Drug for treating leukopenia, preparation method thereof and use thereof

Abstract

The present invention relates to a drug for treating leukopenia, its preparation method and use thereof. The drug of the invention is prepared from 200-30 parts by weight of Folium Epimedii, 100-160 parts by weight of Fructus Psoraleae, 60-120 parts by weight of Radix Aconiti Lateralis Preparata (Processed), 200-300 parts by weight of Fructus Lycii, 200-300 parts by weight of Radix Astragali, 200-300 parts by weight of Caulis Spatholobi, 200-300 parts by weight of Radix Rubiae, 100-160 parts by weight of Radix Angelicae Sinensis, 200-300 parts by weight of Rhizoma Phragmitis, 100-160 parts by weight of Radix Ophiopogonis and 100-160 parts by weight of Radix et Rhizoma Glycyrrhizae. The drug of the invention comprises chemical substances with weight ratios as follows: Leucine:guanosine:psoralenoside:isopsoralenoside:calycosin-7-glucoside:liquiritin:icariin A:1,3-dihydroxyl-2-hydroxymethylanthraquinone:epimedin A:epimedin B:epimedin C:icariin:1,3,6-trihydroxy-2-methylanthraquinone:glycyrrhizic acid=(0.13-0.27):(0.04-0.11):(0.11-0.34):(0.09-0.34):(0.05-0.11):(0.16-0.26):(0.09-0.12):(0.17-0.35):(0.11-0.16):(0.17-0.26):(0.49-0.59):1.00:(0.16-0.24):(0.08-0.14).

Claims

1. A drug for treating leukopenia, which comprises 200-30 parts by weight of Folium Epimedii, 100-160 parts by weight of Fructus Psoraleae, 60-120 parts by weight of Radix Aconiti Lateralis Preparata (Processed), 200-300 parts by weight of Fructus Lycii, 200-300 parts by weight of Radix Astragali, 200-300 parts by weight of Caulis Spatholobi, 200-300 parts by weight of Radix Rubiae, 100-160 parts by weight of Radix Angelicae Sinensis, 200-300 parts by weight of Rhizoma Phragmitis, 100-160 parts by weight of Radix Ophiopogonis, and 100-160 parts by weight of Radix et Rhizoma Glycyrrhizae, wherein said drug comprises chemical substances with weight ratios as follows:Leucine:guanosine:psoralenoside:isopsoralenoside:calycosin-7-glucoside:liquiritin:icariin A:1,3-dihydroxyl-2-hydroxymethylanthraquinone:epimedin A:epimedin B:epimedin C:icariin:1,3,6-trihydroxy-2-methylanthraquinone:glycyrrhizic acid=(0.13-0.27):(0.04-0.11):(0.11-0.34):(0.09-0.34):(0.05-0.11):(0.16-0.26):(0.09-0.12):(0.17-0.35):(0.11-0.16):(0.17-0.26):(0.49-0.59):1.00:(0.16-0.24):(0.08-0.14).

2. The drug for treating leukopenia of claim 1, comprising 240 parts by weight of Folium Epimedii, 120 parts by weight of Fructus Psoraleae, 80 parts by weight of Radix Aconiti Lateralis Preparata (Processed), 240 parts by weight of Fructus Lycii, 240 parts by weight of Radix Astragali, 240 parts by weight of Caulis Spatholobi, 240 parts by weight of Radix Rubiae, 120 parts by weight of Radix Angelicae Sinensis, 240 parts by weight of Rhizoma Phragmitis, 120 parts by weight of Radix Ophiopogonis, and 120 parts by weight of Radix et Rhizoma Glycyrrhizae.

3. The drug for treating leukopenia of claim 1, wherein said drug comprises chemical substances with weight ratios as follows: Leucine:guanosine:psoralenoside:isopsoralenoside:calycosin-7-glucoside:liquiritin:icariin A:1,3-dihydroxyl-2-hydroxymethylanthraquinone:epimedin A:epimedin B:epimedin C:icariin:1,3,6-trihydroxy-2-methylanthraquinone:glycyrrhizic acid=(0.13-0.22):(0.06-0.10):(0.14-0.26):(0.12-0.23):(0.07-0.11):(0.19-0.25):(0.10-0.12):(0.18-0.28):(0.11-0.14):(0.21-0.25):(0.50-0.54): 1.00: (0.19-0.23):(0.09-0.11).

4. The drug for treating leukopenia of claim 1, wherein said chemical substances have the following relative retention time (RRT) according to ultra performance liquid chromatography (UPLC):Leucine 0.120-0.124, guanosine 0.163-0.167, psoralenoside 0.576-0.582, isopsoralenoside 0.593-0.599, calycosin-7-glucoside 0.638-0.644, liquiritin 0.661-0.667, icariin A 0.788-0.792, 1,3-dihydroxyl-2-hydroxymethylanthraquinone 0.887-0.894, epimedin A 0.944-0.952, epimedin B 0.964-0.968, epimedin C 0.980-0.981, icariin 1.000, 1,3,6-trihydroxy-2-methylanthraquinone 1.055-1.058, glycyrrhizic acid 1.138-1.144.

5. A pharmaceutical preparation, comprising the drug of claim 1, further comprising one or more pharmaceutically acceptable carriers, in which the drug accounts for 0.1-99.9% (weight percentage), the rest being pharmaceutically acceptable carriers.

6. The pharmaceutical preparation of claim 5, wherein said pharmaceutical preparation is an oral solution or granule.

7. A method of preparing a drug for treating leukopenia according to claim 1, comprising steps of: Step (1): Taking 200-30 parts by weight of Folium Epimedii, 100-160 parts by weight of Fructus Psoraleae, 60-120 parts by weight of Radix Aconiti Lateralis Preparata (Processed), 200-300 parts by weight of Fructus Lycii, 200-300 parts by weight of Radix Astragali, 200-300 parts by weight of Caulis Spatholobi, 200-300 parts by weight of Radix Rubiae, 100-160 parts by weight of Radix Angelicae Sinensis, 200-300 parts by weight of Rhizoma Phragmitis, 100-160 parts by weight of Radix Ophiopogonis and 100-160 parts by weight of Radix et Rhizoma Glycyrrhizae, standby; Step (2): Adding water into the above formulated amount of 10 kinds of prepared slices of Chinese crude drugs (not including Folium Epimedii) and heating to boiling, then adding the formulated amount of Folium Epimedii, continuing heating to boiling, carrying out timing extraction, then filtering to get the filtrate for the first time, adding water to the residues for a second-time extraction, filtering to get the filtrate for the second time, combining the two-time filtrates, concentrating to get the extract I; Step (3): Slowly adding 85-95% ethanol into the extract I at the ratio of extract I:ethanol=1:1-2, adding while stirring, so that extract I can be evenly dispersed, then adding 85-95% ethanol to alcohol content of 60-80%, adding while stirring, standing, then recovering ethanol, to obtain Shengbai extract.

