High-performance Liquid Chromatography System Research Articles

Partial Validation of High Performance Liquid Chromatography for Analysis of Isoniazid in Rat Plasma

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis. Nicotinic acid derivatives such as isoniazid have the strongest anti-tuberculosis properties. For pharmacokinetics studying of isoniazid (INH), a method is needed to determine the levels of INH in plasma. Objective: The aim of this research is to partial validate of high performance liquid chromatography (HPLC) for analysis of INH in rat plasma. Method: For the preliminary study, rat plasma was used. The HPLC system used is a stationary phase C18 with length 250mm and temperature of 30°C, mobile phase hexane sulphonate acid 20mM pH 2.47–methanol (65:35). The analytical parameters in partial validated were linearity, lower limit of quantification (LLOQ), precision, accuracy, and recovery. Results: The results of linearity test of INH showed r value of 0.9996. LLOQ of this method was 0.1258μg/mL. The resulting accuracy and precision value met FDA requirements with a percent recovery ranging from 96.57–107.99%. Conclusion: The HPLC system was a valid method for analysis of INH in rat plasma.

Validation of a rapid and sensitive HPLC/MS method for measuring sucrose, fructose and glucose in plant tissues

Sucrose and its constituent components, glucose and fructose, are major determinants of sweetness in fruits and cooking quality in plant storage organs, such as potato tubers. Accurate monitoring of these sugars provides a predictive assessment of plant product quality. High Performance Liquid Chromatography (HPLC) coupled to a variety of detectors has been the method of choice for selective detection and quantitation of sugars in plant tissues. However, sugar extraction can be time-consuming to obtain a clean and concentrated sample suitable for accurate quantitation. Herein, we took advantage of a modern HPLC system coupled to a highly sensitive high-resolution high-accuracy mass spectrometer to develop a rapid method for measuring glucose, fructose, and sucrose in plant tissues. The method was validated in mature potato tubers and strawberry fruits and it proved to be highly accurate and robust with minimal sample care requirements.

Simultaneous removal of cationic methylene blue and anionic reactive red 198 dyes using magnetic activated carbon nanoparticles: equilibrium, and kinetics analysis.

For the simultaneous adsorption of cationic dye (methylene blue, MB) and anionic dye (reactive red 198, RR198) from aqueous solution, magnetic activated carbon (MAC) nanocomposite as a promising adsorbent was prepared and used. The concentration of MB at different time intervals was determined using a UV-Vis spectrophotometer while the concentration of RR198 was determined using a high performance liquid chromatography (HPLC) system. The effect of solution pH, contact time, adsorbent amount, and dye concentration were investigated. Also, both kinetic and isotherm experiments were studied. The optimum pH was 10 and 5.5 for adsorption of MB and RR198, respectively, and the equilibrium status was achieved after 120 min. The adsorption kinetics was controlled by the pseudo-second order kinetic model more than pseudo-first order. The best-fitted isotherms were Freundlich and Langmuir models for MB and RR198, respectively. The higher values of Freundlich adsorption capacity (Kf) for MB in comparison with RR198 refer to MAC affinity to remove cationic dyes more than anionic dyes. Apparently, there was no substantial change in the adsorption efficiency among the 10 adsorption-desorption cycles. Overall, MAC can be considered as an effective and efficient viable adsorbent for cationic and anionic dyes removal from industrial wastewaters.

