Abstract Some drug combinations, also known as multi-component formulations, have shown promising results due to their combined mechanism of action in the body, which consists of a mixture of medications (paracetamol, tramadol, metoclopramide, and domperidone). In addition to providing greater therapeutic efficacy due to their cumulative or synergistic effects, these drug combinations are also characterized by their lower production cost and the possibility of administering them in a single dose. The aim of this work is to develop novel, simple, accurate, and selective chromatographic methods (reversed-phase high-performance liquid chromatography) for the determination/quantification of these drugs. These methods can be used as analytical tools in routine analyses in quality control laboratories. In this study, RP-HPLC method was used, where the separation was performed on an ULTISIL L10 column (250 mm × 4.6 mm, 5 µm) using a mobile phase composed of a buffer solution and acetonitrile in a ratio of (80:20, v/v) at a flow rate of 1.5 mL min −1 . Detection was performed at a wavelength of 283 nm. In the proposed RP-HPLC method, the peaks were sharp and well separated, with retention times of 2.11, 4.37, 5.72, and 8.77 min, respectively, for paracetamol (PARA), tramadol (TRA), metoclopramide (METO), and domperidone (DMO). Linearity was obtained within the concentration ranges of 25–150, 10–60, 10–30, and 10–30 μg mL −1 , respectively, for paracetamol, tramadol, metoclopramide, and domperidone. The method was validated according to ICH guidelines for linearity, accuracy, precision, selectivity and robustness. This improved method is used to identify drugs in lab-mixtures containing a combination of four drugs. This method is simple and accurate and can be used for routine drug analysis in compounded dosage forms in quality control laboratories.
Contents of yeast cell wall polysaccharides including mannan and beta -glucan derived from yeast cell wall in yeast cultures (YC) were assessed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with the hydrolytic monosaccharides as markers. The sample hydrolyzed with hydrochloric acid solution in an autoclave and neutralized with sodium hydroxide solution and derivatized with 5-methyl-2-phenyl-1,2-dihydropyrazol-3-one (PMP). Then three main monosaccharides (mannose, glucose and galactose) derivatives were separated using a C18 column (50 mm & times; 2.1 mm, 1.8 mu m) with an isocratic elution of 0.02 mol L -1 ammonium acetate (82%) and acetonitrile (18%) as the mobile phases, and were detected using mass spectrometry with multiple reaction monitoring mode. Within the investigated concentration ranges (0.1-5 mu g mL -1 ), the monosaccharides exhibited good linearity with R 2 values greater than 0.99; the limit of detection (LOD) and limit of quantification (LOQ) was not more than 0.03 and 0.1 mu g mL -1 in the hydrolysate; Spike recoveries ranging from 82.56 to 105.58% were obtained with relative standard deviations (RSDs) between 1.42 and 10.30%. The quantitative results of mannose and glucose released from the yeast cell wall polysaccharides (YCWP) were corrected in the YC sample according to the mass ratios of mannose to galactose in the raw materials and that of glucose to mannose in the yeast respectively, and based on the corrected results the contents of YCWP, mannan and beta -glucan, in YC samples were assessed.
Shengmai Yin (Codonopsis Radix formula) (SMY), a classic prescription for replenishing Qi and nourishing Yin, is composed of Codonopsis Radix, Ophiopogon Radix, and Schisandra chinensis Fructus. However, the underlying compatibility mechanism of this formula remains unclear. A sensitive and reliable HPLC-MS/MS method was established for the simultaneous determination of twelve bioactive components in rat plasma, including syringin, tangshenoside I, lobetyolin, schisandrol A, gomisin D, schisandrol B, methylophiopogonanone A, gomisin G, methylophiopogonanone B, schisantherin A, schisandrin A, and schisandrin B. The method was successfully applied to the pharmacokinetic study of SMY, and the compatibility mechanism was elucidated through comparative analysis of the pharmacokinetic profiles among different compatibility groups. Plasma samples were pretreated by protein precipitation with acetonitrile. Quantification was performed in multiple reaction monitoring mode using positive and negative electrospray ionization, with carbamazepine and nimodipine as internal standards. The method exhibited good linearity ( r 2 >= 0.9864), and the lower limits of quantification (LLOQs) ranged from 0.0100 to 1.00 ng mL -1 , which were significantly lower than previously reported values. All validation parameters were within acceptable ranges. Pharmacokinetic behaviors after compatibility were significantly different from those of the single herb, demonstrating that the synergistic and attenuation effects might coexist in SMY. These findings provide a solid foundation for elucidating the compatibility mechanism of SMY.
