Search

Abstract Dasatinib, a potent oral multi-targeted kinase inhibitor against Src and Bcr-Abl, can decrease inflammatory response in sepsis. A simple and cost-effective method for determination of an effective dose dasatinib was established. This method was validated in human plasma, with the aim of reducing the number of animals used, thus, avoiding ethical problems. Dasatinib and internal standard lopinavir were extracted from 180 uL of plasma using liquid-liquid extraction with methyl tert-butil ether, followed by liquid chromatography coupled to triple quadrupole mass spectrometry in multiple reaction monitoring mode. For the pharmacokinetic study, 1 mg/kg of dasatinib was administered to mice with and without sepsis. The method was linear over the concentration range of 1-98 ng/mL for DAS in mice and human plasma, with r2>0.99 and presented intra- and interday precision within the range of 2.3 - 6.2 and 4.3 - 7.0%, respectively. Further intra- and interday accuracy was within the range of 88.2 - 105.8 and 90.6 - 101.7%, respectively. The mice with sepsis showed AUC0-t = 2076.06 h*ng/mL and Cmax =102.73 ng/mL and mice without sepsis presented AUC0-t = 2128.46 h*ng/mL. Cmax = 164.5 ng/mL. The described analytical method was successfully employed in pharmacokinetic study of DAS in mice.

Copanlisib is a pan phosphatidylinositol 3-kinase (PI3K) inhibitor approved for follicular lymphoma. In this paper, we present the data of development and validation of a high-performance liquid chromatography (HPLC) method for the quantitation of copanlisib in mice plasma as per the FDA regulatory guideline. The method involves the extraction of copanlisib along with internal standard (IS, enasidenib) from mice plasma (100 µL) using ethyl acetate as an extraction solvent. The chromatographic resolution of copanlisib and the IS was achieved on a Hypersil Gold C18 column maintained at 40 °C using a binary gradient mobile phase [10 mM ammonium formate (pH 4.0) and acetonitrile]. The flow-rate was 0.8 mL/min. For the detection of copanlisib and the IS, the photo-diode array detector was set at λmax 310 nm. Copanlisib and the IS eluted at 6.60 and 7.80 min, respectively with a total run time of 10 min. The calibration curve was linear over a concentration range of 50 to 5000 ng/mL for copanlisib (r2³ 0.998). The results of intra- and inter-day accuracy and precision studies were within the acceptable limits. Copanlisib was stable on bench-top, in auto-sampler, up to three freeze/thaw cycle and long-term storage at -80 °C. The application of the validated method was shown in a mice pharmacokinetic study.

[Display omitted] • RP-HPLC method was developed for the quantitative estimation of raloxifene from mice plasma. • The RT of raloxifene and internal standard was found to be 2.258 min and 5.575 min, respectively. • Tamoxifen citrate is used as the internal standard. • The developed method applied successfully to pharmacokinetics. Raloxifene hydrochloride (RLX) belongs to the Selective estrogen receptor modulator (SERM) class of drugs, exhibiting diverse affinity to estrogen receptors. The drug is known for its anti-cancer and anti-osteoporosis activity. The current study employs a Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method that is sensitive and time-efficient for quantitative evaluation of RLX from mice plasma, using tamoxifen citrate as an internal standard (IS). The protein precipitation technique was used to extract the RLX and IS. RLX was separated using a C8 column with an isocratic elution of a mobile phase consisting of acetonitrile and Milli-Q water (pH 3) at a 1 mL/min flow rate. The ultraviolet (UV) detection of the analyte and IS were carried out at 287 nm. The linearity plot was constructed in the range of 100–3200 ng/mL with a correlation coefficient of r2 ≥ 0.9996. The inter-day and intra-day accuracy and precision were found to be within acceptable limits. Stability studies revealed that the analyte remained stable. The validated RP-HPLC method was successfully applied to quantify RLX after oral administration in mice, and the pharmacokinetic (PK) parameters were established. The present RP-HPLC method could be a beneficial tool for the fast and reliable estimation of future PK interactions, PK analysis of formulations, and therapeutic drug monitoring. [ABSTRACT FROM AUTHOR]