DISTRIBUTION AND SOURCES OF POLYCYCLIC AROMATIC HYDROCARBONS IN AQUATIC SEDIMENT FROM CAN GIO COASTAL WETLAND, HOCHIMINH CITY

The distribution of fifteen polycyclic aromatic hydrocarbons (PAHs) indicated from USEPA as priority pollutants was studied in surface sediments (0 - 20 cm) of coastal wetland area of Can Gio district, Hochiminh City. PAHs were recovered from the sediments by solvent extraction and then analyzed by means of high performance liquid chromatography system. Total concentrations of the ∑PAHs in the range 5 – 38 ng/g dw were detected, and a distinct spatial trend was observed. The contents of Nap, Ace, Flu, Phe and dBA were below detection limit. Diagnostic ratios such as benzo[a]anthracene/chrysene and fluoranthene/pyrene were achieved to evaluate the emission sources of PAHs. These ratios indicated an anthopogenic source (pyrolysis) of PAHs for sediments. Furthermore PAHs were associated mainly with fine particle sediments. Although the PAHs contents were below Vietnamese standard but potential risk to ecosystem needs further study since the high percentages of carcinogenic PAHs.

Suppression of Cisplatin-Induced Vomiting by Cannabis sativa in Pigeons: Neurochemical Evidences.

Cannabis sativa (CS, family Cannabinaceae) has been reported for its anti-emetic activity against cancer chemotherapy-induced emesis in animal models and in clinics. The current study was designed to investigate CS for potential effectiveness to attenuate cisplatin-induced vomiting in healthy pigeons and to study the impact on neurotransmitters involved centrally and peripherally in the act of vomiting. High-performance liquid chromatography system coupled with electrochemical detector was used for the quantification of neurotransmitters 5-hydroxytryptamine (5HT), dopamine (DA) and their metabolites; Di-hydroxy Phenyl Acetic acid (Dopac), Homovanillic acid (HVA), and 5-hydroxy indole acetic acid (5HIAA) centrally in specific brain areas (area postrema and brain stem) while, peripherally in small intestine. Cisplatin (7 mg/kg i.v.) induce emesis without lethality across the 24 h observation period. CS hexane fraction (CS-HexFr; 10 mg/kg) attenuated cisplatin-induced emesis ∼ 65.85% (P < 0.05); the reference anti-emetic drug, metoclopramide (MCP; 30 mg/kg), produced ∼43.90% reduction (P < 0.05). At acute time point (3rd h), CS-HexFr decreased (P < 0.001) the concentration of 5HT and 5HIAA in the area postrema, brain stem and intestine, while at 18th h (delayed time point) CS-HexFr attenuated (P < 0.001) the upsurge of 5HT caused by cisplatin in the brain stem and intestine and dopamine in the area postrema. CS-HexFr treatment alone did not alter the basal neurotransmitters and their metabolites in the brain areas and intestine except 5HIAA and HVA, which were decreased significantly. In conclusion the anti-emetic effect of CS-HexFr is mediated by anti-serotonergic and anti-dopaminergic components in a blended manner at the two different time points, i.e., 3rd and 18th h in pigeons.

Evaluation of solid‐phase extraction procedures for the quantitation of venlafaxine in human saliva by high‐performance liquid chromatography

In recent years, the use of human saliva for diagnostic purposes has evoked great interest. Thus, the aim of this study was to choose the optimal solid-phase extraction cartridges and extraction solvents for the quantitation of venlafaxine in saliva. Blank saliva samples spiked with venlafaxine concentrations between 25 and 750ng/mL were analyzed using five solid-phase extraction columns (C18, C8, Strata-X, Strata-X-C, and Strata-X-AW), washing solvents (deionized water, phosphate buffer at pH 5.5, and their mixtures with methanol), and elution solvents (methanol, acetonitrile, and their mixtures with 25% ammonia). A high-performance liquid chromatography system was used to quantify venlafaxine in saliva. The results of this study revealed that nine of 25 procedures enabled quantitation of venlafaxine in the tested concentration range. The procedure that used a C18 cartridge, a mixture of methanol and deionized water as the washing solvent, and methanol as the elution solvent was the most effective and allowed quantitation of all venlafaxine concentrations with an acceptable recovery. In contrast, the Strata-X-C cartridge could not detect venlafaxine at the lowest concentration (25ng/mL). The data acquired from the high-performance liquid chromatography system were confirmed by a multivariate data analysis.

