A kind of separation and purification method of rebaudioside c
A technology of separation and purification, glycosidase, applied in the field of food additive production, which can solve problems such as unsuitable for industrial scale-up production, difficulty in solvent recovery, short service life, etc., and achieve the effects of convenient recycling, easy solvent recovery, and convenient operation
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Embodiment 1
[0056] (1) Take 1200g of mother liquor sugar in batch 1, its RC content is 10.0%, add appropriate amount of water to make it 200mg / mL, load the sample with a pump, and flow through a 30cm×50cm chromatographic column (built-in filler Stevia-AX-2) , was eluted with 60% methanol, and the RC component was collected by HPLC detection, concentrated by nanofiltration membrane, and dried to obtain 344 g of dried RC component; wherein the purity of RC was 32%.
[0057] (2) Take the dried RC component obtained in step (1), add 2752 mL of ethanol-acetone-water (10%:70%:20%), dissolve at 70°C, stir and crystallize for 12 hours, filter and dry to obtain primary crystals 136g; the purity of RC is 60%.
[0058] (3) Add 2800 mL of ethanol-methanol-water (10%: 40%: 50%) to the primary crystal obtained in step (2) for secondary recrystallization test, the dissolution temperature is 30°C, let stand for 36h, filter and dry , 78 g of secondary crystals were obtained; the RC purity was 95.7%.
Embodiment 2
[0060] (1) Take 700g of mother liquor sugar in batch 2, whose RC content is 25.1%, add appropriate amount of water to make it 10mg / mL, load the sample with a pump, and flow through a 30cm×50cm chromatographic column (built-in filler Stevia-AX-2) , eluted with 50% methanol, detected and collected RC components by HPLC, concentrated by nanofiltration membrane, and dried to obtain 326 g of dry matter of RC components; wherein the purity of RC was 50%.
[0061] (2) Take the dried RC component obtained in step (1), add 3260 mL of ethanol-acetone-water (30%: 40%: 30%), dissolve at 30°C, stir and crystallize for 36 hours, filter, and dry to obtain primary crystals 144g; the purity of RC is 80%.
[0062] (3) Add 1160 mL of ethanol-methanol-water (25%: 30%: 45%) to the primary crystal obtained in step (2) for secondary recrystallization test, the dissolution temperature is 70°C, let stand for 12h, filter and dry , 108 g of secondary crystals were obtained; the RC purity was 97.5%.
Embodiment 3
[0064] (1) Take 1300g of mother liquor sugar in batch 3, its RC content is 15.4%, add appropriate amount of water to make it 100mg / mL, load the sample with a pump, and flow through a 30cm×50cm chromatographic column (built-in filler Stevia-AX-2) , was eluted with 55% methanol, detected by HPLC to collect RC components, concentrated by nanofiltration membrane, and dried to obtain 442 g of dry matter of RC components; wherein the purity of RC was 41%.
[0065] (2) Take the dried RC component obtained in step (1), add 3575 mL of ethanol-acetone-water (20%: 60%: 20%), dissolve at 50°C, stir and crystallize for 24 hours, filter, and dry to obtain primary crystals 185g; RC purity 72%.
[0066] (3) Add 1852 mL of ethanol-methanol-water (25%: 40%: 35%) to the primary crystal obtained in step (2) for secondary recrystallization test, the dissolution temperature is 60°C, let stand for 24h, filter and dry , 133.2 g of secondary crystals were obtained; the RC purity was 96.2%.
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