8. The preparation method of claim 7, wherein said step (2): Adding water into the formulated amount of 10 kinds of prepared slices of Chinese crude drugs (not including Folium Epimedii) and heating at 75-100° C. to boiling, then adding the formulated amount of Folium Epimedii, continuing heating at 75-100° C. to boiling, extracting for 0.5-1.5 h, then filtering to get the filtrate for the first time, adding water to the residues for a second-time extraction, filtering to get the filtrate for the second time, combining the two-time filtrates, concentrating to relative density of 1.24-1.27(25+5° C.) to get the extract I; Wherein said step (3): Slowly adding 90% ethanol into the extract I at the ratio of extract I:ethanol=1:1-1.2, adding while stirring, so that the extract can be evenly dispersed, then adding 90% ethanol to alcohol content of 70%, adding while stirring, standing for 48-92 h for alcohol precipitation, filtering, recovering ethanol, concentrating to obtain the Shengbai extract.

9. A method of preparing an oral solution for promoting leucocytes, comprising steps of: Adding purified water and steviosin to the drug prepared according to claim 7, stirring, adjusting pH value to 5.0-6.0, heating to boiling for 30-50 min, refrigerating, taking the supernatant, adjusting pH value to 6.5-7.5, adjusting the volume to required level, filtering, filling, sterilizing.

10. The method of preparing the oral solution of claim 9, wherein said pH regulator is selected from sodium hydroxide solution or sodium bicarbonate solution.

11. A method for treating a person suffering from leukopenia comprising administering a therapeutically effective amount of the composition of claim 1.

12. The drug for treating leukopenia of claim 3, wherein said drug comprises chemical substances with weight ratios as follows: Leucine:guanosine:psoralenoside:isopsoralenoside:calycosin-7-glucoside:liquiritin:icariin A:1,3-dihydroxyl-2-hydroxymethylanthraquinone:epimedinA:epimedinB:epimedin C:icariin:1,3,6-trihydroxy-2-methylanthraquinone: glycyrrhizic acid=(0.14-0.20):(0.06-0.09):(0.19-0.24):(0.17-0.23):(0.09-0.10):(0.23-0.25):(0.10-0.12):(0.20-0.23):(0.12-0.13):(0.22-0.24):(0.51-0.52):1.00:(0.20-0.23):(0.09-0.11).

13. The method of claim 9, further comprising, adding purified water and steviosin to the drug, stirring, adjusting pH value to 5.0-5.5, heating to boiling for 30 min, refrigerating for 48 h, taking the supernatant, adjusting pH value to 7.0-7.3, adjusting the volume to required level, filtering, filling, then sterilizing.

Description

DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is the HPLC chromatogram based on HPLC-ESI-MS of Shengbai oral solution of the invention;

(2) FIG. 2 is the total ion chromatogram (positive ion mode) based on HPLC-ESI-MS of Shengbai oral solution of the invention;

(3) FIG. 3 is the total ion chromatogram (negative ion mode) based on HPLC-ESI-MS of Shengbai oral solution of the invention;

(4) FIG. 4 is the HPLC chromatogram of the drug of the invention (14 common peaks).

DETAILED DESCRIPTION OF THE INVENTION

(5) The specific embodiments of the invention are described in combination with the attached drawings, but are not construed as a limitation of the invention.

(6) Unless otherwise expressly stated, the terms “include” or “including” throughout the specifications and claims will be construed to include the elements or components stated, without excluding other elements or components.

Embodiment 1

(7) Step (1): Taking 240 g of Folium Epimedii, 120 g of Fructus Psoraleae, 80 g of Radix Aconiti Lateralis Preparata (Processed), 240 g of Fructus Lycii, 240 g of Radix Astragali, 240 g of Caulis Spatholobi, 240 g of Radix Rubiae, 120 g of Radix Angelicae Sinensis, 240 g of Rhizoma Phragmitis, 120 g of Radix Ophiopogonis and 120 g of Radix et Rhizoma Glycyrrhizae, standby;

(8) Step (2): Adding water into the above formulated amount of 10 kinds of prepared slices of Chinese crude drugs of the invention first (not including Folium Epimedii) and heating at 75-80° C. to boiling, then adding formulated amount of Folium Epimedii, continuing heating at 75-80° C. to boiling, extracting for 1 h, then filtering to get the filtrate once, adding water to the residues for a second-time extraction, filtering to get the filtrate once again, combining the two-time filtrates, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain extract I;

(9) Step (3): Slowly adding 90% ethanol into extract I of the invention at the ratio of extract I:ethanol=1:1-1.2, adding while stirring, so that extract I could be evenly dispersed, and then adding 90% ethanol to alcohol content of 70%, adding while stirring, standing for 72 h for alcohol precipitation, filtering, recovering ethanol, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain the Shengbai extract, which is the drug of the invention;

(10) Step (4): Taking 100 g of the Shengbai extract (extract density 1.26), adding purified water and steviosin based on the production process, stirring, adjusting pH value to 5.3 with sodium hydroxide, heating to boiling for 30 min, refrigerating for 48 h, taking the supernatant, adjusting pH value to 7.0 with sodium hydroxide, adjusting the volume to required level, filtering, filling, then sterilizing.

