Migraine is a complex neurological condition that often involves a severe headache characterized by moderate to severe throbbing. Lasmiditan a novel ditan approved by USFDA in 2011 to treat migraine. This contemporary research elaborates the systematic development and validation of Reverse Phase-High Performance Liquid Chromatography bio analytical method for estimating Lasmiditan in human plasma employing analytical quality by design. Bio analysis employing protein precipitation was performed to extract Lasmiditan from human plasma using Dolutegravir as the internal standard. Design of Experiments was incorporated employing central composite design, and the methodology was assessed for critical analytical attributes by varying the critical quality attributes using Design Expert Software Version 13.0. The chromatographic conditions optimized were symmetry C18 (150 × 4.6mm, 5μ) column, mobile phase 0.1% orthophosphoric acid: acetonitrile (70:30 (v/v) with a flow rate of 1.0mL/min at detection wavelength of 260nm and column temperature of 30°C. Validation studies indicated linearity between 20 and 800ng/mL (r2 = 0.999), whereas accuracy and precision portrayed a 98.79% recovery. Succinctly, current work exemplifies the significant merits of analytical quality by design approach toward the holistic process in bioanalytical method validation. Furthermore, the method was evaluated using the Whiteness Assessment Tool (Blue Applicability Grade Index), yielding a high score of 82.5, indicating superior performance in terms of analytical efficiency, environmental friendliness and operator safety. Thus, the developed method can be applied for estimation of preclinical and clinical samples of Lasmiditan with improved accuracy and precision.

Per- and polyfluoroalkyl substances (PFAS) are an emerging group of environmental contaminants widely used in consumer products. Public unease regarding the levels of these compounds in the environment has called for methods to keep up with emerging PFAS and regulations. There is a need for analytical methods that are efficient and robust that can be readily used and easily accessed in commercial laboratories. This work discusses the development of an efficient, sensitive and reliable liquid chromatography tandem mass spectrometry method for the analysis of 40 PFAS in aqueous matrices (10min run time). Initial calibration demonstrated excellent linearity for all PFAS analytes with r 2 values greater than 0.997. Limit of detections (0.2-100ng/L) and limit of quantifications (1-350ng/L) show the ability for the method to reach necessary levels to stay up to date with federal and state regulations. Two solid phase extraction techniques were compared based on recovery accuracy and precision. Recoveries ranged from 92.0% to 105.0% and 67.0% to 109.0% for the automated and manual setups, respectively. Evidence was found to support improvements in sample recovery using an automated setup. The validated method was used to analyze samples from six natural water sources in the Baltimore-Metro area, where seven of the forty PFAS compounds were found in at least one sample.

A novel and validated reverse-phase high-performance liquid chromatography (RP-HPLC) approach was established for the concurrent measurement of cyproconazole (CYP) isomers and tebuconazole (TBZ) in suspension concentrate (SC) agrochemical formulations. The approach employed a C18 column with a gradient elution of 0.1% formic acid in water and methanol, attaining baseline resolution of CYP isomer-1, isomer-2 and TBZ without requiring chiral columns or sample pretreatment. Linearity was confirmed within the range of 80-120% of the target concentration, with a R2 value of 0.999. Accuracy varied from 101.2% to 102.8%, with a %RSD of less than 0.8%. LC-MS analysis verified analyte identity through distinctive ion transitions (CYP: m/z 292, TBZ: m/z 308). This is the initial method for isomer-specific detection of CYP using RP-HPLC within a formulation matrix. The methodology is straightforward, resilient and appropriate for standard quality assurance and regulatory adherence in agrochemical manufacturing.

Itraconazole is an oral triazole antimycotic drug. Bioequivalence studies are cornerstones for the approval of generic drug development globally. The present study describes a simple, sensitive and economical LC-MS/MS method for the determination of itraconazole and its metabolite in human plasma. Itraconazole (drug), hydroxyitraconazole (metabolite), and atorvastatin as internal standard were added to plasma samples. The standard curve of both the drug and metabolite covering the 4-320ng/mL concentration range, was linear (R2 = 0.999). Also, a limit of quantification of 4ng/mL of a sample size of 0.4mL is achieved which is comparable or even better than the reported methods. The applicability of this method was proven by analyzing true samples obtained after the administration of 100mg itraconazole of test (Noxifunge®) and reference (Sporanox®) to healthy volunteers. The 90% confidence intervals of the logarithmically transformed AUC0-72, AUC0-∞, and Cmax of the two formulations both for the drug and the metabolite are within the accepted levels proposed by FDA and EMA. Therefore, the presented method is suitable for bioavailability, pharmacokinetic, and bioequivalent studies in humans.

Natural deep eutectic solvents (NaDESs) was efficient and green for componential extraction of traditional Chinese medicine compared to conventional organic solvents. In this study, a total of nine NaDESs were synthesized and screened, and Proline-Urea (Pro-Ur) was screened as extract solvent for experiments. Five bioactive components (neochlorogenic acid, chlorogenic acid, rutin, isoquercitrin and kaempferol-3-O-rutinoside) were selected as target components. The optimal process was as follows: NaDES content of 60%, solid to liquid ratio of 80mg/mL, extract temperature of 70°C and extract time of 40min. The predicted comprehensive value of extraction yield was 0.4364. The IC50 of Betaine-Urea on PC12 cells was ~0.3% (v/v).

