HPLC-UV Method For Simultaneous Determination Research Articles

Development and validation of a reversed-phase HPLC-UV method for simultaneous determination of levosulpiride and omeprazole in human plasma: Applicability of the method for evaluation of pharmacokinetic drug-drug interactions.

Levosulpiride and omeprazole are co-prescribed for gastrointestinal disorders associated with depression and anxiety. Objective of the study was to develop a sensitive, robust and simple method for simultaneous analysis of levosulpiride and omeprazole in human plasma and applicability of the method in determination of pharmacokinetics drug-drug interaction. In the presented study, a reversed-phase HPLC-UV method was developed for the simultaneous determination of levosulpiride and omeprazole using pantoprazole as the internal standard. Experimental conditions were optimized and the developed method was validated as per standard guidelines (USP and ICH). Furthermore, the developed method was applied for evaluation of pharmacokinetics drug-drug interaction between levosulpiride (50 mg) and omeprazole (40 mg) in healthy human volunteers. Sharpsil C8 column (4.6 × 250 mm, 5 μm), Ultisil C8 column (4.6 mm × 150 mm, 5 μm) and Agilent C18 column (4.6 × 250 mm, 5 μm) were evaluated as stationary phase. The best resolution was achieved with Agilent C18 (4.6 x 250 mm, 5 μm) column and was selected for further study. The mobile phase consisted of a mixture of acetonitrile and phosphate buffer (pH 7.2) in 60:40 by volume, and was pumped at a flow rate of 1 mL/min. Detector wavelength was set at 280 nm. Levosulpiride and omeprazole were extracted from human plasma with ethyl acetate and dichloromethane (4:1, v/v). The calibration curves for both levosulpiride (5-150 ng/mL) and omeprazole (10-1500 ng/mL) were linear. The lower limit of quantification and limit of detection for levosulpiride were 5 and 2 ng/mL, while for omeprazole these were 10 and 3 ng/mL, respectively. Pharmacokinetics analysis showed that co-administration of omeprazole increased the AUC and Cmax of levosulpiride, while the clearance was reduced. Both the changes were insignificant. Similarly, no significant change in the pharmacokinetic parameters of omeprazole was observed with co-administration of levosulpiride.

Development and Validation of a HPLC-UV Method for Simultaneous Determination of Lutein and β -Carotene in Food Formulated with Lutein Diester and β-carotene

A simple and rapid method has been developed for determination of lutein and β-carotene in food formulated with lutein diester beadlet, microencapsulated β-carotene as ingredients by reverse-phase high performance liquid chromatograph. This method combines extraction and saponification to one step reaction and lack of evaporation. Dimethyl sulfoxide (DMSO) was selected as solvent to release lutein diester, β-carotene from lutein diester beadlet and microencapsulated β-carotene, then add acetone-ethanol-20%KOH/methanol (10:5:4, v/v/v) to saponify lutein diester as well as extract lutein and β-carotene directly at the same time. The prepared sample solution was subjected to a HPLC analysis using Athena C30 column eluting with Methanol / methyl tert-butyl ether (MTBE) mobile phase. The precision, accuracy, linearity, specificity and ruggedness of the developed method were validated following AOAC guidance for single laboratory validation procedure. The recovery of lutein and β-carotene are 94.80%-104.76% and 98.94%-104.69% respectively. The injection repeatability: RSD(r) =1.8% for lutein, RSD(r) =5.5% for β-carotene. This method developed can be used for the quantitation of free lutein and β-carotene in products adding with lutein diester and β-carotene in quality control.