Embodiment 2

(11) Step (1): Taking 200 g of Folium Epimedii, 100 g of Fructus Psoraleae, 60 g of Radix Aconiti Lateralis Preparata (Processed), 200 g of Fructus Lycii, 200 g of Radix Astragali, 200 g of Caulis Spatholobi, 200 g of Radix Rubiae, 100 g of Radix Angelicae Sinensis, 200 g of Rhizoma Phragmitis, 100 g of Radix Ophiopogonis and 100 g of Radix et Rhizoma Glycyrrhizae, standby;

(12) Step (2): Adding water into the above formulated amount of 10 kinds of prepared slices of Chinese crude drugs of the invention first (not including Folium Epimedii) and heating at 80-85° C. to boiling, then adding formulated amount of Folium Epimedii, continuing heating at 80-85° C. to boiling, extracting for 1 h, then filtering to get the filtrate once, adding water to the residues for a second-time extraction, filtering to get the filtrate once again, combining the two-time filtrates, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain extract I;

(13) Step (3): Slowly adding 90% ethanol into extract I of the invention at the ratio of extract I:ethanol=1:1-1.2, adding while stirring, so that extract I could be evenly dispersed, and then adding 90% ethanol to alcohol content of 70%, adding while stirring, standing for 72 h for alcohol precipitation, filtering, recovering ethanol, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain the Shengbai extract.

Embodiment 3

(14) Step (1): Taking 300 g of Folium Epimedii, 160 g of Fructus Psoraleae, 120 g of Radix Aconiti Lateralis Preparata (Processed), 300 g of Fructus Lycii, 300 g of Radix Astragali, 300 g of Caulis Spatholobi, 300 g of Radix Rubiae, 160 g of Radix Angelicae Sinensis, 300 g of Rhizoma Phragmitis, 160 g of Radix Ophiopogonis and 160 g of Radix et Rhizoma Glycyrrhizae, standby;

(15) Step (2): Adding water into the above formulated amount of 10 kinds of prepared slices of Chinese crude drugs of the invention first (not including Folium Epimedii) and heating at 90-100° C. to boiling, then adding formulated amount of Folium Epimedii, continuing heating at 90-100° C. to boiling, extracting for 1 h, then filtering to get the filtrate once, adding water to the residues for a second-time extraction, filtering to get the filtrate once again, combining the two-time filtrates, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain extract I;

(16) Step (3): Slowly adding 90% ethanol into extract I of the invention at the ratio of extract I:ethanol=1:1-1.2, adding while stirring, so that extract I could be evenly dispersed, and then adding 90% ethanol to alcohol content of 70%, adding while stirring, standing for 72 h for alcohol precipitation, filtering, recovering ethanol, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain the Shengbai extract.

Embodiment 4

(17) Step (1): Taking 240 g of Folium Epimedii, 120 g of Fructus Psoraleae, 80 g of Radix Aconiti Lateralis Preparata (Processed), 240 g of Fructus Lycii, 240 g of Radix Astragali, 240 g of Caulis Spatholobi, 240 g of Radix Rubiae, 120 g of Radix Angelicae Sinensis, 240 g of Rhizoma Phragmitis, 120 g of Radix Ophiopogonis and 120 g of Radix et Rhizoma Glycyrrhizae, standby;

(18) Step (2): Adding water into the above formulated amount of 10 kinds of prepared slices of Chinese crude drugs of the invention first (not including Folium Epimedii) and heating at 75-80° C. to boiling, then adding formulated amount of Folium Epimedii, continuing heating at 75-80° C. to boiling, extracting for 0.5 h, then filtering to get the filtrate once, adding water to the residues for a second-time extraction, filtering to get the filtrate once again, combining the two-time filtrates, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain extract I;

(19) Step (3): Slowly adding 95% ethanol into extract I of the invention at the ratio of extract I:ethanol=1:1-1.2, adding while stirring, so that extract I could be evenly dispersed, and then adding 95% ethanol to alcohol content of 75%, adding while stirring, standing for 36 h for alcohol precipitation, filtering, recovering ethanol, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain the Shengbai extract.

Embodiment 5

(20) Step (1): Taking 240 g of Folium Epimedii, 120 g of Fructus Psoraleae, 80 g of Radix Aconiti Lateralis Preparata (Processed), 240 g of Fructus Lycii, 240 g of Radix Astragali, 240 g of Caulis Spatholobi, 240 g of Radix Rubiae, 120 g of Radix Angelicae Sinensis, 240 g of Rhizoma Phragmitis, 120 g of Radix Ophiopogonis and 120 g of Radix et Rhizoma Glycyrrhizae, standby;

(21) Step (2): Adding water into the above formulated amount of 10 kinds of prepared slices of Chinese crude drugs of the invention first (not including Folium Epimedii) and heating at 100° C. to boiling, then adding formulated amount of Folium Epimedii, continuing heating at 100° C. to boiling, extracting for 1.5 h, then filtering to get the filtrate once, adding water to the residues for a second-time extraction, filtering to get the filtrate once again, combining the two-time filtrates, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain extract I;

(22) Step (3): Slowly adding absolute ethanol into extract I of the invention at the ratio of extract 1:ethanol=1:1-1.2, adding while stirring, so that extract I could be evenly dispersed, and then adding absolute ethanol to alcohol content of 80%, adding while stirring, standing for 60 h for alcohol precipitation, filtering, recovering ethanol, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain the Shengbai extract.

Embodiment 6. A Shengbai Oral Solution

(23) Taking 100 g of Shengbai extract (extract density 1.26) of embodiment 2, adding purified water and 0.2% steviosin, stirring, adjusting pH value to 5.3 with sodium bicarbonate, heating to boiling for 30 min, refrigerating for 48 h, taking the supernatant, adjusting pH value to 7.0 with sodium bicarbonate, adjusting the volume to required level, filtering, filling, then sterilizing.

Embodiment 7. A Shengbai Oral Solution

(24) Taking 100 g of Shengbai extract (extract density 1.26) of any of embodiments 1-5, adding purified water and 0.2% steviosin, stirring, adjusting pH to 5.5 with sodium bicarbonate, heating to boiling for 30 min, refrigerating for 48 h, taking the supernatant, adjusting pH to 7.3 with sodium hydroxide, adjusting the volume to required level, filtering, filling, then sterilizing.

Embodiment 8. A Shengbai Oral Solution

(25) Taking 100 g of Shengbai extract (extract density 1.26) of any of embodiments 1-5, adding purified water and 0.5% white granulated sugar, stirring, adjusting pH to 6.0, heating to boiling for 30 min, refrigerating for 48 h, taking the supernatant, adjusting pH to 7.5, adjusting the volume to required level, filtering, filling, then sterilizing.

