Development and validation of an HPLC-DAD analytical method for quantitative and simultaneous determination of cefoperazone and prednisolone in intramammary ointment

A nanostructured o-hydroxyazobenzene porous organic polymer as an effective sorbent for the extraction and preconcentration of selected hormones and insecticides in river water

High performance liquid chromatography (HPLC) coupled with a diode array detector (HPLC-DAD) for the identification and quantification of carotenoids, namely all-trans lutein, zeaxanthin, β-cryptoxanthin, α-carotene, and β-carotene, in biological samples such as human serum and breast milk, has been developed and validated. Good chromatography separation was achieved using a binary mobile phase system on a YMC C30 column (150 × 2.1 mm, 3 µm) at 30 °C. Owing to the smaller column particle size and diameter of the column, the separation was achieved in 18 min, which is significantly reduced from the typical 30–40 min of other methods. The diode array detector (DAD) acquisition was set at a wavelength of 445 nm; 3D spectra ranging from wavelengths of 240–600 nm were also recorded. Peaks were identified by matching their retention time and spectra with those of standards. Quantification was achieved by internal standard calibration using echinenone as the internal standard. Good linearity was obtained for each compound (R2 > 0.9999). The method quantification limits (MQLs) for serum and breast milk were 10 ng/mL and 5 ng/mL, in matrix, respectively. A spike recovery study and standard reference material (SRM) from the National Institute of Standards and Technology (NIST) 968e analysis has proven that the method has a high degree of accuracy, precision, and robustness. The stability study showed that the carotenoid standard and sample extracts could be stored in a chilled autosampler at 8 °C up to 48 h without being comprised, which provides guidance on re-test time frames. The freeze/thaw process was found to be detrimental to carotenoids, and should always be avoided. Most importantly, UV standardization of the stock standard is to be performed prior to each assay, and simply taking the values on Certificate of Analysis (CoA) for calculation of the standard concentration is not recommended.

This work describes a simple approach to overcome challenges in emergency toxicological analysis, using liquid-liquid extraction and high-performance liquid chromatography coupled with a diode-array detector (HPLC-DAD). A rapid procedure has been developed, for the extraction and detection of 19 analytes from the following drug classes: analgesics, benzodiazepines, antidepressants, anticonvulsants and drugs of abuse. These substances are relevant in the context of emergency toxicology in Brazil. The method has been validated according to international guidelines by establishing parameters such as lower limit of quantification, sensitivity, linearity, accuracy and precision. The intra and inter-day precision values, at the lowest concentration levels, have always been less than 20% considering its relative standard deviation. As for accuracy values, these have also been satisfactory (above 81.3%). This method was successfully applied in 201 blood samples from patients with suspected poisoning of the Poison Control Center of São Paulo (PCC-SP), Brazil. Finally, the developed method has shown to be relevant for emergency toxicology due to its high sensitivity and it could be also very useful in both fields of clinical and forensic toxicology.

Viticis Fructus (VF) was named Manjingzi as a commonly used traditional Chinese medicine (TCM) targeting various pains and inflammation for more than 2000 years. To guarantee the quality of Viticis Fructus, a simple, quick and eco-friendly Beta/ZSM-22 zeolites-based-mixed matrix solid-phase dispersion method (B/Z-MMSPD) was established for simultaneous extraction and determination of eight compounds (two phenolic acids, two iridoid glycosides, vanillin and three flavonoids) with different polarities from Viticis Fructus by high performance liquid chromatography coupled with a diode array detector (HPLC-DAD). Beta and ZSM-22 were mixed as the sorbent. Water, tetrahydrofuran and methanol were blended with certain ratio as the eluent. Several parameters including types of sorbents, mass ratio of Beta to ZSM-22, mass ratio of matrix to sorbent, grinding time, types, concentration and volume of eluent were optimized. The recoveries of eight analytes were within the range of 95.0%–105% (RSDs ≤ 4.13%). The limits of detection and limits of quantitation ranged from 0.5 to 5.5 μg/g and from 1.5 to 16 μg/g, respectively. Compared to the traditional extract methods, it was a simple, rapid, efficient and green method. The results demonstrated that a simple, rapid, efficient and green B/Z-MMSPD was developed for the simultaneous extraction and determination of eight target analytes with different polarities for quality control of Viticis Fructus.

