Simultaneous Determination Of Organic Acids Research Articles

Development and validation of a UHPLC-ESI-MS/MS method for the simultaneous determination of organic acids and phenolic compounds in Filipendula vulgaris, Polygonum divaricatum, Hypericum linarioides, and Rheum ribes

This research focused on investigating biologically active and pharmaceutically important phytochemicals using reliable and reproducible analytical techniques specifically designed for traditional medicinal plants. Considering the complex chemical diversity present in plant extracts, reliable identification methods are crucial. The goal of this study was to develop and validate a robust UHPLC-ESI-MS/MS method for the qualitative and quantitative analysis of organic acids (OAs) and phenolic compounds (PCs). With gradient elution of water containing 0.1 % formic acid and acetonitrile containing 0.1 % formic acid as the mobile phase, OAs and PCs were separated chromatographically on a C18 reversed-phase column. Through the use of an electrospray ionization source operating in negative ionization mode, these compounds were identified on a triple quadrupole tandem mass spectrometer. The validation results clearly indicate that the method exhibited linearity with a squared correlation coefficient (R2 ≥ 0.9986), demonstrating sensitivity (LOQ: 1.08 µg/L-10.79 µg/L), precision (RSD%≤2.73), and trueness (RE% ≤ 1.53) for the OAs and PCs found in the plants. The developed UHPLC-ESI-MS/MS method was successfully applied to the qualitative and quantitative analysis of OAs and PCs in Turkish plants, which are commonly used in traditional medicine. Our study identified a total of 12 OAs and PCs in F. vulgaris, 12 in P. divaricatum, 9 in H. linarioides, and 11 in R. ribes. In particular, gallic acid had the highest content in F. vulgaris (925.67 µg/g extract) and R. ribes (1358.78 µg/g extract), while H. linarioides (730.66 µg/g extract) and P. divaricatum (4003.02 µg/g extract) had relatively high levels of quinic acid. In conclusion, the developed UHPLC-ESI-MS/MS method allows the simultaneous analysis and quantification of 2 OAs and 33 PCs in plants and proves to be a valuable tool for evaluating the efficacy of traditional medicinal plants.

A Rapid HPLC Method for the Simultaneous Determination of Organic Acids and Furans: Food Applications

Concerns over the potential adulteration of commercially produced foods give rise to a requirement for a simple and fast analytical method capable of quantifying potential adulterants. This work demonstrates a simple HPLC method tailored to detect major organic acids and furans within ingredients in commercial food products, for example, pomegranate molasses, balsamic vinegar, and apple cider vinegar. The relative importance of this method is in its simplicity and its use of an environmentally friendly aqueous mobile phase under isocratic conditions, providing results in a less than 20 min runtime. The chromatographic separation was achieved using an Acclaim® OA, 5 µm, 120 Å (4.0 × 250 mm) column; a UV-DAD detector set at 210 nm; and a 200 mM Na2SO4 mobile phase with 0.55 mL/L methanosulfonic acid as a pH modifier. The method was then validated by quantifying the concentration of acetic acid, formic acid, citric acid, and hydroxymethyl furfural (HMF) in pomegranate molasses, balsamic vinegar, and apple cider vinegar commercial products. The concentration of acetic acid and HMF in balsamic vinegar was 80.380 mg/mL (±1.272 mg/mL) and 2.153 mg/mL (±0.021 mg/mL), respectively. The apple cider vinegar was composed only of acetic acid with a concentration of 44.139 mg/mL (±0.053 mg/mL). The concentrations of citric acid and HMF were 123.425 mg/mL (±2.502 mg/mL) and 11.382 mg/mL (±0.582 mg/mL), respectively, in pomegranate molasses. Furthermore, this method is also capable of determining various organic acids and furans in biomass: levulinic acid, formic acid, furfurals, diformylfuran, and gamma-valerolactone.