Embodiment 9. A Shengbai Oral Solution

(26) Taking 100 g of Shengbai extract (extract density 1.26) of any of embodiments 1-5, adding purified water and 0.5% white granulated sugar, stirring, adjusting pH to 5.0, heating to boiling for 30 min, refrigerating for 48 h, taking the supernatant, adjusting pH to 6.5, adjusting the volume to required level, filtering, filling, then sterilizing.

Embodiment 10. A Shengbai Granule

(27) Taking 100 g of Shengbai extract of any of embodiments 1-5, 24 g of cyclodextrin, 16 g of sodium carboxymethyl starch, 21 g of lactose, 0.67 g of steviosin, 9.5 g of citric acid (effervescing agent), 3.1 g of sodium bicarbonate and 6.2 g of sodium carbonate, fully mixing well, conducting dry granulation and granule screening to obtain Shengbai granules.

Embodiment 11. A Shengbai Capsule

(28) Taking 100 g of Shengbai extract of any of embodiments 1-5, adding 110 g of calcium carbonate and 20 g of starch, mixing, drying, grinding into fine powders, screening, mixing well, encapsulating into 1000 capsules.

Embodiment 12. A Shengbai Chewable Tablet

(29) Taking 100 g of Shengbai extract of any of embodiments 1-5, modifying with dextrin, then conducting spray drying, grinding into fine powders, adding sucrose, dextrin, mannitol, flavoring agents and other excipients, carrying out spray granulation, drying at 60° C., adding appropriate amount of magnesium stearate, tabletting into 1000 chewable tablets.

Embodiment 13. A Shengbai Suspension

(30) Taking 100 g of Shengbai extract of any of embodiments 1-5, 16 g of sodium hydroxide, 50 g of sodium citrate, 29 g of citric acid, 400 ml of simple syrup, 10 ml of 4% ethylparaben solution, adding distilled water to 100 ml to obtain the suspension.

Embodiment 14. A Shengbai Granule

(31) Taking 10 g of Shengbai extract of any of embodiments 1-5, 40 g of sucrose, adding 50% ethanol (wetting agent) to prepare into granules according to the method of preparing granules.

Embodiment 15. A Shengbai Tablet

(32) Taking 100 g of Shengbai extract of any of embodiments 1-5, 1 g of microcrystalline cellulose, adding 95% ethanol to prepare into tablets according to the method of preparing tablets.

Embodiment 16. Preparation of Dry Extract Powder

(33) (1) Spray Drying

(34) Step (1): Taking 240 g of Folium Epimedii, 120 g of Fructus Psoraleae, 80 g of Radix Aconiti Lateralis Preparata, 240 g of Fructus Lycii, 240 g of Radix Astragali, 240 g of Caulis Spatholobi, 240 g of Radix Rubiae, 120 g of Radix Angelicae Sinensis, 240 g of Rhizoma Phragmitis, 120 g of Radix Ophiopogonis and 120 g of Radix et Rhizoma Glycyrrhizae, standby;

(35) Step (2): Adding water into the above formulated amount of 10 kinds of prepared slices of Chinese crude drugs of the invention first (not including Folium Epimedii) and heating to 75-80° C., then adding formulated amount of Folium Epimedii, continuing heating to boiling, extracting for 1 h, then filtering to get the filtrate once, adding water to the residues for a second-time extraction, filtering to get the filtrate once again, combining the two-time filtrates, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain extract I;

(36) Step (3): Slowly adding 90% ethanol into extract I at the ratio of extract I:ethanol=1:1-1.2, adding while stirring, so that extract I could be evenly dispersed, and then adding 90% ethanol to alcohol content of 70%, adding while stirring, standing for 72 h for alcohol precipitation, filtering, recovering ethanol, concentrating to relative density of 1.10-1.12 (50° C.) to obtain extract II;

(37) Step (4): Adding 80 g of β-cyclodextrin and 80 g of dextrin to extract II, stirring at 60° C. for 30 min, carrying out spray drying, with inlet air temperature at 150-170° C., outlet air temperature at 75-90° C., atomizer speed at 280 Hz, liquid inlet speed at 12 Hz, obtaining dry extract powder I of 400 g±5%, with moisture of 4-6%.

(38) (2) Microwave Vacuum Drying

(39) Step (1): Taking 240 g of Folium Epimedii, 120 g of Fructus Psoraleae, 80 g of Radix Aconiti Lateralis Preparata, 240 g of Fructus Lycii, 240 g of Radix Astragali, 240 g of Caulis Spatholobi, 240 g of Radix Rubiae, 120 g of Radix Angelicae Sinensis, 240 g of Rhizoma Phragmitis, 120 g of Radix Ophiopogonis and 120 g of Radix et Rhizoma Glycyrrhizae, standby;

(40) Step (2): Adding water into the above formulated amount of 10 kinds of prepared slices of Chinese crude drugs of the invention first (not including Folium Epimedii) and heating to 75-80° C., then adding formulated amount of Folium Epimedii, continuing heating to boiling, extracting for 1 h, then filtering to get the filtrate once, adding water to the residues for a second-time extraction, filtering to get the filtrate once again, combining the two-time filtrates, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain extract I;

(41) Step (3): Slowly adding 90% ethanol into extract I at the ratio of extract I:ethanol=1:1-1.2, adding while stirring, so that extract I could be evenly dispersed, and then adding 90% ethanol to alcohol content of 70%, adding while stirring, standing for 72 h for alcohol precipitation, filtering, recovering ethanol, concentrating to relative density of 1.24-1.27 (50° C.) to obtain extract II;

(42) Step (4): Adding 80 g of β-cyclodextrin and 80 g of dextrin to extract II, stirring at 60° C. for 30 min, drying with the microwave vacuum drying oven, with the drying temperature at 50-60° C., obtaining dry extract powder I of 400 g±5%, with moisture of 4-6%.

Embodiment 17. Granules (Dry Granulation)

(43) Step (1): Adding 1 g of steviosin (flavoring agent) and 20 g of sodium carboxymethyl starch (disintegrant) to the dry extract powder I of embodiment 15, fully mixing well, carrying out dry granulation at 18-60° C. and granule screening to get the granules, packing into 9 g/bag to obtain the product.

