Low-protein diet supplemented with inulin lowers protein-bound toxin levels in patients with stage 3b-5 chronic kidney disease: a randomized controlled study.

Introduction: Objective: this study aimed to evaluate whether low-salt low-protein diet (LPD) supplemented with 10 g of inulin could lower serum toxin levels in patients with chronic kidney disease (CKD), thereby providing evidence for adjusting dietary prescriptions of inhospital patients and outpatient nutrition consultants. Methods: we randomized 54 patients with CKD into two groups. Dietary protein intake compliance was evaluated using a 3-day dietary diary and 24-h urine nitrogen levels. The primary outcomes were indoxyl sulfate (IS) and p-cresyl sulfate (PCS), and secondary outcomes included inflammation marker levels, nutritional status, and renal function. We assessed 89 patients for eligibility, and a total of 45 patients completed the study, including 23 and 22 in the inulin-added and control groups, respectively. Results: PCS values decreased in both groups after intervention: inulin-added group, ∆PCS -1.33 (-4.88, -0.63) µg/mL vs. LPD group, -4.7 (-3.78, 3.69) µg/mL (p = 0.058). PCS values reduced from 7.52 to 4.02 µg/mL (p < 0.001) in the inulin-added group (p < 0.001). Moreover, IS decreased from 3.42 (2.53, 6.01) µg/mL to 2.83 (1.67, 4.74) µg/mL after adding inulin; ∆IS was -0.64 (-1.48, 0.00) µg/mL, and a significant difference was observed compared with the control group (p = 0.004). The inflammation index decreased after intervention. Conclusion: dietary fiber supplementation may reduce serum IS and PCS levels and modulate their inflammatory status in predialysis CKD patients.

Introducción: Objetivo: este ensayo aleatorizado doble ciego comparó el efecto de una dieta baja en proteínas (LPD) con o sin suplementos orales de 10 g de inulina en los niveles de PBUT en pacientes con ERC en prediálisis durante 12 semanas. Métodos: clasificamos aleatoriamente a 54 pacientes con ERC en dos grupos. El cumplimiento de la ingesta dietética de proteínas se evaluó utilizando un diario dietético de 3 días y nitrógeno en orina de 24 horas. Los resultados primarios fueron IS y PCS y los resultados secundarios incluyeron niveles de marcadores de inflamación, estado nutricional y función renal. Evaluamos la elegibilidad de 89 pacientes y 45 completaron la intervención, incluidos 23 y 22 en los grupos de inulina añadida y de control, respectivamente. Resultados: el sodio urinario promedio de 24 horas fue de 86 mmol/día y la ingesta promedio de proteínas fue de ~0,7 g/kg/día. Los valores de PCS exhibieron una tendencia decreciente en ambos grupos después de la intervención: grupo con inulina añadida, ∆PCS -1.33 (-4.88, -0.63) µg/mL vs. grupo LPD, -4.7 (-3.78, 3.69) µg/mL) (p =0,058). Los valores de PCS se redujeron de 7,52 a 4,02 µg/mL (p < 0,001) con inulina (p < 0,001). Además, IS disminuyó de 3,42 (2,53, 6,01) µg/mL a 2,83 (1,67, 4,74) µg/mL después de agregar inulina; El ∆IS fue -0,64 (-1,48; 0,00) µg/mL y se observó una diferencia significativa en comparación con el grupo control (p =0,004). Conclusión: la suplementación con fibra dietética puede reducir las toxinas de unión a proteínas séricas en pacientes con ERC en prediálisis y modular su estado inflamatorio.

Investigation on Antioxidant Activity and Different Metabolites of Mulberry (Morus spp.) Leaves Depending on the Harvest Months by UPLC-Q-TOF-MS with Multivariate Tools.

The changes in active components in mulberry leaves harvested in different months and their antioxidant activities were investigated. Ultra-high-performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) with multivariate statistical tools was used to investigate the chemical constituents in the extracts of mulberry leaves. The results indicated that mulberry leaves were rich in phenolic acids, flavonoids, organic acids, and fatty acid derivatives. In addition, 25 different compounds were identified in the different batches of mulberry leaves. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was measured to evaluate the in vitro antioxidant activities of mulberry leaves. Among the four batches, batch A, harvested in December, exhibited the strongest DPPH radical-scavenging activity, while batch B, harvested in March, showed the weakest activity. This was related to the total phenolic content in the mulberry leaves of each batch. The optimal harvest time of mulberry leaves greatly influences the bioactivity and bioavailability of the plant.