Validation of HPLC-UV method for simultaneous determination of quercetin and luteolin from chartamus tinctorius L in solid lipid nanoparticles incorporated in floating gel in situ formulation

Safflower extract (SE) is known to have various benefits in the drug and food industries. The main ingredients, like quercetin and luteolin, show vigorous antioxidant activites but have low solubility and stability to air, light and heat. Solid lipid nanoparticles (SLNs) are suitable carrier to overcome the stability issues and improve the solubility of SE. In aiming to deliver via oral route, the absorption can be further enhanced through floating rafts system (GFRS). Here, we developed a novel validated RP-HPLC method for determining simultaneous quercetin and luteolin to support formulation development. Effective chromatographic separation was carried out using a reversed-phase column C18 (150 × 4.6 mm, 5 µm) with isocratic method; 0.1% v/v trifluoro acetic acid (TFA) in water and acetonitrile (35 %v/v: 65 %v/v). The method was validated in four different media; methanol, FaSSGF, FeSSGF, and PBS pH 7,4. The results indicate the linearity of the method with a 0.998 correlation coefficient (R). Additionally, all media's LLOQ values ranged between 0.12 µg/mL and 0.31 µg/mL. Notably, the procedure was precise and accurate with the necessary dilution integrity, indicating the methods' validity. Finally, the method was successfully applied to determine quercetin and luteolin in SE, SLNs containing SE, and floating in situ gel containing SLNs of SE. Consequently, the delivery approach developed here could be an appropriate solution for the oral delivery of SE. Based on these findings, additional in vivo investigations on an animal model should be conducted to evaluate the efficacy of this approach.

Development of HPLC-UV Method for Simultaneous Determination of Corticosteroid Co-Administered Immune Therapy.

Prednisolone (PDS) has recently been utilized to treat a variety of medical disorders, including autoimmune illnesses and cancer. It is also used to treat coronavirus disease 2019 infection-related respiratory problems. Because it may induce health problems including gastrointestinal lesions and ulceration, it has to be used alongside other drugs like esomeprazole (ESM), which acts as a proton pump antagonist to reduce the probability of ulceration. As a result, the goal of this research is to create an environmentally safe and sensitive high-performance liquid chromatography (HPLC) approach for determining PDS and ESM in their binary combination and spiked human plasma. C8 column (100 × 4.6mm, 5μm) and gradient mobile phase elution were used to separate the studied drugs with ultraviolet recognition at 290nm. Caffeine was utilized as an internal standard to adjust the sample variance. Plasma, caffeine, ESM and PDS all had tR values of 1.4, 3.5, 6.3 and 7.3, respectively. The suggested method's greenness features were evaluated using three greenness evaluation tools: green analytical procedure index, analytical greenness metric approach and analytical eco-scale, and the findings were approved and satisfied. Validation parameters were evaluated in accordance with US-FDA recommendations in order to meet the global desires for biological analysis technique, acceptable limits were obtained.

Development and validation of a novel HPLC-UV method for simultaneous determination of azathioprine metabolites in human red blood cells

A rapid, specific and accurate high-performance liquid chromatography with tunable ultraviolet detection method was developed to simultaneously determine azathioprine metabolites, 6-thioguanine nucleotides (6-TGN) and 6-methyl mercaptopurine riboside (6-MMPr) in human red blood cells. Erythrocyte lysate sample was precipitated by perchloric acid under the protection of dithiothreitol, with 6-TGN and 6-MMPr being acid hydrolyzed to produce 6-thioguanine (6-TG) and 6-methymercaptopurine (6-MMP). A Waters Cortecs C18 column (2.1 × 150 mm, 2.7 μm) was used for chromatographic separation with a water (containing 0.01 mol/L ammonium acetate and 0.2% acetic acid)/methanol linear gradient at a flow rate of 0.45 mL/min in a 5.5 min. UV detection wavelengths were 340 nm for 6-TG, 303 nm for 6-MMP and the IS (5-bromouracil). The calibration curves fitted a least squares model (weighed 1/x2) from 0.15 to 15 μmol/L for 6-TG (r2 = 0.9999) and from 1 to 100 μmol/L for 6-MMP (r2 = 0.9998). This method was validated according to the FDA bioanalytical method validation guidance and ICH M10 bioanalytical method validation and study sample analysis guidance for industry, and successfully utilized in ten IBD patients receiving azathioprine therapy.