Embodiment 18. Granules (One-Step Granulation)

(44) Step (1): After screening, adding the dry extract powder I of embodiment 15.1 g of steviosin (flavoring agent) and 20 g of sodium carboxymethyl starch (disintegrant) respectively into the fluid bed granulator of the fluid bed drying granulating machine, with the inlet hot air temperature at 70-80° C., to make the solid materials boil, premix and preheat for about 20 minutes. After qualified preheating (i.e. material temperature over 60° C.), spraying the purified water into the granulator through a spray gun for boiling drying granulation, then collecting dry granules. When spraying, the inlet air temperature: 70-90° C., the outlet air temperature: 60-70° C., peristaltic pump speed: 50-150 rpm. After spraying, continuing supplying air (with the temperature maintained at 60-70° C.) for about 20-30 minutes to control the granule moisture within the range of not more than 4%. Collecting and screening to get the granules, packing into 9 g/bag to obtain the product.

Embodiment 19. Tablets

(45) Step (1): After screening, adding the dry extract powder I of embodiment 15.1 g of steviosin (flavoring agent) and 20 g of sodium carboxymethyl starch (disintegrant) respectively into the fluid bed granulator of the fluid bed drying granulating machine, with the inlet hot air temperature at 70-80° C., to make the solid materials boil, premix and preheat for about 20 minutes. After qualified preheating (i.e. material temperature over 60° C.), spraying the purified water into the granulator through a spray gun for boiling drying granulation, then collecting dry granules. When spraying, the inlet air temperature: 70-90° C., the outlet air temperature: 60-70° C., peristaltic pump speed: 50-150 rpm. After spraying, continuing supplying air (with the temperature maintained at 60-70° C.) for about 20-30 minutes to make the granule moisture not more than 4%. Collecting and screening granules, adding 3% magnesium stearate and 3 g of sodium bicarbonate, mixing well, tabletting into 0.9 g/tablet to obtain the product.

Embodiment 20. Capsules

(46) Step (1): After screening, adding the dry extract powder I of embodiment 15.1 g of steviosin (flavoring agent) and 20 g of sodium carboxymethyl starch (disintegrant) respectively into the fluid bed granulator of the fluid bed drying granulating machine, with the inlet hot air temperature at 70-80° C., to make the solid materials boil, premix and preheat for about 20 minutes. After qualified preheating (i.e. material temperature over 60° C.), spraying the purified water into the granulator through a spray gun for boiling drying granulation, then collecting dry granules. When spraying, the inlet air temperature: 70-90° C., the outlet air temperature: 60-70° C., peristaltic pump speed: 50-150 rpm. After spraying, continuing supplying air (with the temperature maintained at 60-70° C.) for about 20-30 minutes to make the granule moisture not more than 4%. Collecting and screening granules, adding 3 g of sodium bicarbonate, mixing well, encapsulating into 0.9 g/capsule to obtain the product.

Embodiment 21. Pills

(47) Step (1): Adding sodium carboxymethyl starch (disintegrant) to the dry extract powder I of embodiment 15, fully mixing well, adding purified water to make soft materials, then preparing into concentrated pills, drying to 0.8 g/pill to obtain the product.

Embodiment 22. Preparation of Fluid Extract

(48) Step (1): Taking 240 g of Folium Epimedii, 120 g of Fructus Psoraleae, 80 g of Radix Aconiti Lateralis Preparata, 240 g of Fructus Lycii, 240 g of Radix Astragali, 240 g of Caulis Spatholobi, 240 g of Radix Rubiae, 120 g of Radix Angelicae Sinensis, 240 g of Rhizoma Phragmitis, 120 g of Radix Ophiopogonis and 120 g of Radix et Rhizoma Glycyrrhizae, standby;

(49) Step (2): Adding water into the above formulated amount of 10 kinds of prepared slices of Chinese crude drugs of the invention first (not including Folium Epimedii) and heating to 75-80° C., then adding formulated amount of Folium Epimedii, continuing heating to boiling, extracting for 1 h, then filtering to get the filtrate once, adding water to the residues for a second-time extraction, filtering to get the filtrate once again, combining the two-time filtrates, concentrating to relative density of 1.24-1.27 (25±5° C.) to obtain extract I;

(50) Step (3): Slowly adding 90% ethanol into extract I at the ratio of extract I:ethanol=1:1-1.2, adding while stirring, so that extract I could be evenly dispersed, and then adding 90% ethanol to alcohol content of 70%, adding while stirring, standing for 72 h for alcohol precipitation, filtering, recovering ethanol, concentrating to relative density of 1.24-1.27 (50° C.) to obtain extract II.

Embodiment 23. Syrups

(51) Step (1): Taking 650 g of sucrose, boiling with water to make syrup, mixing well with the fluid extract II of embodiment 22, boiling, cooling to 40° C., adding 2 g of sodium benzoate, adjusting pH to 6-7, adding water to 1000 ml, stirring well, standing, filtering, filling to get the product.

Embodiment 24. Pastes

(52) Step (1): Adding appropriate amount of sucrose into the fluid extract II of embodiment 22 (200 g of sucrose per 100 g of extract II), heating to melt, mixing well, concentrating to relative density of 1.30-1.35 (25° C.), filling to get the product.

Embodiment 25. Analysis of Chemical Constituents of Shengbai Oral Solution Fingerprint

(53) 1. Instruments

(54) Chromatograph: Dionex Ultimate 3000 RSLC (HPG) HPLC system (automatic sampler, dual trinary pump, column oven, in-line degassing unit and DAD detector);

(55) Chromatographic column: Kromasil 100-5-C18 column (250 mm×4.6 mm, 5 μm).

(56) Mass spectrometer: Thermo Scientific Q Exactive Series system (sample injection system, HESI-II ion source, mass analyzer, TraceFinder data processing system).

(57) 2. Reagents

(58) Acetonitrile: Chromatographically pure, supplied by Fisher Chemical;

(59) Methanol: Chromatographically pure, supplied by Tianjin Kemiou Chemical Reagent Co., Ltd.;

(60) Water: Watsons distilled water.

(61) 3. Chromatographic Conditions

(62) Using octadecylsilane chemically bonded silica as filler (column length 25 cm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile as mobile phase A and 0.1% formic acid as mobile phase B, conducting gradient elution based on the following table; flow rate 1.0 ml/min; column temperature 30° C.; detection wavelength 270 nm; sample size 10 μL; sample plate temperature 25° C.

