Simultaneous Determination Of Salicylic Acid Research Articles

Simultaneous determination of shikimic acid, salicylic acid and jasmonic acid in wild and transgenic Nicotiana langsdorffii plants exposed to abiotic stresses

The presence and relative concentration of phytohormones may be regarded as a good indicator of an organism's physiological state. The integration of the rolC gene from Agrobacterium rhizogenes and of the rat glucocorticoid receptor (gr) in Nicotiana langsdorffii Weinmann plants has shown to determine various physiological and metabolic effects. The analysis of wild and transgenic N. langsdorffii plants, exposed to different abiotic stresses (high temperature, water deficit, and high chromium concentrations) was conducted, in order to investigate the metabolic effects of the inserted genes in response to the applied stresses. The development of a new analytical procedure was necessary, in order to assure the simultaneous determination of analytes and to obtain an adequately low limit of quantification. For the first time, a sensitive HPLC-HRMS quantitative method for the simultaneous determination of salicylic acid, jasmonic acid and shikimic acid was developed and validated. The method was applied to 80 plant samples, permitting the evaluation of plant stress responses and highlighting some metabolic mechanisms. Salicylic, jasmonic and shikimic acids proved to be suitable for the comprehension of plant stress responses. Chemical and heat stresses showed to induce the highest changes in plant hormonal status, differently affecting plant response. The potential of each genetic modification toward the applied stresses was marked and particularly the resistance of the gr modified plants was evidenced. This work provides new information in the study of N. langsdorffii and transgenic organisms, which could be useful for the further application of these transgenes.

Simultaneous determination of salicylic, 3-methyl salicylic, 4-methyl salicylic, acetylsalicylic and benzoic acids in fruit, vegetables and derived beverages by SPME–LC–UV/DAD

Salicylic and benzoic acid are phenolic acids occurring in plant cells, thus they can be present in fruit and vegetables at various levels. They possess anti-inflammatory and antimicrobial properties, however they may induce symptoms and health problems in a small percentage of the population. Therefore, a low phenolic acid diet may be of clinical benefit to such individuals. In order to achieve this goal, the concentration of these substances in different food and beverages should be assessed. The present work describes for the first time a new method, based on solid phase microextraction (polydimethylsiloxane–divinylbenzene fiber) coupled to liquid chromatography with UV diode array detection, for the simultaneous determination of salicylic acid, 3-methyl salicylic acid, 4-methyl salicylic acid, acetylsalicylic acid and benzoic acid in selected fruit, vegetables and beverages.All the aspects influencing fiber adsorption (time, temperature, pH, salt addition) and desorption (desorption and injection time, desorption solvent mixture composition) of the analytes have been investigated. An isocratic separation was performed using an acetonitrile-phosphate buffer (pH 2.8; 2mM) mixture (70:30, v/v) as the mobile phase. The estimated LOD and LOQ values (μg/mL) were in the range 0.002–0.028 and 0.007–0.095. The within-day and day-to-day precision values (RSD%) were between 4.7–6.1 and 6.6–9.4, respectively.The method has been successfully applied to the analysis of fava beans, blueberries, kiwi, tangerines, lemons, oranges and fruit juice (lemon and blueberry) samples. The major advantage of the method is that it only requires simple homogenization and/or centrifugation and dilution steps prior to SPME and injection in the LC system.

Simultaneous Determination of Salicylic Acid, Jasmonic Acid, Methyl Salicylate, and Methyl Jasmonate from Ulmus pumila Leaves by GC-MS.

Salicylic acid, jasmonic acid, methyl salicylate, and methyl jasmonate are important phytohormones and defensive signaling compounds, so it is of great importance to determine their levels rapidly and accurately. The study uses Ulmus pumila leaves infected by Tetraneura akinire Sasaki at different stages as materials; after extraction with 80% methanol and ethyl acetate and purification with primary secondary amine (PSA) and graphitized carbon blacks (GCB), the contents of signal compounds salicylic acid, jasmonic acid, methyl salicylate, and methyl jasmonate were determined by GC-MS. The results showed that the level of salicylic acid, jasmonic acid, methyl salicylate, and methyl jasmonate increased remarkably in U. pumila once infected by T. akinire Sasaki, but the maximums of these four compounds occurred at different times. Salicylic acid level reached the highest at the early stage, and jasmonic acid level went to the maximum in the middle stage; by contrast, change of content of methyl salicylate and methyl jasmonate was the quite opposite.