Phytochemical analysis of Rosa hybrida cv. ‘Jardin de Granville’ by HPTLC, HPLC-DAD and HPLC-ESI-HRMS: Polyphenolic fingerprints of six plant organs

Potatoes in the diet contribute significantly to antioxidant daily intake worldwide. The influence of different domestic cooking conditions, boiling, microwaving, and baking, on total phenolics (TP), antioxidant capacity, phenolic composition, and tryptophan content was studied using eight commercial potato varieties. The antioxidant capacity was detected by the methods of oxygen radical absorbance capacity assay (ORAC) and the 2,2-diphenyl-1-picrylhydrazyl free radical (DPPH(*)) assay. The phenolic composition and tryptophan content were determined using high-performance liquid chromatography coupled with a diode array detector (HPLC-DAD), whereas phenolics and tryptophan were identified by means of HPLC-mass spectrometry, HPLC-DAD, and authentic standards. Antioxidant capacity was influenced by potato variety and cooking conditions; however, cooked potatoes retained 68-97% ORAC value depending on cooking procedure and variety. Chlorogenic acid and its isomers dominated the phenolic composition of each variety involved in this study. ORAC and TP were highly and positively correlated (r = 0.9119). Norkotah ranked highest in chlorogenic acid content and antioxidant value. Principal component analysis showed different cooking processes did not influence the trend of the antioxidant profile of the eight potato varieties, but specific compounds exert influence on the antioxidant capacity. The results imply that the potato varieties rich in antioxidant components could be good antioxidant sources as activities are not greatly affected by different cooking conditions.

The antioxidant profile of 23 native Andean potato cultivars has been investigated from a human nutrition perspective. The main carotenoid and tocopherol compounds were studied using high-performance liquid chromatography coupled with a diode array detector (HPLC-DAD) and a fluorescence detector, respectively, whereas polyphenols (including anthocyanins in colored tubers) were identified by means of both HPLC-mass spectrometry and HPLC-DAD. Antioxidant profiling revealed significant genotypic variations as well as cultivars of particular interest from a nutritional point of view. Concentrations of the health-promoting carotenoids, lutein and zeaxanthin, ranged from 1.12 to 17.69 microg g(-1) of dry weight (DW) and from 0 to 17.7 microg g(-1) of DW, with cultivars 704353 and 702472 showing the highest levels in lutein and zeaxanthin, respectively. Whereas beta-carotene is rarely reported in potato tubers, remarkable levels of this dietary provitamin A carotenoid were detected in 16 native varieties, ranging from 0.42 to 2.19 microg g(-1) of DW. The amounts of alpha-tocopherol found in Andean potato tubers, extending from 2.73 to 20.80 microg g(-1) of DW, were clearly above the quantities generally reported for commercial varieties. Chlorogenic acid and its isomers dominated the polyphenolic profile of each cultivar. Dark purple-fleshed tubers from the cultivar 704429 contained exceptionally high levels of total anthocyanins (16.33 mg g(-1) of DW). The main anthocyanin was identified as petanin (petunidin-3-p-coumaroyl-rutinoside-5-glucoside). The results suggest that Andean potato cultivars should be exploited in screening and breeding programs for the development of potato varieties with enhanced health and nutritional benefits.

Extraction optimization of accelerated solvent extraction for eight active compounds from Yaobitong capsule using response surface methodology: Comparison with ultrasonic and reflux extraction

Anthemis secundiramea is an endemic species of the Asteraceae family, native to northeastern Algeria. Despite its unique presence in this region, a comprehensive phytochemical study focusing on its phenolic content has not yet been conducted. In this work, we obtained four leaf extracts of this species using ultrasonic extraction with different solvents.To quantify the phenolic compounds present in the aerial parts (leaves) of Anthemis secundiramea, we conducted several analyses to determine their phenolic composition and antioxidant activity. Two qualitative and quantitative colorimetric methods, employing specific reagents, were used. The results confirmed that this plant is indeed rich in phenolic compounds. Antioxidant activities were evaluated using two different assays: the free radical scavenging test (DPPH) and the ferric reducing antioxidant power test (FRAP).Additionally, the phenolic profiles of the extracts were determined by high-performance liquid chromatography coupled with a diode-array detector (HPLC/DAD). Among the various extracts studied, the methanol/water extract exhibited the highest antioxidant capacity (IC50: 1.42 ± 0.01) and contained the highest levels of total phenols (109.159 ± 12.213 mg GAE/g) and total flavonoids (95.44 ± 3.93 mg QE/g). The HPLC/DAD analysis also revealed that the leaves of Anthemis secundiramea are particularly rich in catechin, cinnamic acid, and coumaric acid.This study highlights the antioxidant potential of Anthemis secundiramea, emphasizing its richness in bioactive compounds. Further exploration is needed to fully understand the role of these bioactive compounds in herbal drug development, potentially paving the way for new natural antioxidant sources.