Simultaneous determination of organic acids and sugars in fruit juices by High performance liquid chromatography: characterization and differentiation of commercial juices by principal component analysis

ABSTRACT: The present research presents an analytical methodology based on High Performance Liquid Chromatography (HPLC) and Principal Component Analyses (PCA) for simultaneous quantification and analytical differentiation of organic acids and sugars in commercial fruit juice samples (orange, grape, apple and tangerine). In addition to the development of the method that generated suitable validation paramters for quantitative analytical applications, the analysis of fourteen commercial samples and the use of Principal Component Analysis indicated the relationship between the constituents and the very constitutional chemical nature of the juice. In general, in grape and apple juices, ascorbic acid was not quantified and the citric acid content was very low; however, the concentrations of fructose and glucose were the highest in both juices. In orange and mandarin orange juices, the content of all analytes, except acorbic acid, did not differ statistically from each other. However, these differed significantly from the others, mainly in relation to sucrose content. Finally, the apple juice samples differed according to the content of fructose and malic acid, the predominant constituint of the apple. Results showed that the simultaneous chromatographic method associated with principal component analysis generated important information about characteristics of commercial juices, with the potential to be used in systems of quality control and identification of adulterations.

Determination of citric and oxalic acid in fungi fermentation broth through HPLC-DAD and solid-phase extraction

This work describes a method for the simultaneous determination of organic acids in fungi fermentation broths. The sample preparation technique and the chromatographic conditions were optimized to enable the monitoring of the fermentation process. The determination of the citric and oxalic acid was done with a high resolution liquid chromatograph adapted to a detector with diode arrangement, an IC-Pak Ion-Exclusion 7μm (7.8 x 300 mm) column and a mobile phase of 0.001N H2SO4 with isocratic elusion. The validation parameters show efficiency, adequate linearity, and standard deviation values (%RSD) from 0.018% and 4.650%. the quantification limits (LDC) were 50.76 mg/L for citric acid and 20.18 mg/L for oxalic acid and the detection limits (LDD) were 0.6 mg/L for both acids. The method was applied in the analysis of organic acids in fermentation broths of Aspergillus niger.

Simultaneous Determination of Organic Acids, Inorganic Anions, and Alditols in Wine with Valve-Switching Ion Chromatography

A valve-switching ion chromatography method coupled with conductivity and pulsed amperometric detection has been set up for the simultaneous determination of organic acids, inorganic anions and alditols in wine. The system, based on valve-switching technique, consisted of two pumps, two analytical columns and a trap column connected in series via one six-port valve. In addition, the optimal valve-switching time and chromatographic conditions were explored to obtain a good linearity, precision, accuracy, and sensitivity with a mean correlation coefficient of > 0.99, repeatability of 0.62–6.18% for eight replicates, and the average spiked recovery of 96.66%. The new method was successfully applied in the analysis of six kinds of wine from different regions.

Determination of organic acids in the root exudates of Cr-hyperaccumulator Leersia hexandra Swartz using high performance liquid chromatography

High performance liquid chromatography (HPLC) was used for the separation and quantification of the low molecular-mass organic acids in the root exudates of Cr-hyperaccumulator Leersia hexandra Swartz. Eight organic acids, oxalic, tartaric, formic, malic, lactic, acetic, maleic and citric acids, were separated on an XSelect HSS T3 column (250 mm×4.6 mm, 5 μm) within 13 min with the mobile phase of 40 mmol/L potassium dihydrogen phosphate-phosphorous acid at pH 2.40, a flow-rate of 1.0 mL/min, a detection wavelength of 205 nm and a column temperature of 25℃. This is a simple and highly sensitive method for the separation of the eight organic acids with well-shaped peaks. The limits of detection (LODs) for the eight acids ranged from 0.12 mg/L to 12.32 mg/L. Under the optimized conditions, tartaric, malic, lactic, maleic and citric acids in the root exudates were detected with high linearities, acceptable recoveries and excellent precisions. The contents of these acids calculated in the root exudates were (130.90±1.44) μg/g (root dry weight (DW)) for tartaric acid, (1031.34±4.38) μg/g (root DW) for malic acid, (65.54±1.01) μg/g (root DW) for lactic acid, (0.96000±0.00367) μg/g (root DW) for maleic acid and (201.50±1.13) μg/g (root DW) for citric acid. The proposed HPLC method is quite suitable for the simultaneous determination of organic acids in the root exudates of Leersia hexandra Swartz, and can be used in other plant root exudates as well.