Preparation of MIL-53(Fe)@polydopamine@Fe3O4 magnetic composite for magnetic solid-phase extraction of sulfonylurea herbicides in environmental water

Polydopamine (PDA) and MIL-53(Fe) modified Fe3O4 particles (MIL-53(Fe)@PDA@Fe3O4 magnetic composite) were prepared by simple one-pot solvothermal method. The obtained composite was introduced to rapidly extract four kinds of sulfonylurea herbicides (SUHs) from environmental water samples by magnetic solid-phase extraction (MSPE). Then the herbicides were analyzed by a high performance liquid chromatographic system equipped with a photodiode array detector. The mobile phase was a mixture of acetonitrile and water containing 0.01% (v/v) trifluoroacetic acid, and the detection wavelength was 233 nm. Significant extraction parameters were optimized to improve the extraction efficiency. Under the optimum conditions (5 mL desorption solvent of acetone, 4.5 min extraction time, 60 mg adsorbent dosage, 0.5 g NaCl and the pH 3 of the solution), the developed method showed good linearities with correlation coefficients (r) no less than 0.9980. The limits of detection (LODs, S/N=3) of the four SUHs were 0.28-0.77 μg/L. The method was successfully used to determine four SUHs in three kinds of environmental water samples with satisfactory recoveries ranging from 78.8% to 109.7%. Therefore, the MIL-53(Fe)@PDA@Fe3O4 magnetic composite is efficient and has good potential for the extraction of SUHs.

Determination of Pharmaceuticals by Dynamic Cell Membrane Chromatography Coupled with High-Performance Liquid Chromatography

ABSTRACTAs a biological affinity chromatographic method, cell membrane chromatography (CMC) using a silica stationary phase covered with specific cell membrane has been used in screening active components. The innovation of this work is that the bioactive cell membrane and the chromatographic packing are mixed and absorbed for the first time to form the pre-column. The pre-column was placed in front of a C18 column to create dynamic CMC online high-performance liquid chromatography (HPLC) system. The retention behavior and dynamic changes of pharmaceuticals were studied for this system. The results indicate that the retention time of the drug was increased and the symmetry factor reached the analytical level after the addition of the dynamic cell membrane pre-column. Therefore, the dynamic CMC coupled with HPLC system may be a potentially rapid and efficient drug analysis approach for the interaction of drug molecule and receptor on red blood cell membranes.

Eco-friendly synthesis of metal nanoparticles using ginger and garlic extracts as biocompatible novel antioxidant and antimicrobial agents

Ginger and garlic plants were used for the preparation of water and ethanolic extracts. The silver, copper, iron and zinc nanoparticles were eco-friendly synthesized using ginger and garlic extracts and their antioxidant and antibacterial activities were evaluated. The growth of nanoparticles was complemented with characterization using TEM and UV–Vis spectroscopic analysis. Transmission electron microscopy revealed the presence of mono-dispersed metals nanoparticles and silver nanoparticles of ginger have the smallest particle size around 10.10–18.33 nm. The investigated samples were evaluated as antibacterial agents against Staphylococcus aureus, Klebsiella pneumoniae, Candida albicans, Bacillus subtilis, Erwinia carotovora and Proteus vulgaris and as antifungal agents against C. albicans. The results of antimicrobial tests indicate that these nanoparticles possess significant activities compared to the results obtained by antibiotic standard. In addition, the total polyphenols, fractionation of phenols, flavonoids by high-performance liquid chromatography systems and antioxidant activity were evaluated.Graphical Comparison of the DPPH and ABTS antioxidant results.

ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF DILTIAZEM HYDROCHLORIDE IN BULK AND PHARMACEUTICAL DOSAGE FORM BY RP- HPLC

A simple, specific, precise and accurate Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method was developed and validated for the estimation of Diltiazem Hydrochloride in bulk and pharmaceutical dosage form. The chromatographic determination was performed on isocratic high performance liquid chromatography system of Agilent model no.1220. The separation was conducted by using column of Zorbax [C8 (5µ, 4.6 mm×250)] with mobile phase consisting of buffer and Acetonitrile in the ratio of (60:40). The mobile phase was delivered at the flow rate of 1.0ml/min. The eluent was monitored at wavelength 240 nm and found a sharp and symmetrical peak with retention time of 4.66min. The method was validated for linearity, accuracy, precision, specificity, robustness. Recovery of Diltiazem Hydrochloride was found to be in the range of 98%-102%. The method was found to be linear over the concentration range 50-150 µg/ml with coefficient correlation r2 = 0.995. After developing method, validation parameters were carried out successfully and obtained results were complied with USP monograph.

Enantioselective and simultaneous determination of lactate and 3-hydroxybutyrate in human plasma and urine using a narrow-bore online two-dimensional high-performance liquid chromatography system.

For the enantioselective and simultaneous analysis of lactate and 3-hydroxybutyrate, a validated online two-dimensional high-performance liquid chromatography system using 4-nitro-7-piperazino-2,1,3-benzoxadiazole as a fluorescent derivatization reagent has been developed. For the reversed-phase separation in the first dimension, a Capcell Pak C18 ACR column (1.5×250mm, particle size 3μm) was used, and the target fractions were isolated by their hydrophobicity. In the second dimension, a polysaccharide-coated enantioselective column, Chiralpak AD-H (2.0×250mm, 5μm), was used. The system was validated by the calibration curve, intraday precision, interday precision, and accuracy using standards and real human samples, and satisfactory results were obtained. The present method was applied to human plasma and urine, and in the plasma, trace amounts of d-lactate (8.4μM) and l-3-hydroxybutyrate (1.0μM), besides high levels of l-lactate (860.9μM) and d-3-hydroxybutyrate (59.4μM), were successfully determined. In urine, trace levels of d-lactate (3.7μM), d-3-hydroxybutyrate (2.3μM), and l-3-hydroxybutyrate (3.3μM) in addition to a relatively large amount of l-lactate (15.4μM) were observed. The present online two-dimensional high-performance liquid chromatography system is useful for the simultaneous determination of all the lactate and 3-hydroxybutyrate enantiomers in human physiological fluids, and further clinical applications are ongoing.

Detection of monohydroxylated polycyclic aromatic hydrocarbons in urine and particulate matter using LC separations coupled with integrated SPE and fluorescence detection or coupled with high-resolution time-of-flight mass spectrometry.

A high-performance liquid chromatographic (HPLC) method with integrated solid-phase extraction for the determination of 1-hydroxypyrene and 1-, 2-, 3-, 4- and 9-hydroxyphenanthrene in urine was developed and validated. After enzymatic treatment and centrifugation of 500 μL urine, 100 μL of the sample was directly injected into the HPLC system. Integrated solid-phase extraction was performed on a selective, copper phthalocyanine modified packing material. Subsequent chromatographic separation was achieved on a pentafluorophenyl core-shell column using a methanol gradient. For quantification, time-programmed fluorescence detection was used. Matrix-dependent recoveries were between 94.8 and 102.4%, repeatability and reproducibility ranged from 2.2 to 17.9% and detection limits lay between 2.6 and 13.6 ng/L urine. A set of 16 samples from normally exposed adults was analyzed using this HPLC-fluorescence detection method. Results were comparable with those reported in other studies. The chromatographic separation of the method was transferred to an ultra-high-performance liquid chromatography pentafluorophenyl core-shell column and coupled to a high-resolution time-of-flight mass spectrometer (HR-TOF-MS). The resulting method was used to demonstrate the applicability of LC-HR-TOF-MS for simultaneous target and suspect screening of monohydroxylated polycyclic aromatic hydrocarbons in extracts of urine and particulate matter.

High-Density Lipoprotein Function in Cardiovascular Disease and Diabetes Mellitus.

