Asarum heterotropoides (AH) is a notable medicinal plant in traditional Chinese medicine (TCM), but distinguishing and assessing its quality from various origins is challenging due to similar morphology and chemical profiles, using traditional chemical analysis methods. In this study, we developed an effective approach to identify AH from different origins by combining high-performance liquid chromatography (HPLC) fingerprint with chemometrics. The contents of three characteristic constituents (Asarinin, sesamin and methyleugenol) were selected for quantitive analysis, it found that the content of asarinin, sesamin and methyleugenol in Jilin province ranged from 3.393 ± 0.003-3.704 ± 0.004 mg/g, 1.081 ± 0.037-1.182 ± 0.008 mg/g, and 2.429 ± 0.010-2.968 ± 0.186 mg/g, respectively, whereas the range in Liaoning province was 1.275 ± 0.025-1.947 ± 0.013 mg/g, 0.180 ± 0.012-0.905 ± 0.055 mg/g, and 1.089 ± 0.006-1.624 ± 0.064 mg/g. Moreover, the constituents in the methanol extract of AH were analyzed by ultra-performance liquid chromatography coupled to a quadrupole exactive orbitrap mass spectrometer (UPLC-Q-Exactive MS) operating in positive electrospray ionization (ESI+) mode with MS/MS acquisition. A total of 22 compounds were identified, including eight associated with various diseases such as alzheimer's disease, cardiovascular disease, bronchial spasm and chronic inflammatory disease, suggesting their potential therapeutic benefits. This research establishes a theoretical basis for quality control of AH and proposes a framework for source identification methods for other medicinal and economic plants.

A rapid, simple and highly sensitive stability-indicating high-performance liquid chromatography (HPLC) method was developed and validated for the estimation of asciminib in human plasma. Plasma samples (100 μL) were processed by solid-phase extraction. Chromatographic separation was performed using a CAPCELL PAK C18 MGII column (4.6 × 250 mm, 5 μm); the mobile phase consisted of 0.5% potassium dihydrogen phosphate (KH2PO4, pH 3.5): acetonitrile: methanol (50:45:5, v/v/v) at a flow rate of 0.5 mL/min. Detection was carried out at a wavelength of 250 nm. The developed method showed good linearity over the concentration range of 10-5000 ng/mL (correlation coefficient: 0.9999). The method was then validated according to guidelines published by the US Food and Drug Administration. The coefficients of variation for intra- and interday assays for asciminib were less than 13.9%, whereas accuracies for intra- and interday assays were within 8.9%. The limit of quantification was 10 ng/mL. The mean (minimum - maximum) plasma trough concentration of asciminib in 29 Japanese patients with chronic myeloid leukemia who received repeated administration of 40 mg twice daily was 367 (101-807) ng/mL. The validated method can be used as a routine method to support pharmacokinetic studies of asciminib.

With the rising interest in therapeutic potential of microdosing lysergic acid diethylamide (LSD), accurate quantification and stability analysis are imperative for both scientific research and end-user safety. Currently, high throughput quantification of LSD in clinical microdosing formulations and community sourced microdosing samples remain a challenge due to the absence of fast analytical methods. This study aimed to develop a robust, stability indicating, isocratic reverse-phase-high performance liquid chromatography (RP-HPLC) method for the accurate and precise quantification of LSD. Importantly, the method was capable of separating LSD from its primary-degradation product, iso-LSD. Adhering to International Conference on Harmonization guidelines, the method was validated for various parameters such specificity, linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ) and robustness. The developed method proved its efficacy in quantifying LSD even in the presents of degradants induced by various recommended stress conditions. This method was applied to quantify LSD in illicit microdosing samples obtained from New Zealand's unique community drug checking service and found a significant discrepancy between user-estimated and actual LSD levels. This highlights the method's critical importance in providing reliable data for the safety and quality control of LSD microdosing in pharmaceutical and illicit formulations and highlights potential risks for users in using non-pharmaceutically prepared microdosing formulations.