Metabolomics analysis of differential chemical constituents and α-glucosidase inhibiting activity of Phyllanthus urinaria L. root, stem, leaf and fruit.

The differential chemical constituents of the different Phyllanthus urinaria L. (PUL) parts were investigated by UPLC/Q-TOF MS-based metabolomics. A total of 69 compounds were tentatively identified in the whole plant extract and 35 of them were common to root, stem, leaf and fruit parts. And four compounds were selected as biomarkers for leaves, fruits, stems and roots, respectively. The four PUL parts all had good α-glucosidase inhibitory activities and the activities of fruit, root and stem extracts were about fivefold higher than the leaf part. The hierarchical cluster analysis and heat map were used to explore the relationship between the α-glucosidase inhibitory activities and chemical constituent differences of four PUL parts.

Study of Chemical Compositions and Anticancer Effects of Paris polyphylla var. Chinensis Leaves.

Paris polyphylla var. chinensis (Franch.) Hara is a perennial herb belonging to the Trilliaceae family. Ultraperformance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF MS) was used to detect the composition of different fractions of Paris polyphylla var. chinensis leaves. Meanwhile, the extracts of different fractions were evaluated for their cytotoxic activities against four selected human cancer cell lines and one human normal epithelial cell line based on the MTT assay method. Multivariate statistical analysis was performed to screen differential compounds and to analyze the distributions between different fractions. Finally, more than 60 compounds were obtained and identified from the different fractions of Paris polyphylla var. chinensis leaves, and the chloroform and n-butanol extracts showed significant cytotoxic effects on these four cancer cells. Several compounds were preliminarily identified from different fractions, including 36 steroidal saponins, 11 flavonoids, 10 ceramides, 8 lipids, 6 organic acids, and 8 other compounds. Various compounds were screened out as different chemical components of different fractions, which were considered as a potential substance basis for the cytotoxicity of Paris polyphylla var. chinensis leaves.

Mechanochemical-Assisted Extraction and Hepatoprotective Activity Research of Flavonoids from Sea Buckthorn (Hippophaë rhamnoides L.) Pomaces.

Pomaces of sea buckthorn berry were usually side-products during the processing of juice. Due to a lack of an economical and effective extraction method, it was typically recognized as waste. For the purpose of resource utilization, the mechanochemical-assisted extraction (MCAE) method was applied to develop an ecofriendly extraction method and product with better pharmacology activity. The parameters were investigated through response surface methodology (RSM) design experiments. The processing conditions were optimized as follows: amount of Na2CO3 40%, ball-to-material rate 29:1 g/g, milling speed 410 rpm, milling time 24 min, extraction temperature 25 °C, extraction time 20 min and the solid-to-solution ratio 1:10 g/mL. Under these conditions, the yields of flavonoids from sea buckthorn pomaces were 26.82 ± 0.53 mg/g, which corresponds to an increase of 2 times in comparison with that extracted by the heat reflux extraction method. Meanwhile, the hepatoprotective activity of sea buckthorn pomaces extracts was studied by the liver injury induced by ip injection of tetracycline. Biochemical and histopathological studies showed that biomarkers in serum and liver of nonalcoholic fatty liver disease (NAFLD) mice were significantly ameliorated when sea buckthorn flavonoids extracted by MCAE were used. Altogether, these results demonstrate that, as a green and efficient extraction, MCAE treatment could increase the extraction yield of sea buckthorn flavonoids, meanwhile it could exhibit significant activity of improving liver function. This research provided a new way to use pomaces of sea buckthorn as a functional food. It also has great value on the comprehensive utilization of nature's resources.

Components identification and isomers differentiation in pigeon pea (Cajanus cajan L.) leaves by LC-MS.