Abstract Medicinal herbs have long been an integral part of traditional healthcare systems and are increasingly gaining global recognition for their therapeutic value. This review explores the chemical complexity of herbal plants, with a particular emphasis on their volatile organic compounds (VOCs) content, which contribute significantly to their pharmacological and industrial relevance. It delves into the therapeutic roles and ethnopharmacological significance of herbal VOCs, while also examining their economic impact. This review provides a critical overview of the chemical diversity, biosynthetic pathways, and functional roles of VOCs. The review further outlines the chromatographic techniques for chemical profiling and fingerprinting, essential for quality control, standardization, and regulatory compliance of herbal products that led to the plants-based drug discovery. A detailed overview of sample preparation and extraction methods, including emerging green and nano-enabled technologies is provided. Analytical chromatographic techniques and sensor-based methods, are discussed alongside advances in miniaturized systems. The application of statistical, chemometric, and machine learning tools for data interpretation is reviewed, emphasizing their role in enhancing the relevance of volatile profiling. The review identifies a growing trend toward integrative approaches that combine advanced analytical methods with robust statistical tools to ensure reproducibility, standardization, and regulatory compliance of herbal products. Notably, significant challenges remain, including variability in VOC composition due to geographical, genetic, and processing factors. We propose that future research should focus on data fusion strategies, AI-driven metabolomics, and precision phytomedicine to translate volatile profiling into clinically relevant outcomes. This review thus provides a comprehensive synthesis that bridges traditional phytochemistry with cutting-edge analytical science, setting the foundation for sustainable innovation in herbal therapeutics.
HPLC is an analytical method that provides precise, accurate, and simultaneous quantification of one or more drugs in the formulation, ensuring an accurate dose of active ingredients. The current research work focuses on developing and validating a reversed-phase high-performance liquid chromatography (RP-HPLC) procedure that can quantify three drugs, i.e., Empagliflozin, Pitavastatin, and Metformin HCl, simultaneously. The validation of the developed method was performed as per the Q2 (R1) guidelines of the International Council for Harmonization (ICH). The analytical procedure was performed on a Shimadzu HPLC system, having an Agilent C18 column (250 mm & times; 4.6 mm, 5 mu m). The chromatography method used an isocratic mobile phase comprising a solution A having pH 3.5 +/- 0.05 (consisting of KH 2 PO 4 solution with the ion-pairing agent i.e., sodium 1-heptanesulfonate), and acetonitrile mixed in a 55:45 V/V ratio, pumped at a flow speed of 1.20 mL min -1 . Wavelength was set at 260 nm (3.4 min) and 220 nm (12 min) to detect the peaks of the drugs at column oven temperature maintained at 25 degrees C. The method was characterized by determining and evaluating different values, including linearity, precision, robustness, system suitability, solution stability, and specificity. The approximate retention time for Metformin HCl was 2.0 min, for Empagliflozin was 4.1 min, and for Pitavastatin was 8.5 min. The results of the research showed that all three drugs were effectively distinguished with excellent resolution. The value of linearity was determined by using concentrations ranging between 5 and 7.5 mu g mL -1 for Empagliflozin, 0.4-0.6 mu g mL -1 for Pitavastatin, and 100-150 mu g mL -1 for Metformin HCl. The values of R 2 were 0.9999, 0.9999, and 0.9991, for Pitavastatin, Empagliflozin, and Metformin HCl, respectively. The method was proven to be accurate, with recovery values of 99.70-100.08% for Pitavastatin, 100.55-101.23% for Empagliflozin, and 98.15-101.76% for Metformin HCl. The results of the study demonstrated that the developed procedure was linear, precise, rapid, accurate, robust, and found to be applicable for the simultaneous determination of Metformin HCl, Pitavastatin, and Empagliflozin.
