (nâ¯=â¯20â¯day) precision studies showed a coefficient of variation (CV) of <7% at several concentrations across the measurement range. To determine accuracy recovery studies were performed at several concentrations spanning the measurement range. Recoveries for both compounds were between 98 and 103%. Additionally, busulfan was compared with an existing assay and showed excellent correlation. Experiments were conducted to rule out matrix effects, carryover and interference from endogenous substances. The validated clinically reportable range (CRR) and assay precision will allow this assay to be used clinically to monitor and adjust Mel and Bu levels to ensure better therapeutic outcomes and also to support clinical trials aimed at better defining therapeutic ranges.
Subject(s)
Alkylating Agents/blood , Busulfan/blood , Immunosuppressive Agents/blood , Melphalan/blood , Tandem Mass Spectrometry/methods , Chromatography, High Pressure Liquid/methods , DNA/chemistry , HumansABSTRACT
A methodology is proposed, based on Raman spectroscopy coupled with multivariate analysis, to determine the Limit of Detection (LOD) and Limit of Quantification (LOQ) for therapeutic drug monitoring in human serum, using the examples of Busulfan, a cell cycle non-specific alkylating antineoplastic agent, and, Methotrexate, a chemotherapeutic agent and immune system suppressant. In this study, ultrafiltration is employed to fractionate spiked human pooled serum to efficiently recover the drug in the filtrate prior to performing Raman analysis. The drug concentration ranges were chosen to encompass the recommended therapeutic ranges and toxic levels in patients. Raman spectra were collected from the filtrates in the liquid form, using an inverted backscattering microscopic geometry, using 532 nm as source. Finally, prediction models were built by using Partial Least Squares Regression (PLSR) and LOD and LOQ were calculated directly from the linear prediction models. The LOD calculated for Busulfan is 0.0002 ± 0.0001 mg mL-1, 30-40 times lower than the level of toxicity, enabling the application of this method in target dose adjustment of Busulfan for patients undergoing, for example, bone marrow transplantation. The LOD and LOQ calculated for Methotrexate are 7.8 ± 5 µM and 26 ± 5 µM, respectively, potentially enabling high dose monitoring. The promising results obtained from this study suggest the potential of Raman spectroscopy for therapeutic drug monitoring of drugs in bodily fluids.
Subject(s)
Busulfan/blood , Methotrexate/blood , Antineoplastic Agents/blood , Drug Monitoring/methods , Humans , Least-Squares Analysis , Limit of Detection , Multivariate Analysis , Myeloablative Agonists/blood , Spectrum Analysis, Raman/methodsABSTRACT
OBJECTIVE: Busulfan, frequently used as a conditioning regimen for hematopoietic stem cell transplantation, has a narrow therapeutic range and wide intra-and interpatient variabilities. Therefore, therapeutic drug monitoring of busulfan is necessary to ensure that the drug concentrations of patients are within a targeted therapeutic range. In this study, we developed a simple and accurate method for measuring busulfan concentrations using liquid chromatography tandem mass spectrometry (LC-MS/MS). METHODS: Separation and detection of busulfan was performed using T3 column equipped with LC-MS/MS. Busulfan was isolated from 50 µL human plasma after mixing with busulfan-2H8 (internal standard) solution, calibrator, and quality-control material. The sample was eluted and gradated with a mobile phase composed of ammonium acetate, formic acid, and water or methanol. The busulfan concentration was quantified using a six-point standard curve. Busulfan and busulfan-2H8 were detected in positive-ion multiple-reaction-monitoring mode. According to the Clinical and Laboratory Standards Institute guideline, we verified the precision, linearity, limit of detection (LOD), limit of quantification (LOQ), and carryover. RESULTS: Busulfan and busulfan-2H8 were detected at m/z 264.1>151.1 and 272.2>159.1. The total run time was 3 min. Both intra-and inter-assay coefficients of variation were <3%. The calibration curve was linear at 25-5,000 ng/mL. The LOD and LOQ were 2.5 ng/mL and 25 ng/mL, respectively. The recoveries ranged from 92.0-104.8% and the carryover was-0.02%. CONCLUSIONS: Our method for busulfan reduces total run time and has excellent analytical performance. It will be a useful method for therapeutic drug monitoring of busulfan in clinical laboratories.
Subject(s)
Busulfan/blood , Chromatography, Liquid/methods , Drug Monitoring , Tandem Mass Spectrometry/methods , Humans , Quality Control , Reference Standards , Reproducibility of ResultsABSTRACT
BACKGROUND: Pharmacokinetics (PK) is the study of the absorption, distribution, metabolism, and excretion of drugs. Many chemotherapeutic agents have a sensitive PK index, in which a small margin in blood concentrations is the difference between nontherapeutic, therapeutic, and adverse outcomes. OBJECTIVES: This article will provide an overview of evidence-based approaches to the collection of PK samples, monitoring of PK levels, and the resulting management of patients undergoing PK testing. METHODS: A case study involving busulfan, an alkylating agent used in the pre-stem cell transplantation setting, will highlight the cross-contamination of samples while a drug is being infused through a central venous catheter with PK sample collection from a proximal peripherally inserted central catheter. The influence of false elevations in drug concentrations on PK-guided dose adjustments will also be emphasized. FINDINGS: Imprecise blood collections or cross-contamination of samples may lead to inaccurate drug concentration results and, subsequently, undesired low or high drug dosage calculations.