Simultaneous determination of salicylic acid and salicylamide in biological fluids

A new methodology for the simultaneous determination of salicylic acid and salicylamide in biological fluids is proposed. The strong overlapping of the fluorescence spectra of both analytes makes impossible the conventional fluorimetric determination. For that reason, the use of fluorescence decay curves to resolve mixtures of analytes is proposed; this is a novel technique that provides the benefits in selectivity and sensitivity of the fluorescence decay curves. In order to assess the goodness of the proposed method, a prediction set of synthetic samples were analyzed obtaining recuperation percentages between 98.2 and 104.6%. Finally, a study of the detection limits was done using a new criterion resulting in values for the detection limits of 8.2 and 11.6 μg L −1 for salicylic acid and salicylamide respectively. The validity of the method was tested in human serum and human urine spiked with aliquots of the analytes. Recoveries obtained were 96.2 and 94.5% for salicylic acid and salicylamide respectively.

Development and validation of a stability-indicating HPLC method for simultaneous determination of salicylic acid, betamethasone dipropionate and their related compounds in Diprosalic Lotion ®

Diprosalic Lotion ® is an anti-inflammatory drug product that contains salicylic acid and betamethasone dipropionate as active pharmaceutical ingredients (APIs). A reversed-phase high performance liquid chromatography (RP-HPLC) method was developed for simultaneous determination of salicylic acid, betamethasone dipropionate, and their related compounds in Diprosalic Lotion ®. A 150 mm × 4.6 mm I.D. YMC J'sphere ODS-H80 column at 35 °C and UV detection at 240 nm was used. A gradient elution was employed using 0.05% (v/v) methanesulfonic acid solution and acetonitrile as mobile phases. A total of thirty three compounds from Diprosalic Lotion ® samples were separated in 38 min. The stability-indicating capability of this method has been demonstrated by the adequate separation of all the impurities and degradation products in expired stability samples of Diprosalic Lotion ®. The method was validated as per the current ICH guidelines.

A novel application of Onyx™ monolithic column for simultaneous determination of salicylic acid and triamcinolone acetonide by sequential injection chromatography

A novel and fast simultaneous determination of triamcinolone acetonide (TCA) and salicylic acid (SA) in topical pharmaceutical formulations by sequential injection chromatography (SIC) as an alternative to classical high performance liquid chromatography (HPLC) has been developed. A recently introduced Onyx™ monolithic C18 (50 mm × 4.6 mm, Phenomenex ®) with 5 mm monolithic precolumn were used for the first time for creating sequential injection chromatography system based on a FIAlab ® 3000 with a six-port selection valve and 5.0 mL syringe pump in study. The mobile phase used was acetonitrile/water (35:65, v/v), pH 3.3 adjusted with acetic acid at flow rate 0.9 mL min −1. UV detection provided by fibre-optic DAD detector was set up at 240 nm. Propylparaben was chosen as suitable internal standard (IS). There is only simple pre-adjustment of the sample of topical solution (dilution with mobile phase) so the analysis is not uselessly elongated. Parameters of the method showed good linearity in wide range, correlation coefficient >0.999; system precision (relative standard deviation, R.S.D.) in the range 0.45–1.95% at three different concentration levels, detection limits (3 σ) 1.00 μg mL −1 (salicylic acid), 0.66 μg mL −1 (triamcinolone acetonide) and 0.33 μg mL −1 (propylparaben) and recovery from the pharmaceutical preparations in the range 97.50–98.94%. The chromatographic resolution between peaks of compounds was more than 4.5 and analysis time was 5.1 min under the optimal conditions. The advantages of sequential injection chromatography against classical HPLC are discussed and showing that SIC can be a method of option in many cases.

Simultaneous determination of salicylic acid and acetylsalicylic acid in aspirin delayed-release tablet formulations by second-derivative UV spectrophotometry

A rapid, simple assay procedure was developed for simultaneous analysis of aspirin (ASA) and salicylic acid (SA) in aspirin delayed-release tablet formulation by ‘zero crossing’ second-derivative UV spectrophotometry. The zero-order absorption spectra and second derivative spectra of ASA and SA were recorded in diluting solution acetonitrile–formic acid (99:1). The accuracy of the method was demonstrated by the determination of ASA and SA in five tablets formulations (each 20 tablets of the same batch) by the described method and by high performance liquid chromatographic method, and the results were in good agreement.

Kinetic determination of salicylic acid, diflunisal and their mixture based on lanthanide-sensitized luminescence

The combined use of kinetic and equilibrium measurements, obtained with the aid of the stopped-flow mixing technique, allowed the development of a simple, fast direct method for the simultaneous determination of salicylic acid and diflunisal in untreated serum samples that can be readily applied to routine analyses. The method is based on the formation of ionassociation complexes between the cationic surfactant cetyltrimethylammonium bromide and the ternary anionic chelates formed by salicylate and diflunisal with terbium(III) and EDTA in a basic medium. The intramolecular energy transfer from salicylate or diflunisal to terbium(III) gives rise to luminescence emission by the lanthanide ion. The calibration graphs thus obtained are linear over a wide concentration range and the detection limits are in the nanogram per millilitre region. Withinassay and between-assay relative standard deviation ranged from 2.6% to 3.6% and from 1.3% to 8.5%, respectively. Mixtures of salicylic acid and diflunisal in ratios between 6:1 and 1:12 can be satisfactorily resolved. The proposed method was applied to the direct determination of these compounds in serum with recoveries of 92.7–109.0% for salicylic acid and 86.4–110.9% for diflunisal.