A molecularly imprinted polymer (MIP) was synthesized for the selective extraction of oleanolic acid (OA), employing OA as the template molecule, acrylamide as the functional monomer, ethylene glycol dimethacrylate as the cross-linking agent, azobisisobutyronitrile as the initiator, and chloroform as the porogenic solvent. The characterization of the obtained MIPs was evaluated by scanning electron microscopy, Fourier transform infrared spectroscopy, Brunauer-Emmett-Teller (BET) analysis and X-ray photoelectron spectroscopy. The MIPs reached adsorption equilibrium within 2hours to OA, with high adsorption capacity of 124.68mg/g. Subsequently, these MIPs were packed in empty solid-phase extraction (SPE) cartridge to enrichment OA from Ligustrum lucidum fruit extract. The parameters of SPE were optimized as follows: loading 0.5ml of a 1.0mg/ml OA reference solution, washing with 0.5ml of toluene, and eluting with 4ml of methanol. Under these conditions, the enrichment rate of OA from L. lucidum extract reached 76.0%. Additionally, an high-performance liquid chromatography method for determining OA content was validated. This study provides an effective approach for the separation and enrichment of OA from complex matrices and also provides practical feasibility for the separation and enrichment of other triterpenoids.

In this paper, a new headspace gas chromatography (HSGC) method has been developed for the determination of ethylene oxide (EO) and 1,4-Dioxane (Dioxane) in bulk lots of polyethylene glycols (PEG). PEG samples are dissolved in dimethyl sulfoxide, heated in a headspace oven maintained at 90°C for 10min and then injected into the GC system for analysis. Analytes are separated through a thermal gradient elution from 36 to 240°C on an Agilent DB-624 column (60m × 0.53mm diameter, film thickness 3.00μm). The carrier gas is helium and detector is flame ionization detector. Total run time of the new HSGC method is about 18min. In the new HSGC method, other common residual solvents present in PEGs such as methanol (MeOH), methyl formate (MF) and acetyl aldehyde (Acetal) are also sufficiently separated from EO and Dioxane peaks. In the HSGC methods prescribed in USP monograph <228> and EP monograph <2.4.25>, EO co-elutes with MF. The new HSGC method was successfully validated according to current ICH/VICH guidelines and was found to be specific, linear, accurate, precise, robust and sensitive. This HSGC method is fast and quality control (QC) friendly and suitable for routine analysis for the determination of EO and Dioxane in bulk batches of PEGs in QC laboratories.

We developed a reversed-phased high-performance liquid chromatographic method combining ultraviolet detection and integrated pulsed amperometric detection for the simultaneous quantification of dopamine, 5-hydroxyindolacetic acid, homovanillic acid, serotonin, 3,4-dihydroxyphenylacetic acid, norepinephrine and epinephrine. All target components were completely separated in a C18 column with isocratic elution of 5% acetonitrile solution containing 8mM HClO4 and 0.20mM 1-octanesulfonic acid as an ion pairing reagent. This method showed limits of detection of 0.03-0.1ng and limits of quantification of 0.10-0.3ng with linear regression coefficients of 0.9998-1.0000. All inter-day and intra-day precision values were below 9.58%, and the average recoveries were 93.71-109.82% for mouse striatum samples. The mean levels of the seven components in striatal brain tissue in a mouse model of Parkinson's disease decreased by 2-12 times compared to those of a control group. In particular, the decrease in dopamine, 5-hydroxyindolacetic acid and 3,4-dihydroxyphenylacetic acid was statistically significant (P < 0.05). Orthogonal partial least squares discriminant analysis confirmed that the seven components are useful biomarkers. The significance of the developed method lies in its ability to simultaneously analyze seven biogenic amines related to Parkinson's disease by combining two detectors, offering a simple and cost-effective approach for clinical and biological labs.

In this research study, a simple, rapid, sensitive, and cost-effective Reverse Phase High Performance Liquid Chromatography method was developed and validated for determination of Cetirizine and Levocetirizine in human plasma. The drugs were separated on an ACE Generix100-5 C18 RP (250 × 4.6mm, 5μm) column, using Acetonitrile and Distilled water pH7 (59:41v/v) as mobile phase with a flow rate of 1.5mL/min, retention time of 3.39 ± 0.02min, detection wavelength of 230nm, and column oven temperature at 26°C. The drug was extracted from human plasma using Methanol, the percent recovery was 97%. The LOD and LOQ for Cetirizine and Levocetirizine were 0.5ng and 1ng. The method was linear over the concentration range of 0.5-1000ng /mL for Cetirizine and Levocetirizine. The method was applied for analysis of drugs in plasma following oral administration of Cetirizine (10mg) and Levocetirizine (5mg) in smokers and nonsmokers of Pashtun ethnicity belonging from different districts of Khyber Pakhtunkhwa, Pakistan. The difference between Cetirizine plasma levels in smokers and non-smokers was significant with p -value 0.0279.