A valid and reliable method based on ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry using electrospray ionization was established to identify chemical constituents in the ethanol extract of pigeon pea leaves. A total of 58 compounds were detected both in positive and negative modes. Among them, 42 compounds, including 16 flavones, 1 flavonol, 5 flavanones, 9 isoflavones, 1 coumarin, 1 lactone, 6 stilbenes, 2 chalcones, and 1 other compound, were unambiguously identified or tentatively assigned in view of the retention time, the molecular formula, as well as the fragmentation patterns. Moreover, eight sets of isomers were differentiated by the ion trap mass spectrometry based on the fragment ion differences or the abundance differences of the same fragment ions. The energy-resolved mass spectrometry in light of the relative abundance of characteristic fragment ions was adopted in the study.

Components characterisation of Berchemia lineata (L.) DC. by UPLC-QTOF-MS/MS and its metabolism with human liver microsomes.

Investigations were performed on the determination of the main components in Berchemia lineata (L.) DC. (BL) and its metabolism with human liver microsomes (HLM). A total of 35 compounds were detected in BL extracts and 25 of them including 6 naphthopyrones, 10 flavonoids, 2 phenolic acids, 2 phenols, 4 fatty acids and 1 quinone were unambiguously or tentatively identified by UPLC-QTOF-MS/MS. Among them, naphthopyrones were first identified in BL extracts and labelled in chromatography. In addition, the weak inhibitory effects of BL extracts (IC50＝149.25 µg/mL) and rubrofusarin-6-O-α-L-rhamnosyl-(1-6)-O-ß-D-glu-copyranside (the main component of BL extracts, M0; IC50＝82.14 µM) on CYP3A4 were also proved using testosterone as specific probe drug. The main metabolic pathway of M0 by HLM was hydroxylation in its aglycone, the metabolite was tentatively identified as 10-hydroxy-rubrofusarin-6-O-α-L-rhamnosyl-(1-6)-O-ß-D-glucopyranside. Components characterisation and the metabolism with HLM could help the further development and application of BL.

Simultaneous Quantification and Pharmacokinetic Study of Nine Bioactive Components of Orthosiphon stamineus Benth. Extract in Rat Plasma by UHPLC-MS/MS.

Orthosiphon stamineus Benth. (OS) is a traditional folk medicine for the treatment of kidney stones and other urinary tract diseases. In this study, a rapid and sensitive Ultra high-performance liquid chromatography (UHPLC)-MS/MS approach was established and validated for the simultaneous quantification of nine bioactive components in rat plasma. The nine components from OS extract detected in rat plasma were danshensu, protocatechuic acid, caffeic acid, rosmarinic acid, salvianolic acid A, salvianolic acid B, cichoric acid, sinensetin and eupatorin. After liquid-liquid extraction with ethyl acetate, the plasma samples were subjected to a triple quadrupole mass spectrometer employing electrospray ionization (ESI) technique and operating in multiple reaction monitoring (MRM) with both positive and negative ion modes. The standard curves showed good linear regression (r > 0.9915) over the concentration range for the nine analytes. The inter-day and intra-day precision and accuracy were found to be within 15% of the nominal concentration. The recovery and stability of nine compounds were all demonstrated to be within acceptable limits. The approach was successfully applied to investigate the pharmacokinetic analysis of the nine bioactive components after oral administration of OS extract in rats.

Qualitative and quantitative analysis on the chemical constituents in Orthosiphon stamineus Benth. using ultra high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry.

A sensitive and efficient method was established and validated for qualitative and quantitative analysis on the chemical constituents in Orthosiphon stamineus Benth. (O. stamineus) using ultra high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry. Based on the retention time and MS spectra, 61 compounds were detected by using ultra high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry. 52 chemical structures in the O. stamineus extracts including 26 phenolic acids, 11 flavonoids, 6 diterpenoids, 4 fatty acids and 5 tanshinones were tentatively identified without the time-consuming process of isolation. Moreover, five chemical constituents (Danshensu, Caffeic acid, Rosmarinic acid, Sinensetin and Eupatorin) were quantified in three different batches of O. stamineus samples by the developed ultra high-performance liquid chromatography coupled with electrospray ionization triple-quadrupole mass spectrometry method in 10 min. The method validation of the five compounds was performed with acceptable linearity (R2, 0.9930-0.9997), precision (RSD, 1.87-10.36%), repeatability (RSD, 0.59-4.87%) and recovery (105.30-110.53%, RSD ≤ 13.90%).