Synchronous fluorimetric determination of salicylic acid and diflunisal in human serum using partial least-squares calibration

The simultaneous determination of salicylic acid and diflunisal in human serum has been accomplished by synchronous fluorimetry, in combination with partial least-squares multivariate calibration. The total luminescence information of the analytes has been used to optimize the spectral data set for the calibration, by analysis of the three-dimensional excitation-emission matrices. The synchronous spectrum, maintaining a constant difference of Δ λ = 128 nm between the emission and excitation wavelengths, has been selected as optimum to perform the determination. The method is based on the fluorescence of these compounds in chloroform containing 1% (v/v) acetic acid. Serum samples are treated with trichloroacetic acid to remove the proteins, and both analytes are extracted into chloroform−1% (v/v) acetic acid prior to the determination. For concentrations ranging from 60–240 μg ml −1 of each drug, analytical recoveries range from 96% to 103% for salicylic acid and from 97% to 105% for diflunisal.

Automatic continuous on‐line monitoring of salicylic acid and acetylsalicylic acid in pharmaceuticals

An asymmetrical FIA merging-zones manifold based on the dual injection of two sample microvolumes was developed for the simultaneous determination of salicylic acid and acetylsalicylic acid in pharmaceutical preparations at a sampling frequency of 30/h. The complex formed between the Fe(III) reagent continuously introduced in the system and salicylic acid was monitored photometrically at 520 nm. One of the sample plugs was prehydrolysed on injection into an NaOH stream and was circulated through a longer channel than the other plug. This yielded two FIA peaks corresponding to salicylic acid and the overall content, respectively. The proposed manifold was successfully used to control the dissolution test of a pharmaceutical preparation.

Determination of salicylic acid and its metabolites in urine by derivative synchronous spectrofluorimetry

The simultaneous determination of salicylic acid in binary and/or ternary mixtures and its two main urinary metabolites is proposed. Mixtures of salicylic, salicyluric and gentisic acids are resolved by synchronous spectrofluorimetry, in combination with first-derivative measurements. The urine is extracted with diethyl ether in acid medium. Salicylic and salicyluric acids are re-extracted into glycine-sodium hydroxide buffer solution of pH 11.6 and determined at that pH, and salicylic and gentisic acids are re-extracted into boric acid-sodium hydroxide buffer solution of pH 8.5 and determined at pH 6.

High-performance liquid chromatographie determination of vinca-alkaloids in plasma and uterine

Salicylic and benzoic acid are phenolic acids occurring in plant cells, thus they can be present in fruit and vegetables at various levels. They possess anti-inflammatory and antimicrobial properties, however they may induce symptoms and health problems in a small percentage of the population. Therefore, a low phenolic acid diet may be of clinical benefit to such individuals. In order to achieve this goal, the concentration of these substances in different food and beverages should be assessed. The present work describes for the first time a new method, based on solid phase microextraction (polydimethylsiloxane–divinylbenzene fiber) coupled to liquid chromatography with UV diode array detection, for the simultaneous determination of salicylic acid, 3-methyl salicylic acid, 4-methyl salicylic acid, acetylsalicylic acid and benzoic acid in selected fruit, vegetables and beverages.All the aspects influencing fiber adsorption (time, temperature, pH, salt addition) and desorption (desorption and injection time, desorption solvent mixture composition) of the analytes have been investigated. An isocratic separation was performed using an acetonitrile-phosphate buffer (pH 2.8; 2 mM) mixture (70:30, v/v) as the mobile phase. The estimated LOD and LOQ values (μg/mL) were in the range 0.002–0.028 and 0.007–0.095. The within-day and day-to-day precision values (RSD%) were between 4.7–6.1 and 6.6–9.4, respectively.The method has been successfully applied to the analysis of fava beans, blueberries, kiwi, tangerines, lemons, oranges and fruit juice (lemon and blueberry) samples. The major advantage of the method is that it only requires simple homogenization and/or centrifugation and dilution steps prior to SPME and injection in the LC system.