Cardiovascular disease (CVD) is the most common cause of death worldwide.1 Major CVD risk factors are hypertension, smoking, physical inactivity, abnormal glucose levels/diabetes mellitus, and dyslipidemia. Among these, dyslipidemia characterized by a low level of HDL-C (high-density lipoprotein cholesterol) is strongly and inversely correlated with CVD risk,2–4 and low HDL-C levels is part of the atherogenic dyslipidemia complex associated with diabetes mellitus.5 These observations triggered intense interest in increasing HDL-C levels for therapeutic intervention of CVD.6,7 However, several recent lines of evidence now suggest that the association between HDL-C levels and CVD status may be indirect or more complicated than previously recognized.7 Thus, human genetic studies demonstrate that genetically altered HDL-C levels do not necessarily translate to an altered risk of CVD.8–12 Phase III clinical trials of drugs that elevate HDL-C, such as niacin and CETP (cholesteryl ester transfer protein) inhibitors, also have largely failed to reduce CVD events in statin-treated subjects with established CVD.13–15 For several CETP inhibitors, improvement in CVD prevention was unsuccessful because of off-target effects (torcetrapib) or lack of efficacy (dalcetrapib and evacetrapib).14,16–18 The exception is the recent REVEAL trial (Randomized Evaluation of the Effects of Anacetrapib Through Lipid-Modification), which included more subjects and was performed for longer than previous CETP inhibitor trials. The REVEAL demonstrated that the CETP inhibitor anacetrapib exhibited beneficial effects on cardiovascular outcomes on top of those of statin therapy, although the risk reduction was moderate19,20 and might have been due, at least in part, to a reduction in non-HDL-C rather than elevated HDL-C.21 Considering these cumulative observations, it remains uncertain whether increased HDL-C directly impacts atherosclerosis and the …

Analysis of phosphatidylethanolamine, phosphatidylcholine, and plasmalogen molecular species in food lipids using an improved 2D high-performance liquid chromatography system

Phospholipids are an important class of lipids in cell membranes and food. Several high-performance liquid chromatography (HPLC) methods have been developed to analyze phospholipids at the molecular species level. We developed a two-dimensional HPLC system with a charged aerosol detector and mass spectrometry (MS) to analyze phosphatidylethanolamine (PE), phosphatidylcholine (PC), and their plasmalogens (pls) extracted from food materials. Accordingly, the phospholipid molecular species can be analyzed in a single step despite using smaller samples. We confirmed that chromatogram peaks from soybean lecithin are mostly baseline separated, assigned, and quantified (24 molecular species for PE and 27 for PC). In addition, it was confirmed that chromatograms of lipids extracted from chicken breast meat include plasmalogen peaks. The PE fraction in lipids extracted from chicken breast meat contained 17 types of ethanolamine plasmalogens, corresponding to approximately 57% of the total by weight. The PC fraction contained only four choline plasmalogens, corresponding to approximately 11% of the total weight. The composition of the pls-PC molecular species differed from that of pls-PEs. The polyunsaturated fatty acids connected at the sn-2 positions of the pls-PEs consisted of 20.5% 20:4 fatty acid and were independent of the carbon chain at the sn-1 position. However, the 18:1 fatty acid at the sn-2 position was dependent on the carbon chain at the sn-1 position.

A platform for automated nanomole-scale reaction screening and micromole-scale synthesis in flow.

The scarcity of complex intermediates in pharmaceutical research motivates the pursuit of reaction optimization protocols on submilligram scales. We report here the development of an automated flow-based synthesis platform, designed from commercially available components, that integrates both rapid nanomole-scale reaction screening and micromole-scale synthesis into a single modular unit. This system was validated by exploring a diverse range of reaction variables in a Suzuki-Miyaura coupling on nanomole scale at elevated temperatures, generating liquid chromatography-mass spectrometry data points for 5760 reactions at a rate of >1500 reactions per 24 hours. Through multiple injections of the same segment, the system directly produced micromole quantities of desired material. The optimal conditions were also replicated in traditional flow and batch mode at 50- to 200-milligram scale to provide good to excellent yields.