A simple, selective and rapid LC-ESI-MS/MS method has been developed and validated for the quantification of ivosidenib in mice plasma using warfarin as an internal standard (I.S.) as per regulatory guideline. Sample preparation was accomplished through a simple protein precipitation process. Chromatography of ivosidenib and the I.S. was achieved on an Atlantis dC18 column using an isocratic mobile phase comprising 0.2 % formic acid in water and acetonitrile (25:75, v/v) delivered at a flow rate of 1.0 mL/min. LC-MS/MS was operated under the multiple reaction-monitoring mode (MRM) using the electrospray ionization technique in positive ion mode and the transitions of m/z 583.1→186.1 and m/z 309.2→251.3 were used to quantitate ivosidenib and the I.S, respectively. The total chromatographic run time was 2.0 min. Linearity was established in the concentration range of 1.10-3293 ng/mL (r2>0.99). The intra- and inter-day accuracy and precision for ivosidenib in mice plasma were in the range of 5.72-9.91 and 5.90-10.7 %, respectively. Ivosidenib was found to be stable on bench-top for 6 h, up to three freeze-thaw cycles, in in-injector for 24 h and for one month at -80 °C. The applicability of the validated method has been demonstrated in a mice pharmacokinetic study. Following intravenous (2 mg/kg) and oral (5 mg/kg) administration of ivosidenib to mice, concentrations were quantifiable up to 24 and 48 h, respectively. The bioavailability was 61 %.

We developed and validated a simple, sensitive, selective, and reliable LC–MS/MS–ESI method for the direct quantitation of lumefantrine (LFN) enantiomers [(−)‐LFN and (+)‐LFN] in mice plasma as per regulatory guideline. LFN enantiomers and carbamazepine (internal standard) were extracted from mice plasma using Strata X SPE (solid‐phase extraction) cartridges. Good resolution between enantiomers was achieved on a Chiralpak IA‐3 column using an isocratic mobile phase (0.1% of diethyl amine in methanol), which was delivered at a flow rate of 0.8 mL/min. Detection and quantitation were performed using multiple reaction monitoring mode following the transitions m/z 530.27 → 512.30 and 237.00 → 194.00 for LFN enantiomers and the internal standard, respectively, in the positive‐ionization mode. The proposed method provided accurate and reproducible results over the linearity range of 2.39–895 ng/mL for each enantiomer. The intra‐ and inter‐day precisions were in the range of 1.03–6.14 and 6.36–8.70 and 2.03–4.88 and 5.82–11.5 for (−)‐LFN and (+)‐LFN, respectively. Both (−)‐LFN and (+)‐LFN were found to be stable under different stability conditions. The method was successfully used to delineate stereoselective pharmacokinetics of LFN enantiomers in mice after an oral administration of rac‐LFN (20 mg/kg). The pharmacokinetic results indicated that the disposition of LFN enantiomers was stereoselective in mice. [ABSTRACT FROM AUTHOR]

Isorhapontigenin (ISO), a novel derivative of stilbene compound, possessess good pharmacological activities such as antiviral, antioxidant, and anticancer. The purpose of this study is to investigate the pharmacokinetic of ISO in mice plasma, after oral administration of three doses (40, 80, and 160 mg/kg). Isorhapontigenin was detected by liquid chromatography–tandem mass spectrometry (LC-MS/MS), and a reliable LC-MS/MS assay for ISO in mice plasma was developed. The method was linear over a concentration range of 5–2000 ng/ml, and the lower limit of quantification was 5 ng/ml. The results indicated that there was a linear relationship between AUC(0–t), AUC(0–∞), and Cmax of ISO and dosages. ISO underwent quick absorption and elimination in mice. [ABSTRACT FROM AUTHOR]