An HPLC method for the determination of salicylic acid, phenacetin and paracetamol in serum, with indications; two case-reports of intoxication

Acetylsalicylic acid, paracetamol and phenacetin are the most frequently used non-narcotic analgesics in The Netherlands. Two new case reports, an intoxication by salicylate ointment and a paracetamol overdose, are described. Indications and methods for the determination of these compounds are discussed. We describe a newHplc method for the simultaneous determination of salicylic acid, paracetamol and phenacetin in serum. Some other analgesics, like aminophenazone, phenazone and dipyrone can be determined by this method at the same time, 100Μl of serum is mixed with 600Μl methanol, centrifuged and an aliquot is injected into theRP-5C18Hplc column. A methanol—buffer pH=3.0 (40 + 50) mixture is used as mobile phase. Salicylamide can be used as internal standard. Coefficients of variation of 3.4% for salicylic acid, 1.9% for paracetamol and 3.4% for phenacetin are found and recoveries from calf's serum are 98–101% (n=10 per drug at 50 mg/l serum). In neonatology the sample volume can be decreased from 100Μl to 10Μl. This method has proved to be reliable and suitable in clinical pharmacy and toxicology.

An automated method for the simultaneous determination of salicylic acid and salicylazoiminopyridine in commercial salicylazosulphapyridine

An automated method for simultaneous determination of salicylic acid and salicylazoiminopyridine in salicylazosulphapyridine is presented. The components are separated from salicylazosulphapyridine on a two-layer anion and cation exchanger with a yield of almost 100%. The effluent is analyzed simultaneously for the fluorescence of salicylate and the absorbance of salicylazoiminopyridine in an Auto-Analyzer II system. The system is run at 40 samples per h with a relative standard deviation of 0.2%. The method gives, at the content of 1% of each component, a reproducibility (s r) of 3–4%. Five commercial products from four suppliers were analyzed and found to contain 0–0.9% salicylic acid and 0–3.6% coloured impurities, expressed as salicylazoiminopyridine. Some anomalies of the automatic systems are presented and discussed. An automated method for colorimetric assay of salicylazosulphapyridine is briefly described.

Simultaneous Determination of Salicylic Acid and Salicyluric Acid in Plasma and Urine by High Pressure Liquid Chromatography

Abstract A sensitive, rapid, and specific high pressure liquid chromatographic (HPLC) assay was developed for the determination of salicylic (SA) and salicyluric (SU) acids in plasma and urine. The compounds are extracted into ethyl ether at acid pH, evaporated, and reconstituted prior to instrumental separation. Overall recovery of both compounds is 90 ± 5%, and the sensitivity limits are 150 ng of SU and 300 ng SA per ml of biological fluid. The assay was used for the determination of both compounds in plasma and urine of man following oral doses of 40 mg/kg of sodium salicylate.

Determination of traces of salicylic acid in acetylsalicylic acid bulk, tablets and other preparations containing phenacetin and caffeine; Simultaneous determination of salicylic acid, and acetylsalicylic acid with additional components in pharmaceutical preparations

A GLC method for the determination of traces of salicylic acid in acetylsalicylic acid bulk, tablets and other preparations containing phenacetin and caffeine after methylation with methyl iodide and potassium carbonate on a column of 3 % OV-17 on Varaport 30, 70–80 mesh, AW/DMCS is reported. A column-chromatographic separation of salicylic acid from acetylsalicylic acid is necessary prior to methylation on account of the slight hydrolysis of the latter. A ferric chloride-urea reagent holds the salicylic acid back on a Celite 545 column. Further, salicylic acid, acetylsalicylic acid, phenacetin, caffeine, barbital, phenobarbital, bromodiethylacetylcarbamide and codein could also be determined simultaneously through GLC using this methylation technique. The chromatogram of such a mixture is presented.

Simultaneous determination of salicylic acid and mononitrosalicylic acids using a dual-wave-length spectrophotometer

Simultaneous determination of salicylic acid and mononitrosalicylic acids using a dual-wave-length spectrophotometer

Rapid Simultaneous Determination of Salicylic Acid and Aspirin by GC I: Analysis of Synthetic Aspirin—Salicylic Acid Mixtures and of Single-Component Aspirin Tablets

A rapid, sensitive GC method for the simultaneous analysis of salicylic acid and aspirin in admixtures and in single-component acetylsalicylic acid (aspirin) tablets is presented. The procedure involves conversion of the two drugs to their methyl esters with diazomethane in tetrahydrofuran solution and subsequent isothermal elution of the components from a 5% OV-210 on Diatoport S glass column. Quantitation is effected by means of an electronic digital integrator with methyl o-methoxybenzoate as the internal standard. The average total drug recovery of 16 synthetic mixtures was 99.70%, with a standard deviation of ±0.58%, while the mean total recovery using the USP procedure was 99.40 ± 0.68%. When applied to the analysis of commercial single-component compressed and enteric coated aspirin tablets, the proposed technique gave results which were, in the majority of cases, in excellent agreement with the USP values for both aspirin and salicylic acid content. With a few formulations, the salicylic acid content was found to be slightly higher with the GC procedure than with the method described in the USP.