High-performance liquid chromatographic detection of selected phenolic acidsin wine

The aim of this study was to develop a method for identification and quantification of phenolic acids in different wine samples. The simple reversed-phase HPLC-UV method for simultaneous determination of p-coumaric and ferulic acid was developed. The method was validated and working range, linearity, repeatability, accuracy, limit of quantitation LOQ and limit of detection LOD were determined. The linearity of the method was tested in concentration ranges 0.1-1 mg L-1 and 1-10 mg L-1. The correlation coefficients (r2) were greater than 0.996 and quality coefficients (QC) ≤ 5%. Detection limit for both compounds was 0.01 mg L-1. The method is precise (RSD

Novel HPLC-UV method for simultaneous determination of valsartan and atenolol in fixed dosage form; Study of green profile assessment

Aim of this work was to develop the first simple, rapid, green, economical and selective HPLC method for simultaneous quantification of the cited drugs in their challenging binary mixture. The work was motivated by the global trends towards sustainable chemistry in designing eco-friendly mobile system without affecting the analysis parameters. The proposed method was subjected to a greenness profiles using some metrics as Eco-scale. Materials and methods. This was accomplished under the following chromatographic conditions: HPLC column Discovery C18 (4.6 mm i.d. × 150 mm, 5 μm), column temperature 30 °C, flow rate 1.0 mL/min, mobile phase composed of 20% acetonitrile, 80% of 0.16% ammonium acetate and 0.2% of 1.5 M tetramethylammonium hydroxide (V/V) and signal monitoring at a wavelength of 225 nm and 237 nm. Results. A conventional mixture of acetonitrile and 0.16% ammonium acetate was tried in different ratios, but the drugs were not well separated. The shortest aliphatic chain cationic ion pair reagent tetramethylammonium hydroxide should not be exchanged with other type similar with this, like tetramethylammonium hydrogen sulfate, it did not work to our experiments. Increasing salt concentration, ammonium acetate, more than 0.2%, pushes the peak of atenolol closer to dead volume, which is negative. Atenolol in their methods for multicomponent mixtures elutes in dead volume, or when retained longer, much stronger, hydrophobic mobile phase should be used if valsartan should be seen in same chromatogram at dissent time. The 237 nm can be chosen as compromise signal for nearly equal peaks height with high sensitivity is not essential. The 225 nm signal shows much higher sensitivity for atenolol and less increase for valsartan peaks, which can be used when higher sensitivities will be essential. Linearity was examined and proven at different concentration levels in the range of working concentration of valsartan (0.16–0.96 mg/mL) and atenolol (0.2–1.20 mg/mL). The high value of recoveries obtained for valsartan and atenolol indicates that the proposed method was found to be accurate. The results of proposed method found to be an excellent green analysis with a score of 84. Conclusion. A new fast, simple and green, but selective, accurate, precise and robust HPLC-UV method for simultaneous determination of valsartan and atenolol in newly formulated dosage form was developed and many possible variations of the same were suggested. The developed method for the simultaneous quantification of valsartan and atenolol in their challenging binary mixture offers simplicity essential for quality control of a large number of samples in short time intervals, which is necessary for routine analysis. The method was subjected to greenness profile assessment.

A bio-analytically validated HPLC-UV method for simultaneous determination of doripenem and ertapenem in pharmaceutical dosage forms and human plasma: a dual carbapenem regimen for treatment of drug-resistant strain of Klebsiella pneumoniae.

The emergence of strains resistant to certain antibiotics is turning into an important issue worldwide that threatens global health with the increasing incidence of carbapenemase-producing Klebsiella pneumoniae (KPC). Thus, successful doripenem–ertapenem (DOR–ERTA) combination is highly recommended in treatment of bacteremic ventilator-associated pneumonia due to Klebsiella pneumoniae. Hence, a fast and highly-sensitive HPLC-UV method was developed for the estimation of the cited drugs simultaneously in their pure form, pharmaceutical dosage forms and in simulated synthetic mixtures. The DOR–ERTA mixture was successfully separated within 6 min on a reversed-phase ODS column using an isocratic elution; a mobile phase mixture consists of 0.05 M phosphate buffer (pH 3.0 adjusted by 85% ortho-phosphoric acid) : acetonitrile : methanol (86 : 12 : 2; % v/v/v). The proposed method was optimized and validated according to ICH guidelines, where the calibration graph was constructed from 0.04 to 50 μg mL−1 and from 0.05 to 50 μg mL−1 with low detection limits reached 1.7 and 1.4 ng mL−1 for DOR and ERTA respectively. The proposed method showed higher sensitivity than several previous methods, which allowed an effective estimation of the DOR and ERTA in human plasma after a simple extraction method with high recovery results ranged from 96.30% ± 1.55 to 97.90% ± 1.45 and without any interference from plasma components.

