Biomedical Chromatography最新文献

Preparative chromatographic enantioseparation is now the preferred technique for obtaining milligram quantities of pure enantiomers in the initial phase of development of a therapeutic compound. Supercritical fluid chromatography offers several advantages over liquid chromatography and was therefore selected for the preparative enantioseparation of a new potential anti-inflammatory molecule. Approximately 10 mg of each of the two enantiomers was successfully prepared using a Chiralpak AD-H (tris-3,5-dimethylphenylcarbamate of amylose) polysaccharide-based stationary phase with 40% of ethanol as a co-solvent, using a stacked injection mode. A peak distortion was observed during volume overloading, which may be due to the mixed-stream injection method.

The purpose of this research was to establish and validate a reverse phase HPLC method for the determination of Elagolix impurities in pharmaceutical dosage form. Mobile phase A, consisting of 10 mM sodium dihydrogen phosphate (pH 6.0) and acetonitrile in a 95:5 v/v ratio, and mobile phase B, containing 85:10:5 v/v/v of acetonitrile, Milli-Q water, and methanol, were used to achieve the method's specificity in the analytical column Kromasil 100-C18 (250 mm × 4.6 mm, 5 μm). The gradient program includes (%B/Time [min]: 36/0, 36/10, 38/15, 85/55, 85/65, 36/67, and 36/75). The flow rate is 0.8 mL/min. The overall run duration is 75.0 min, the injection volume is 10.0 μL, and the detection is at 210 nm in UV. The samples were subjected to hydrolysis, oxidation, and heat conditions in order to facilitate their forced degradation. The procedure was validated and determined with the standards of ICH guidelines. From the LOQ to a concentration level of 200%, the linearity of the technique was ascertained. An accuracy range of LOQ to 150% was established for the method, and the average recovery was acceptable. Design of experiments, part of the quality by design idea, was used to prove the method's reliability.

To facilitate clinical therapeutic drug monitoring (TDM) of polymyxin B (PB) and polymyxin E (PE), we developed and validated a simple LC–MS/MS method for simultaneous determination of PB (including polymyxin B1 (PB1), polymyxin B2 (PB2), polymyxin B3 (PB3) and isoleucine-polymyxin B1 (ile-PB1)) and PE (including polymyxin E1 (PE1) and polymyxin E2 (PE2)) in human plasma. PB or PE was extracted from 20.0 μL plasma using a 5% (v/v) formic acid acetonitrile solution and separated on a BEH-C18 column (2.1 × 100 mm, 1.7 μm) with a mobile phase consisting of 0.8% formic acid aqueous solution and 0.2% formic acid acetonitrile solution. Gradient elution was performed over 5.5 min at a flow rate of 0.250 mL/min. Quantitative analysis was conducted in positive ion scanning mode by electrospray ionization and multiple reaction monitoring. The method validation was conducted based on bioanalytical method validation guidance, including specificity, calibration curve, precision, accuracy, recovery, matrix effect, stability and dilution integrity and all of the results satisfied the requirements. The method was simple, robust and high-throughput and is currently being used to provide a TDM service to enhancing therapeutic efficacy and safety use of the PB and PE.

Improvement of strategies to treat heart failure (HF) has been a longstanding global goal and challenge. Shen-Fu formula (SF), as a classic herbal preparation, has demonstrated efficacy in treating HF in clinical settings. However, further understanding of the therapeutic mechanisms of SF is required. In this study, metabolomics and 16S rDNA sequencing were used to analyze the effects of SF on metabolic profiling and gut microbiota in HF rats. After 4 weeks of SF treatment, the cardiac function of HF rats showed improvement, with a significant increase in ejection fraction and fractional shortening, as well as a significant decrease in left ventricular volume and mass. Metabolomics study revealed that SF regulates the levels of substances related to energy metabolism, primarily involving lysophosphatidylcholines and polyunsaturated fatty acids. In addition, we found that SF regulates the structure of the microbial community in HF rats and modulates the balance between probiotic and pathogenic bacteria. Furthermore, the SF combination exhibited a superior effect that was better than the use of each herb separately. These results demonstrate the potential of SF therapy in the management of HF and highlight the role of SF in regulating fatty acid metabolism and gut microbiome during HF.

Enantiomers of a chiral active pharmaceutical ingredient (API) often exhibit different physicochemical, pharmacokinetic, and biological properties. Therefore, enantioseparation becomes a critical aspect of pharmaceutical development. Sertraline, one of the most widely prescribed antidepressant medications, requires purification from its chiral impurities, and this is recommended and essential for its quality control. This perspective highlights the current established research on the separation and quantification of sertraline's chiral impurities, with a focus on instrumental and crystallization-based techniques.

Aconiti Lateralis Radix praeparata (also known as Fuzi in Chinese) has been extensively used in clinic. However, the toxicity issues limit the therapeutic range of Fuzi. Thus, a rapid and sensitive HPLC-MS/MS method was developed to simultaneously quantify and compare six active/toxic constituents in decocting extracts from four different processed Fuzi products and in rat plasma after oral administration of its decocting extracts. The selectivity, linearity, sensitivity, precisions, accuracy, matrix effects, extraction recoveries, and stability were validated. The comparative analysis of six alkaloids in decocting extracts between the four kinds of Fuzi products were conducted by the validated HPLC-MS/MS. Principal component analysis (PCA) and orthogonal partial least-squares discriminate analysis (OPLS-DA) were adopted to compare the differences of decocting extracts from the different processed Fuzi products. Besides, selecting Heishunpian (HSP) as the representative of the processed Fuzi products, we investigated the pharmacokinetic behaviors of these major alkaloids after oral administration. The developed HPLC-MS/MS method to simultaneously analyze these aconitine-type alkaloids in decocting extracts, and its pharmacokinetic behavior after oral administration may pave a way for quality control of Fuzi and monitoring the safety and rationality of clinical prescriptions.