Subject(s)
Antineoplastic Agents, Alkylating/blood , Busulfan/blood , Drug Monitoring/methods , Neoplasms/blood , Neoplasms/drug therapy , Aged , Antineoplastic Agents, Alkylating/pharmacokinetics , Antineoplastic Agents, Alkylating/therapeutic use , Busulfan/pharmacokinetics , Busulfan/therapeutic use , Humans , Male , Neoplasms/nursing , Vascular Access DevicesABSTRACT
This study aimed to predict the area under the curve (AUC) of the initial busulfan dose using a test dose with the sparse sampling scheme in adult patients who underwent hematopoietic cell transplant. A test dose of 0.8 mg/kg busulfan was used 2 days before twice-daily intravenous busulfan-based conditioning regimens were administered. The AUC and the clearance (CL) were calculated for both the test dose and the first dose (AUCT , CLT , AUC1, and CL1 ) by noncompartmental analysis. The sparse sampling schemes of the test dose were developed by Bayesian method based on the population pharmacokinetic model. The optimal sparse sampling schemes were determined by evaluating the mean prediction error, the root mean square error, the absolute mean prediction error, and Bland-Altman plot. The mean AUC1 was 7.20 ± 1.48 mg ⢠h/L, which ranged from 4.70 to 9.46 mg ⢠h/L. The AUC1 was below the therapeutic concentration of 7.38 mg ⢠h/L in 45% (9 of 20) of the patients. The CLT of 3.05 ± 0.56 mL/min/kg was not significantly different with the CL1 of 3.03 ± 0.69 mL/min/kg (P = .901). A sampling scheme at 2 and 6 hours after the test dose was developed to predict the AUCT (mean prediction error of 1.64%, root mean square error of 6.17%, and absolute mean prediction error of 4.94%). Additionally, the Bland-Altman plot showed that the 2-sampling scheme provided an acceptably accurate prediction of the AUC1 . A test dose with a 2-sampling scheme was sufficient to personalize the initial busulfan dosing in hematopoietic cell transplant recipients.
Subject(s)
Busulfan/pharmacokinetics , Hematologic Neoplasms/therapy , Hematopoietic Stem Cell Transplantation/methods , Transplantation Conditioning/methods , Adult , Area Under Curve , Bayes Theorem , Blood Specimen Collection/methods , Busulfan/administration & dosage , Busulfan/blood , Cyclophosphamide/administration & dosage , Cyclophosphamide/blood , Cyclophosphamide/pharmacokinetics , Deoxycytidine/administration & dosage , Deoxycytidine/analogs & derivatives , Deoxycytidine/blood , Deoxycytidine/pharmacokinetics , Drug Administration Schedule , Drug Dosage Calculations , Drug Monitoring/methods , Etoposide/administration & dosage , Etoposide/blood , Etoposide/pharmacokinetics , Female , Hematologic Neoplasms/blood , Hematologic Neoplasms/metabolism , Humans , Male , Middle Aged , Vidarabine/administration & dosage , Vidarabine/analogs & derivatives , Vidarabine/blood , Vidarabine/pharmacokinetics , GemcitabineABSTRACT
Currently, there is an urgent need to establish the optimal dosing of TREO in conditioning prior to hematopoietic stem cell transplantation, especially in children. For that purpose, pharmacokinetic analyses are ongoing within clinical phase II and III trials. In this paper, HPLC methods for determination of prodrug treosulfan and/or its biologically active epoxides in human plasma or serum are reviewed for the first time, including the spectrum of analytes being quantified, detection type, and derivatization methodology. The major focus is addressed to the stability of TREO and its monoepoxide related with different strategies of patients' blood processing, e.g. blood pH lowering to different values, no pH adjustment; centrifugation of blood immediately after collection or within a few hours later. This issue is crucially important for the robust bioanalysis because the epoxytransformation of TREO is a nonenzymatic, highly pH and temperature-dependent reaction. In-depth analysis of the literature results demonstrates that some methodologies of blood treatment could produce the systematic underestimation of TREO concentrations. Consequently, the drug clearance and volume of distribution will be overestimated, which might false the association of the drug exposure with the regimen-related toxicity and clinical outcomes. The paper indicates the deficiencies of the blood processing strategies and offers hints for their refinement. The provided information ought to be important in the current investigations of the personalized TREO pharmacokinetics.