Rapid Determination for Benzoic Acid, Sorbic Acid, Phenyllactic Acid, Phenylalanine, and Saccharin Sodium in Vinegar by High-Performance Liquid Chromatography–UV

Preservatives and sweeteners are always added in the food production process, which can extend the life and flavor of food. Benzoic acid and saccharin sodium are commonly used as preservatives and sweeteners respectively. Phenyllactic acid as a natural preservative, mainly produced by phenylalanine fermentation, may exist in fermented food. The objective of the current study is to develop a reverse-phase HPLC-UV method for simultaneous determination of benzoic acid, sorbic acid, phenyllactic, phenylalanine, and saccharin sodium in vinegar. They were separated on Sapphiresil TM (C18) column (250 × 4.6 mm, 5 μm) with a mobile phase consisting of methyl alcohol:ammonium acetate buffer (pH 6.44; 0.02 M) (20:80, v/v) ran in isocratic mode at a flow rate of 1 mL/min. Complete baseline separation of five analytes was achieved in 0.9975, the method was validated with recoveries in the 96.93 to 118.42% range, and RSD was 0.39–6.21%. The detection limits of phenylalanine, benzoic acid, saccharin sodium, sorbic acid, and phenyllactic acid were 0.25, 0.01, 0.01, 0.025, and 0.05 mg/L. This method could be used for determining of benzoic acid, sorbic acid, phenyllactic acid, phenylalanine, and saccharin sodium in vinegar.

Development and validation of an improved HPLC-UV method for simultaneous determination of lamotrigine and oxcarbazepine and its active metabolite 10,11-dihydro-10-hydroxycarbazepine in human blood plasma and comparison with an UHPLC-MS/MS method

Lamotrigine (LTG) and oxcarbazepine (OXC) are first-line drugs for epilepsy treatment. Their large pharmacokinetics variabilities and relations between efficacy and toxicity and blood plasma concentration require routine monitoring for dose adjustment. In this study, we developed and validated a simple, accurate, and reliable method for simultaneous determination of LTG, OXC and 10,11-dihydro-10-hydroxycarbazepine (MHD) in human blood plasma by high-performance liquid chromatography-ultraviolet detection (HPLC-UV) with a simple one-step protein precipitation using methanol (1% acetic acid) and 15 min elution time under isocratic elution at 1 mL/min. Calibration range was 2.4 to 120 mg/L for LTG, OXC, and MHD. The intra-day and inter-day bias were − 8.84 to 4.18%, and the imprecision was less than 8.08% for all analytes. The internal standard (fluconazole) normalized recovery was 96.30 to 107.69% for LTG, 98.51 to 111.04% for MHD, and 95.04 to 109.86% for OXC. A total of 186 LTG samples and 25 MHD samples were used to evaluate the agreement between HPLC-UV and ultra-performance liquid chromatography-mass spectrometry (UHPLC-MS/MS) by Passing-Bablok regression and Bland-Altman plot. The mean bias and the 95% limits of agreement (95% LOA) of the two measurements were 0.575 mg/L and − 1.238 to 2.387 mg/L for LTG (n = 186) and − 1.222 mg/L and − 8.271 to 5.827 mg/L for MHD (n = 25), which indicated the UV method was comparable with the MS method for LTG and MHD analysis.