Simultaneous analysis of ten low-molecular-mass organic acids in the tricarboxylic acid cycle and photorespiration pathway in Thalassiosira pseudonana at different growth stages.

A method using high-performance liquid chromatography coupled with tandem mass spectrometry was developed for the simultaneous determination of organic acids in microalgae. o-Benzylhydroxylamine was used to derivatizethe analytes, and stable isotope-labeled compounds were used as internal standards for precisequantification. The proposed method was evaluated in terms of linearity, recovery, matrix effect, sensitivity, and precision. Linear calibration curves with correlation coefficients >0.99 were obtained over the concentration range of 0.4-40 ng/mL for glycolic acid, 0.1-10 ng/mLfor malic acid and oxaloacetic acid, 0.02-2 ng/mLfor succinic acid and glyoxylic acid, 4-400 ng/mLfor fumaric acid, 20-2000 ng/mLfor isocitric acid, 2-200 ng mL-1 for citric acid, 100-10000 ng mL-1 for cis-aconitic acid, and 1-100 ng mL-1 for α-ketoglutaric acid.Analyte recoveries were between 80.2 and 115.1%, and the matrix effect was minimal. Low limits of detection (0.003-1 ng/mL) and limits of quantification (0.01-5 ng/mL) were obtained except cis-aconitic acid. Variations in reproducibility for standard solution at three different concentrations levels were <9%. This is the first report of the simultaneous analysis of ten organic acids in microalgae, which promotes better understanding of their growth state and provides reference value for high-yield microalgae cultures.

Simultaneous determination of 14 organic acids in Shenfu injection by hydrophilic interaction chromatography-tandem mass spectrometry

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.

Simultaneous Determination of Organic Acids in Chinese Liquor by GC-MS Method

Simultaneous Determination of Organic Acids in Chinese Liquor by GC-MS Method

Determination of organic acids in rice wine by ion-exclusion chromatography

An ion-exclusion chromatographic method for the simultaneous determination of organic acids in rice wine was developed. An IC-Pak Ion Exclusion column (300 mm x 7.8 mm, 7 microm) was used at 50 degrees C. The mobile phases were H2SO4 (phase A) and acetonitrile (phase B) (98:2, v/v) at a flow rate of 0.5 mL/min. The gradient elution program was as follows: 0-40 min, 0.01 mol/L H2SO4 to 0.02 mol/L H2SO4; 40-50 min, 0.01 mol/L H2SO4. The injection volume was 10 microL. The detection wavelength was set at 210 nm. The results showed that oxalic acid, maleic acid, citric acid, tartaric acid, malic acid, ascorbic acid, succinic acid, lactic, fumaric acid, acetic acid, propionic acid, isobutyric acid and butyric acid were completely separated and determined in 30 min. The linear correlation coefficients were above 0.999 7 in the range of 0.001- 1.000 g/L. Under the optimized conditions, the recoveries of organic acids in rice wine were in the range of 93.4% - 103.8% with the relative standard deviations (RSDs, n = 5) of 0.1% - 1.5%. This method is feasible, convenient, fast, accurate and applicable for the quantitative analysis of the organic acids in rice wine.

Simultaneous determination of organic acids and saccharides in lactic acid fermentation broth from biomass using high performance liquid chromatography

Abstract: A high performance liquid chromatographic method for the simultaneous determination of organic acids and saccharides in lactic acid fermentation broth from biomass was developed. A Bio-Rad Aminex HPX-87H column was used at 55 degrees C. The mobile phase was 5 mmol/L sulfuric acid solution at a flow rate of 0.6 mL/min. The samples were detected by a refractive index detector (RID). The results showed that six organic acids and three saccharides in fermentation broth were completely separated and determined in 17 min. The linear correlation coefficients were above 0.999 8 in the range of 0.15-5.19 g/L. Under the optimized conditions, the recoveries of the organic acids and saccharides in Rhizopus oryzae fermentation broth at two spiked levels were in the range of 96.91%-103.11% with the relative standard deviations (RSDs, n = 6) of 0.81%-4.61%. This method is fast and accurate for the quantitative analysis of the organic acids and saccharides in microbial fermentation broths.