Aiweixin Oral Liquid (AWX) is a classic Chinese patent medicine formulation for the treatment ofcoronary artery disease, with potential medicinal, environmental, agricultural, and commercial value.Multiple active components are clinically used to treat cardiovascular diseases such as coronary arterydisease and angina pectoris. This study utilized Ultra-High Performance Liquid Chromatography-Tandem Electrostatic Field Orbitrap Mass Spectrometry (UPLC-Orbitrap-MS) and HeadspaceSolid-Phase Microextraction Gas Chromatography-Mass Spectrometry (HS-SPME-GC-MS) to analyzethe chemical composition of non-volatile and volatile metabolites in the AWX residue. A total of85 chemical components with non-volatile content exceeding 0.2% were identified in the AWX residue,including flavonoids, organic acids, polyphenols, terpenoids, and others. Among volatile components,50 chemical constituents with a content of 0.1% or higher were identified, involving monoterpenes,hydrocarbons, esters, phenols, aldehydes, ketones, alcohols, and others. This study preliminarilyexplored the main pharmacologically active substances in the AWX residue, providing a reference forsubsequent quality control and pharmacodynamic research.
Dual COX-2/5-LOX inhibitors represent a promising strategy for safer and more effective treatment of chronic inflammation-related diseases. Building on our previous work, a set of thirteen structurally diverse compounds ( 1-13 ) that act as inhibitors of these enzymes was analyzed to investigate their lipophilicity and retention behaviour using reversed-phase high-performance liquid chromatography (RP-HPLC). Some of these compounds were designed by introducing polar pharmacophores into the structure of conventional COX inhibitors. Therefore, it is highly important to assess the impact of these structural modifications on lipophilicity. Among several chromatographic systems tested, the C18 column with an acetonitrile/phosphate buffer mobile phase was identified as optimal, showing the strongest correlation between the experimental retention data and the calculated lipophilicity ( logD ). The retention parameter logkw proved to be the most reliable indicator of lipophilicity. It ranged from 1.40 to 3.21, with the highest values for bulky di- tert -butylphenols and the lowest for polar hydroxamic/urea derivatives. PCA analysis highlighted five molecular descriptors with the highest influence on retention ( CATS2D_05_AL, CATS2D_02_AA, MaxssNH, NssNH , and P_VSA_charge_6 ). Interpretation of these descriptors could be used as a starting point for further chemical modifications of tested compounds.
Felodipine is recommended as the first-line medication for the treatment of hypertension, and most of the formulations currently used in clinical practice are sustained-release tablets. In this study, a highly sensitive, rapid and cost-effective UPLC-MS/MS method was developed and validated for the determination of felodipine in clinical study. After liquid-liquid extraction for sample preparation, felodipine was separated on a Hedera ODS-2 column with isocratic elution utilizing methanol and 2 mM ammonium acetate buffer containing 0.2% formic acid, and detected by multiple reaction monitoring of m/z 384.0 -> 338.1 for felodipine and m/z 430.1 -> 372.2 for mifepristone (internal standard) in the positive ion mode. The entire chromatographic separation was completed within a short runtime (4 min per sample), allowing rapid analysis of large batches of biological samples. The calibration curve demonstrated good linearity ( r >= 0.9934) over the concentration range of 0.07540-10.05 ng mL (-1 )for felodipine with acceptable precision and accuracy. The extraction recovery was higher than 84.8% and the matrix effect ranged from 102.6 to 109.8% for analyte. Felodipine was also stable in human plasma and processed samples under various storage conditions. The present method was successfully applied to a pharmacokinetic study following a single oral administration of 5 mg felodipine sustained-release tablets in healthy Chinese subjects.