3,14,19-triacetyl andrographolide (ADA), a derivative of andrographolide, has emerged as a promising anti-inflammatory compound. In the present study, we established a rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry method for the quantification of ADA in rat plasma. ADA and celecoxib (internal standard, IS) were separated using an ACQUITY UPLC BEH C18 chromatographic columns with a gradient mobile phase of acetonitrile (containing 0.1% formic acid)/water (containing 0.1% formic acid) at a flow rate of 0.3 mL/min. Quantification was performed using the multiple reaction monitoring transitions to m/z 499.1 → 439.1 for ADA, m/z 381.9 → 361.9 for celecoxib, with plasma samples undergoing liquid-liquid extraction. The method effectively detected ADA in plasma and exhibited excellent linearity (R 2 > 0.9921) over concentrations ranging from 5 to 2000 ng/mL. Both within-run and between-run precision (%RSD) were less than 6.88%, and accuracy ranged from 94.86% to 107.61%. Matrix effects were 87.53% and 112.78%, with recoveries ranging from 98.60% to 104.39%. ADA was proven to be stable under different storage conditions (4°C and -70°C) in plasma. The validated method was successfully applied to determine ADA concentrations in rat plasma. The results demonstrated that ADA exhibited a noticeable improvement in bioavailability compared with andrographolide.

An Ultra Performance Liquid Chromatography - Mass Spectrometry/Mass Spectrometry (UPL-MS/MS) method for simultaneous determination of 8, 11, 12, 20- hydroxyeicosatetraenoic acids (HETEs) and 13 s-hydroxyoctadecadienoic acid (13 s-HODE) in plasma was developed and optimized. The chromatographic separation of 8, 11, 12, 20-HETEs and 13 s-HODE was achieved on a Peptide Bridged Ethylene Hybrid Particles (BEH) C18 column (2.1 mm × 150 mm, 1.7 μm) with mobile phase A (Water containing 0.1% formic acid) and mobile phase B (acetonitrile containing 0.1% formic acid). HETEs and 13 s-HODE were quantified using multiple reaction monitoring mode. The developed UPLC-MS/MS method was applied to determine the plasma samples of 32 paraquat (PQ) poisoning patients, 20 diquat (DQ) poisoning patients and 38 healthy subjects. The results showed that the 8, 11, 12, 20-HETEs and 13 s-HODE have good linearity (R2 > 0.99) from 0.1-500 ng/mL. The levels of HETEs and 13 s-HODE were dramatically increased in PQ and DQ poisoning patients. There were high correlation between 8, 11, 12, 20-HETEs and 13 s-HODE. Orthogonal partial least squares discrimination analysis showed that three groups were distributed in different areas and separated well. In conclusion, a sensitive UPLC-MS/MS method had been developed and validated for determination of 8, 11, 12, 20-HETEs and 13 s-HODE in plasma of PQ and DQ patients.

Pine bark extract (PBE) is a material used as a health food. Its functionality is attributed to the antioxidant effect of its contained procyanidins, and quantification of the procyanidin content is important to monitor its quality. However, conventional single-column High-Performance Liquid Chromatography (HPLC) methods often lack sufficient separation for procyanidins in complex matrices like PBE, leading to inaccurate quantification. In order to improve the accuracy of analysis for procyanidins B1 (PCB1) and B3 (PCB3) contained in PBE, we developed a highly accurate analytical method employing heart-cutting HPLC using columns with multiple properties and performed a detailed validation. The PCB1 and PCB3 contained in commercial health foods ranged from 0.43-2.95 mg/g, with a relative standard deviation of 0.77-6.17%. The spike and recovery tests showed that the recovery of beverages ranged from 98 to 106% at low and high concentrations, and for tablets, the recovery ranged from 95 to 118%. The repeatability relative standard deviation ranged from 1.10 to 3.57%, and the Horwitz Ratio ranged from 0.26-1.89. The intermediate relative standard deviation ranged from 2.62 to 6.83%. Single-laboratory validation of the analytical method employing heart-cutting HPLC showed that it is sufficiently accurate for analyzing PCB1 and PCB3 in commercial health foods containing PBE.

Sensitive and specific eco-friendly chromatographic approaches were developed to quantitatively determine Dacarbazine along with its United States Pharmacopeia (USP) related impurities simultaneously. Method I was reveresed phase HPLC (RP-HPLC) by which separation of the ternary mixture was achieved. Zorbax Eclipse Plus octadecyl column as stationary phase, a mobile phase formed of double distilled water and methanol at a ratio of (70: 30, v/v) and 0.6 mL/min as a flow rate were used for isocratic separation. Ultra violet (UV) detection was adjusted at 230 nm. Method II is high performance thin layer chromatography (HPTLC), where the separation was performed on silica gel plates using green developing system composed of butanol: ethyl acetate: 0.05 M ammonium acetate (2: 4: 0.4, by volume) with UV detection at 280 nm. Green behavior of the developed techniques was evaluated. The International Council for Harmonisation (ICH) guidelines were considered for the proposed approaches validation. Moreover, Dacarbazine was successfully determined in Dacarbazine medac® powder for injection by the proposed techniques. No significant difference aroused between the results of the developed techniques and the results aroused from the official USP spectrophotometric method. Furthermore, the proposed methods had the merits of being more friendly towards the environment, sensitive and time saver than other reported HPLC methods.