Development and validation of an HPLC-UV method for simultaneous determination of sildenafil and tramadol in biological fluids: Application to drug-drug interaction study

The introduction of sildenafil (SDF) to treat erectile dysfunction has solved a widespread condition with negative on the quality of life. Recently, the co-administration of tramadol (TMD) with SDF to manage premature ejaculation has illegally increased and thus drug-drug interaction studies of these drugs became of great importance. Although certain biological functions have been altered upon co-administration of the two drugs, methods for their determination in vivo to understand their interactions have yet to be published. Herein, therefore, an HPLC method with photometric detection was developed for the determination of a binary mixture of TMD and SDF in rabbit plasma after oral administration. In this study, a reversed-phase chromatography was performed at room temperature on a C18 column with a mobile phase composed of 10 mM Na2HPO4 solution (pH 7.5): acetonitrile (45:55, v/v) at a flow rate of 0.8 mL min−1 using caffeine (CAF) as an internal standard. The detector was set at 220 nm. The total analysis time was 6 min. Calibration graphs were linear in the concentration ranges of 0.1–10 and 0.05–10 μg mL−1 with a detection limit of 0.05 and 0.02 μg mL−1 for TMD and SDF, respectively. The method was validated in terms of accuracy, precision, limit of detection and quantitation, recovery, and stability as per US FDA bioanalytical guidelines. In addition, the metabolites N-desmethylsildenafil (UK-103,320) and O-desmethyltramadol were quantified in rabbit plasma after 2 h of oral administration using LC–MS/MS. The simultaneous administration of TMD with SDF has affected peak plasma concentration (Cmax), Tmax, area under the concentration-time curve (AUC), and the elimination rate constant (Kel) of SDF. The present study is the first to give valuable insights into the drug-drug interaction and the pharmacokinetic implications associated with the co-administration of SDF and TMD.

A Rapid and Precise Liquid Chromatographic Method for Simultaneous Determination of Alpha Lipoic Acid and Docetaxel in Lipid-Based Nanoformulations

Combinational drug delivery successfully merges the benefits of nanotechnology and combination therapy by providing diversity to improve the carrier properties and better control over tailoring them as per the need of cancer treatment. A combination of conventional chemotherapeutic agent; docetaxel (DTX) and antioxidant agent; alpha lipoic acid (ALA) which acts by preventing metastasis may fulfill idealness of control and targeted drug delivery against breast cancer. The objective of the current study is to develop a reverse-phase HPLC-UV method for simultaneous determination of DTX and ALA in lipid-based nanoformulations. DTX and ALA were separated on Intersil® ODS (C18) column (250 × 4.6 mm, 5 μm) with a mobile phase consisting of acetonitrile: sodium acetate buffer (pH 3.5; 10 mM) (65:35% v/v) run in isocratic mode at a flow rate of 1 mL/min. The developed method was validated as per ICH guidelines. The method showed linearity in the concentration range of 1-15 μg/mL for DTX and 2-30 μg/mL for ALA. It can detect minimum 200 ng/mL of DTX and 500 ng/mL of ALA. The method was further successfully applied in lipid-based formulation characterization. In conclusion, a simple, accurate and precise reverse-phase HPLC-UV method was established for simultaneous determination of DTX and ALA in nanoformulations.

HPLC-UV method for simultaneous determination of irbesartan, candesartan, gliquidone and pioglitazone in formulations and in human serum

HPLC-UV method for simultaneous determination of irbesartan, candesartan, gliquidone and pioglitazone in formulations and in human serum

Novel HPLC-UV Method for Simultaneous Determination of Fat-soluble Vitamins and Coenzyme Q10 in Medicines and Supplements.

A precise, accurate and rapid HPLC-UV method for simultaneous determination of fat-soluble vitamins (vitamin D3, E-acetate, K1, β-carotene, A-palmitate) and coenzyme Q10 was developed and validated according to ICH guidelines. Optimal chromatographic separation of the analytes in minimal analysis time (8 min) was achieved on a Luna C18 150 × 4.6 mm column using a mixture of acetonitrile, tetrahydrofuran and water (50:45:5, v/v/v). The described reversed phase HPLC method is the first published for quantification of these five fat-soluble vitamins and coenzyme Q10 within a single chromatographic run. The method was further applied for quantification of the analytes in selected liquid and solid dosage forms, registered as nutritional supplements and prescription medicines, which confirmed its suitability for routine analysis.

