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Chemical component separation and identification method for Parisforrestii (Takht.) H. Li antitumor activity part PFE-PT3

A technology of PFE-PT3 and long column double building, which is applied in the direction of material separation, medical preparations containing active ingredients, plant/algae/fungus/moss components, etc., and can solve problems such as chemical components that have not been reported yet

Active Publication Date: 2015-12-02
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

The patent "a herbal drug for treating cancer and its preparation method and application" (publication number: CN104706896A) discloses that the PFE-PT3 part of Parisia polyphylla has significant anticancer activity, but its chemical composition has not been reported yet

Method used

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  • Chemical component separation and identification method for Parisforrestii (Takht.) H. Li antitumor activity part PFE-PT3

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Experimental program
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Effect test

Embodiment 1

[0039] The preparation of PFE-PT3 of the present invention and the separation of contained chemical composition thereof:

[0040] Preparation of PFE-PT3: dry rhizomes (4.4kg) of Parisforrestii (Takht.) H.Li, crushed, soaked in 95% ethanol (13L) for 7 days, filtered, and the filter residue was further extracted. A total of 4 extractions were performed, the filtrates were combined, and the solvent was recovered to obtain 550 g of extract (code PFE-PT1). PFE-PT1 was made into a suspension with water (2L), extracted with ethyl acetate (2L×3), and the ethyl acetate was recovered to obtain 19 g of the ethyl acetate extraction fraction (code PFE-PT2). The mother liquor extracted with ethyl acetate was further extracted with n-butanol (2L×3), and n-butanol was recovered to obtain 290 g of n-butanol extraction fraction (code PFE-PT3).

[0041] Separation of chemical components contained in PFE-PT3:

[0042] Take PFE-PT3 with a weight of 180g and use chloroform-methanol mixed solvent ...

Embodiment 2

[0045] The structural identification of PFE-PT3 chemical composition of the present invention:

[0046] Papaya saponin I, white solid, molecular formula C 44 h 70 o 16 , CAS Registry No. 50773-41-6. 1 HNMR (C 5 D. 5 N, 500MHz) δ: 6.28 (1H, brs), 5.92 (1H, d, J = 1.4Hz), 5.29 (1H, d, J = 5.1Hz), 1.76 (3H, d, J = 6.2Hz), 1.12 (3H, d, J = 7.0Hz), 1.03 (3H, s), 0.81 (3H, s), 0.68 (3H, d, J = 5.6Hz). 13 CNMR (C 5 D. 5 N,125MHz)δ:140.6,121.7,109.5,109.1,101.8,100.0,86.6,82.6,81.0,77.9,77.7,77.6,77.3,76.8,76.6,74.0,72.7,72.4,69.4,66.7,62.8,62.4, 61.2, 56.5, 50.1, 41.8, 40.3, 39.7, 38.8, 37.4, 37.0, 32.2, 32.1, 31.7, 31.5, 30.5, 30.0, 29.1, 21.0, 19.3, 18.6, 17.2, 16.2, 14.9. ESIMSm / z853[M-H] - . Its NMR data are basically consistent with the data in the literature [Wang Yu, Gao Wenyuan, Yuan Lichun, Liu Xinqiao, Wang Shujun, Chen Cui. Study on the chemical composition of Dianzhonglou. Chinese Herbal Medicine, 2007,38(1):17-20].

[0047] Papaya saponin II, white solid, mol...

Embodiment 3

[0057] Inhibition of tumor cell growth by the n-butanol extraction fraction (PFE-PT3) of the extract of Pachyphylla lanceolata:

[0058] 1. Segment the PFE-PT3.

[0059] Take PFE-PT3 with a weight of 180g and use chloroform-methanol mixed solvent gradient (10:1, 5:1, 3:1, 1:1) to elute and divide through silica gel column chromatography to obtain Fr.A (14g), Four parts of Fr.B (101g), Fr.C (45g) and Fr.D (12g). Take 30g of part Fr.B and pass through C 18 Reverse-phase silica gel column chromatography (methanol-water, 10:90 → 100:0), to obtain Fr.B 1 (1.0g), Fr.B 2 (0.5g), Fr.B 3 (0.8g), Fr.B 4 (4.6g), Fr.B 5 (16.1g) and Fr.B 6 (1.0 g) 6 portions.

[0060] 2. Take a small amount of sample from the heavy part, and use column chromatography and semi-preparative high performance liquid chromatography to quickly obtain the chemical composition.

[0061] Take Fr.B 3 A portion of 150 mg was subjected to LH-20 gel column chromatography (methanol elution), followed by silica ...

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Abstract

The invention provides a Parisforrestii (Takht.) H. Li antitumor activity part PFE-PT3 and an extraction and identification method for chemical components in the Parisforrestii (Takht.) H. Li antitumor activity part PFE-PT3. By means of a separation method of chromatographic column chromatography and semi-preparation high performance liquid chromatography and an identification method of a mass spectrum and a nuclear magnetic resonance spectrum, polyphyllin I, polyphyllin II, polyphyllin III, polyphyllin V, polyphyllin H, pennogenin-3-O-alpha-L-rhamnopyranosyl-(1-<>Z4)-[alpha-L-rhamnopyranosyl-(1->2)]-beta-D-glucopyranoside (PGRR), methyl polyphyllin I, methyl polyphyllin V, PariposideA and beta-ecdysone are separated from and identified in PFE-PT3. The main chemical components of PFE-PT3 are illuminated for the first time, the chemical components can be easily and rapidly obtained from PFE-PT3 through the method, and the method can be used for establishing the quality control standard of PFE-PT3.

Description

Technical field: [0001] The invention belongs to the technical field of extraction and separation of natural products, and in particular relates to a method for separating and identifying chemical components of PFE-PT3, an anticancer active part of Parisia lanceolata. Background technique: [0002] Parisforrestii (Takht.) H.Li] is a plant of the genus Parisforrestii (Takht.) in the family Trillium. Its rhizome is used as medicine. Nameless swollen poison, venomous snake bite, sore throat, mumps, tonsillitis and big head plague (Zhu Zhaoyun. Yunnan Natural Medicine Illustrated Book. Volume 6. Kunming: Yunnan Science and Technology Press, 2010:127). The patent "a herbal drug for treating cancer and its preparation method and application" (publication number: CN104706896A) discloses that the PFE-PT3 part of Parisia polyphylla has significant anticancer activity, but its chemical composition has not been reported yet. Clarifying the chemical substance basis of PFE-PT3 is of gre...

Claims

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Application Information

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IPC IPC(8): A61K36/896A61P35/00G01N30/88
Inventor 王跃虎牛红梅杨珺罗吉凤梅任强杨永平李恒
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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