A simple and sensitive high performance liquid chromatography with fluorescence detection (HPLC–FD) has been developed for simultaneous quantification of doxorubicin (DOX) and its dipeptide conjugate prodrug (PDOX) in mice plasma. The chromatographic separation was carried out on an Amethyst C18–H column with gradient mobile phase of 0.1% formic acid and 0.1% formic acid in acetonitrile at a flow rate of 1.0 mL/min. The excitation and emission wavelengths were set at 490 and 550 nm, respectively. The method was comprehensively validated. The limits of detection were low up to 5.0 ng/mL for DOX and 25.0 ng/mL for PDOX. And the limits of quantification were low up to 12.5 ng/mL for DOX and 50 ng/mL for PDOX, which were lower than those for most of the current methods. The calibration curves showed good linearity (R2>0.999) over the concentration ranges. The extraction recoveries ranged from 84.0% to 88.2% for DOX and from 85.4% to 89.2% for PDOX. Satisfactory intra-day and inter-day precisions were achieved with RSDs less than 9.1%. The results show that the developed HPLC–FD method is accurate, reliable and will be helpful for preclinical pharmacokinetic study of DOX and PDOX.

A novel, fit-for-purpose, highly sensitive, analytical UPLC-PDA methodology was developed and fully validated, according to ICH, FDA and EMA guidelines, for the rapid and accurate quantification of trans -crocin 4 (TC4) and crocetin (CRC) in mice plasma and brain after i.p. administration. A PDA based methodology shows a wider applicability as it is cost effective and can be easily and seamlessly adopted by the pharma industry. The separation of the analytes was performed on a C18 Hypersil Gold column with 2.5 min run time, employing the internal standard (ISTD) methodology. The two methods were successfully applied for the determination of CRC and TC4 in mouse plasma and brain after i.p . administration of TC4 (50 mg/kg) in a time range of 0–240 min. Due to the selection of i.p. administration route, the first-pass metabolism and/or gastric hydrolysis were bypassed, a fact that enhanced the bioavailability of TC4. Furthermore, TC4 was found to be capable of crossing the Blood Brain Barrier (BBB) and build up levels in the mouse brain, regardless of its highly hydrophilic character. CRC was not detected in any plasma or brain sample, although it has been reported that TC4 quickly hydrolyzes to CRC after p.o. administration. Therefore i.p. administration could be used in the case of TC4 for the accurate determination of its biological role. Overall, the developed methodology offers important information about the bioavailability of TC4 in mouse plasma and for the first time, demonstrates the ability of TC4 to penetrate the BBB and localize inside the brain. [ABSTRACT FROM AUTHOR]

A sensitive and rapid LC–MS/MS method was developed and validated for the simultaneous quantitation of enzalutamide, N -desmethylenzalutamide (active metabolite of enzalutamide), apalutamide, darolutamide and ORM-15341 (active metabolite of darolutamide) in mice plasma as per regulatory guidelines. The analytes and the internal standard (I.S.: apalutamide- d 3 ) were extracted from 50 μL mice plasma by simple protein precipitation using acetonitrile, followed by chromatographic separation using an Atlantis C 18 column with an isocratic mobile (0.2% formic acid:acetonitrile; 30:70, v/v) at a flow rate of 0.8 mL/min within 2.5 min. Detection and quantitation was done by multiple reaction monitoring on a triple quadrupole mass spectrometer following the transitions: m / z 465 → 209, 451 → 195, 478 → 450, 399 → 178, 397 → 194 and 481 → 453 for enzalutamide, N -desmethylenzalutamide, apalutamide, darolutamide, ORM-15341 and the I.S. respectively. The calibration curves were linear from 1.07 to 2000 ng/mL with r 2 ≥0.99 for all the analytes. The intra- and inter-batch accuracy and precision (% CV) across quality controls varied from 88.5–111% and 1.13–13.1, 85.4–106% and 3.15–14.3, respectively for all the analytes. All the analytes were found to be stable under different conditions. The method was applied to an intravenous pharmacokinetic study in mice. [ABSTRACT FROM AUTHOR]

