Quantitative analysis method and separation method of glucosides from Auranthus chinensis

Abstract

The invention relates to a method for quantitatively analyzing the content of clematis glycosides (kinsenosides) in clematis plants or dried products of clematis plants. A method for quantitatively analyzing aurosides (kinsenoside) of the present invention, comprising carrying out proton nuclear magnetic resonance analysis (1H? NMR) on a sample containing aurosides, wherein fumaric acid is used as an internal standard; And for the resulting 1H? Integrating the signal peak at the δ2.88 chemical shift in the NMR spectrum, the area representing the aureus glucoside of the following formula 1 is obtained, and the integration of the signal peak at the δ6.50 chemical shift is obtained to represent the anti-butene The area of ​​the diacid, and the ratio of the area of ​​the auroside 1 to the area of ​​the fumaric acid is used to calculate the content of the auroside 1 in the sample.

Description

technical field

[0001] The invention relates to a method for quantitatively analyzing the content of clematis glycosides (kinsenosides) in clematis plants or dried products of clematis plants. The present invention also provides a method for isolating clematis kinsenosides from clematis plants or dried products of clematis plants. Background technique

[0002] Anoectohilus formosanus can be found in Taiwan and Ryukyu. Anologia glucosides have been proven to have the ability to protect the liver, resist hyperlipidemia, and anti-inflammation, and are important active ingredients in clematis plants and their derivative health care products.

[0003] Anologia glucoside (kinsenoside) itself lacks a chromophoric group, so the detection sensitivity of this compound by UV detector is very poor. In the past, HPLC-RI was used as a quantitative analysis method in literature reports, but the RI detector itself has problems such as temperature sensitivity, unstable baseline, and poor s...

Examples

Embodiment 1

[0037] Fresh clematis plants were shredded and extracted three times with methanol at room temperature by cold immersion, the three methanol extracts were combined, the solvent was evaporated under vacuum and the solvent was concentrated to obtain a crude extract. Clematis clematis capsules (pcs) and tea bags (bags) are extracted in the same way after removing the packaging.

[0038] Take 100 mg of the aforementioned crude extract and dissolve it in 3 mL of secondary deionized water, and perform sample pretreatment with DiaionHP-20 column (4.0 × 30 cm) chromatography, and the eluents are H 2 O (200mL, AF-I), H 2 O-MeOH (7:3 volume, 300mL, AF-II) and H 2 O-MeOH (1:1 volume, 300 mL, AF-III). AF-II was collected, 0.5 mg fumaric acid internal standard was added and concentrated.

[0039] Add AF-II to 0.6mLD 2 After O was completely dissolved, proton NMR analysis was carried out ( 1 HNMR), and recorded with BrukerAVANCEIII400MHz 1 HNMR spectrum. 100 scans were recorded for e...

Embodiment 2

[0058] The AF-II eluate of Example 1 was collected and concentrated, and then separated again with DiaionHP-20 reverse chromatography column (4.0×30em). Dissolve the concentrate in H 2 O, and then filled in the column (the ratio of sample weight to column filling is 1:100), with H 2 After O (100ml) was used as the eluent to remove highly polar compounds, H 2 O:MeOH (8:2) (300ml) was extracted, and one bottle was collected every 50mL. Repeat embodiment 1 to collecting every bottle of eluate 1 HNMR analysis (no fumaric acid was added), the 3rd to 4th bottle 1 The HNMR spectrum is shown in Figure 4 . Figure 4 The signal peak at the chemical shift of figure 1 The compound 1 in the same, proves wherein the 3~4 bottle after concentration promptly obtains the aureoside (kinsenoside) compound with following chemical formula 1:

[0059]

technical field

[0001] The invention relates to a method for quantitatively analyzing the content of clematis glycosides (kinsenosides) in clematis plants or dried products of clematis plants. The present invention also provides a method for isolating clematis kinsenosides from clematis plants or dried products of clematis plants. Background technique

[0002] Anoectohilus formosanus can be found in Taiwan and Ryukyu. Anologia glucosides have been proven to have the ability to protect the liver, resist hyperlipidemia, and anti-inflammation, and are important active ingredients in clematis plants and their derivative health care products.

[0003] Anologia glucoside (kinsenoside) itself lacks a chromophoric group, so the detection sensitivity of this compound by UV detector is very poor. In the past, HPLC-RI was used as a quantitative analysis method in literature reports, but the RI detector itself has problems such as temperature sensitivity, unstable baseline, and poor s...

Examples

Embodiment 1

[0037] Fresh clematis plants were shredded and extracted three times with methanol at room temperature by cold immersion, the three methanol extracts were combined, the solvent was evaporated under vacuum and the solvent was concentrated to obtain a crude extract. Clematis clematis capsules (pcs) and tea bags (bags) are extracted in the same way after removing the packaging.

[0038] Take 100 mg of the aforementioned crude extract and dissolve it in 3 mL of secondary deionized water, and perform sample pretreatment with DiaionHP-20 column (4.0 × 30 cm) chromatography, and the eluents are H 2 O (200mL, AF-I), H 2 O-MeOH (7:3 volume, 300mL, AF-II) and H 2 O-MeOH (1:1 volume, 300 mL, AF-III). AF-II was collected, 0.5 mg fumaric acid internal standard was added and concentrated.

[0039] Add AF-II to 0.6mLD 2 After O was completely dissolved, proton NMR analysis was carried out ( 1 HNMR), and recorded with BrukerAVANCEIII400MHz 1 HNMR spectrum. 100 scans were recorded for e...

Embodiment 2

[0058] The AF-II eluate of Example 1 was collected and concentrated, and then separated again with DiaionHP-20 reverse chromatography column (4.0×30em). Dissolve the concentrate in H 2 O, and then filled in the column (the ratio of sample weight to column filling is 1:100), with H 2 After O (100ml) was used as the eluent to remove highly polar compounds, H 2 O:MeOH (8:2) (300ml) was extracted, and one bottle was collected every 50mL. Repeat embodiment 1 to collecting every bottle of eluate 1 HNMR analysis (no fumaric acid was added), the 3rd to 4th bottle 1 The HNMR spectrum is shown in Figure 4 . Figure 4 The signal peak at the chemical shift of figure 1 The compound 1 in the same, proves wherein the 3~4 bottle after concentration promptly obtains the aureoside (kinsenoside) compound with following chemical formula 1:

[0059]

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