A simple, sensitive, and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was successfully developed and validated to determine navitoclax and doxorubicin in rat plasma. Ketoconazole and daunorubicin were employed as internal standards to ensure accurate quantification and method consistency. The sample preparation involved a straightforward protein precipitation technique, which facilitated efficient extraction of the analytes from the plasma matrix. The resulting supernatant was subjected to drying using a vacuum concentrator and later reconstituted before being injected into the LC-MS system. Separation was carried out using a SHIMADZU Shim-pack GIST C-18 column with mobile phase, consisting of a carefully balanced mixture of acetonitrile and water with 0.1% formic acid, with a flow rate of 0.5 mL/min. This composition ensured efficient elution and minimized matrix effects, contributing to the method's robustness and reproducibility. The developed method was comprehensively validated according to regulatory guidelines, assessing various parameters including specificity, selectivity, sensitivity, calibration curve performance, precision, accuracy, stability and dilution integrity. This bioanalytical method has the potential to be extended to various clinical settings, enabling the monitoring drug metabolism, monitoring potential drug-drug interactions and understanding the pharmacokinetic profile and adverse effect potential of the combination therapy.

Polymer monoliths are stationary-phase materials for liquid chromatography and solid-phase extraction. Their porous structure, tuneability and simple synthesis enable tailoring to specific analysis requirements in analytical chemistry. Typically, polymer monoliths are used to separate larger biomolecules. Due to their lower binding capacity, the applications of polymer monoliths for the chromatographic separation of small molecules remain limited. However, recent literature has shown that polymer monoliths have the potential for the extraction of small molecules. In this research, butyl methacrylate-co-ethylene glycol dimethacrylate polymer monoliths were synthesized using localized UV polymerization in capillaries. The performance of reversed-phase polymer monoliths in automated in-line solid-phase extraction-mass spectrometry was demonstrated by the analysis of endocannabinoids from neat standard mixes and spiked cell culture media without prior sample preparation. The synthesized monoliths exhibited a binding capacity of 1896 pmol. Furthermore, we showed the repeatability of the monolith synthesis, with a variance in permeability of 19%. The system's stability is demonstrated through the analysis of multiple batches, comparing different monoliths and reusing the same monolith repeatedly, resulting in relative standard deviations (RSDs) below 20% for all extracted compounds. This automated method with hyphenated mass spectrometry improves throughput over previous manual monolithic extractions for small molecules.

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 4-320 ng/mL concentration range, was linear (R2 = 0.999). Also, a limit of quantification of 4 ng/mL of a sample size of 0.4 mL 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 100 mg 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.

Antidiabetic drugs, including metformin hydrochloride, are typically used to treat diabetes mellitus. Current methods for evaluating these drug combinations often involve the use of harmful solvents and specific conditions, leading to environmental risks and substantial toxic waste. White analytical chemistry proposes an innovative solution to this problem by developing environmentally friendly, cost-effective and user-friendly chromatographic methods. A synchronous chromatographic assay method for multiple combined pharmaceutical dosage forms of antidiabetic drugs was developed using a single set of chromatographic conditions and safe organic solvents. Method development was carefully carried out using a design of experiments approach to reduce organic solvent waste. The Plackett-Burman design was used in the screening phase to identify critical method variables and responses, facilitated by Minitab 18 software. The Box-Behnken design was then used to optimize these critical variables. A synchronous chromatographic assay of multiple combined pharmaceutical dosage forms was conducted under optimized chromatographic conditions. Finally, a thorough evaluation of the proposed and existing chromatographic methods was conducted using green and white analytical chemical metrics.

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.

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 80 mg/mL, extract temperature of 70°C and extract time of 40 min. The predicted comprehensive value of extraction yield was 0.4364. The IC50 of Betaine-Urea on PC12 cells was ~0.3% (v/v).

