Optimisation of Ethanol-Reflux Extraction of Saponins from Steamed Panax notoginseng by Response Surface Methodology and Evaluation of Hematopoiesis Effect

Abstract

The present study aims to optimize the ethanol-reflux extraction conditions for extracting saponins from steamed Panax notoginseng (SPN). Four variables including the extraction time (0.5–2.5 h), ethanol concentration (50–90%), water to solid ratio (W/S, 8–16), and times of extraction (1–5) were investigated by using the Box-Behnken design response surface methodology (BBD-RSM). For each response, a second-order polynomial model with high R2 values (>0.9690) was developed using multiple linear regression analysis and the optimum conditions to maximize the yield (31.96%), content (70.49 mg/g), and antioxidant activity (EC50 value of 0.0421 mg/mL) for saponins extracted from SPN were obtained with a extraction time of 1.51 h, ethanol concentration of 60%, extraction done 3 times, and a W/S of 10. The experimental values were in good consistency with the predicted ones. In addition, the extracted SPN saponins could significantly increase the levels of blood routine parameters compared with the model group (p < 0.01) and there was no significant difference in the hematopoiesis effect between the SPN group and the SPN saponins group, of which the dose was 15 times lower than the former one. It is suggested that the SPN saponins extracted by the optimized method had similar functions of “blood tonifying” at a much lower dose.

Analyzing Active Constituents and Optimal Steaming Conditions Related to the Hematopoietic Effect of Steamed Panax notoginseng by Network Pharmacology Coupled with Response Surface Methodology

“…Repeat the extraction process three times, and the extract solution was combined, centrifuged, filtered, and concentrated to obtain the saponins of SPN which are related to hematopoietic effect. According to the published articles [5,11], HPLC analysis was performed on an Agilent 1260 HPLC system (Agilent Technologies) consisting of a G1311B Pump, a G4212B DAD detector, and a G1329B autosampler. A Vision HT C18 column (250 mm × 4:6 mm, 5 μm) was adopted for the analysis.…”

“…. The analysis of variance (ANOVA) was used to estimate the statistical significance of factors and their interaction [11]. For each term in the models, a large F value and a small P value would imply a more significant effect on the respective response variable.…”

“…In addition, previous studies showed that most of the saponins in PN tend to be stable after 6 h of steaming at 120°C [4]. Moreover, the PN sample steamed at 120°C for 2 h was considered to be the best condition for extracting saponins by reverse phase extraction with ethanol [11]. Furthermore, there is also a lack of researches on the hematopoietic activities of SPN.…”

“…Furthermore, prevailing evidence demonstrated that Box-Behnken design response surface methodology (BBD-RSM) is widely used in the researches of the extraction, preparation, and processing of TCM, which is a set of mathematical and statistical techniques used to establish models for evaluating multiple parameters and their interactions with quantitative data and statistically and effectively optimizing the complex extraction procedures [11]. In the present study, we first used network pharmacology to predict the active ingredients related to the hematopoiesis effect of SPN and verified it by pharmacological experiments in vivo.…”

Innovative Extraction Techniques and Hyphenated Instrument Configuration for Complex Matrices Analysis

“…Other important papers explore exhaustively by means of innovative equipment such as UPLC-MS (Ultra Performance Liquid Chromatography-Mass Spectrometry) [ 10 ], HSCCC (High-Speed Counter-Current Chromatography) [ 11 ], NIR (Near Infrared spectroscopy) [ 12 ], and UPLC-qTOF MS/UPLC-QqQ MS [ 13 , 14 ] plants and their derived products. Lastly, some research articles were devoted not only to the recovery and full characterization of plant metabolites, but also to the assessment of their biological activity against a panel of pharmacologically relevant targets (acetylcholinesterase, tyrosinase, α-amylase, sirtuin 1, hematopoiesis and hemostasis, skin-whitening ability) [ 15 , 16 , 17 , 18 , 19 ].…”