The bioactive compounds of leaves, stems, fruits, and seeds of Casuarina glauca from Djerba, Tunisia, was investigated using maceration and ultrasound-assisted extraction (UAE) techniques. The aqueous extracts were analyzed through high-performance liquid chromatography coupled with a diode-array detector (HPLC-DAD) to identify and quantify bioactive compounds. A total of fifteen compounds were identified, including eight phenolic acids, six flavonoids, and one phenylethanoid glycoside. The HPLC-DAD analysis revealed that catechin was the most abundant compound, particularly in the leaf's extracts, with a concentration of 139.7mg/mL. Significant variations were observed in the phenolic profiles depending on the extraction technique and the analyzed plant part. Total phenolic content (TPC) and total flavonoid content (TFC) were quantified, indicating that with the use of the UAE method, the leaves extract exhibited the highest level of TPC and TFC (17.53mg GAE/g dry weight [DW] and 15.43mg QE/g DW, respectively). Notably, the aqueous leaf extract obtained using UAE exhibited the highest antioxidant activity, as determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric-reducing antioxidant power (FRAP) assays. These results underscore the potential of C. glauca extracts from Tunisia for the first time as valuable sources of natural antioxidants, with promising applications in green pharmacy, complementary medicine, and preventive therapies.

This paper reports, for the first time, the development of a modified Quick, Easy, Cheap Effective, Rugged and Safe (QuEChERS) combined with a dispersive SPE (d-SPE) based clean-up procedure as a new and powerful strategy for the simultaneous and efficient isolation of two different antidepressants, fluoxetine and clomipramine, and their active metabolites in human urine samples. A univariate experimental design with four independent variables such as sample volume, extraction solvent, buffered salts and clean-up step, was performed and used to investigate the effect of process variables on the extraction efficiency. Good linearity was achieved at the studied concentration range (0.1-5.0 µg mL-1), with correlation coefficients (R2) higher than 0.9961. Low detection limits, ranging between 0.060 and 0.092 μg mL-1 were obtained for all analytes, whereas the lowest quantification limit was 0.1 μg mL-1, corresponding to the lowest concentration of the standard curve. The method also showed good results for accuracy, with values ranging from 91% to 105%. Intra- and inter-day precision, expressed as the relative standard deviation (RSD), were also satisfactory (<10%). Consistent recoveries of antidepressants ranging from 86% to 109% were observed when urine samples were fortified at three concentrations, namely 0.1, 2.5 and 5.0 μg mL-1 In order to evaluate the proposed method for clinical use, the QuEChERS/UHPLC-PDA method was applied to analysis of 12 urine samples from depressed patients.

HPLC-DAD method for simultaneous determination of gallic acid, catechins, and methylxanthines and its application in quantitative analysis and origin identification of green tea

ABSTRACTEstrogen hormones are present in the human body at varying concentrations, either naturally or through ingestion. At regulated levels, these compounds perform vital functions in the body. However, elevated concentrations are related to deregulation of several processes, such as abnormal cell proliferation, which can lead to the development of cancer (breast and ovarian) and other diseases (endometriosis). In this work, an analytical methodology is proposed for the determination of 17β‐estradiol (E1), estrone (E2), and 17α‐ethinylestradiol (EE2) in human urine. Hollow fiber‐microporous membrane liquid–liquid extraction (HF‐MMLLE) is used with natural deep eutectic solvents (NADESs). This technique was associated with a 96‐well plate system followed by high‐performance liquid chromatography coupled with a diode array detector (HPLC‐DAD). The optimized conditions consisted of using thymol and camphor (1:1 v/v) as extraction solvent; an extraction time of 100 min; acetonitrile as desorption solvent; a desorption time of 30 min; and a sample pH adjusted to 12. Limits of quantification (LOQs) of 25 ng mL−1 for E2 and 50 ng mL−1 for E1 and EE2, and limits of detection (LOD) of 7.5 ng mL−1 for E2 and 15 ng mL−1 for E1 and EE2 were obtained. Intraday and interday precisions were lower than 8.7% and 21.6%, respectively. Relative recoveries were examined in two samples and ranged from 68.2% to 131.8%. The method was applied in samples from healthy volunteers, and concentrations from 75.6 to 182.7 ng mL−1 for E2 were found. Finally, the method was evaluated by two sustainable metrics to examine the green aspects of the experimental approach.

A novel integrated platform enabling simultaneous microextraction and chemical analysis on-chip

Simultaneous determination of eight flavonoids in propolis using chemometrics-assisted high performance liquid chromatography-diode array detection.