A sensitive, specific, selective and rapid LC-ESI–MS/MS method has been developed and validated for the quantification of apalutamide in mice plasma using apalutamide- d 3 as an internal standard (I.S.). Sample preparation was accomplished through a simple protein precipitation process. Chromatography of apalutamide and the I.S. was achieved on an Atlantis dC 18 column using an isocratic mobile phase comprising 0.2% formic acid in water and acetonitrile (20:80, v/v) delivered at a flow rate of 0.8 mL/min. LC–MS/MS was operated under the multiple reaction-monitoring mode (MRM) using the electrospray ionization technique in positive ion mode and the transitions of m / z 478 → 450 and m / z 481 → 453 were used to measure the derivative of apalutamide and the I.S, respectively. The total chromatographic run time was 2.5 min and the elution of apalutamide and I.S. occurred at 1.10 and 1.09 min, respectively. Method validation was performed as per regulatory guidelines and the results met the acceptance criteria. Linearity was established in the concentration range of 1.02–2030 ng/mL ( r  > 0.995) for apalutamide. The intra- and inter-day accuracy and precision for apalutamide in mice plasma were in the range of 2.11–8.44 and 2.51–6.09%, respectively. Apalutamide was found to be stable under various stability conditions. This novel method has been applied to a pharmacokinetic study in mice. [ABSTRACT FROM AUTHOR]

A sensitive and rapid LC-MS/MS method was developed and validated for the simultaneous quantitation of five selective KCNQ channel openers, namely ICA-27243, ML-213, PF-05020182, SF-0034 and flupirtine in mice plasma as per regulatory guideline. The analytes and the internal standard (IS; flupirtine-d4) were extracted from 50 µL mice plasma by liquid-liquid extraction, followed by chromatographic separation using an Atlantis C18 column with an isocratic mobile phase comprising 0.2% formic acid: acetonitrile (20:80, v/v) at a flow rate of 0.6 mL min−1 within 2.5 min. Detection and quantitation was done by multiple reaction monitoring on a triple quadrupole mass spectrometer following the transitions: m/z 268.9 → 140.8, 258.1 → 95.1, 367.2 → 269.1, 322.2 → 248.2, 305.7 → 196.4 and 309.1 → 196.1 for ICA-27243, ML-213, PF-05020182, SF-0034, flupirtine and the IS, respectively, in the positive ionization mode. The calibration curves were linear from 1.00 to 2008 ng mL−1 for all the analytes with r2 ≥ 0.99. The intra- and inter-batch accuracy and precision (% CV) across quality controls varied from 90.0 to 113 and 2.64 to 13.0; 93.8 to 114 and 3.15 to 14.9%, respectively, for all the analytes. Analytes were found to be stable under different stability conditions. The method was applied to a pharmacokinetic study in mice. [ABSTRACT FROM AUTHOR]

A sensitive, specific, selective and rapid LC-ESI–MS/MS method has been developed and validated for the quantification of defactinib in mice plasma using 13 C 3, 15 N-tofacitinib as an internal standard (I.S.). Sample preparation was accomplished through a liquid–liquid extraction process. Baseline chromatographic resolution of defactinib and the I.S. was achieved on an Atlantis dC 18 column using an isocratic mobile phase comprising 0.2% formic acid in water and acetonitrile (25:75, v/v) delivered at a flow rate of 0.5 mL/min. Defactinib and the I.S. eluted at ∼1.59 and 0.99 min, respectively. The total chromatographic run time was 2.50 min. A linear response function was established in the concentration range of 0.13–106 ng/mL. Method validation was performed as per regulatory guidelines and the results met the acceptance criteria. The intra- and inter-day accuracy and precision were in the range of 5.57–13.3 and 8.63–12.1%, respectively. Defactinib was found to be stable under various stability conditions. This novel method has been applied to a pharmacokinetic study in mice. [ABSTRACT FROM AUTHOR]