Determination of 2,4-Dichlorophenoxyacetic acid (2,4-D) in rat serum for pharmacokinetic studies with a simple HPLC method.

2,4-Dichlorophenoxyacetic acid (2,4-D) is a chlorophenoxy herbicide used worldwide. We describe a high-performance liquid chromatography (HPLC) method with UV detection for the determination of 2,4-D in female and male rat serum. This allows to observe the change of serum 2,4-D concentration in rats with time and its pharmacokinetics characteristics with a simple, rapid, optimized and validated method. The serum samples are pretreated and introduced into the HPLC system. The analytes are separated in a XDB-C18 column with a mobile phase of acetonitrile (solvent A) and 0.02 M ammonium acetate (containing 0.1% formic acid) (solvent B) using a gradient elution at a flow rate of 1.0 mL/min. The wavelength for UV detection was set at 230 nm. Calibration curve for 2,4-D was constructed over a range of 0.1–400 mg/L. The method was successfully applied to study the pharmacokinetics of 2,4-D in rats in this study. After oral administration of 300 mg/kg and 60 mg/kg 2,4-D, the mean Cmax values were 601.9 and 218.4 mg/L, the AUC0→∞ values were 23,722 and 4,127 mg×h/L and the clearance (Cl) were 1.10 and 0.02 L/(h×kg), respectively. The developed method was found to be specific, precise, reproducible and rapid.

Levels of malondialdehyde in the gastric juice: Its association with Helicobacter pylori infection and stomach diseases.

Helicobacter pylori (H.pylori) infection causes elevation of lipid peroxidation product malondialdehyde (MDA) and this association may be due to the bacterium causing reactive oxygen species-mediated damage to DNA in the gastric epithelium. The aim of this study was to investigate the gastric juice MDA levels in relation to H.pylori infection and associated gastric diseases. Gastric juice samples were obtained from 117 patients undergoing endoscopy, and gastric juice MDA levels were determined by high-performance liquid chromatography (HPLC) system. We compared the MDA levels between patients with and without H.pylori infection and assessed the differences of MDA levels between chronic gastritis, gastric intestinal metaplasia, and gastric cancer postsurgical resection. Malondialdehyde levels in gastric juice were significantly higher in chronic gastritis patients with H.pylori infection than in those without H.pylori infection (P<.0001). In patients without H.pylori infection, patients with gastric intestinal metaplasia and gastric cancer postsurgical resection had significantly higher gastric juice MDA level than patients with chronic gastritis. As a whole, patients with gastric intestinal metaplasia and gastric cancer postsurgical resection also had significantly higher MDA levels in gastric juice as compared to patients with chronic gastritis (P<.01). However, the difference of gastric juice MDA levels between gastric intestinal metaplasia and gastric cancer postsurgical resection was not significant. Malondialdehyde in gastric juice could be used as a potential diagnostic biomarker for H.pylori infection and associated gastric diseases. The gastric juice MDA levels increased proportionally with the severity of gastric diseases.

A Validated Stability-indicating RP-HPLC Method for Analysis of Azelaic Acid in Pharmaceuticals

