Acta Chromatographica

Development of ultrasound-assisted extraction followed by solid-phase extraction followed by dispersive liquid–liquid microextraction followed by gas chromatography for the sensitive determination of diazinon in garden parsley as vegetable samples

Mohammad Rezaee — 2022-08-24

A new pretreatment method termed ultrasound-assisted extraction (UAE) which is combined with solid-phase extraction which is combined with dispersive liquid-liquid microextraction (SPE-DLLME) followed by gas chromatography-flame ionization detector (GC-FID) analysis has been developed for the determination of diazinon in garden parsley as vegetable samples. The analyte was extracted from garden parsley sample using ultrasound-assisted extraction followed by solid-phase extraction followed by dispersive liquid-liquid microextraction. Various parameters that affect the efficiency of the extraction techniques have been optimized. The calibration plot was linear in the range of 5.0-1000 µg kg^-1 with detection limit of 1.0 µg kg^-1 for diazinon in garden parsley samples. The results confirm the suitability of the UAE-SPE-DLLME-GC-FID as a sensitive method for the analysis of the targeted analyte in garden parsley samples.

Quantitative evaluation and chromatographic fingerprinting for the quality assessment of Pudilan tablet

Mengya Lu — 2022-10-11

An easy, quick, and sensitive approach adopting ultra-performance liquid chromatography (UPLC) equipped with diode array detector was used to analyze and systematically evaluate the quality of Pudilan tablets manufactured by 12 distinct pharmaceutical companies. In this research, 15 peaks were chosen as the common peaks to assess the similarities for different batches (S1–S43) of Pudilan tablet samples. In comparison with the control fingerprint, similarity values for 43 batches of samples exceeded 0.922. In addition, by analyzing the reference substances of epigoitrin, caffeic acid, chlorogenic acid, acetylcorynoline, baicalin and baicanshialein, the chromatogram of the 6 reference substances was established. The recoveries for the reference substances which demonstrated good regression in the linear range (r²>0.999) were in the range of 98.3 to 101.1%. The results demonstrated that the established method was highly accurate, efficient and reliable. This study provides a valid, dependable and pragmatic method to evaluate the quality of Pudilan tablet.

An improved ultra-high performance liquid chromatography-tandem mass spectrometry method determining hispidulin and homoplantaginin in rat plasma and associated pharmacokinetic studies

Xianqin Wang — 2022-09-06

Flavonoids are the most abundant components in Salvia plebeia, with significant pharmacological antioxidant and hepatoprotective properties. Hispidulin and homoplantaginin are the main flavonoid components in S. Plebeia. In this study, we established an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to determine hispidulin and homoplantaginin levels in rat plasma samples, which were precipitated using a simple acetonitrile-methanol (9:1, v/v) method. We used a UPLC HSS T3 (100 mm × 2.1 mm, 1.7 μm diameter) chromatographic column, an acetonitrile-water (containing 0.1% formic acid) mobile phase, and a gradient elution flow rate of 0.4 mL/min in an elution time of 4 min. We used electrospray ionization (ESI) detection in positive ion mode, and multiple reaction monitoring mode (MRM) for quantitative analysis: m/z 301 → 286 for hispidulin, m/z 463 → 301 for homoplantaginin, and m/z 465 → 303 for internal standard (IS). In animal studies, 24 rats were orally administered hispidulin and homoplantaginin (5 mg/kg) and received sublingual intravenous injections (1 mg/kg) at two different doses. Thus, we had four animal groups with six rats/group. Differences in hispidulin and homoplantaginin pharmacokinetics in rat serum were evaluated. The calibration curve showed good linearity in the 0.5–1000 ng/mL range, with r > 0.99. Precision, accuracy, recovery, matrix effects, and stability results all met standard biological sample detection requirements. Combined, our UPLC-MS/MS method and pharmacokinetic studies showed hispidulin bioavailability was much higher than homoplantaginin at 17.8% and 0.1%, respectively.

Simultaneous determination of multiple components in Fuke Yangrong pills by HPLC

Hua Wang — 2022-08-29

An HPLC method was established for simultaneous quantification of paeoniflorin, hesperidin and ligustilide in Fuke Yangrong pills . 0.5 g powdered Fuke Yangrong pills was extracted with 25 mL 50% (v/v) ethanol for 60 min by ultrasonication. With acetonitrile-0.1% formic acid as mobile phase, gradient elution was carried out using Agilent ZORBAX Eclipse Plus C18 column (250 mm × 4.6 mm, 5.0 μm) at 1.0 mL/min. Detection wavelength was set at 230 nm for paeoniflorin, 280 nm for hesperidin and ligustilide. The temperature was 30 °C. There was a good linearity between the peak area and the concentration of each component to be measured within their linear range (r≥0.9994). The average recoveries were between 98.6% - 102.6% with RSDs no more than 2.93%. This method was validated to be suitable for the quality control of Fuke Yangrong pills.