Cefixime is a third-generation oral cephalosporin commonly used to treat infections of the upper and lower respiratory tract, middle ear, sinuses, urinary tract, and gonorrhea. Currently, various dosage forms of cefixime are available on the market, produced by many local and multinational pharmaceutical companies. This study aims to systematically develop and optimize cefixime trihydrate orodispersible tablets (400 mg) using a meticulous Design of Experiments approach and to evaluate their in vivo pharmacokinetics by a validated RP-HPLC method. A central composite design was employed, with croscarmellose and mannitol as the input variables. The effects of these variables were assessed on tablet breaking force (hardness), disintegration time, and cumulative drug release. The optimization was performed using desirability, both numerically and graphically, and the optimized formulation was subsequently tested for accelerated stability. A robust RP-HPLC method was validated for plasma linearity, accuracy, and precision, and applied for the pharmacokinetic evaluation of cefixime in the optimized formulation. Micromeritics results showed variation in flowability and compressibility across the formulation blends, ranging from excellent to slightly poor characteristics. The tablet hardness was greatly affected by the relative amount of mannitol, which also improved drug dissolution by enhancing drug wettability in the medium. Higher levels of croscarmellose significantly reduced tablet disintegration time. Numerical and graphical optimization was conducted to find a formulation with suitable hardness, minimal disintegration time, and maximum dissolution within 15 min. Fourier transform infrared spectra and differential scanning calorimetry thermograms indicated no chemical interactions, and stability studies predicted a shelf life of approximately 33.36 months for the optimized formulation. The validated method showed linearity in human plasma ( R 2 = 0.9991) and acceptable accuracy and precision. In vivo pharmacokinetic evaluation showed improved performance compared to existing immediate-release formulations, with increased C max , decreased T max , and higher AUC. Overall, this study suggests that high-dose orodispersible tablets can be systematically developed without trial and error by using an appropriate design space, enabling the industry to create robust, well-designed formulations that align with ICH quality guidelines.
This study was predicated on the valuable medicinal properties of Crocus sativus L. and the fact that its residue remains rich in multiple active components. A comprehensive analytical method was established, combining ultraviolet spectrophotometry and high-performance liquid chromatography in order to achieve comprehensive utilization of this byproduct. This method employs Ultraviolet spectrophotometry to rapidly determine total crocin at a wavelength of 440 nm. Concurrently, an efficient and stable HPLC method was developed and validated using a C18 column with acetonitrile-0.2% phosphoric acid aqueous solution as the mobile phase for gradient elution. The employment of wavelength switching detection, in conjunction with simultaneous baseline separation and qualitative and quantitative analysis of crocin-I, crocin-II, and picrocrocin, has been achieved within a 30-min timeframe. The established method underwent systematic validation, demonstrating excellent linearity (r(2) > 0.999) within the corresponding linear ranges, high precision (RSD <2.0%), and reliable accuracy (mean recovery rates: 101.00-102.17%). This method was applied to analyse 10 batches of medicinal residue, enabling precise quantification of key active components. Concurrently, HPLC fingerprint profiles were successfully constructed for all ten batches. The results of the similarity analysis (all >0.995) demonstrated a high degree of consistency in chemical composition across batches. The integrated strategy established in this study enables comprehensive evaluation of C. sativus L. residues quality across three dimensions: total quantity, individual components, and overall chemical characteristics. This provides a foundation for the subsequent implementation of quality control measures in related industries, facilitating the development and reuse of resources.