HPLC-UV method for simultaneous determination of MK-1775 and AZD-7762 in both acetonitrile-aqueous solution and mouse plasma.

A sensitive and precise method is described for the simultaneous determination of two small molecule kinase inhibitors: MK-1775 (MK) and AZD-7762 (AZD), in acetonitrile (ACN)-aqueous solution and in mouse plasma. A Nova-Pak C18 reversed phase column (3.9mm×150mm, 4μm, 60Å) was utilized in the separation using an isocratic mobile phase of 0.1% v/v triethylamine in phosphate buffer (pH=7.4): acetonitrile (ACN) (60:40, v/v), at a flow rate of 0.8mL/min. Detection wavelength was set at 310nm for both MK and AZD, and 431nm for the internal standard sunitinib (SUN). The developed method was validated following the ICH guidelines and it was shown to be accurate, precise and linear in the range of 41ng/mL to 8333ng/mL for both drugs in the ACN-aqueous solution and from 83ng/mL to 8333ng/mL for both drugs in mouse plasma samples. For the first time, the presented data suggest the suitability of this method for the simultaneous separation and quantification of MK and AZD in both ACN aqueous solution as well as in mouse plasma samples.

Development of validated HPLC-UV method for simultaneous determination of Metformin, Amlodipine, Glibenclamide and Atorvastatin in human plasma and application to protein binding studies

A simple, sensitive, fast, and economical HPLC method was developed and validated for simultaneous estimation of two fixed dose combinations frequently prescribed in diabetes (Metformin plus Glibenclamide) and hypertension with dyslipidemia (Amlodipine plus Atorvastatin) in Human plasma for the first time. The validated HPLC method was used to quantify the concentration of selected actives in ultrafiltrate. Optimum separation conditions were obtained with Water’s Novapack Phenyl (150mm×4.6mm, i.d., 5.0 μm) column with mobile phase consisting of 0.1% Phosphoric acid (pH 3.0) and acetonitrile (ACN) in gradient mode with column oven temperature maintained at 30°C and elution monitored by a UV detector at 227nm. Protein precipitation was employed to extract the selected analyte form human plasma. The recoveries were more than 90% for all analytes in cold aqueous 10% trichloroacetic acid (TCA) and acetonitrile. The optimized HPLC-UV was validated in the calibration range of 10–10,000ngmL−1 for Metformin, 25–5000ngmL−1 for amlodipine, 50–10,000ngmL−1 for glibenclamide and 10–5000ngmL−1 for atorvastatin. The mean relative error was least when weighing of 1/×2 was applied for calibration curve. The accuracy of samples for six replicate measurements at LLOQ level was within limit. The precision and accuracy of samples for six replicate measurements at LLOQ level was within limit. The validated method was applied for quantitation of selected analytes in ultrafiltrate from protein binding experiments. A four to five fold increase in unbound fraction was observed when spiked to human serum albumin. Further the unbound fraction of highly albumin bound drugs was increased nearly to double when incubated with Gly-HSA as compare to HSA.

Simultaneous determination of mycophenolate and its metabolite mycophenolate-7-o-glucuronide with an isocratic HPLC-UV-based method in human plasma and stability evaluation

Objectives: Mycophenolic acid (MPA) is an immunosuppressive agent which is commonly used in a fixed dose regime in solid organ transplantation. For clinical trials and therapeutic drug monitoring measuring plasma concentrations is necessary. Also, stability issues have to be addressed.Methods: We describe an isocratic, RP-based HPLC-UV method for simultaneous determination of MPA and its major metabolite Mycophenolic acid 7-o Glucuronide (MPAG) in human plasma. Pre-analytics included protein precipitation with acetonitrile. The method was validated according to EMA/FDA guidelines. Patient lithium-heparin plasma and blood was used for evaluation of short-term (72 hours at room temperature = RT) and long-term stability (2 years at −80 °C) without acidification.Results: Linearity was assessed in the concentration range of 0.5–40.0 μg/mL for MPA and 5.0–350.0 μg/mL for MPAG, respectively. For MPA coefficient of variation was <7.0% (lower limit of quantification = LLOQ: 10.8%), for MPAG <9.6% (LLOQ: 10.6%). Bias ranged between −1.9 and +1.5% for MPA and for MPAG between −4.3 and −0.3%. The method showed agreement with a reference method for both analytes. MPA remained stable for 7 h (−1.6 to +8.4% change to the initial concentration) and MPAG for 24 h (−1.8 to −11.5% change) at RT in lithium heparin blood. After 2 years of storage at −80 °C MPA, MPAG concentrations and 95% CIs remained within ±15% of the initial value.Conclusion: The presented assay is applicable for clinical studies. Blood samples were stable for 7 hours at RT and plasma for 2 years stored at −80 °C.

