ABSTRACT
Abstract The objective of the present study is to develop and validate a simple, selective and accurate hydrophilic interaction liquid chromatography - a high performance liquid chromatography incorporating an evaporative light scattering detector (HILIC-HPLC-ELSD) method for simultaneously determining glucosamine hydrochloride and chondroitin sulfate in dietary supplements. The chromatographic separation was carried out on a ZIC-HILIC column (150 mm x 4.6 mm x 5µm) in isocratic system mode with a mobile phase of acetonitrile, 30 mM ammonium formate and water (77:20:3, v/v/v) at pH 4.5, a column temperature of 35°C, a flow rate of 1 mL.min-1, and an injection volume of 5 µL. An evaporative light scattering (ELS) detector was used. Effective separation was achieved by means of analyte resolution of more than 1.5 with an analysis run time of approximately 20 minutes. The linearity of glucosamine hydrochloride and chondroitin sulfate ranged from 0.4 to 2.5 mg.mL-1. The limits of the detection and quantification of glucosamine hydrochloride were 20 and 80 mg.mL-1 respectively, while for chondroitin sulfate they were 80 and 400 mg.mL-1. All validation parameters satisfied the acceptance criteria in accordance with International Conference on Harmonisation (ICH) guidelines. The method was successfully applied to the assay of commercial dietary supplement samples
Subject(s)
Chromatography, High Pressure Liquid/methods , Chromatography, Liquid/methods , Dietary Supplements/analysis , Validation Study , Glucosamine/agonistsABSTRACT
Aryloxypropanolamine is an essential structural scaffold for a variety of β-adrenergic receptor antago-nists such as metoprolol. Molecules with such a structural motif tend to degrade into α, β-hydroxypropanolamine impurities via a radical-initiated oxidation pathway. These impurities are typically polar and nonchromophoric, and are thus often overlooked using traditional reversed phase chromatography and UV detection. In this work, stress testing of metoprolol confirmed the generation of 3-isopropylamino-1,2-propanediol as a degradation product, which is a specified impurity of metoprolol in the European Pharmacopoeia (impurity N). To ensure the safety and quality of metoprolol drug products, hydrophilic interaction chromatography (HILIC) methods using Halo Penta HILIC column (150 mm×4.6 mm, 5 μm) coupled with charged aerosol detection (CAD) were developed and optimized for the separation and quantitation of metoprolol impurity N in metoprolol drug products including metoprolol tartrate injection, metoprolol tartrate tablets, and metoprolol succinate extended-release tablets. These HILIC-CAD methods were validated per USP validation guidelines with respect to speci-ficity, linearity, accuracy, and precision, and have been successfully applied to determine impurity N in metoprolol drug products.
ABSTRACT
To study the substances in fudosteine, one synthetic by-product and five forced degradation products were detected by hydrophilic interaction chromatography (HILIC). Quadrupole-time-of-flight mass spectrometry (Q-TOF MS) was used for accurate mass determination and product ion scanning. Five related substances were identified in the products of mass spectra fragmentations elucidation, and verified further according to synthetic process and stress testing results. The results obtained are valuable for fudosteine manufacturing process control and quality assurance.
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This study was aimed to identify the photodegradation impurities (PD1 and PD2) in melphalan hydrochloride and establish a method for determining PD1 and PD2.The structures of the photodegradation impurities were inferred by LC-MS/MS.The impurities were confirmed by comparing with synthesized impurities.An HILIC method was established to determine PD1 and PD2.The method was carried out on an Atlantis HILIC column(4.6 mm × 150 mm,3 μm) with the mobile phase consisting of 0.1 mol/L ammonium formate (adjusted to pH 3.0 with formic acid) and acetonitrile (13∶ 87) at the flow rate of 1.0 mL/min.The column temperature was 35 ℃.The detection wavelength was 260 nm.PD1 and PD2 were characterized as 4-amino-L-phenylalanine hydrochloride and 4-(2-chloroethyl) amino-L-phenylalanine hydrochloride,respectively.Melphalan hydrochloride,PD1 and PD2 were separated completely under the HILIC condition.The established HILIC method can be used to determine the photodegradation impurities.
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OBJECTIVE: To develop an intergrated urinary metabonomic strategy based on RP-UPLC-MS and HILIC-UPLC-MS method and to investigated the mechanism of osteoporosis and the prevention effect of glucocorticoid osteoporosis of Gushudan. METHODS: The RP-UPLC-MS and HILIC-UPLC-MS method were developed for rat urine metabolite profiling study among control group, model group and Gushudan treatment group. Principal component analysis (PCA) were used to find the biomarkers, further more, to explore the mechanism of the prevention effect on glucocorticoid osteoporosis of Gushudan. RESULTS: In the mode of RP-UPLC-MS, low polarity metabolites like phenylacetylglycine, N2-succinic acid-L-ornithine were found while relatively high polarity metabolites like betaine, hypoxanthine were found in the mode of HILIC-UPLC-MS. And twenty-two potential biomarkers have been totaly found in the urine of glucocorticoid osteoporosis, primiarily related to amino acid metabolism, energy metabolism, lipid metabolism, intestinal flora metabolism and kidney damage. Gushudan has intervention effects on rats with osteoprosis via the regulation of multiplemetabolic pathways. CONCLUSION: The combination of RP-UPLC-MS and HILIC-UPLC-MS method can give a more comprehensive profiling of endogenous metabolites and it also indicates metabonomic strategy has become a powerful tool to evaluate the overall efficacy of traditional Chinese medicine (TCM) and to calrify the mechanism of TCM.
