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Typhonine glycoside

Puhuang is the dried pollen of Typha angustifolia L., Typha Orientalis Presl or similar plants in the Typhaceae. The content of isorhamnetin-3-o-neohesperidin in Chinese Pharmacopoeia (2005 Edition) is determined by HPLC. The main methods for the determination of isorhamnetin-3-o-neohesperidin in Typhae and its preparations are

HPLC. Under the content determination of Typha in Chinese Pharmacopoeia (2005 Edition):

Chromatographic conditions and system applicability test: octadecyl silane bonded silica gel as filler; Acetonitrile water (15:85) was used as mobile phase; The detection wavelength was 254 nm. Column temperature 40 ℃. The number of theoretical plates shall not be less than 1000 according to the peak of isorhamnetin-3-o-neohesperidin.

Preparation of test solution: take about 0.5g of this product, accurately weigh it, put it into a 50ml measuring bottle, add 45ml of methanol, ultrasonic treatment (power 250W, frequency 20KHz) for 30 minutes, cool it, add methanol to the scale, shake it well, filter it, and take the continuous filtrate

Determination of isorhamnetin-3-o-neohesperidin in Puhuang and its preparations by HPLC. The general mobile phase system is acetonitrile water:

① Hp1050 high performance liquid chromatograph; Chromatographic column C18 (5) μ m,4.6mm × 250mm) Dalian Institute of Chemical Physics, Chinese Academy of Sciences); Mobile phase: acetonitrile water (15:85); Detection wavelength: 254nm; Column greenhouse temperature.

② The high performance liquid chromatograph is hp1050 system of Hewlett Packard Company in the United States; Chromatographic column bondclone 10 C18 (3.9mm) × 300mm）； Mobile phase: water acetonitrile (80:20); detection wavelength 254 nm, bandwidth 4 nm; reference wavelength 550 nm, bandwidth 100 nm; attenuation 8.

Fu Daxu et al. Determined the contents of typhoniflorin and isorhamnetin-3-o-neohesperidin in the total flavone extract of Typha by HPLC and 1100 chromatograph. The chromatographic column is eclipse XdB C18 (250mm × 4.6mm，5 μ m)； Mobile phase: acetonitrile-0.5% acetic acid methanol gradient elution (0min: 15:78:7; 15min: 23:70:7); The detection wavelength is 248 nm. Preparation of total flavonoids extract of Typha: 50g of Typha was extracted with 60% ethanol (500ml) × 2) , 1H each time, the extract was concentrated at 40 ℃, the extract was dissolved with 600ml water and centrifuged. The residue was dissolved with 300ml of water and centrifuged. Combine the centrifugal solution, fix the volume to 1000ml with water, adsorb it on a column of macroporous resin (150ml macroporous resin dry weight 75g) with 1.5 times the crude drug amount, elute with 600ml of water and 450ml of 20% ethanol to remove impurities, then elute with 600ml of 50% ethanol, and the eluent is obtained by vacuum drying at 40 ℃.

Yang Yonghua et al. Used high performance capillary electrophoresis and high performance liquid chromatography to determine the contents of isorhamnetin 232o2, neohesperidin and typhoside in Puhuang. Hpce-3d high performance capillary electrophoresis instrument (Hewlett Packard, USA), quartz capillary column 50 μ m × 56cm (HP), lc-6a high performance liquid chromatograph (Shimadzu company of Japan), C18 column 5mm × 200mm, particle size 5 μ M (Dalian Yite company). HPCE conditions: buffer 0.02mol/l borax-0.05mol/l 10% acetonitrile of sodium dodecyl sulfate (SDS), voltage 20kV, column temperature 30 ℃, detection wavelength 270nm, injection volume 30KPa · s. HPLC conditions: the mobile phase was acetonitrile water (15:85), the column temperature was 25 ℃, and the detection wavelength was 254 nm.

Typhaneoside Reference substance

[name] typhonioside [1]

[CAS No.] 104472-68-6

[detection method] HPLC ≥ 98%

[Specification] 20mg 50mg 100mg 500mg 1g (can be packaged according to customer requirements)

[properties] amorphous powder