English

A sensitive method of the simultaneous determination of organic acids and iridoid glycoside in reduning injection was described by rapid resolution liquid chromatography (RRLC) with ultraviolet (UV) detection under segmental monitoring mode. RRLC separation was achieved by use of a C18 analytical column packed with sorbent of particle size 1.8 μm while the injected sample volume was set to 2 µl. A binary gradient elution program of aqueous phosphoric acid (0.1%, v/v) versus acetonitrile was selected for the quantitative analysis of nine major components. The RRLC-UV method under segmental monitoring mode showed satisfactory linearity, the limit of detection and the limit of quantitation. The method had been successfully applied to the determination of nine major components content in reduning injection samples. The analytical time of the proposed protocol is less than that of conventional HPLC-UV analytical methods, thus RRLC-UV method under segmental monitoring mode could function as an attractive alternative method to be used to evaluate the quality of reduning injection in the process of production. Key words: Reduning injection, organic acids, iridoid glycosides, rapid resolution liquid chromatography- ultraviolet (RRLC–UV).

Ion-Exclusion Chromatography Determination of Organic Acid in Uridine 5'-Monophosphate Fermentation Broth

Simultaneous determination of organic acids using ion-exclusion liquid chromatography and ultraviolet detection is described. The chromatographic conditions are optimized when an Aminex HPX-87H column (300 × 7.8 mm) is employed, with a solution of 3 mmol/L sulfuric acid as eluent, a flow rate of 0.4 mL/min and a column temperature of 60°C. Eight organic acids (including orotic acid, α-ketoglutaric acid, citric acid, pyruvic acid, malic acid, succinic acid, lactic acid and acetic acid) and one nucleotide are successfully quantified. The calibration curves for these analytes are linear, with correlation coefficients exceeding 0.999. The average recovery of organic acids is in the range of 97.6% ∼ 103.1%, and the relative standard deviation is in the range of 0.037% ∼ 0.38%. The method is subsequently applied to obtain organic acid profiles of uridine 5'-monophosphate culture broth fermented from orotic acid by Saccharomyces cerevisiae. These data demonstrate the quantitative accuracy for nucleotide fermentation mixtures, and suggest that the method may also be applicable to other biological samples.

Simultaneous Determination of Organic Acids and Nitrate in Xylem Saps of the Hyperaccumulator Alyssum Murale by RP-HPLC after Solid-Phase Extraction with Nanosized Hydroxyapatite

An analytical approach based on solid-phase extraction (SPE) with nanosized hydroxyapatite and reversed-phase high performance liquid chromatography (RP-HPLC) for the simultaneous determination of organic acids (oxalic, malic, malonic, citric, and fumaric) and nitrate in xylem saps of the hyperaccumulator Alyssum murale is presented in this study. The optimum experimental conditions for the separation and determination of organic acids and nitrate are studied. The interference from nitrate on oxalic acid in RP-HPLC is eliminated by SPE with nanosized hydroxyapatite, and the simultaneous determination of organic acids and nitrate is achieved by RP-HPLC at the optimum chromatographic conditions. The accuracy of the method is confirmed with an average recovery ranging between 95.2% and 99.8%, the relative standard deviations (RSD) are less than 2.0%. This method is successfully applied to determine the organic acids and nitrate in xylem saps of the hyperaccumulator Alyssum murale.

Simultaneous determination of organic acids, amygdalin, and benzoic acid related compounds in pickled Japanese apricot (Umeboshi) by high performance liquid chromatography

Simultaneous determination of organic acids, amygdalin, and benzoic acid related compounds in pickled Japanese apricot (Umeboshi) by high performance liquid chromatography

Otimização e validação de método para determinação de ácidos orgânicos em vinhos por cromatografia líquida de alta eficiência

The organic acids (tartaric, malic, citric, lactic and succinic) are de main components responsible for the acidity in the wine. This method for simultaneous determination of organic acids and interfering peaks in wines can be achieved in 16 min. The sample preceded by a dilution and filtration step. The chromatographic separation required one reversed phase column, isocratic mobile phase (acetonitrila, formic acid in water) and detection wavelength was set at 212 nm. The validation confirmed good repeatability, recovery and application in red and white wines.