A sensitive, specific and rapid LC-ESI–MS/MS method has been developed and validated for the quantification of tideglusib in mice plasma using warfarin as an internal standard (I.S.) as per regulatory guidelines. Sample preparation was accomplished through liquid-liquid extraction process. Chromatographic separation was performed on Atlantis dC 18 column using mobile phase A (acetonitrile) and B (5 mM ammonium acetate in water) in a flow-gradient mode. Elution of tideglusib and the I.S. occurred at ∼2.06 and 1.29 min, respectively. The total chromatographic run time was 3.2 min. A linear response function was established in the concentration range of 20.2–1008 ng/mL. The intra- and inter-day accuracy and precision were in the range of 4.61–12.6 and 6.04–11.8%, respectively. This novel method has been applied to a pharmacokinetic study in mice. [ABSTRACT FROM AUTHOR]

A sensitive and rapid LC–MS/MS method was developed and validated for the simultaneous quantitation of darolutamide and its active metabolite i.e. ORM-15341 in 50 μL mice plasma using bicalutamide as an internal standard (I.S.) as per regulatory guidelines. Sample processing was accomplished through liquid-liquid extraction. Chromatographic separation was achieved using an Atlantis C 18 column with an isocratic mobile phase comprising 0.2% formic acid:acetonitrile (35:65, v/v) at a flow rate of 0.8 mL/min within 2.5 min. Detection and quantitation were done by multiple reaction monitoring on a triple quadrupole mass spectrometer following the transitions: m / z 397 → 202, 395 → 202 and 429 → 255 for darolutamide, ORM-15341 and I.S, respectively in the negative ionization mode. The calibration curve was linear from 0.61–1097 ng/mL for both darolutamide and ORM-15341. The intra- and inter-day precisions were in the range of 1.34–13.8 and 4.85–12.9 and 3.91–13.7 and 6.54–14.2%, for darolutamide and ORM-15341, respectively. Darolutamide and ORM-15341 were found to be stable under different stability conditions. The validated method was applied to a pharmacokinetic study in mice. [ABSTRACT FROM AUTHOR]

A highly sensitive, specific and enantioselective assay has been validated for the quantitation of OTX015 enantiomers [(+)-OTX015 and (−)-OTX015] in mice plasma on LC-MS/MS-electrospray ionization as per regulatory guidelines. Protein precipitation was used to extract (±)-OTX015 enantiomers and internal standard (IS) from mice plasma. The active [(−)-OTX015] and inactive [(+)-OTX015] enantiomers were resolved on a Chiralpak-IA column using an isocratic mobile phase (0.2% ammonia/acetonitrile 20 : 80, v/v) at a flow rate of 1.2 mL/min. The total run time was 6.0 min. (+)-OTX015, (−)-OTX015 and IS eluted at 3.34, 4.08 and 4.77 min, respectively. The MS/MS ion transitions monitored were m/z 492 → 383 for OTX015 and m/z 457 → 401 for IS. The standard curves for OTX015 enantiomers were linear ( r2 > 0.998) in the concentration range 1.03-1030 ng/mL. The inter- and intraday precisions were in the range 2.20-13.3 and 8.03-12.1% and 3.80-14.4 and 8.97-13.6% for (+)-OTX015 and (−)-OTX015, respectively. Both the enantiomers were found to be stable in a battery of stability studies. This novel method has been applied to the study of stereoselective oral pharmacokinetics of (−)-OTX015 and unequivocally demonstrated that (−)-OTX015 does not undergo chiral inversion to its antipode in vivo in mice. [ABSTRACT FROM AUTHOR]