Validated LC-MS/MS method for quantitation of solasodine in rat urine and feces: blocking nonspecific adsorption

titian lu — 2022-08-12

Solasodine, a steroidal alkaloid, is distributed extensively in Solanaceae plants with multiple biological activities such as neuroprotection, antineoplastic and anticonvulsant activies. To investigate its excretion, a reliable LC-MS/MS method for quantitation solasodine in rat urine and feces was established and validated. MTBE could efficiently extract the analyte from urinary and fecal samples with a ngligible matrix effect. Moreover, BSA, an anti-adsorptive additive, was employed to prevent the nonspecific binding of solasodine to containers and tubes. The calibration curve ranged from 4 to 2000 ng/mL. The intra- and inter-day precision and accuracy were within 16.9% and between -11.0 and 8.9%, respectively. This method was applied to the excretion of solasodine following oral and intravenous administration.

Method development and validation for simultaneous determination of Eletriptan hydrobromide and Itopride hydrochloride by using RP-HPLC

Waqar Siddique — 2022-08-23

One of the most effective, rapid, and simple method reversed-phase high-performance liquid chromatography (RP-HPLC) was used for simultaneous development and validation of Eletriptan hydrobromide (ELE HBR) and Itopride hydrochloride (ITP HCL) in combination. The method was validated based on the instructions of United States Pharmacopeia (USP) and International Conference on Harmonization (ICH) guidelines. Separation of both drugs was achieved within approximately 5 minutes by using a mobile phase made up of a 70:30 ratio of phosphate buffer and acetonitrile having a flow rate of 1ml/min. Furthermore, a comprehensive study was conducted on precision, accuracy, linearity, inter-day, intra-day studies, an assay of formulated films, and stability studies of combined prepared film. Linearity ranged between 0.9993, and 0.9965 for ELE HBR and ITP HCL respectively. The accuracy of the developed method was accurate as drug recoveries in both cases of ITP HCL, and ELE HBR falls between (99.87, 99.96, and 99.84 %) to (99.81, 99.12, and 98.44%) respectively having a concentration range of solutions between 10, 30 and 50 ng/ml dilution. Films developed by using both drugs in combination were then validated for assay studies, and it was found that substantial results of 99.05 %, and 99.87 % were found in the case of ITP HCL and ELE HBR respectively. The stability of the solution and mobile phase showed the method's accuracy as the results were 97 % for ITP HCL and 99 % for ELE HBR. In the nutshell, the proposed method developed for simultaneous determination of ITP HCL and ELE HBR was developed and validation and no interaction of any excipient were found.

Simultaneously measure the concentrations of busulfan and phenytoin in human plasma using an HPLC-MS/MS method: Application to the TDM for children underwent hematological stem cell transplantation

Ning Sun — 2022-07-31

In this work, a simple and rapid high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method has been developed and validated to carry out the simultaneous measurement of busulfan (BU) and phenytoin (PHT) in plasma of children. In this method, plasma sample could be prepared by one-step protein precipitation using 1 mL of methanol/water (1:1, v/v). After centrifugation (14500 rpm, 5min, 4^oC), 10 μL of the supernatant was injected into a Hypersil Gold C18 column (150×2.1 mm, 5 μm, Thermo Fisher Scientific) for separation by gradient elution. Quantification was carried out using multiple reactions monitoring (MRM) under positive scan mode. In the method verification, the calibration curves of BU and PHT showed satisfactory linearity (r > 0.99) at the concentration ranging from 0.02 to 20 μg/mL. The accuracy and precision were tested at four concentration levels (including the LLOQ level) with RE ranging from -0.80% to 11.45%, CV between 0.93% and 7.74%. There was no pronounced matrix effect to interfere with the quantitative analysis. Compared to determine BU and PHT using two individual methods, less pre-treatment process, labor and blood sample volume required in this proposed method. Finally, this method has been successfully applied to the therapeutic drug monitoring of BU and PHT for children underwent hematological stem cell transplantation.

UHPLC–Q/Orbitrap/MS/MS fingerprinting of Bai-Hu-Jia-Ren-Shen-Tang Decoction and evaluation of its antioxidant activity in streptozotocin-induced diabetic rats

Libin Wang — 2022-08-26

Bai-Hu-Jia-Ren-Shen-Tang Decoction (BHJRSTD) is one of the oldest classic Chinese medicine prescriptions which used in the field of treatment of diabetes. However, to the best of our knowledge, the ingredients of this prescription have not been identified, and there are very few studies on the anti-diabetic mechanism of this prescription. Therefore, BHJRSTD was detected and identified by ultra-high-performance liquid chromatography coupled with Quadrupole-Exactive Focus Orbitrap MS (UHPLC–Q/Orbitrap/MS/MS). We identified 74 compounds, including flavonoids, alkaloids, chalcones, xanthones, phenols, phenylpropanoids, terpenes, triterpenes, terpenes, amino acid derivatives, etc. Then, Sprague Dawley rats were fed with a high-fat and high-sugar diet for two months and injected with streptozotocin (STZ) to induce type 2 diabetes (T2DM). The diabetic rats were randomized to given metformin (200 mg/kg·d, n=15), BHJRSTD extracts (40 g/kg·d) and BHJRSTD extracts (10 g/kg·d) by gavage for 8 weeks. The results confirmed that BHJRSTD significantly decreased the level of MDA and increased levels of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), it shows that the prescription has significant antioxidant activity in the treatment of T2DM.