Biomedical Chromatography最新文献

Upon the development of biosimilar drugs, the structural, physical–chemical, and functional similarity to the original drug should be ensured. We have proposed a simple and rapid HPLC-UV method for assessing the content of C-terminal Arg¹⁶⁶ in darbepoetin alfa samples, in particular after their isolation from the culture fluid. All the stages of the described method (from the sample extraction to obtaining a chromatographic profile) take about 5 h. The proposed approach will be useful for laboratories developing a technology for producing recombinant darbepoetin alfa, to assess the quality of the product, and for subsequent optimization of the production process.

Artemisia campestris subsp. campestris (tuguft) is a medicinal plant traditionally used in Algerian medicine. This study investigates the chemical composition and bioactivity of its essential oil (ACEO). Gas chromatography–mass spectrometry (GC-MS) analysis identified key compounds, including linalyl acetate (2.92%), geranyl acetate (2.45%), and eucalyptol (1.38%). ACEO demonstrated significant antioxidant activity, with IC₅₀ values of 11.09 μg/mL (DPPH), 15.81 μg/mL (FRAP), and 22.70 μg/mL (β-carotene). It also enhanced peroxidase activity by 82.67 U/g. The anti-inflammatory effects were confirmed with an IC₅₀ of 18.87 μg/mL. Notably, in silico molecular docking revealed that 3-cyclopentyl-N-(2-(3,4-dimethoxyphenyl)ethyl) exhibits strong binding affinity to phosphoinositide 3-kinase gamma, a target for pancreatic cancer therapy, suggesting potential anticancer activity. These findings underscore the therapeutic potential of ACEO, highlighting its antioxidant, anti-inflammatory, and anticancer properties.

The whole plant of Tribulus terrestris (TT) is used to treat hypertension and cardio-cerebrovascular diseases. The harvest time has an important influence on the quality of medicinal plants. Therefore, it is necessary to study the effect of harvest time on the content of components in different parts of TT and compare the antihypertensive effect of different parts of TT at harvest time with a relatively high content of 14 components. Ultraviolet–visible spectrometry (UV–Vis spectrometry) was used to determine the total saponin content in T. terrestris fruit (TTF), T. terrestris stems (TTS), and T. terrestris leaves (TTL) at different harvest times. Within the studied harvest times, the total saponin content was higher in TTL than in TTF or TTS. Ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry analysis revealed significant differences in the spatial and temporal differences in the content of 14 components in the three plant parts. The multivariate statistical analysis identified six components as markers of different TT content in the different parts. The suitable TTF harvest period was from August 11 to September 1, while TTS and TTL were suitable for harvesting from July 21 to September 11. The treatment effects on blood pressure, serum indexes, and thoracic aorta morphology confirmed that the antihypertensive effects of TTL were better than those of TTF or TTS. The study emphasized the effect of harvest time on the content of components in different parts of TT and confirmed that TTS and TTL also had medicinal value.

Proteolysis targeting chimera (PROTAC) has been developed and currently enjoys widespread interest among the field of pharmaceutical manufacturing in recent decades. NX-5948 is an orally active PROTAC Bruton tyrosine kinase degrader, which shows anti-inflammatory and antitumor activities. In this study, a simple and fast bioanalytical method for the quantification of NX-5948 in beagle dog plasma was developed using liquid chromatography–tandem mass spectrometry (LC-MS/MS). A full method validation was performed according to regulatory guidelines. For the quantification, [M + H]⁺ was formed using an electrospray ionization (ESI) source in the positive ion mode, and selective reaction monitoring (SRM) was employed using a triple quadrupole mass spectrometry. The monitored precursor-to-product transitions were m/z 807.5 > 790.5 for NX-5948 and m/z 812.4 > 452.1 for internal standard. A linear response was obtained at the concentration range of 0.5–200 ng/mL, with correlation coefficient > 0.99. The interday accuracy ranged from −9.03% to 5.99% (RSD < 7.84%), and the intraday accuracy from −5.15% to 8.56% (RSD < 6.90%). NX-5948 was demonstrated to be stable under the present storage conditions. After validation, the method was successfully applied for the quantification of NX-5948 in beagle dog plasma after oral and intravenous administration of the NX-5948.

Pharmacokinetic studies can provide a method for optimizing preparations. In this study, three most abundant compounds from Fritillaria ussuriensis bulb were isolated, and a selective, sensitive new analytical method was developed. This method aims for the simultaneous determination of peimisine-3-O-β-D-glucopyranoside (1), karelinine (2), and peimisine (3) in rat plasma after oral administration of F. ussuriensis bulb powder (FUP), total alkaloid extract (FUE), and its nanodispersion preparation (FUN) using UHPLC–MS/MS. The chromatographic separation was performed on the UHPLC Waters BEH C18 column. The pharmacokinetic results showed that among three groups, 1 and 3 exhibited slower absorption and elimination processes when rats were orally administered with FUP and FUE. Steroidal alkaloids 1 ~ 3 from FUN reached peak concentrations within 3 h. Compared with FUE and FUP, the absorption and elimination rates were accelerated, and the area under the curve (AUC) significantly increased. In this study, the pharmacokinetic characteristics of 1 ~ 2 were first detected and calculated parameters in rat plasma when orally administered in different dosage forms. And a new nanodispersion preparation, FUN, has been developed. FUN has better absorption efficiency and higher bioavailability compared with traditional powder oral administration.