A new electrochemiluminescence (ECL) sensor based on reduced graphene oxide-copper sulfide (rGO-CuS) composite coupled with capillary electrophoresis (CE) was constructed for the ultrasensitive detection of amlodipine besylate (AML) for the first time. In this work, rGO-CuS composite was synthesized by one-pot hydrothermal method and used for electrode modification. The electrochemical and ECL behaviors of the sensor were investigated. More than 5-fold enhance in ECL intensity was observed after modified with rGO-CuS composite. The results can be ascribed to the presence of rGO-CuS composite on the electrode surface that facilitates the electron transfer rate between the electroactive center of Ru(bpy) 3 2+ and the electrode. The ECL sensor was coupled with CE to improve the selectivity and the CE-ECL parameters that affect separation and detection were optimized. Under the optimum conditions, the linear ranges for AML was 0.008–5.0 μg/mL with a detection limit of 2.8 ng/mL (S/N=3). The method displayed the advantages of high sensitivity, good selectivity, wide linear range, low detection limit and fine reproducibility, and was used to analyze AML in mice plasma with a satisfactory result, which holds a great potential in the field of pharmaceutical analysis. [ABSTRACT FROM AUTHOR]

A sensitive, specific and rapid LC-ESI–MS/MS method has been developed and validated for the quantification of ulixertinib in mice plasma using phenacetin as an internal standard (I.S.) as per regulatory guidelines. Sample preparation was accomplished through a protein precipitation procedure with acetonitrile:methanol mixture. Chromatographic separation was performed on Atlantis dC 18 column using a binary gradient using mobile phase A (0.2% formic acid in water) and B (acetonitrile) at a flow rate of 0.60 mL/min. Elution of ulixertinib and I.S. occurred at ∼1.07 and 1.20 min, respectively. The total chromatographic run time was 2.5 min. A linear response function was established in the concentration range of 1.58–2054 ng/mL. The intra- and inter-day accuracy and precisions were in the range of 2.11–11.8 and 5.80–11.4%, respectively. This novel method has been applied to a pharmacokinetic study in mice. [ABSTRACT FROM AUTHOR]

A simple and sensitive high performance liquid chromatography with fluorescence detection (HPLC–FD) has been developed for simultaneous quantification of doxorubicin (DOX) and its dipeptide conjugate prodrug (PDOX) in mice plasma. The chromatographic separation was carried out on an Amethyst C 18 –H column with gradient mobile phase of 0.1% formic acid and 0.1% formic acid in acetonitrile at a flow rate of 1.0 mL/min. The excitation and emission wavelengths were set at 490 and 550 nm, respectively. The method was comprehensively validated. The limits of detection were low up to 5.0 ng/mL for DOX and 25.0 ng/mL for PDOX. And the limits of quantification were low up to 12.5 ng/mL for DOX and 50 ng/mL for PDOX, which were lower than those for most of the current methods. The calibration curves showed good linearity ( R 2 >0.999) over the concentration ranges. The extraction recoveries ranged from 84.0% to 88.2% for DOX and from 85.4% to 89.2% for PDOX. Satisfactory intra-day and inter-day precisions were achieved with RSDs less than 9.1%. The results show that the developed HPLC–FD method is accurate, reliable and will be helpful for preclinical pharmacokinetic study of DOX and PDOX. [ABSTRACT FROM AUTHOR]

A simple, rapid and highly sensitive spectrofluorimetric method was developed for determination of a novel type of dopamine receptor antagonist LE300 in mouse plasma. The method is based on measuring the native fluorescence of LE-300 in methanol at 343 nm after excitation at 280 nm. The fluorescence concentration plot was rectilinear over the range of 3.5-100 ng/mL with a lower detection limit of 1.0 ng/mL and quantification limit of 3.5 ng/mL. The method was statistically validated for linearity, accuracy, precision and selectivity according to the International Conference on Harmonization guidelines. The accuracy and precision results was expressed as % recovery and relative standard deviation (RSD). The accuracy for LE-300 was in the range 95.5-103.6% and RSD values were in the range of 0.21-1.55% of the theoretical value. The method was successfully applied to the analysis of LE-300 in mice plasma. The results were compared statistically with those obtained by the reported method and were found to be in good agreement, which could be applied in a pharmacokinetic study. [ABSTRACT FROM AUTHOR]