(63) TABLE-US-00012 Gradient elution table Time (min) Mobile phase A (%) Mobile phase B (%)  0~10 2.fwdarw.6 98.fwdarw.94 10~31  6.fwdarw.23 94.fwdarw.77 31~40 23.fwdarw.26 77.fwdarw.74 40~56 26.fwdarw.53 74.fwdarw.47 56~60 53.fwdarw.80 47.fwdarw.20 60~70 80.fwdarw.80 20.fwdarw.20 70~75 80.fwdarw.2  20.fwdarw.98
4. Mass Spectrometry Conditions

(64) Atmosphere electrospray ionization source (ESI), positive and negative ion scanning, Full MS-ddms2 scan mode; Scan range: 100-1500 m/z; Resolution: Full MS: 70,000; MS/MS: 17,500; Capillary voltage: 3.0 kv for positive ion scanning mode, 2.5 kv for negative ion scanning mode; Sheath gas pressure: 30 bar; Aux gas pressure: 10 bar; Capillary temperature: 320° C.; Atomized gas temperature: 350° C.; Compound stability: 100%; NCE: 30.

(65) 5. Results and Discussion

(66) 5.1 HPLC-ESI-MS Analysis of Shengbai Oral Solution

(67) Under the condition of LC-MS, obvious [M+H].sup.+ or [M−H].sup.− signals were observed in the liquid chromatogram, the total ion chromatogram (both positive and negative ion scanning modes) of mass spectrometry of test samples. ESI-MS data as shown in Table 12 and FIG. 1-3.

(68) TABLE-US-00013 TABLE 12 ESI-MS data of Shengbai oral solution Time Common HPLC/ESI-MS No. (min) peak Name [M + H].sup.+ [M − H].sup.− m/z 1 2.58 Glutamic acid 148 MS2[148 .fwdarw. 130, 84] 2 2.87 Proline 116 MS2[116 .fwdarw. 70] 3 3.04 5-hydroxymethylfurfural 127 MS2[127 .fwdarw. 109] 4 4.70 Nicotinic acid 124 MS2[124 .fwdarw. 79] 5 6.96 1 Leucine 132 MS2[132 .fwdarw. 86] 6 9.50 Adenosine 268 MS2[268 .fwdarw. 136] 7 9.76 2 Guanosine 284 MS2[284 .fwdarw. 152] 8 21.84 Senkyunolide A 193 MS2[193 .fwdarw. 91] 9 22.44 Songorine 358 MS2[358 .fwdarw. 340] 10 24.54 Fuziline 454 MS2[454 .fwdarw. 436] 11 27.72 p-hydroxy benzaldehyde 123 MS2[123 .fwdarw. 95] 12 29.12 3 Psoralenoside 365 MS2[365 .fwdarw. 203, 159] 13 29.86 4 Isopsoralenoside 365 MS2[365 .fwdarw. 203, 159] 14 30.92 Vanillin 153 MS2[153 .fwdarw. 125] 15 31.32 p-hydroxy-cinnamic acid 165 MS2[165 .fwdarw. 147] 16 31.95 5 Calycosin-7-glucoside 447 MS2[447 .fwdarw. 285] 17 33.20 6 Liquiritin 417 MS2[417 .fwdarw. 255, 135] 18 33.59 Ferulic acid 195 MS2[195 .fwdarw. 177, 145] 19 34.24 Catechin 291 MS2[291 .fwdarw. 245, 227] 20 39.01 Benzoylmesaconine 590 MS2[590 .fwdarw. 540] 21 39.60 7 Icariin A 663 MS2[663 .fwdarw. 355] 22 43.13 Ononin 431 MS2[431 .fwdarw. 269] 23 44.88 8 1,3-dihydroxyl-2-hydroxy 271 MS2[271 .fwdarw. 215] methylanthraquinone 24 46.52 9 Epimedin A 839 MS2[839 .fwdarw. 369] 25 46.94 10 Epimedin B 809 MS2[809 .fwdarw. 369] 26 47.33 11 Epimedin C 823 MS2[823 .fwdarw. 369] 27 48.13 12 Icariin 677 MS2[677 .fwdarw. 369, 313] 28 50.24 13 1,3,6-trihydroxy-2- 271 MS2[271 .fwdarw. 255] methylanthraquinone 29 52.04 Psoralen 187 MS2[187 .fwdarw. 143] 30 53.11 Isopsoralen 187 MS2[187 .fwdarw. 143] 31 53.95 14 Glycyrrhizic acid 823 MS2[823 .fwdarw. 453] 32 56.38 Ligustilide 191 MS2[191 .fwdarw. 173] 33 56.80 Icarisid I 531 MS2[531 .fwdarw. 369, 313] 34 58.73 Baohuoside I 513 MS2[513 .fwdarw. 366, 323] 35 59.92 Neobavaisoflavone 323 MS2[323 .fwdarw. 267] 36 60.73 Bavachin 325 MS2[325 .fwdarw. 269] 37 61.38 Bavachinin 339 MS2[339 .fwdarw. 283] 38 63.09 Isobavachalcone 325 MS2[325 .fwdarw. 269]

Embodiment 26. Determination of Common Peaks of Shengbai Oral Solution

(69) 1. Instruments and Materials

(70) Chromatograph: USA Waters e2695 LC system (Waters PDA detector, Empower 3 workstation);

(71) Chromatographic column: Kromasil 100-5 C18 column (250 mm×4.6 mm, 5 μm);

(72) Reagents:

(73) Ethanol, analytically pure, supplied by Beijing Chemical Works;

(74) Acetonitrile, chromatographically pure, supplied by Tianjin Fuyu Fine Chemical Co., Ltd.;

(75) Formic acid, supplied by Tianjin Kemiou Chemical Reagent Co., Ltd.;

(76) Purified water, supplied by Hangzhou Wahaha Group Co., Ltd.;

(77) Adenosine (lot number CHB170802), supplied by Chengdu Chroma-Biotechnology Co., Ltd.;

(78) Calycosin-7-glucoside (lot number 111920-201505), supplied by National Institutes for Food and Drug Control;

(79) Liquiritin (lot number 11160-201106), supplied by National Institutes for Food and Drug Control;

(80) Glycyrrhizic acid (lot number 110731-201418), supplied by National Institutes for Food and Drug Control;

(81) Epimedin A (lot number MUST-16072304), supplied by Chengdu Must Biotechnology Co., Ltd.;

(82) Epimedin B (lot number MUST-15121410), supplied by Chengdu Must Biotechnology Co., Ltd.;

(83) Epimedin C (lot number 111780-201503), supplied by National Institutes for Food and Drug Control;

(84) Icariin (lot number 110737-201516), supplied by National Institutes for Food and Drug Control;

(85) Psoralen (lot number MUST-17092820), supplied by Chengdu Chroma-Biotechnology Co., Ltd.;

(86) Isopsoralen (lot number MUST-18062910), supplied by Chengdu Chroma-Biotechnology Co., Ltd.;

(87) Psoralenoside (lot number DST180723-923), supplied by Chengdu Desite Biotechnology Co., Ltd.;

(88) Isopsoralenoside (lot number DST180723-924), supplied by Chengdu Desite Biotechnology Co., Ltd.

