The powder of WAE (yield: 30

The powder of WAE (yield: 30.43%) was kept at 4?C for further experiments. Characterization of the extracts Total polysaccharidesThe contents of total polysaccharides in WIE and WAE were determined using Cobimetinib (racemate) phenolCsulfuric acid method. using spectrophotometric and chromatographic approaches. In addition, the in vivo immunomodulatory effect of WIE, WAE and UFP of Astragalus were comprehensively compared in cyclophosphamide (Cy)-induced immunosuppressive mice. Results The compositions and contents of main active ingredients (polysaccharides, saponins and flavonoids) in WIE were determined to be more abundant than those in WAE. In Cy-induced immunosuppressive mice, oral administered with low dosage of WIE (equalled to 1 1.0?g herb/kg/day) for Cobimetinib (racemate) 18 consecutive days significantly improved the immune-related responses (body weight, number of peripheral white blood cells, thymus and spleen indexes, splenocyte proliferations, natural killer cell activity, splenic lymphocyte subset, and serum levels of immunoglobulins G and M). The potency of three Astragalus preparations on immunomodulation was observed to be WIE??UFP? ?WAE. Conclusions WIE maximally retained the chemical integrity of astragalus, and offered better restorative performance than UFP and WAE. It can be further developed as fresh strategy for reasonable use of medicinal/edible herb-derived product (draw out) for pharmaceutical and healthcare applications. Electronic supplementary material The online version of this article (10.1186/s13020-019-0234-0) contains supplementary material, which is available to authorized users. (Fisch.) Bge. var. (Bge.) Hsiao) and its ultrafine powder (UFP) with D90? ?45?m were both purchased from Guangjitang CSPC Pharm Group (Guizhou, China). The plant sample was authenticated from the related author and the voucher specimen (HQ-2017001) was deposited in Institute of Chinese Medical Sciences, University or college of Macau. For Cobimetinib (racemate) WIE, the air-dried and powdered Astragalus (400?g) was gradient extracted with 95% ethanol (4?L), 50% ethanol (4?L) and water (4?L) at 60?C for 1?h for each. The filtered components were combined and concentrated under rotate reduced pressure to remove ethanol. The concentrated draw out was then lyophilized having a Virtis Freeze Dryer (The Virtis Organization, New York, USA). The powder of WIE (yield: 31.27%) was kept at 4?C for further experiments. For WAE, the air-dried and powdered Astragalus (400?g) was extracted thrice with water (4?L) at 60?C for 1?h for each. The combined draw out was filtered, concentrated and then lyophilized having a Virtis Freeze Dryer. The powder of WAE (yield: 30.43%) was kept at 4?C for further experiments. Rabbit Polyclonal to SFRS17A Characterization of the components Total polysaccharidesThe material of total polysaccharides in WIE and WAE were identified using phenolCsulfuric acid method. Briefly, a 2?mL of glucose remedy (0C50?g/mL) or sample remedy (1?mg/mL) was mixed with 1?mL of 6% phenol remedy, and then incubated at 60?C for another 15?min after addition of 5?mL concentrated sulfuric acid. After chilling, the absorbance was measured at 490?nm. The content of total polysaccharides in WIE and WAE were determined using glucose as standard. Total saponins and Astragaloside IVThe total saponins in WIE and WAE were identified using Vanillin (glacial acetic acid) assay. Briefly, 1?mL of WIE or WAE remedy (1?mg/mL in water) was loaded onto an activated SepPak C18 Cartridges (Waters Corp., Milford, USA) and then washed with 2?mL of water. The adsorbed saponins were eluted with 1?mL methanol into a glass tube. After evaporation, the residue was dissolved in 0.2?mL 5% vanillin in glacial acetic acid solution and 0.8?mL perchloric acid. Subsequently, the combination was incubated at 60?C for 15?min followed by addition of 5?mL glacial acetic acid after cooling. The absorbance was measured at 560?nm. The material of total saponins in WIE and WAE were determined using Astragaloside IV as requirements. The content of Astragaloside IV in WIE and WAE was determined by a Waters Alliance HPLC Cobimetinib (racemate) system coupled with a Waters ACQUITY QDa Mass Detector (Waters Corp., Milford, USA). Samples were eluted on an Agilent Extend C-18 analytical column (150?mm??2.1?mm I.D., 5?m) at 25?C with mobile phases of water-acetonitrile-formic acid (65:35:0.1, v/v/v), at a flow rate of 1 1.0?mL/min. Between.

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