Background: Moxifloxacin and linezolid are key antibiotics used for multidrug-resistant tuberculosis (MDRTB) and other infections, including those caused by gram-positive bacteria like Mycobacterium tuberculosis and Methicillin-Resistant Staphylococcus Aureus (MRSA). Accurate and simultaneous quantification of these antibiotics in human plasma is essential for therapeutic drug monitoring and pharmacokinetic studies. The existing methods often focus on single drugs or require expensive Liquid Chromatography-Mass Spectrometry (LC-MS/MS) unavailable in resource limited settings like Pakistan. Methods: An isocratic HPLC-UV method was developed using an Agilent 1100 system with a C18 column (250 & times; 4.6 mm), mobile phase of 0.1% formic acid:acetonitrile (75:25 v/v) at 1 mL min (-1) , and detection at 292 nm (moxifloxacin) and 262 nm (linezolid). Sample preparation was done through protein precipitation with acetonitrile and liquid-liquid extraction. The method was validated following European Medicines Agency (EMA) guidelines for selectivity, linearity, within-run and between-run accuracy and precision, and stability at room temperature, freeze-thaw and long-term stability for 6 months. Results: The method was selective for moxifloxacin and linezolid with no interfering peaks at retention times (4.4 min for moxifloxacin, 9 min for linezolid). The method was linear with r (2) >= 0.999 over the concentration range of moxifloxacin (0.2-12 mg L (-1) ) and linezolid (0.5-30 mg L (-1) ). The lower limit of quantification for moxifloxacin was 0.2 mg L (-1) and for linezolid was 0.5 mg L (-1) as determined by back calculation. The mean percentage relative recovery for accuracy was 91.6-111.8% for moxifloxacin and 88-97.9% for linezolid while RSD% for precision was <= 13.7% for moxifloxacin and <= 12.4% for linezolid. Both analytes remained stable at room temperature for 24 h, after four freeze and thaw cycles and also after storage at -20 degrees C for 6 months. Conclusion: This validated, cost-effective HPLC-UV method enables reliable simultaneous quantification of moxifloxacin and linezolid in plasma, supporting pharmacokinetic research within resource constrained settings like Pakistan.
The use of fingolimod as a treatment for multiple sclerosis relapses is crucial in reducing disease activity, reducing relapses, and slowing down neurological impairment. A basic and efficient isocratic reverse-phase HPLC technique was designed and optimized for the quantitative analysis of Fingolimod under controlled chromatographic conditions in support of its critical clinical value. The separation was done under ambient temperature in a C18 column (250 & times; 4.6 mm, 5 & micro;m) using a mobile phase, which consisted of 45% 0.05 M KH2PO4 (pH 6.8) and 55% ethanol to ensure high retention and symmetry of the peaks. The process worked with a flow rate of 1.5 mL min(-1) and UV detection of 210 nm, and 100 & micro;L injection volume. The chromatographic system was allowed to equilibrate for 60 min so that the baseline could stabilize and consistent retention was achieved. An in-vitro dissolution comparison study identified that the test and reference products were very similar with a similarity factor f2 = 81 and a difference factor f1 = 2, which proved bio-relevant performance similarity. The excellent acceptance value (AV) of 2.4 was obtained by content uniformity testing which was within the pharmacopeial range. The method's linearity was excellent in the concentration range of 0.25-1.5 mu g mL(-1) with a r = 0.9999 whereas accuracy and recovery fell within a range of 100.14 percent to 101.07 percent indicating high reliability of the method in routine quantitative work. Forced degradation was studied in the presence of acid, base, oxidative, photolytic, and thermal stresses. Fingolimod was discovered to be unstable in all the degradation pathways tested, and so, it must be protected and not subjected to stressful conditions to avoid degradation. AGREEprep was used to assess the greenness of the method with a score of 0.6 and BAGI was used to evaluate the analytical eco-score with a score of 82.5, which confirmed that the method has a positive environmental footprint compared to other conventional HPLC methods. All in all, the devised method is accurate, precise, stable-indicating, environmentally thoughtful and appropriate to complete quality-assessment tests of Fingolimod preparations in dissolution profiling, assay, and content uniformity analyses.
Chlorogenic acid, caftaric acid, and chicoric acid are the active components ofCichorium glandulosum,and high-performance thin-layer chromatography (HPTLC) is currently widely used in the analysis ofthe quality of medicinal herbs, however, the study of HPTLC on the three phenolic acid components indifferent parts ofC.glandulosumhas not been reported yet. Methods: Samples were extracted byrefluxing in 80% ethanol at a 1:10 material-to-liquid ratio, and petroleum ether-butyl acetate-formicacid-water (1:7:2.5:2) was used as the mobile phase. The mixture was scanned at 330 nm to determinethe amount of the components in the samples. Results: TheRFvalues of chlorogenic acid, caftaric acid,and chicoric acid were 0.39 +/- 0.05, 0.54 +/- 0.05, and 0.78 +/- 0.05, respectively, with linear ranges of0.0966-0.2899, 0.0998-0.2994, and 0.0946-0.2838 mu g/spot, respectively. The correlation coefficientswere 0.9767, 0.9948, and 0.9942, and the recoveries were 102.13, 100.14, and 99.65%, with relativestandard deviations of 3.3, 5.4, and 4.4%, respectively, for chlorogenic acid, caftaric acid, and chicoricacid, respectively. Chlorogenic acid contained the highest content (247.30 mu gg-1) in the whole herb,caftaric acid contained the highest content (122.98 mu gg-1) in the seeds, and chicoric acid contained thehighest content (69.70 mu gg-1) in the stems. Conclusion: Three phenolic acid components (chlorogenicacid, caftaric acid, and chicoric acid) from different parts ofC. glandulosum(whole herb, stem, andseed) can be effectively analyzed qualitatively and quantitatively using HPTLC.