A screening method was developed for the determination of per- and polyfluoroalkyl substances in textiles by combustion and ion chromatography. In this work, a new type of high-temperature combustion absorption device was designed, and the samples were burned, cracked and gasified by temperature programmed heating mode. The produced free fluorine and hydrogen fluoride gases were absorbed by water vapor and completely transformed into inorganic fluoride anion. After condensation and collection, it was separated and determined by ion chromatography. The pre-treatment conditions were optimized including the sample weight, the combustion method, the combustion gas and its flow rate, the water evaporation rate and the condensate collection method. Method validation was performed in negative samples of cotton, wool and polyester, at spiked levels of 5, 50 and 200 mg/kg, respectively. The average recoveries were between 85.7 and 95.0% with relative standard deviations <20% (n = 7) indicating satisfactory accuracy and repeatability. The calibration curve was performed with the correlation coefficients (R2) higher than 0.999 within a linearity range of 0.01-2.0 mg/L. The limit of quantification was 5.0 mg/kg. The developed method was successfully used to determine the total fluorine in market samples. It can meet the screening determination of PFAS in various textiles.

Sodium N-lauroylsarcosinate (NLS) and sodium methyl cocoyl taurate (SMCT) are widely used in sulfate free personal care products. We present a straightforward and highly sensitive High-Performance Liquid Chromatographic (HPLC) method for simultaneous detection and estimation of these substances in commercial personal care products. The HPLC method has a detection limit for NLS of concentrations as low as 1.5 ppm, and that for SMCT of 4.0 ppm, all with a signal-to-noise (S/N) ratio exceeding 3.0. Additionally, the method enables quantification down to 4.5 ppm for NLS, and 12.0 ppm for SMCT, with an S/N ratio surpassing 10.0. This validated method serves as a versatile tool for the accurate quantification of NLS, and SMCT in sulfate free personal care products like shampoos, face washes, body washes, baby shampoos, and toothpastes.

The combination of metformin (MET), teneligliptin (TEN), and pioglitazone (PIO) is newest fixed-dose combination for the treatment type 2 diabetes launched in India. An accurate, rapid, and cost-effective reversed phase ultra-fast liquid chromatography method was developed, validated, and applied for the quantification of MET, TEN, and PIO in bulk and tablet with a very short runtime. Box-Behnken design was implemented to optimize the ultra-fast liquid chromatography conditions. A C18 column (150 × 4.6 mm, 5μ) was utilized with a mobile phase buffer (0.01 M phosphate buffer, pH = 6.2) and acetonitrile in a 51:49 ratio with flow rate of 1.5mL/min at column oven temperature 40°C. Detection was carried out at 255nm by 20 μL injection volume. Retention times were found to be 0.94, 1.36, and 2.07 min, whereas the limit of quantification were 0.209, 0.712, and 57.030 μg/mL for MET, TEN, and PIO, respectively. The linearity of anticipated development was studied in the range of 250-1250 μg/mL (MET, r2 = 0.99974), 10-50 μg/mL (TEN, r2 = 0.99997), and 7.5-37.5μg/mL (PIO, r2 = 0.99999). The relative standard deviation values for method and intermediate precision were <2%. The method was validated as per International Conference on Harmonization guideline for accuracy, precision, linearity, limit of detection, limit of quantification, specificity, robustness, and forced degradation. This is the only study that presents reversed phase ultra-fast liquid chromatography method for the new fixed-dose combination of MET, TEN, and PIO, with the shortest run time of 3.0 min. The proposed method is innovative, efficient, precise, quickest, and economical with high accuracy which makes the study novel.

The rhizome of Dioscorea bulbifera (RDB) has been used in folk and traditional Chinese medicine for the treatment of thyroid diseases and cancer. However, the chemical constituents are not fully clear. In this study, high-performance liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry was successfully used for the comprehensive characterization of chemical constituents in RDB. As a result, a total of 40 compounds were identified in RDB by comparison with the authentic standards or reported references, including 13 flavonoids, 2 diterpenoids, 2 stilbenoids, 12 steroidal saponins, and 11 phenolic derivatives. This study comprehensively elucidated the chemical basis of RDB and provided a robust foundation for the quality control and pharmacology research in the future.

In this report, an ultrasonication and vacuum-assisted headspace solid-phase microextraction procedure followed by gas chromatography-flame ionization detection (UVA-HS-SPME-GC-FID) was proposed for direct extraction of solvent residuals, including benzene, toluene, ethyl benzene, m,p-xylene and o-xylene, in pharmaceutical matrices. A novel robust, reliable and durable nanocomposite solid-phase microextraction (SPME) fiber was prepared by layer-by-layer coating of 3-aminopropyltriethoxysilane-functionalized graphene on a stainless-steel wire. Then, the proposed fiber was used for headspace SPME and trapping of toluene as a residual solvent in solid penicillin, ampicillin and cefazolin vials followed by gas chromatography-flame ionization detection. UVA-HS-SPME-GC-FID achieves better validation parameters, including the limit of detection, limit of quantification, linearity, recovery and repeatability, in comparison with conventional HS-SPME-GC-FID. The UVA-HS-SPME-GC-FID strategy is very effective for quantitatively tracing volatile and semivolatile solvent residuals in various pharmaceutical drugs.