[Display omitted] • A new magnetic graphene oxide functionalized SBA-15 was synthesized. • AMT and NOR were extracted by a dual-template imprinted polymer. • Separetion was carried out based on micro solid phase extraction. • The behavior of variables was accurately modeled and predicted. • The method was applied in mice plasma samples after taking two drugs. In this study, a new magnetic dual-template imprinted polymer modified with functionalized SBA-15 and graphene oxide (Fe 3 O 4 @SBA-15-NH-GO/dtMIP) was synthesized using surface imprinting technique. The synthesized polymer was used for preconcentration of amitriptyline and nortriptyline in plasma of mice (male BALB / cJRj mice) using µ-solid phase extraction (µ-SPE) and simultaneous determination by HPLC technique. The mice plasma was studied at regular intervals after taking two drugs. The polymer was characterized by Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), X-ray energy diffraction spectroscopy (XRD) , vibrating sample magnetometer (VSM), and Nitrogen adsorption/desorption analysis. By studying the response surface methodology (RSM) and desirability function (DF), the optimal experimental condition was determined as 16.72 min for ultrasonic time, 8.11 for pH and 40.51 mg for polymer dosage. The results of the analysis were reported for amitriptyline in mice plasma in the concentration range 6.0–950.0 and the detection limit was achieved as 1.5 ng mL −1 respectively. For nortriptyline the LDR and LOD was reported as 5.0–850.0 and 1.1 ng mL −1 respectively. [ABSTRACT FROM AUTHOR]

A sensitive and simple LC method for the quantification of ginkgolic acids in mice plasma has been developed. Following acetonitrile deproteinization, samples were separated on a SinoChrom ODS-AP C18 column. The mobile phase was 3% ( v/ v) acetic acid water solution–methanol (8:92, v/v) at a flow rate of 1.0 mL min−1. Detection was at 310 nm. Calibration curve was linear over the range of 0.25–50 μg mL−1 with intra- and inter-day precisions (RSD%) of less than 9.5%. The extraction recovery ranged from 87.0 to 90.2% (RSD 2.4–6.4%) for ginkgolic acids. The method was successfully applied to the pharmacokinetic study of ginkgolic acids in mice after oral dosing of 1.0 g kg−1. [ABSTRACT FROM AUTHOR]

Abstract: For the evaluating of the absorption, tumor affinity and metabolism of wilfoside C3N and wilfoside C1N (two main C21 steroidal glycosides from Baishouwu), an LC–MS/MS method was developed. Plasma or tumor homogenate samples were extracted by liquid–liquid extraction with ethyl acetate after internal standard (ginsenoside Rh2) spiked. The separation was performed by a Luna C18 column (3.0μm, 2.0mm×50mm) with gradient elution. The method was fully validated and successfully applied to determine the concentrations of the parent drugs and metabolites after intragastric administration of wilfoside C3N and wilfoside C1N to mice respectively. [Copyright &y& Elsevier]

