Journal of chromatographic science最新文献

A validated high-performance thin-layer chromatography (HPTLC) method was developed for the simultaneous quantification of bergenin and oleanolic acid in Pashanbhed (Bergenia ligulata), Lambadi (Celosia argentea) and Kalipath (Cissampelos pareira) for quality assessment. The developed HPTLC method employed a mobile phase of toluene, ethyl acetate, methanol and formic acid (4:5:1:2 v/v) on silica gel 60F254 plates. The plates were scanned at 254 and 366 nm using a Camag Scanner IV with vision CATS software. The method was validated for linearity, accuracy, precision, specificity, limit of detection and limit of quantification. The linearity range for bergenin and oleanolic acid was 4-20 and 10-50 μg/spot, respectively, with correlation coefficients of 0.9998. The average recoveries for bergenin and oleanolic acid were 98.75-99.94% and 98.62-100.83%, respectively. The bergenin content in the methanolic extract and crude drug of B. ligulata was 4.8% and 1.25% w/w, respectively. The oleanolic acid content in the methanolic extract of C. argentea and C. pareira was 1.98% and 1.55%, while in the crude drug, it was 0.24% and 0.16% w/w, respectively. The developed HPTLC method is simple, accurate and reproducible, making it suitable for routine analysis, standardization and quality control of bergenin and oleanolic acid in various natural herbs and Ayurvedic formulations.

Accurate quantification of all 20 proteinogenic amino acids (AAs) is critical for the quality control of total parenteral nutrition (TPN) formulations containing complex excipient matrices. This study presents a rapid and validated reversed-phase high-performance liquid chromatography (RP-HPLC) method employing pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) and fluorescence detection (FLD) for the simultaneous determination of AAs. Complete baseline separation of all 20 AAs was achieved within 18 minutes, with a total analytical run time of 25 minutes on a conventional HPLC system. The method was validated in accordance with ICH Q2(R2) guidelines, demonstrating excellent linearity (R2 > 0.999), high sensitivity (limits of detection = 0.02-0.16 μmol/L), precision (relative standard deviation< 2%), accuracy (recoveries = 98.2-101.7%), specificity (no matrix interference) and robustness under deliberate minor variations in chromatographic conditions. These findings confirm the method's reliability and suitability for pharmaceutical quality control. This RP-HPLC-FLD method provides a rapid, cost-effective and reproducible alternative to LC-MS for routine quality control of TPN formulations in pharmaceutical laboratories.

To optimize the dyeing process and minimize pollution, it is essential to develop rapid, real-time and accurate analytical techniques for detecting dye forms. Structurally similar reactive dyes, C.I. Reactive Orange 107 (RO107) and Yellow 201 (RY201), exhibit highly overlapping spectra, making the accurate identification and quantification of their six forms (original, activated and hydrolyzed forms for each) extremely challenging. This study aimed to address this issue by developing a novel capillary electrophoresis technique. The optimized background electrolyte consisted of 30.0 mmol/L Na2B4O7·10H2O and 45.0 mmol/L sodium cholate (SC), at pH 8.60 adjusted with 0.40 mol/L H3BO3. With UV detection, the limits of detection (LOD, S/N = 3) and quantification (LOQ, S/N = 10) for the six analytes ranged from 0.9 to 2.9 mg/L and 3.1-9.8 mg/L, respectively, with good linearity (R2 > 0.9965), migration time repeatability and intra-/inter-day peak area precision. The separation mechanism, based on micellar electrokinetic chromatography, utilizes SC micelles as pseudo-stationary phase to enhance migration differences via differential partitioning of analytes with varying hydrophilicity. Practical dyeing experiments demonstrated higher fixation rates for RY201 (25.57%, 29.85%) than for RO107 (17.42%, 18.01%) in both single and combination dyeing, confirming RO107's weaker adsorption capacity on the fabric.

Macamides and macaenes are two kinds of secondary metabolites in Lepidium meyenii (Maca). Geographical origin has an important effect on accumulation of active components in Maca. Hence, it is feasible to differentiate Maca from different origins according to the type and content of macamides and macaenes. Using petroleum ether as solvent, macamides and macaenes were extracted from Maca cultivated in Yunnan province, China (Qujing, Lijiang and Shangri-La) and Cerro de Pasco in central Peru, respectively. Then, these extracts were purified by silica gel column eluted with petroleum ether and analyzed qualitatively and quantitatively by UPLC/MS. Based on the fact that macamides and macaenes are characteristic components in Maca, the contents of them were used as principal components to successfully differentiate these Maca combined with PCA. The result suggests that based on remarkable difference of the content of macamides and macaenes in Maca from these four origins, PCA can be used to differentiate these Maca successfully. This strategy can be applied to evaluate quality and identify origins of other natural products.