(89) Test samples: Prepared according to embodiment 1, supplied by Hubei Monyan Pharmaceutical Co., Ltd., details as shown in Table 13.

(90) TABLE-US-00014 TABLE 13 Test samples for fingerprint study of Shengbai oral solution Name Lot number Specification 160302 20 ml/vial 160303 20 ml/vial 160304 20 ml/vial 170603 20 ml/vial 170902 20 ml/vial 170904 20 ml/vial 171002 20 ml/vial 171202 20 ml/vial 171203 20 ml/vial 171204 20 ml/vial Shengbai oral solution 171205 20 ml/vial 171207 20 ml/vial 180101 20 ml/vial 180301 20 ml/vial 180304 20 ml/vial 151011 10 ml/vial 151106 10 ml/vial 151107 10 ml/vial 151108 10 ml/vial 151109 10 ml/vial 161201 10 ml/vial 161206 10 ml/vial 170207 10 ml/vial 170803 10 ml/vial 170901 10 ml/vial 171007 10 ml/vial 171101 10 ml/vial 171104 10 ml/vial 171107 10 ml/vial 171109 10 ml/vial

(91) Preparation of reference solution An appropriate amount of icariin reference standards were accurately weighed and prepared into a solution of 100 μg/ml, which was shaken well to obtain the reference solution.

(92) Preparation of test solution 2 ml each of 15 lots of Shengbai oral solution (20 ml/vial) and 2 ml each of 15 lots of Shengbai oral solution (10 ml/vial) were accurately measured respectively, transferred into 10 ml flasks, diluted with water to scale, shaken well, filtered to get the subsequent filtrates as the test solutions.

(93) [fingerprint] Detect according to HPLC method (CP 2015, Volume IV, General Chapter 0512).

(94) Using octadecylsilane chemically bonded silica as filler (column length 25 cm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile as mobile phase A and 0.1% formic acid as mobile phase B, conducting gradient elution based on the following table; detection wavelength 270 nm; column temperature 30° C.; flow rate 1.0 ml/min; theoretical plate number not less than 10000 calculated based on icariin peak.

(95) TABLE-US-00015 Table of gradient elution, fingerprint detection of Shengbai oral solution Time(min) Mobile phase A(%) Mobile phase B(%)  0~10 2.fwdarw.6 98.fwdarw.94 10~31  6.fwdarw.23 94.fwdarw.77 31~40 23.fwdarw.26 77.fwdarw.74 40~56 26.fwdarw.53 74.fwdarw.47 56~60 53.fwdarw.80 47.fwdarw.20 60~70 80.fwdarw.80 20.fwdarw.20

(96) Test method 10 μl each of the reference solution and test solution were accurately taken, then injected into the liquid chromatograph to record the chromatogram for 70 minutes. Fingerprint similarity evaluation software was adopted to calculate the results and generate the comparison fingerprints. Results showed that the similarity of 15 lots of Shengbai oral solution (20 ml/vial) was 0.991˜0.999, and that of 15 lots of Shengbai oral solution (10 ml/vial) was 0.987˜0.999, indicating a good reproducibility and relatively stable process of the oral solution.

(97) Common peak marking: Integration time of chromatogram: 0˜70 min; Integration parameters: Peak width of 30, minimum peak area of 1, minimum peak height of 1. Within the fingerprints of 15 lots each of Shengbai oral solution (20 ml/vial) and Shengbai oral solution (10 ml/vial) samples, the chromatographic peaks with good peak patterns, high resolutions and peak areas accounting for 0.8% of the total peak area were selected for marking.

(98) The similarity of 15 lots of Shengbai oral solution (20 ml/vial) was 0.991˜0.999, and that of 15 lots of Shengbai oral solution (10 ml/vial) was 0.987˜0.999, with the average similarity of 0.993, comprehensively considering other factors, the similarity of fingerprint of Shengbai oral solution was provisionally set at the level of not being less than 0.85. The 14 common peaks marked include leucine, guanosine, psoralenoside, isopsoralenoside, calycosin-7-glucoside, liquiritin, icariin A, 1,3-dihydroxyl-2-hydroxymethylanthraquinone, epimedin A, epimedin B, epimedin C, icariin, 1,3,6-trihydroxy-2-methylanthraquinone, glycyrrhizic acid.