This letter responds to recent research on Lingguizhugan decoction (LGZGD) in treating gestational diabetes mellitus (GDM). While acknowledging the significance of their findings regarding PI3K-AKT pathway modulation and oxidative stress reduction, we suggest expanding future research directions. We highlight the importance of incorporating animal models alongside cell studies, considering multiple mechanistic pathways, and integrating traditional Chinese medicine approaches with modern research methodologies. Recent studies on natural antioxidants and traditional Chinese medicines in GDM treatment provide supporting evidence for these recommendations. These perspectives aim to enhance understanding of LGZGD's therapeutic potential and contribute to developing more effective GDM treatments.

For the control of excessive blood loss occurring from major trauma, postpartum bleeding, surgery, or hereditary angioedema, antifibrinolytic compound tranexamic acid (TXM) is highly efficient in controlling blood loss by reversibly binding with on lysine receptor sites. A selective and sensitive method has been developed and validated for the quantitation of TXM in human plasma using high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). Tranexamic acid-D2 (TXM-D2) was used as internal standard (ISTD) to minimize the errors in TXM quantification. TXM quantification was performed with positive polarity mode using a Shimadzu high performance liquid chromatography coupled with AB-SCIEX API-4000 tandem mass spectrometer. A Zorbax Eclipse C18 (150 × 4.6 mm, 5 μ) column was used for the TXM quantification, 8-mM ammonium formate buffer with 0.1% formic acid ionization enhancer was used as a mobile phase-A, and acetonitrile was used as a mobile phase-B. (38:62) v/v portion of mobile phases A and B selected to elute the TXM and TXM-D2 with the flow rate of 0.8 mL/min. An electrospray ionization (ESI) technique was selected for detection of TXM in the human plasma. Linearity was assessed in the concentration range from 75 to 15,000 ng/mL by using least squares of weighting factor linear regression (1/X²).

This research study presents a precise, accurate, and linear liquid-chromatography method for the determination of pemafibrate in tablet dosage forms, despite the presence of a high volume of excipients. Various challenges were addressed through experimental approaches and evidence-based solutions. The method employs a suitable stationary phase, X-Bridge C₁₈ (150 × 4.6 mm, 3.5 μm), and an appropriate isocratic program. Key parameters include a flow rate of 1.0 mL/min, a column temperature of 40°C, an injection volume of 10 μL, and a runtime of 15 min. The wavelength was set at 210 nm because of its high response. Mobile phase optimization, based on experimental results, consists of 0.1% H₃PO₄ buffer and acetonitrile in a 40:60 v/v ratio. The method has been validated according to ICH Q2 (R2) and Ch. P <9101> guidelines, achieving a recovery rate of 99.1% to 100.5% at levels of 50%, 100%, and 150%. Linearity was demonstrated from 25% to 300% concentration levels, with a correlation coefficient (r²) value of 1.000. Precision results showed %RSD values of 1.0 and 1.1. Forced degradation studies indicated sensitivity to acid hydrolysis stress conditions and stability under physical stress conditions.

Jianpihuazhuotiaozhi granules (JPHZTZ) are traditional Chinese medicine formula. An analytical method using ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry was established to characterize the chemical constituents of JPHZTZ and determine its metabolic profile in rat urine and plasma after oral administration. A total of 220 compounds were identified in JPHZTZ extract, including 61 flavonoids, 16 alkaloids, 53 organic acids, 31 terpenoids, and 59 other compounds. Among these compounds, 11 were tentatively identified by comparing the retention times and mass spectral data with the corresponding reference standards and the literature; the other 209 components were tentatively identified based on their mass spectra alone. After the oral administration of JPHZTZ extract to rats, 13 prototypes and 54 metabolites were identified or tentatively characterized based on their retention time and mass spectra. The primary in vivo metabolic reactions that occurred after the administration of JPHZTZ extract included glucuronidation, sulfation, hydroxylation, and methylation. The 13 prototypes and 54 metabolites were identified in rat urine and plasma and were confirmed to be the potential active ingredients of JPHZTZ. Our findings provide a starting point for the further elucidation of the mechanism of action of JPHZTZ.

DM and PM were the two major subtypes in myositis; among which, a unique metabolomic and biomarker profile remains lacking. Serum from 36 diagnosed myositis patients (28 DM and 8 PM) and 29 healthy controls was analyzed using HPLC-Q-TOF-MS/MS. PLS-DA was conducted through MetaboAnalyst 5.0 to identify the differential metabolites. The KEGG analysis was utilized to observe the related metabolic pathways. The potential biomarker value was assessed using ROC analysis. The relationship between the clinical characteristics and the levels of identified differential metabolites was analyzed using R language. PLS-DA showed a clear separation between healthy controls and myositis patients, and 131 differential metabolites were identified. KEGG analysis uncovered multiple disturbed metabolic pathways. Besides, nine differential metabolites were identified between PM and DM patients, which were involved in pentose and glucuronate interconversions. ROC curve analysis revealed the AUC of these identified metabolites is above 0.7. Among them, indoxyl sulfate, oleamide, and palmitoylethanolamide presented moderate or strong correlation with clinical characteristics. Metabolomics presents a different spectrum between myositis patients and healthy controls, PM and DM patients. Besides, indoxyl sulfate, oleamide, and palmitoylethanolamide may be potential biomarkers in distinguishing PM from DM.