Cinnamomi and poria composition, method to prepare same and uses thereof

A method for identifying a composition of Cinnamomi and Poria comprising:

• a) extracting the composition of Cinnamomi and Poria by an appropriate organic solvent;
• b) using appropriate standard for liposoluble matter in the composition of Cinnamomi and Poria, as internal control;
• and
• c) performing high pressure liquid chromatography using a C18 column.

A method for obtaining a composition of Cinnamomi and Poria comprising steps of:

• a) obtaining, pruning, washing and cutting the plant parts:
• stem of Cinnamomum cassia Presl (Fam. Lauraceae), fungus of Poria cocos (Schw.) Wolf (Fam.
• Polyporaceae), Root of Paeonia suffruticosa Andr. (Fam. Ranunculaceae). and fruit of Prunus persica (L.) Batsch or Prunus davidiana (Carr.) Franch. (Fam. Rosaceae);
• b) drying the said plants to form 5 medicinal materials:
• Ramulus Cinnamomi, Poria Cortex, Moutan Radicis, Radix Paeonize Alba and Semen Persicae;
• c) Smashing Ramulus Cinnamomi, Semen Persicae and Moutan Radicis into coarse powders and chopping Radix Paeonize Alba into slice;
• d) Sterilizing the Poria Cortex before granulating 50% of its formula weight into fine powder and filtering the powder;
• e) putting full amount of powder of Cortex Moutan through a process of hot reflux in water and collecting its distillate;
• (residue and fluid reserved)
• f) filtering and vacuum drying said distillate to obtain crude Paeonol;
• g) dissolving the curde paeonol into 95% alcohol;
• Slowly adding the solution into saturated water solution of β-cyclodextrin while agitating it at thermostatic 80° C. to form a mixture A;
• h) exhausted filtering mixture A;
• washing the residue with anhydrous alcohol and letting it dried, a clathrate A is obtained;
• i) distilling full amount of Ramulus Cinnamomi in water for four hours and collecting its volatile matter;
• (residue and fluid reserved)
• j) dissolving the volatile matter into 95% alcohol;
• slowly adding the solution into saturated water solution of β-cyclodextrin while agitating it at thermostatic 45° C. to form a mixture B;
• k) exhausted filtering mixture B;
• washing the residue with anhydrous alcohol and letting it dried, a clathrate B is obtained;
• l) mixing residues from step (f) and (j) with full amount of Radix Paeonize Alba, Semen Persicae, 50% of Poria Cortex and 90% alcohol;
• extracting the mixture, filtering the extract and recovering alcohol from the filtered extract;
• (residue reserved)
• m) adding water in residue from step (m), distilling it and filtering the water extract;
• n) mixing water extract from step (n), alcohol extract from step (m), fluid from step (f) and step (j);
• enriching the mixture to form a creamed extractive;
• o) mixing the creamed extractive with Poria Cortex powder from step (e);
• grinding the mixture into fine powder after vacuum drying it to form a granule;
• p) mixing the fine powder with some 60% alcohol and starch gum;
• Granulating the powder to 30 meshes;
• q) mixing certain amount of silicon dioxide with clathrate A from step (i) and clathrate B from step (1);
• and
• r) mixing the mixture from step (r) with the granule from step (p) to obtain a final granule—the composition of Cinnamomi and Poria.

A method for determination of total paeoniflorin and total paeonol in a Cinnamomi and Poria composition comprising steps of:

• a) preparing the assay comprising the steps of:
• I. accurately weighing a suitable amount of standard paeoniflorin;
• dissolving it in chromatographically pure methanol to form a standard solution containing 0.9 mg. of standard paeoniflorin per 1 ml;
• II. dilute said standard solution of paeoniflorin to 18.0 μg./ml standard solution before using;
• III. accurately weighing a suitable amount of standard paeonol;
• dissolving it in chromatographically pure methanol to form a standard solution containing 0.3 mg. of standard paeonol per 1 ml;
• IV. dilute said standard solution of paeonol to 6.0 ug./ml standard solution before using;
• V. accurately weighing approximately 0.5 g. of granular of Cinnamomi and Poria composition;
• and dissolving it with 20 ml. water to form a water solution;
• VI. putting said solution in ultrasound for 10 minutes for dispersing;
• VII. extracting the solution in a separating funnel with 30 ml. analytical pure chloroform each time for 5 times;
• obtaining a solution A by combining the extracts;
• VIII. distilling solution A on a water bath of 70° C. to a suitable volume;
• mixing it with analytically pure chloroform to a constant volume of 50 ml in a measuring flask;
• VII. accurately sucking 5 ml. of the 50 ml. solution from step (VI);
• mixing it with analytically pure chloroform to a constant volume of 50 ml;
• and filtrating the diluted solution with 0.45 μm. filtration membrane to obtain a sample solution I;
• VIII. extracting the water solution left from step (V) with analytical pure water saturated n-butanol 30 ml. each time for 5 times;
• combining the extracts;
• IX. mixing the combination from step (VIII) with analytical pure water saturated n-butanol to a constant volume of 200 ml in a measuring flask to obtain solution B;
• IX. distilling to dry 50 ml solution B on a water bath;
• dissolving the residue with chromatographically pure methanol to a constant volume of 25 ml in a measuring flask;
• and
• X. accurately sucking 5 ml. of the 25 ml. solution from step (IX);
• mixing it with chromatographically pure methanol to a constant volume of 25 ml;
• filtrating the diluted solution with 0.45 um. filtration membrane to obtain sample solution II.
• b) using methanol solution of paeoniflorin (18.0 μg/ml) and methanol solution of paeonol (6.0 ug/ml) obtained from steps (II) and (IV) as the standard solutions;
• and
• c) performing HPLC assay under following conditions:
• I. spectrum column:
• ALLtech-426 C18 5 μm., 4.6×250 mm;
• II. detector:
• ALLtech UVIS-201;
• pump:
• ALLtech-426HPLC pump;
• injection valve:
• 7725i with 10 μl volume;
• III. column temperature:
• room temperature;
• flow rate 1 ml/min;
• IV. injection volume:
• 10 μl;
• V. detective wavelength:
• 230 nm (paeoniflorin) and 274 (paeonol);
• VI. mobile phase:
• methanol-water
• 35:65 (paeoniflorin)
• 60:40 (paeonol);
• and
• VII. number of theoretical plates:
• base on peak of paeoniflorin, the 20 number of theoretical plates should be no less than 5000;
• base on peak of paeonol, the number of theoretical plates should be no less than 1500.
• d). performing HPLC assay with following steps:
• I. injecting 10 μl. sample solution I (for paeonol) or sample solution II (for paeoniflorin) into HPLC to perform chromatography under given circumstance;
• and
• II. measuring area of the peaks to calculate the relative components of paeoniflorin and paeonol in the Cinnamomi and Poria composition.

A method for obtaining a composition comprising:

• distilling a required amount of Cortex Moutan with water through a process of hot reflux and collecting its distillate, filtering and drying said distillate after its cooling down to obtain crude Paeonol;
• mixing the residues with required amount of Ramulus Cinnamomi, Radix Paeonize Alba, Semen Persicae and 50% of Poria Cortex, adding alcohol to the mixture and extract, extracting the mixture, filtering the extract to obtain an alcohol extract;
• adding water to residue from the alcohol extraction, extracting and filtering it to obtain a water extract;
• Mixing the alcohol extract, water extract and solution from distilling of Cortex Moutan, enriching the mixture to a creamed extract;
• and granulating the rest 50% of Poria Cortex into fine powder, mixing the fine powder with the creamed extract, granulating the mixture after vacuum drying, mixing the granule with crude Paeonol;
• and filling the mixture in to capsules to form the product.