Camelliaside A is a rare kaempferol glycoside with proved neuroprotective and antioxidant effects. In the present study, a rapid and precise HPLC-UV method for its quantitation was developed and validated. Separations were carrier over a C 18 column with a gradient a gradient elution with 0.5% trifluoroacetic acid and acetonitrile as the mobile phase. Statistical parameters were calculated to prove the methods' applicability. The method was used to determine this secondary metabolite's quantity in two closely related Astragalus species - A. glycyphyllos L. and A. glycyphylloides DC. The results showed great differences with 0.13% of camelliaside A found in the first, and only 0.01% of it in the second examined taxon. This is the first report on the chemical differences in the bioaccumulation of this metabolite in both plants.
In this study, a novel hydrophilic interaction liquid chromatography (HILIC) technique was established for the concurrent analysis of four active ingredients; chlorpheniramine maleate (CM), dextromethorphan hydrobromide (DHB), ephedrine hydrochloride (EHC) and guaifenesin (GUA). The developed method was characterized by its simplicity, accuracy and reliability. It has been successfully applied to quantify these compounds both in their pure forms and within combined pharmaceutical formulations containing all four active pharmaceutical ingredients APIs. A high-purity silica L3 column provided sharp and well-resolved peaks. Separation was achieved using an isocratic mixture of methanol and a 0.05 M acetate buffer at pH 4.0 in a ratio of 95.5:4.5 (v/v) with detection at lambda max 258 nm. This method demonstrated better separation than conventional reversed-phase HPLC techniques. The method validation followed the requirements outlined in ICH Q2 (R2) and USP. The results showed high linearity, with correlation coefficients of 0.998 for GUA, 0.996 for EHC, 0.996 for DHB and 0.998 for CM in the concentration range of 50-750, 7.5-112.5, 6.25-93.75 and 4.0-60.0 mu g mL -1 for GUA, EHC, DHB and CM respectively. LODs of the method were 2.70, 1.00, 0.76 and 0. 33 mu g mL -1 and LOQs were 8.10, 3.00, 2.28 and 0.99 mu g mL -1 for GUA, EHC, DHB and CM respectively. The method showed robust performance and minimal matrix interference and it was suitable for routine quality control of multi-ingredient pharmaceutical formulations containing the quaternary mixture of the cited antitussive APIs specifically as syrups.
Ardisia Japonica (AJ) is a traditional herbal medicine commonly used by the Zhuang and Yao people to relieve cough and asthma. The purpose of this study was to establish a high-performance liquid chromatography (HPLC) fingerprint of AJ and determine the content of 10 chemical components. Firstly, the fingerprints of 10 batches of AJ samples were obtained by HPLC. Secondly, cluster analysis, principal component analysis, and partial least squares discriminant analysis were used to evaluate the quality of AJ. The results showed that 18 common chromatographic peaks were calibrated in the HPLC fingerprint of 10 batch samples, and the similarity evaluation was greater than 0.9. 10 components were identified as gallic acid, norbergenin, protocatechuic acid, bergenin, chlorogenic acid, (-)-epigallocatechin-3-gallate, myricitrin, (-)-epigallacatechin-3-O-gallate, quercitrin, and quercetin. The results of cluster analysis and partial least squares discriminant analysis were consistent, indicating that all 10 batches of samples could be divided into two categories. Further analysis revealed that seven components played a crucial role in the quality control of AJ. In conclusion, the HPLC fingerprint and content determination method established in this study is stable and reliable, providing a basis for the quality evaluation of AJ.