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@#A two-dimensional preparative liquid chromatographic method was developed for purification of constituents from water extract of Armillaria luteo-virens using reversed phase preparative liquid chromatography(RPLC)coupled with hydrophilic interaction liquid chromatography(HILIC). Seven compounds were isolated from fruit bodies of Armillaria luteo-virens, and their structures were identified on the basis of physicochemical and spectroscopic analyses. These compounds were identified as pyroglutamic acid(1), uridine(2), 2′-deoxyuridine(3), uracil(4), guanosine(5), inosine(6), and adenosine(7). All the compounds were purified and identified from fruit bodies of Armillaria luteo-virens for the first time.
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Organic acids are widely distributed in plants and related products, and participate in a wide range of metabolic pathways (e.g. tricarboxylic acid cycle), showing diverse pharmacological activities. As a widely used Chinese patent medicine, its adverse reactions are often reported. Therefore, we should further clarify the chemical components of Shenfu injection, and prepare strict quality standards to ensure the safety and effectiveness of its clinical use. Shenfu injection is prepared from red ginseng (steamed roots of Panax ginseng) and black prepared lateral roots of Aconitum carmichaelii (Heishunpian) by using modern extraction process, and organic acids are regarded as one of its main components. In current study, a hydrophilic interaction chromatography (HILIC) coupled with mass spectrometric method (HILIC-LC-MS) was developed and validated for the simultaneous determination of 14 organic acids, including cinnamic acid, ferulic acid, 4-hydroxylbenzoic acid, L-(+)-lactic acid, adipic acid, fumaric acid, caffeic acid, succinic acid, maleic acid, malonic acid, D-malic acid, (-)-shikimic acid, D-tartaric acid, and quinic acid in Shenfu injection. Satisfactory retention and separation were achieved for all organic acids on HILIC chromatographic column. Except cinnamic acid (231 μg•L⁻¹), lactic acid (113 μg•L⁻¹) and malonic acid (32.5 μg•L⁻¹), the limit of quantitation for the remaining 11 compounds were less than 10 μg•L⁻¹. D-Malic acid, malonic acid, quinic acid, L-(+)-lactic acid, and cinnamic acid were observed to have higher contents in Shenfu injection (>1.89 mg•L⁻¹), whereas caffeic acid and adipic acid were undetectable in all batches. Above all, the developed method is suitable for the simultaneous determination of organic acids in Shenfu and some other traditional Chinese medicine injections.
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A novel bioanalytical method was developed and validated for the quantitative determination of darunavir (DRV) in rat plasma by employing hydrophilic interaction chromatography and tandem mass spectrometry (HILIC–MS/MS) with supported liquid extraction (SLE). Irbesartan (IRB) was used as an internal standard (IS). The analyte in rat plasma (200 mL) was isolated through SLE using ethyl acetate as the eluting solvent. The chromatographic separation was achieved on Luna-HILIC (250 mm*4.6 mm, 5 μm) column with a mobile phase of 0.1% of formic acid in water:acetonitrile (5: 95, v/v), at a constant flow rate of 1.0 mL/min. The MS/MS ion transitions for DRV (548.1-392.0) and IS (429.2-207.1) were monitored on an ion trap mass spectrometer, operating in the multiple reaction monitoring (MRM) mode. The lower limit of quantitation (LLOQ) was 0.2 ng/mL and quantitation range was 0.2–5000 ng/mL. The method was validated for its selectivity, sensitivity, carryover, linearity, precision, accuracy, recovery, matrix effect and stability. The method was successfully applied to pharmacokinetic study in rats.
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Objective:To establish an HILIC-HPLC-ELSD method for the determination of allantoin in Rhizoma Dioscoreae. Methods:The separation was performed on a Waters XBridge HILIC column(150 mm × 4. 6 mm,5μm),the mobile phase was ace-tonitrile-water(85:15)with the flow rate of 0. 8 ml·min-1 ,and the nebulizer pressure was 40psi,the drifting tube temperature was 80℃ and the nebulizer temperature was 50℃. Results:The linearity of allantoin was good within the range of 1. 5-7. 5 μg( r =0. 998 1). The regress equation was Y=1. 67X+2. 35,the detection limit was 25 ng,and the average recovery of allantoin in Rhizoma Dioscoreae was 99. 9% with RSD of 0. 72%(n=9). Conclusion:The method is simple and reliable,which can be used in the deter-mination of allantoin in Rhizoma Dioscoreae.