The study is designed to develop a simple, rapid, selective and robust stability indicating reversed phase-high performance liquid chromatography method for quantitative analysis of azelaic acid in pharmaceutical preparations. The chromatographic separation and estimation of azelaic acid was carried out using a Waters high performance liquid chromatography system employing Kromasil 100-5C18 column (250×4.6 mm; 5 µm particle size) as a stationary phase. The mobile phase comprised of 75 volumes of sodium di-hydrogen orthophosphate (pH 3.5; 50 mM) and 25 volumes of acetonitrile, eluted at a flow rate of 1.2 ml/min. The eluents were monitored at 206 nm using 2487 dual wavelength ultra violet detector. The method was developed and validated in terms of stability as per International Conference on Harmonisation and Center for Drug Evaluation and Research guidelines. A linear relationship between peak area and concentration of azelaic acid was observed in a concentration range of 5-400 µg/ml (correlation coefficient, r2= 0.998). The method showed acceptable levels of precision (%RSD ≤2), accuracy (>96 % recovery), robustness (<10 % content difference) and stability (>96 % recovery) over varied environmental and laboratory conditions. The method was successfully applied for the determination of azelaic acid, extracted from three different batches of Aziderm® cream, which yielded a recovery of >97 %, indicated its applicability in routine analysis of azelaic acid in pharmaceutical preparations.

Determination of Eupatilin in Folium artemisiae Argyi and Its Inhibitory Effect on Hepatoma Cells.

Aim:The aim of this study is to establish a method for determination of eupatilin in Folium artemisiae Argyi and observe the inhibitory effect of Folium artemisiae Argyi extract on human hepatoma SMMC-7721 cells.Methods:High-performance liquid chromatograph system with 2910 pump, 2930 UV detector, and N2000 workstation was used for determination of eupatilin in Folium artemisiae Argyi. Human hepatoma SMMC-7721 cells were cultured and cell proliferation was measured using the MTT assay. The expression protein levels of p53, Topo II, and bcl-2 were detected using Western blotting.Results:Eupatilin exhibited a linearity range of 0.5–3.0 μg/mL and a recovery of 100.72%, relevant standard derivation = 2.28%. Folium artemisiae Argyi extract had marked cytostatic and cytotoxic effects on SMMC-7721 cells, inhibited the SMMC-7721 colony formation in a dose-dependent manner. Folium artemisiae Argyi extract already possessed delayed effect after treating SMMC-7721 cells for 8 h, which became obvious at 12 h from treatment. After drug withdrawal, cells still tended to apoptosis. Folium artemisiae Argyi extract could inhibit p53, Topo II, and bcl-2 expressions in tumor cells. The present method for determination of eupatilin is simple, fast, accurate, sensitive, and reproducible.Conclusion:Hepatoma SMMC-7721 cells are quite sensitive to Folium artemisiae Argyi extract, which may be associated with its suppression of p53, Topo II, and bcl-2 expressions.SUMMARY The study aimed to establish a method for determination of eupatilin in Folium artemisiae Argyi and observe the inhibitory effect of Folium artemisiae Argyi extract on human hepatoma SMMC-7721 cells. The results suggested that the present method for determination of eupatilin is simple, fast, accurate, sensitive, and reproducible. Hepatoma SMMC-7721 cells are quite sensitive to Folium artemisiae Argyi extract, which may be associated with its suppression of p53, Topo II, and bcl-2 expressions. Abbreviations used: HPLC: High-performance liquid chromatograph; OD: Optical density; RSD: Relevant standard derivation; IC50: Inhibitory 50% concentration.

Analysis of traditional Chinese medicine components by high performance liquid chromatography with diode array detection based on double qualitative principles

The double qualitative principle is a new composite qualitative method based on retention time and the characteristic peaks of the absorption spectra. Using a self-designed and assembled diode array detector (DAD), a high performance liquid chromatography (HPLC) system was constructed. The illegal additive auramine O in six kinds of herbal slices and the active ingredient schisandrin in Jujube kernel Tianma capsules were separated and qualitative analyzed using the HPLC-DAD system. The results showed that there were similar peaks in the chromatograms of pollen typhae and Jujube kernel Tianma capsules when comparing the target analytes. However, the probabilities of the targets were excluded by comparing the absorption spectra. The application results indicated that, based on the double qualitative principle of retention time/absorption spectrum, the interference of impurities in the samples could be well eliminated and the false positives could be avoided. This provides a reference method for the study of traditional Chinese medicine components.