Cigarettes are a product that contains thousands of dangerous chemicals that can disrupt the function of body organs and even threaten life. Nicotine is the most dangerous compound of cigarettes due to its addictive effect, and one way to reduce nicotine dependence is to use citronella oil. This study began with a quality test of citronella oil from Jatijejer Village using Thin Layer Chromatography (TLC), Stahl Distillation, and Gas Chromatography-Mass Spectrometry (GC-MS) instruments. Next, an in silico study was conducted to assess drug-likeness, bond energy, and bond interactions. The results of the quality test showed that citronella oil was proven to have good quality, and there were 5 spots indicating the presence of essential oils through TLC testing with a total essential oil content of 0.82% w/v, which was determined by the Stahl distillation method. The chemical content of citronella oil from Jatijejer Village has a content profile and chromatogram pattern that is quite similar to PT.N, as proven by GC-MS testing. The results of in silico testing show that one of the chemical contents, namely Geraniol (-6.3 kcal/mol), has a lower binding energy value (Delta G) than nicotine and can bind to important amino acids, so that it has the potential to replace nicotine. Based on the results of the drug-likeness analysis, the compound has also met Lipinski's rule of five and Veber's rule but does not meet the Ghose Filter Law. Meanwhile, based on the interaction, the chemical content of (1R)-cis-Verbenol and Geranial dimethyl acetal has the most appropriate %similarity to nicotine, and the chemical content of Chavibetol has the most appropriate %similarity to varenicline.
A selective UPLC-MS/MS assay was established and fully validated to quantify segetalin A, segetalin B, and hypaphorine in mouse plasma, supporting subsequent pharmacokinetic evaluation. Plasma samples were prepared by protein precipitation using methanol, with only 10 mu L of plasma required. A UPLC HSS T3 column was used for separation with an 0.1% formic acid-acetonitrile as mobile phase, with elution completed within 3.5 min. Multiple reaction monitoring coupled with electrospray ionization was used for quantitative analysis. Mice received segetalin A, segetalin B, and hypaphorine intravenously (5 mg kg(-1)) or orally (10 mg kg(-1)), and the pharmacokinetics of all three compounds were evaluated. The UPLC-MS/MS method showed excellent linearity in mouse plasma: 2.0-2,100 ng mL(-1) for segetalin A, 2.0-2,080 ng mL(-1) for segetalin B, and 2.2-2,280 ng mL(-1) for hypaphorine (all R-2 > 0.995). It was used to evaluate the pharmacokinetics of all three compounds after intravenous and oral dosing in mice. The oral bioavailability of segetalin A, segetalin B, and hypaphorine was determined to be 3.18, 1.17, and 63.44% in mice, respectively.
High-performance liquid chromatography (HPLC) is a critical analytical technique in the pharmaceutical sector, valued for its accuracy, speed and efficiency in drug quantification. This study aimed to develop and validate a novel reversed-phase HPLC (RP-HPLC) method for quantifying triamcinolone base (TRb) in solid pharmaceutical forms, using consistent chromatographic conditions for both dosage determination and dissolution studies. The method was validated in accordance with regulatory guidelines and demonstrated excellent precision, accuracy, and robustness. The mobile phase consisted of methanol and water (50:50), with a flow rate of 1.5 mL min(-1), detection at 254 nm and a column temperature of 30 degrees C. Key performance parameters included selectivity, linearity (R = 0.9998 across 2-12 mu g mL(-1)), precision (RSD < 2%), accuracy (98-102% recovery in the presence of two placebo matrices), and robustness (seven variables evaluated without significant impact). The retention time was 4.61 min. The method proved effective for both dosage quantification and formulation dissolution studies. Applying the same methodology across various stages of pharmaceutical development enhances standardization, traceability, and data reliability. As such, this RP-HPLC method is highly recommended for quality control laboratories seeking practical, sustainable, and scientifically robust analytical solutions.
