A New Simplified and Stability Indicating Liquid Chromatography Method for Routine Analysis of Isoflavones Aglycones in Different Complex Matrices

Yatsu, Francini K.J.; Pedrazza, Graziele Pereira Ramos; Argenta, Débora Fretes; Barreto, Fabiano; Nemitz, Marina Cardoso; Teixeira, Hélder Ferreira; Koester, Letícia Scherer; Bassani, Valquíria Linck

doi:10.1007/s12161-014-9827-5

Cited by 7 publications

(5 citation statements)

References 30 publications

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“…3A). The λmax spectra corresponding to each peak are shown in figure 3B (HPLC-DAD); the mass spectrum with the characteristic molecular ions is shown in Figure 3C, which is consistent with those already reported for this type of molecules [8,15,18]. To evaluate the specificity of the HPLC method, samples of Trifolium pratense extract and cyclodextrins were injected in the equipment and the purity of the peaks was checked.…”

Section: Specificitysupporting

confidence: 83%

“…The matrix effect evaluation was performed as described by Watanabe et al [14] and Yatsu et al [15] by comparing the slopes of calibration curves of the isoflavone aglycone standards (FOR, BIO, DAID, and GEN) in the solvent and the matrix. Three analytical curves were obtained on three consecutive days by plotting the peak areas versus the concentration of isoflavone aglycone standards (0.5, 1.0, 4.0, 8.0, 12.0, 16.0, and 20.0 µg/mL) in acetonitrile 50% (v/v) and the matrix solution.…”

Section: Assessment Of the Matrix Effectmentioning

confidence: 99%

“…When the analyte is incorporated into a complex matrix compared to a standard or reference solution, the matrix effect on the analyte quantification is a critical parameter that should be evaluated during the validation of an analytical method. This effect can result in suppression or enhancement of the response, which may impair the correct quantification of analytes [15]. The results for the matrix effect (ME% <2.24) for each aglycone isoflavone are shown in Table 3.…”

Section: Assessment Of the Matrix Effectmentioning

confidence: 99%

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Stability-Indicating HPLC Method for isoflavones aglycones analysis from Trifolium pratense L. extract

Martiny¹,

Waszczuk²,

Kaiser³

et al. 2020

Drug Anal. Res.

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The purpose of this study was to develop and validate a fast HPLC stability-indicating method for simultaneously quantifying the four main isoflavones in Trifolium pratense. Validation procedures followed the ICH requirements for complex matrices. The stability-indicating tests were performed by exposing the isoflavones to conditions of forced degradation and further analysis for verifying the formation of degradation products and their possible interferences in the HPLC analysis. The major isoflavones of Trifolium pratense proved to be stable against acid and oxidative media, thermodegradation, and photodegradation. However, they proved to be unstable in alkaline media, even for short periods of exposure like 2h. In this condition, in addition to the peaks corresponding to isoflavones, the HPLC analysis showed the presence of three additional peaks which were eluted at different retention times to the reference substances, without interfering in the quantification of the four analytes of interest, formononetin, biochanin A, daidzein and genistein. The method was validated following ICH guidelines showing to be specific, linear, precise, accurate, and robust.This first report concerning a stability-indicating method revealed that the proposed HPLC method reliably quantify the isoflavones and separate them from the degradation products in a short time of analysis.

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Section: Specificitysupporting

confidence: 83%

Section: Assessment Of the Matrix Effectmentioning

confidence: 99%

Section: Assessment Of the Matrix Effectmentioning

confidence: 99%

See 1 more Smart Citation

Stability-Indicating HPLC Method for isoflavones aglycones analysis from Trifolium pratense L. extract

Martiny¹,

Waszczuk²,

Kaiser³

et al. 2020

Drug Anal. Res.

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show abstract

“…The Soxhlet method has also been used for the extraction of isoflavones from solid samples of soybeans, soy products, and legumes. The extraction is performed using 90% aqueous MeOH (Klejdus and others ; Leuner and others ) or 80% aqueous ethanol (Xiao and others ; Yatsu and others ) as the solvents. Klejdus and others () proposed a modified Soxhlet extraction method consisting of a 2‐step temperature program.…”

Section: Sample Preparationmentioning

confidence: 99%

Analysis of Isoflavones in Foods

Bustamante-Rangel

Delgado-Zamarreño

Pérez‐Martín

et al. 2018

Comp Rev Food Sci Food Safe

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In recent years the nutritional and bioactive properties of foods are being intensively investigated with a view to control, in addition to food quality, their possible influence on human health. Because of this, there is a growing demand for rapid, selective, sensitive, and validated methods for analysis and quantification. Bioactive plant compounds include those with weak estrogenic activity (phytoestrogens), among which are the isoflavones. Some of the beneficial activities that have been attributed to isoflavones are anticarcinogenic activity, the prevention of cardiovascular disease, the improvement of bone health, and antioxidant activity. The objective of this work is to provide an updated review of the methods used in sample preparation and subsequent analysis for the determination of isoflavones in food samples, including both soybean and soy products, as well as other foods with low isoflavone contents. The review focuses on the most common sample preparation techniques used during the last 10 years, including both conventional solvent extraction and other more recent extraction techniques. Separation and detection methods, including current trends in liquid chromatography analysis, such as the use of monolithic columns or ultra-high-pressure liquid chromatography, are also discussed.

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“…Most methods for determining isoflavones in soy and related products, however, are based on LC, using absorbance detection [20-23], MS detection [23-25], or tandem MS (MS/MS) detection [26]. The LC absorbance methods in the recent literature for soy isoflavones are based on an external standard quantitation approach [20-22], which may lead to biases during the sample preparation, separation, and detection. One method describes an internal standard approach using formononetin [23], which may be naturally present in soy and cause erroneous quantitative results in some products.…”

Section: Introductionmentioning

confidence: 99%

Liquid chromatography with absorbance detection and with isotope-dilution mass spectrometry for determination of isoflavones in soy standard reference materials

et al. 2016

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Two independent analytical approaches, based on liquid chromatography with absorbance detection and liquid chromatography with mass spectrometric detection, have been developed for determination of isoflavones in soy materials. These two methods yield comparable results for a variety of soy-based foods and dietary supplements. Four Standard Reference Materials (SRMs) have been produced by the National Institute of Standards and Technology to assist the food and dietary supplement community in method validation and have been assigned values for isoflavone content using both methods. These SRMs include SRM 3234 Soy Flour, SRM 3236 Soy Protein Isolate, SRM 3237 Soy Protein Concentrate, and SRM 3238 Soy-Containing Solid Oral Dosage Form. A fifth material, SRM 3235 Soy Milk, was evaluated using the methods and found to be inhomogeneous for isoflavones and unsuitable for value assignment.

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A New Simplified and Stability Indicating Liquid Chromatography Method for Routine Analysis of Isoflavones Aglycones in Different Complex Matrices

Cited by 7 publications

References 30 publications

Stability-Indicating HPLC Method for isoflavones aglycones analysis from Trifolium pratense L. extract

Stability-Indicating HPLC Method for isoflavones aglycones analysis from Trifolium pratense L. extract

Analysis of Isoflavones in Foods

Liquid chromatography with absorbance detection and with isotope-dilution mass spectrometry for determination of isoflavones in soy standard reference materials

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