• i.p. administration of TC4 resulted in alterations to the mice metabolic profiles. • MVA allowed sufficient clustering between the treated and the untreated animals. • ML-(s)PLS–DA, was the only effective approach to detect any interior clustering. • Indication of a preliminary sex-related effect on the metabolome. • Annotation of 25 Metabolites including two neuroactive agents. Trans -crocin 4 (TC4) is an important carotenoid constituent of saffron showing potential activity against Alzheimer's Disease (AD) due to its antioxidant and antiamyloidogenic properties. Metabolomics is an emerging scientific field that enhances biomarker discovery and reveals underlying biochemical mechanisms aiming towards the early subclinical diagnosis of diseases. To date, there are no reports on the changes induced to mice plasma metabolome after TC4 administration. We report a novel untargeted UHPLC-ESI HRMS metabolomics strategy to determine the alteration of the metabolic fingerprint following i.p. administration of TC4 in male and female mice. Blood samples from fiftysix mice treated with TC4 as well as from control animals were analyzed with UHPLC-ESI HRMS. Statistical evaluation of the results was achieved by multivariate analysis (MVA), i.e., principal component analysis (PCA), Partial Least Squares–Discriminant Analysis (PLS-DA) in order to discover the variables that contributed to the discrimination between treated and untreated groups which were identified by online database searching (e.g., Metlin, HMDB, KEGG) aided by chemometric processing, e.g., covariance searching etc. Due to the high variability imposed by various factors, e.g., sex of the animals participating in the study, administration dose and time-points of sacrifice, multilevel sparse PLS-DA analysis, e.g., splitting variation to each individual component, has been employed as a more efficient approach for such designs. This methodology allowed the identification of the time sequence of metabolome changes due to the administration of TC4, whereas a sex-related effect on the metabolome is indicated, denoting that the administration in both sexes is indispensable in order to acquire safe conclusions as reliable metabolome pictures. The results demonstrated a number of annotated metabolites playing a potential role in neuroprotection while they are closely related to AD. Moreover, five additional annotated metabolites were involved in the steroid biosynthesis pathway while two of them may be considered as putative neuroprotective agents. [ABSTRACT FROM AUTHOR]

We developed and validated a simple, sensitive, selective, and reliable LC-MS/MS-ESI method for the direct quantitation of lumefantrine (LFN) enantiomers [(-)-LFN and (+)-LFN] in mice plasma as per regulatory guideline. LFN enantiomers and carbamazepine (internal standard) were extracted from mice plasma using Strata X SPE (solid-phase extraction) cartridges. Good resolution between enantiomers was achieved on a Chiralpak IA-3 column using an isocratic mobile phase (0.1% of diethyl amine in methanol), which was delivered at a flow rate of 0.8 mL/min. Detection and quantitation were performed using multiple reaction monitoring mode following the transitions m/z 530.27 → 512.30 and 237.00 → 194.00 for LFN enantiomers and the internal standard, respectively, in the positive-ionization mode. The proposed method provided accurate and reproducible results over the linearity range of 2.39-895 ng/mL for each enantiomer. The intra- and inter-day precisions were in the range of 1.03-6.14 and 6.36-8.70 and 2.03-4.88 and 5.82-11.5 for (-)-LFN and (+)-LFN, respectively. Both (-)-LFN and (+)-LFN were found to be stable under different stability conditions. The method was successfully used to delineate stereoselective pharmacokinetics of LFN enantiomers in mice after an oral administration of rac-LFN (20 mg/kg). The pharmacokinetic results indicated that the disposition of LFN enantiomers was stereoselective in mice., (© 2020 John Wiley & Sons, Ltd.)

Copanlisib is a pan phosphatidylinositol 3-kinase (PI3K) inhibitor approved for follicular lymphoma. In this paper, we present the data of development and validation of a high-performance liquid chromatography (HPLC) method for the quantitation of copanlisib in mice plasma as per the FDA regulatory guideline. The method involves the extraction of copanlisib along with internal standard (IS, enasidenib) from mice plasma (100 μL) using ethyl acetate as an extraction solvent. The chromatographic resolution of copanlisib and the IS was achieved on a Hypersil Gold C 18 column maintained at 40 °C using a binary gradient mobile phase [10 mM ammonium formate (pH 4.0) and acetonitrile]. The flow-rate was 0.8 mL/min. For the detection of copanlisib and the IS, the photo-diode array detector was set at λ max 310 nm. Copanlisib and the IS eluted at 6.60 and 7.80 min, respectively with a total run time of 10 min. The calibration curve was linear over a concentration range of 50 to 5000 ng/mL for copanlisib (r 2 ≥ 0.998). The results of intra- and inter-day accuracy and precision studies were within the acceptable limits. Copanlisib was stable on bench-top, in auto-sampler, up to three freeze/thaw cycle and long-term storage at -80 °C. The application of the validated method was shown in a mice pharmacokinetic study., Competing Interests: Conflict of interest: The authors are scientists at Jubilant Biosys Ltd., (Copyright © 2020 by the authors.)