A simple, sensitive and reproducible high-performance liquid chromatographic (HPLC) method with fluorescence detection was developed and validated for the simultaneous quantification of mixed tocotrienols (α-, γ- and δ-) in human plasma. The method involves a straightforward sample preparation step, where plasma samples are deproteinized using a mixture of acetonitrile and tetrahydrofuran (3:2, v/v), followed by direct injection into the HPLC system. Separation was achieved using a methanol mobile phase at a flow rate of 1.5 mL/min, with fluorescence detection at excitation and emission wavelengths of 296 nm and 330 nm, respectively. The method demonstrated excellent linearity over concentration ranges of 12.7 ng/mL to 2.54 μg/mL for α-tocotrienol, 19.2 ng/mL to 3.84 μg/mL for γ-tocotrienol and 4.6 ng/mL to 0.923 μg/mL for δ-tocotrienol, with quantification limits of 12.7 ng/mL, 19.2 ng/mL and 4.6 ng/mL, respectively. Recovery rates ranged from 85.0% to 111.2%, and both intra-day and inter-day precision showed relative standard deviations below 15%. The method was validated for accuracy, precision and stability under various conditions, including freeze-thaw cycles and long-term storage. This HPLC method offers a rapid, sensitive and reliable approach for quantifying tocotrienols in human plasma, making it suitable for preclinical and clinical studies, particularly in pharmacokinetic and bioavailability research.

Terminalia arjuna is a widely recognized medicinal tree known for its diverse pharmacological properties, particularly its cardioprotective effects. This study aimed to quantify two key bioactive triterpenoids, arjungenin and arjunic acid, in T. arjuna bark samples collected from 16 different geographical locations. A validated High-Performance Liquid Chromatography method was developed, demonstrating high precision, accuracy and sensitivity for the simultaneous quantification of these compounds. Statistical analyses, including Hierarchical Cluster Analysis, Principal Component Analysis and heat map visualization, were employed to classify populations based on their phytochemical composition. The results revealed significant variations in arjungenin and arjunic acid content among different populations, with the highest concentrations observed in samples from Amangarh tiger reserve (Bijnor) and Gorakhpur Forest division (Pharenda). These findings were further confirmed by ANOVA and Tukey's post hoc test. Additionally, variations in yield and total tannin content were observed, suggesting that environmental factors play a crucial role in secondary metabolite biosynthesis. The identification of high-yielding populations highlights the potential for targeted conservation efforts and the selection of superior genetic resources for pharmaceutical and nutraceutical applications. This study provides a comprehensive phytochemical assessment of T. arjuna populations and establishes a reliable analytical method for quality control and standardization. The findings contribute valuable insights into the influence of environmental factors on metabolite production, paving the way for future research on genetic and biochemical pathways regulating triterpenoid biosynthesis in T. arjuna. Moreover, flash chromatography was employed to isolate arjunic acid, and its structure was further validated using 1H-NMR and 13C-NMR spectroscopy, reinforcing the analytical accuracy of the method.

Various spices and their oleoresins are being marketed since long and it has multibillion market. One of the most often used spices is black pepper (Piper nigrum), which gets its pungent flavour from volatile chemical components, essential oils, and an alkaloid called piperine. Piperine is also known for huge therapeutic benefits and most people are using daily in food products also. Most significantly, piperine is recognised as a bioavailability enhancer by stopping CYP enzyme activity. Therefore, our aim is to develop black pepper extract and isolation of piperine from black pepper. Various analytical techniques including, UV-Visible Spectroscopy, Fourier Transform Infrared Spectroscopy, Differential Scanning Calorimetry, Thermogravimetric Analysis, Proton Nuclear Magnetic Resonance, Carbon-13 Nuclear Magnetic Resonance, High-Resolution Mass Spectrometry were used to characterize piperine. NMR confirms its structure and molecular weight was confirmed by HRMS. Purified piperine has shown HPLC purity >99.5%. DSC has shown the melting peaks of 130.12°C. Piperine have great commercial potential as reference materials for regular quality control in the herbal industries for black pepper-based products.