The synthesis of optically active compounds requires determination of ee, er, and enantiomeric purity. The aim of the present paper is to review the synthesis of several chiral derivatizing reagents (CDRs) in a rational manner, which were successful for the separation and isolation of enantiomers of a variety of active pharmaceutical ingredients and other important and useful racemates. Besides, the application of (i) certain enantiomerically pure amines, either directly or by incorporating each of them as chiral auxiliary in difluorodinitrobenzene or cyanuric chloride moieties to construct the CDR, (ii) (S)-ketoprofen and (S)-levofloxacin as chiral platforms, and (iii) a few isothiocyanates, have been suitably included. Attention is drawn to the use of water micellar mobile phase as the “green” RP-HPLC method and the use of simple achiral derivatization with ninhydrin, particularly. Synthesis of CDRs and their application for enantioseparation of racemates and detagging of certain chromophoric reagent components for obtaining native enantiomers are other interesting features included herein. The methods can be easily used to determine and control enantiomeric purity with advantages over a variety of commercial chiral phases. This comprehensive review not only highlights innovative methodologies for enantioseparation but also underscores their practical applications in controlling and ensuring the enantiomeric purity of pharmaceutical compounds.

Aromatherapy based on essential oil (EO) has been widely used for alleviating pain and intense where no compound EO reports its application on pharmacological effects. In order to explore the active pharmaceutical ingredients (API) and mechanism of a compound EO, a blend of Artemisia argyi, Boswellia carterii, Commiphora myrrha, Cinnamomum cassia, Zingiber oj-jicinale, and Ilex pubescens EO, in treating neck and shoulder pain (NSP). Network pharmacology hyphenated with mice model was employed to investigate. Gas chromatography–mass spectrometry (GC–MS) was applied for the identification of constituents in compound EO. Lastly, transdermal absorption of compound EO was studied before verifying analgesic and anti-inflammatory effects in mice. Totally, 75 compounds were tentatively identified through GC–MS, predicting 46 potential analgesic targets. Moreover, 11 core targets were obtained through network topology screening. Animal test resulted that the compound EO had significantly stronger anti-inflammatory and analgesic effects compared to single EO. Multiple API in compound EO affected on targets and exerted therapeutic effects on NSP through multiple pathways. Afterwards, eucalyptol, camphor, and borneol from compound EO exhibited a sustained-release effect, which provide scientific basis to illustrate the application of compound EO in clinical.

Codonopsis pilosula (Franch.) Nannf. is a traditional herb for treating immunosuppression. C. pilosula boiling powder (CP-BP) contains particles of a small size made from C. pilosula decoction pieces (CP-DP). It is still unclear how changes in particle size during the decoction process affect the dissolution of various chemical components in C. pilosula. Herein, an ultra-high-performance liquid chromatography–quadrupole-Exactive Orbitrap mass spectrometry technique was established to characterize the components of CP-BP and CP-DP decoctions. The contents of the components were evaluated based on the relative peak area, extract yield, and alcohol solubility rate. A total of 71 compounds were finally identified, and their content in the CP-BP decoction was generally higher than that in the CP-DP decoction. Alkaloids had the highest average content, whereas terpenoids were the most affected by changes in particle size. In addition, immunosuppression was used as model to investigate whether these changes have practical significance. The results of network pharmacology suggested that the phosphoinositide 3-kinase (PI3K)–Akt pathway may be a potential pathway of C. pilosula for treating immunosuppression. The results of molecular docking indicated that compounds with large content variations have good docking affinity with key targets (epidermal growth factor receptor [EGFR], prostaglandin-endoperoxide synthase 2 [PTGS2], and peroxisome proliferator-activated receptor gamma [PPARG]). These results provide an important reference for further development and use of C. pilosula.

Given the genetic and clinical overlap observed between schizophrenia and depression, the present study was to identify the similarities and differences in serum metabolic profiles between patients with schizophrenia and depression. Global metabolomics research methods based on UHPLC-QTOF-MS/MS were performed. A total of 113 and 118 differential metabolites were screened and identified in depression and schizophrenia groups, respectively, as compared to health control; among those, 94 differential metabolites were shared by both. Pathway analysis indicated arginine and proline metabolism, alanine, aspartate, and glutamate metabolism were two significant metabolic pathways both in depression and schizophrenia groups as compared with health control groups, respectively. Similarly, 77 differential metabolites were identified between depression and schizophrenia groups, in which, serum N-acetylglutamine and isovalerylglycine levels showed significant differences between patients with depression and schizophrenia with p values less than 0.001 and without significant outliers. Sphingolipid metabolism was identified as a significant metabolic pathway distinguishing between depression and schizophrenia groups based on pathway analysis. Conclusively, common alterations in arginine and proline metabolism, alanine, aspartate, and glutamate metabolism were observed in patients with schizophrenia and depression; whereas differences in serum N-acetylglutamine and isovalerylglycine levels as well as sphingolipid metabolism were discovered between the two categories of patients.