Determination of Organic Acids in Red Wine and Must on Only One RP-LC-Column Directly After Sample Dilution and Filtration

A new method for simultaneous determination of organic acids in red wine and must by liquid chromatography was studied. The determination of organic acids in wines can be achieved in less than 13 min, preceded only by a simple sample dilution and filtration step. With this method, the chromatographic separation of eight organic acids and interfering peaks present in red wine, required only one reversed phase column (Waters Atlantis dC18 column, 4.6 × 150 mm ID, 5 μm). As mobile phase, isocratic acetonitrile–0.01 mol L−1 KH2PO4 at pH 2.7 5:95 (v/v) at a flow rate of 0.8 mL min−1 was used. Detection wavelength was set at 210 nm except for ascorbic acid which was detected at 243 nm. Application to red wine and must confirmed good repeatability and showed a wide variation range for concentrations of organic acids.

Simultaneous determination of organic acids in beverages by capillary zone electrophoresis

A rapid, reliable and simple capillary zone electrophoresis method for the determination of organic acids in beverages was developed. The complete separation of oxalic, formic, tartaric, malic, succinic, maleic, glutaric, pyruvic, acetic, lactic, citric, butyric, benzoic, sorbic, ascorbic and gluconic acids can be achieved in less than 3.5 min with a simple electrolyte composed by phosphate as the carrier buffer (7.5 mM NaH 2PO 4 and 2.5 mM Na 2HPO 4), 2.5 mM TTAOH as electroosmotic flow modifier and 0.24 mM CaCl 2 as selectivity modifier, adjusting the pH at 6.40 constant value. Injection was performed in hydrodynamic mode (30 s) and the detection mode was UV direct at 185 nm. The running voltage was −25 kV at thermostated temperature of 25 °C. The method developed has been applied to several beverage samples with only a simple dilution and filtration treatment of the sample. The proposed method is fast because the separation time decrease two, four or, even, six times the separation times of the previous reported CZE methods. It is also simple and cheap due to a low consumption of chemicals and samples. These reasons permit it to be considered adequate for routine analysis of organic acids in beverage samples.

Determination of glycerophosphate and other anions in dentifrices by ion chromatography

Simple, reliable and sensitive analytical methods to determine the anions, such as fluoride, monofluorophaosphate, glycerophosphate related to anticaries are necessary for basic investigations of anticaries and quality control of dentifrices. A method for the simultaneous determination of organic acids, organic anions and inorganic anions in the sample of commercial toothpaste is proposed. Nine anions (fluoride, chloride, nitrite, nitrate, sulfate, phosphate, monofluorophaosphate, glycerophosphate and oxalic acid) were analyzed by means of ion chromatography using a gradient elution with KOH as mobile phase, IonPac AS18 as the separation column and suppressed conductivity detection. Optimized analytical conditions were further validated in terms of accuracy, precision and total uncertainty and the results showed the reliability of the IC method. The relative standard deviations (RSD) of the retention time and peak area of all species were less than 0.170 and 1.800%, respectively. The correlation coefficients for target analytes ranged from 0.9985 to 0.9996. The detection limit (signal to noise ratio of 3:1) of this method was at low ppb level (<15 ppb). The spiked recoveries for the anions were 96–103%. The method was applied to toothpaste without interferences.

Ion chromatographic determination of some organic acids, chloride and phosphate in coffee and tea

An ion chromatographic method for the simultaneous determination of organic acids and inorganic ions is described. Acetic, malic, ascorbic, citric, malic and succinic acids, chloride and phosphate were determined in coffee and tea samples. The separation is performed on an anion-exchange column operated at 40 °C within 25 min by an isocratic elution with 0.6 mM aqueous potassium hydrogenphthalate (pH 4.0) solution containing 4% (v/v) acetonitrile as eluent and determination by conductivity detection. The method does not need a special sample treatment and was successfully applied to the analysis of black, green and oolong tea samples. Also, green and roasted coffee samples from the varieties arabica and robusta were analyzed.