Study On The Preparation Technique And Quality Control Of Shizhu Ginseng Chewable Tablets

To prepare two extracts of total saponins and polysaccharides from the roots of Shizhu Ginseng, orthogonal experiment design and single factor test were used in optimizing the extraction and purification technique by taking the content of total saponins and total carbohydrates as the indices, respectively. The process for preparation of total saponin extract was as follows:finely pulverized Shizhu Ginseng roots were refluxed with 12 times of 70% ethanol for 2 h, and the extraction was repeated three times. The content of the total saponins in total solid was 13.3%. The process for preparation of polysaccharide extract was as follows:the dregs of Shizhu Ginseng roots after saponins had been extracted were refluxed with 20 times of water for 2 h, and the extraction was repeated four times. The combined extracts were filtered, concentrated to 1:8 (ratio of solid to liquid) and precipitated with 85% ethanol for 24 h under 4℃. The precipitates were collected to yield polysaccharide extract after washing and drying. The content of the total carbohydrates in total solid was 3.0%.The prescription and technique process were optimized based on the criterion of hardness and taste. The amounts and ratios of diluents, correctant and lubricant of excipients were investigated. The dosage of Shizhu Ginseng chewable tablet was determined by taking American Ginseng Buccal Tablets as a reference formulation. The product was prepared by mixing the extracts of Shizhu Ginseng total saponins (163 g) and polysaccharides (67 g) with mannitol (320 g), microcrystalline cellulose (50 g), saccharin sodium (1.2 g) and magnesium stearate (3.0 g). Appropriate amount of 75% ethanol was added as the binder. Finally, Shizhu Ginseng chewable tablets were produced by compression after wet granulation.A TLC method was developed for identification of ginsenoside Rg1, Re and Rb1 in Shizhu Ginseng, its extracts and chewable tablets. An HPLC method was developed for simultaneous determination of ginsenoside Rg1, Re, Rb1, Rc, Rb2 in Shizhu Ginseng, its extracts and chewable tablets. The analysis was performed on a Kromasil C18 column (4.6 mm×250 mm,5μm) using gradient elution with the mobile phase of acetonitrile-water and the detection at 203 nm. The flow rate was 1 mL/min, the column temperature was maintained at 25℃. A good linearity was found over the concentration range of 19.8～198μg/mL,20.6-206μg/mL,33.0～330μg/mL, 18.0～180μg/mL and 13.0～130μg/mL for Ginsenoside Rg1, Re, Rb1, Rc and Rb2 respectively. The fully validated HPLC method is applicable to the quality evaluation of Shizhu Ginseng, its extracts and chewable tablets.An ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometric (UPLC-ESI-MS/MS) method was developed for the qualitative identification of saponins in Shizhu Ginseng. The analysis was performed on an AcQuity UPLCTM BEH C18 column using gradient elution with a mobile phase of 0.1% formic acid and acetonitrile over 30 min. A tandem quadrupole mass spectrometer operating in scan mode was used to get total ion chromatogram. The molecular weights of the 31 constituents were concluded on the basis of their positive and negative ion electrospray mass spectra, which showed precursor ions ([M+Na]+and [M-H]-). The possible structures of 29 constituents were deduced on-line by careful studies on their MS and MS/MS spectra and/or by comparison their spectra with reference substances or literatures.A UPLC-ESI-MS/MS method determining of notoginsenoside R1, ginsenoside Rg1, Re, Rf, Rg2, Rb1, Rb2, Rb3, Rc, Rd, and malonyl ginsenoside Rg1, Re, Rb1, Rb2, Rb3, Rc and Rd in different parts of Shizhu Ginseng was developed. The analysis was carried out on an AcQuity UPLCTM BEH C18 column using gradient elution with a mobile phase of 0.1% formic acid and acetonitrile over 10 min. The detection was performed by means of electrospray ionization mass spectrometry in negative ion mode with selective ion monitoring (SIR). The results showed a significant variation in the contents of saponins among the different parts of Shizhu Ginseng. Malonyl ginsenosides existed in all parts of Shizhu Ginseng, and the contents of malonyl ginsenoside Rb1, Rb2, Rb3, Rc and Rd were high. The result provided a scientific data for further utilization of Shizhu Ginseng.