Terbutaline sulfate is a pediatric β2-adrenergic agonist that acts as a bronchodilator by stimulating β2-adrenoceptors in bronchial smooth muscle. A novel and reliable high-performance liquid chromatography method was developed to identify process- and degradation-related impurities in terbutaline sulfate oral syrup. The separation was achieved using an Inertsil ODS-3 column (250 × 4.6 mm, 5 μm) maintained at 30°C, with a mobile phase composed of buffer and acetonitrile delivered at a flow rate of 1.0 mL/min under a linear gradient elution mode. The injection volume and sample concentration were set at 40 μL and 500 μg/mL, respectively. The drug and its impurities demonstrated good linearity, with determination coefficients greater than 0.999. Recovery rates at all levels ranged from 95% to 105%. Forced degradation studies were conducted under acidic, alkaline, oxidative, thermal and photolytic forced degradation conditions. Under photolytic forced degradation, Imp-B and three furfural-related degradation impurities were formed. The validated stability-indicating HPLC method, which has been verified in accordance with International Conference on Harmonisation guidelines, is suitable for both stability studies and routine quality control.

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 precise, fast and efficient technique utilizing reverse-phase liquid chromatography was established for the simultaneous estimation of Remogliflozin, Metformin and Vildagliptin, in a commercial pharmaceutical preparation. The method utilized a HiQ Sil C18 analytical column having a size of 250 × 4.6 mm and particle size utilized was 5 μm. The mobile phase composition comprised a mixture of potassium dihydrogen orthophosphate buffer (pH 4.7), methanol and acetonitrile in a ratio of 20:55:25 v/v/v. The flow rate was adjusted to 1.2 mL/min. Three UV wavelengths were finalized for Remogliflozin (276 nm), Metformin (227 nm) and Vildagliptin (213 nm), but the detection wavelength utilized for HPLC was 210 nm (isosbestic point). After validation, it was observed that the method exhibited linearity, as evidenced by correlation coefficients of 0.9994, 0.9997 and 0.9980 for Remogliflozin, Metformin and Vildagliptin, respectively. The recovery values for the three compounds were found 103.42%, 100.39% and 99.97%. Additionally, the relative standard deviation for six replicates consistently remained below 2%. This HPLC method was effectively employed for the quantitative analysis of the target drugs in tablet formulations.

The objective was to analyze the consistency of Aster tataricus L. f. decoction and its formula granule using fingerprint similarity, quantitative multi-component analysis, and chemical pattern recognition. A. tataricus L. f. decoction and its formula granule were prepared. Similarity evaluation, cluster analysis, principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were performed to evaluate the consistency of the chemical composition of A. tataricus L. f. decoction and its formula granule. OPLS-DA was used to screen key compounds. A total of nine key shared peaks were obtained from the fingerprint profile analysis. The fingerprint similarity of A. tataricus L. f. decoction and its formula granule ranged from 0.906 to 0.995. Cluster analysis suggested a similarity in the chemical composition of A. tataricus L. f. decoction and its formula granule. The common contribution of PC1 and PC2 was 58.1%. OPLS-DA analysis resulted in a R2X, R2Y, and Q2Y of 0.711, 0.35, and 0.168, respectively. The OPLS-DA method proposed neochlorogenic acid (Peak 1) and chlorogenic acid (Peak 4) as the key compounds. The method, which incorporates high-performance liquid chromatography fingerprinting and multi-component chemical pattern recognition techniques, demonstrated the consistency of A. tataricus L. f. formula granule with the decoction.

Deoxythioguanosine (dTG), incorporated into DNA as DNA-TG, is considered a more relevant thiopurine metabolite than erythrocyte thioguanine nucleotides, as erythrocytes are not the drug target. We established a simple and sensitive liquid chromatography-tandem mass spectrometry method to detect dTG in human blood nucleated cell DNA and validated its clinical applicability in Chinese patients with inflammatory bowel disease to improve individualized thiopurine treatment. dTG was released from 1 μg DNA in a single hydrolysis step, separated by the X-Terra RP18 column and quantified by tandem mass spectrometry with 2'- Deoxyguanosine-13C,15N2 as the internal standard. The calibration curve covered eight concentration levels (0.125-25 ng/mL) with the correlation coefficients r2 ≥ 0.99. The intra- and inter-assay variability was achieved less than 9.9%. This method was employed in a study where a total of 125 inflammatory bowel disease patients with a median DNA-TG level of 279.7 fmol/μg DNA (interquartile range 183.5 ~ 410.6 fmol/μg DNA). Nucleoside diphosphate-linked moiety X-type motif 15 variant carriers (*2, *3, *5, *6) showed significantly higher DNA-TG/dose and DNA-TG/erythrocyte thioguanine nucleotides ratios than wild-type patients (P = 0.0023, P = 0.0001). In summary, this method was simple and stable to detect dTG in DNA, which may act as a promising biomarker predicting the toxicity and efficacy in patients treated with thiopurine.

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.