Validated HPLC-UV Method for Simultaneous Determination of Some Anti-Inflammatory and Analgesic Drugs

In the presented work identification and quantification of some anti-inflammatory and analgesic drugs, namely aceclofenac, diclofenac, paracetamol and para-aminophenol were carried out by validated HPLC-UV method. The chromatographic separation was achieved on HPLC C18 (100.0 × 2.1 mm, 1.7μm) column using isocratic mobile phase consisting of acetonitrile-phosphate buffer (50: 50, v/v) at a flow rate of 1.0 mL/min. The UV detection was carried out at 275 nm for all the compounds. The elution of aceclofenac, diclofenac, paracetamol and para-aminophenol was occurred at 7.0, 9.2, 2.0 and 4.2 min, respectively. The calibration curves were linear over the concentration range of 1–1000 μg/mL for aceclofenac and paracetamol, and 1–100 μg/mL for diclofenac and para-aminophenol. The developed method was validated according to ICH guidelines. The method was applied in the identification and quantitative determination of these compounds during routine quality control analysis and in stability studies.

A HPLC-UV Method for Simultaneous Determination of Hypocrellin A and B from Fermentation Broth of Shiraia Bambusicola LPHP-89

A rapid, sensitive and reproducible high performance liquid chromatographic (HPLC) method was developed and validated for simultaneous quantification of hypocrellin A, hypocrellin B from Fermentation Broth of Shiraia bambusicola LPHP-89. Isocratic RP-HPLC system with C18 column (4.6 mm × 250 mm i.d., 5 μ particle size) and a detector UV-VWD was employed. The mobile phase consisted of methanol、water and acetic acid (60:40:1, v/v/v) and was pumped at a flow rate of 1.0 mL/min. The injection volume was5 μL. The quantitation wavelength was set at 254 nm.Total run time was 20 min and retention times for hypocrellin A and hypocrellin B were 10.06 and 15.38 min, respectively.

Establishment of a HPLC-UV Method for Simultaneous Determination of DON, 15ACDON, and 3ACDON in Wheat

建立了测定小麦中B型单端孢酶烯族毒素脱氧雪腐镰刀菌烯醇(deoxynivalenol, DON)、3-乙酰脱氧雪腐镰刀菌烯醇(3-acetyldeoxynivalenol, 3-ACDON)和15-乙酰脱氧雪腐镰刀菌烯醇(15-acetyldeoxyni-valenol, 15-ACDON)的高效液相色谱-紫外(HPLC-UV)检测方法。用水提取小麦样品，提取液经无水乙醇等体积沉淀，再结合Oasis HLB固相萃取小柱可取得较好的净化效果。采用乙腈/0.005%磷酸水溶液二元梯度洗脱程序在高效液相色谱-紫外检测器上完成DON、15ACDON和3ACDON的定性定量分析。结果表明，在0.5~15.0 mg L -1 线性范围内，DON、15ACDON和3ACDON的平均加标回收率分别为89.8%、93.4%和92.9%，相对标准偏差分别为2.2%、2.0%和2.5% ( n =3)；检测限分别为12.2、10.5和16.7 μg kg -1 。该方法准确、重现性好，样品净化方法使杂峰干扰少，大幅减少有机溶剂的使用，成本低，适用于小麦中B型单端孢酶烯族毒素的大批量检测。