(99) TABLE-US-00016 TABLE 14 Retention time and peak areas of common peaks, based on fingerprints of 15 lots of Shengbai oral solution (10 ml/vial) 151011 151108 161206 170901 Retention Peak Retention Peak Retention Peak Retention Peak Peak time area time area time area time area  1 5.617 1173709 5.623 1289161 5.636 1052686 5.639 1000671  2 7.615 277414 7.582 479362 7.591 437915 7.587 532143  3 26.822 576309 26.738 964385 26.788 1034371 26.846 979233  4 27.628 451572 27.538 881777 27.588 948575 27.629 723561  5 29.737 335559 29.631 499380 29.682 450756 29.685 666502  6 30.835 948222 30.7 1207170 30.762 1146035 30.763 1459540  7 36.768 504708 36.555 518298 36.457 484178 36.488 643250  8 41.518 1559279 41.214 1666274 41.024 1096298 41.129 1306327  9 44.215 703217 43.943 605134 43.661 734651 43.806 725901 10 44.958 1149848 44.754 899506 44.545 1375417 44.659 1388946 11 45.558 2906515 45.398 2963397 45.224 2900992 45.326 3046505 12 46.436 5055032 46.301 5065248 46.161 5459449 46.251 6015263 13 49.022 830111 48.919 903647 48.832 1121002 48.905 1221697 14 52.912 417637 52.819 714707 52.753 501981 52.804 588391 171101 151107 151109 170207 Retention Peak Retention Peak Retention Peak Retention Peak Peak time area time area time area time area  1 5.671 907823 5.607 1308365 5.665 1296292 5.647 905375  2 7.689 567962 7.593 440752 7.662 424929 7.577 421531  3 26.943 1588178 26.832 1616508 26.719 1457461 26.767 778943  4 27.734 1356727 27.693 1654218 27.514 1306189 27.57 720842  5 29.819 642065 29.799 421802 29.609 438030 29.656 462671  6 30.895 1489566 30.875 1182228 30.699 1252428 30.737 1291460  7 36.681 651198 36.791 501388 36.488 533024 36.415 538764  8 41.344 1235225 41.52 1650979 41.141 1832174 40.975 1166995  9 44.012 685320 44.203 578820 43.82 634395 43.6 747007 10 44.801 1475572 44.952 863136 44.665 892082 44.558 1412713 11 45.431 2873841 45.558 2835329 45.321 3006971 45.267 2993764 12 46.33 5701311 46.442 4798808 46.237 5289941 46.206 5705031 13 48.953 1329727 49.041 918033 48.871 847657 48.892 1191975 14 52.879 503948 52.932 363056 52.758 660790 52.815 489957 171007 171107 151006 161201 Retention Peak Retention Peak Retention Peak Retention Peak Peak time area time area time area time area  1 5.66 1037215 5.745 964074 5.601 812441 5.639 1040209  2 7.601 616167 7.738 630946 7.564 225925 7.6 451479  3 26.801 1433682 26.85 1407628 26.737 964907 26.792 738859  4 27.584 1236266 27.649 1236113 27.544 784411 27.592 686776  5 29.64 670602 29.765 410846 29.671 268446 29.687 355082  6 30.717 1509022 30.855 1484963 30.712 846269 30.765 1088238  7 36.441 622704 36.679 652625 36.693 483579 36.486 445717  8 41.081 1296801 41.344 1136879 41.383 1771575 41.079 1056623  9 43.755 720849 43.993 814135 44.144 691867 43.742 683533 10 44.614 1497869 44.788 1401556 44.91 1228442 44.602 1306433 11 45.288 3076908 45.416 3337063 45.525 2758636 45.267 2675443 12 46.215 6078041 46.315 6521470 46.413 5204994 46.194 5142659 13 48.884 1401572 48.912 1232470 49.005 994617 48.858 994302 14 52.799 600678 52.786 581555 52.913 444311 52.773 478650 170803 171104 171109 Retention Peak Retention Peak Retention Peak Peak time area time area time area  1 5.69 1135512 5.675 1048592 5.61 983199  2 7.624 613218 7.625 676693 7.531 637143  3 26.801 926724 26.805 1515319 26.639 1812181  4 27.588 791699 27.602 1328871 27.427 1717791  5 29.651 457248 29.682 651217 29.515 711661  6 30.732 1487743 30.758 1534456 30.559 1608016  7 36.506 630043 36.466 698890 36.437 711115  8 41.195 1301599 41.069 1246680 41.039 1307561  9 43.908 706629 43.726 754696 43.813 764396 10 44.738 1316850 44.596 1493861 44.665 1509274 11 45.39 2892935 45.275 3139483 45.328 3388547 12 46.302 5654281 46.213 6129244 46.242 6785488 13 48.934 1272587 48.904 1426376 48.854 1509678 14 52.832 622820 52.856 560503 52.788 618662

(100) TABLE-US-00017 TABLE 15 Retention time and peak areas of common peaks, based on fingerprints of 15 lots of Shengbai oral solution (20 ml/vial) 180304 160304 170904 171202 Retention Peak Retention Peak Retention Peak Retention Peak Peak time area time area time area time area  1 5.697 927649 5.681 1172799 5.653 1234410 5.583 1088981  2 7.717 630280 7.678 406901 7.63 402705 7.57 735710  3 26.871 1567341 26.899 774976 26.936 1661819 26.754 2156173  4 27.663 1288934 27.702 703186 27.731 1399018 27.565 1971411  5 29.745 626850 29.777 511912 29.825 593112 29.682 826714  6 30.83 1391701 30.855 1122728 30.942 1574717 30.765 2038606  7 36.644 786220 36.609 552494 36.719 724673 36.537 1001102  8 41.305 1306480 41.262 1640289 41.317 1533342 41.157 1612793  9 43.982 916571 43.95 831715 44.091 827114 43.817 1179174 10 44.788 1622408 44.76 1384529 44.876 1545960 44.669 1989989 11 45.424 3607264 45.392 3096666 45.491 3216434 45.328 4392527 12 46.323 7409928 46.289 5963992 46.371 6553326 46.242 8689524 13 48.919 1893075 48.884 988301 48.945 1484064 48.849 1941169 14 52.801 711753 52.762 488081 52.815 667467 52.743 756871 180101 160302 170603 171002 Retention Peak Retention Peak Retention Peak Retention Peak Peak time area time area time area time area  1 5.633 947272 5.68 1207202 5.665 984746 5.638 1246937  2 7.592 637873 7.676 343332 7.65 477011 7.639 427183  3 26.781 1762466 26.884 1285699 26.825 872813 26.895 1633036  4 27.592 1557906 27.684 1222181 27.628 774143 27.7 1456854  5 29.708 689217 29.745 413077 29.728 491428 29.793 555752  6 30.78 1663270 30.821 1106340 30.805 1257075 30.867 1502960  7 36.573 802278 36.549 516256 36.587 661428 36.632 698487  8 41.196 1439704 41.136 1879976 41.225 1209798 41.29 1409040  9 43.859 904803 43.785 862927 43.908 818495 43.969 901338 10 44.695 1558201 44.641 1286676 44.733 1456617 44.777 1445086 11 45.34 3602280 45.3 3090143 45.371 3206804 45.41 3182719 12 46.25 7204596 46.215 5447871 46.273 6271367 46.302 6289819 13 48.86 1568431 48.84 1059293 48.894 1367738 48.894 1449251 14 52.763 647568 52.732 612373 52.773 521825 52.795 667025 171204 180301 160303 170902 Retention Peak Retention Peak Retention Peak Retention Peak Peak time area time area time area time area  1 5.645 971429 5.594 949550 5.665 1196995 5.648 1120125  2 7.609 666503 7.552 634128 7.65 297249 7.859 427557  3 26.871 1739996 26.77 1740377 26.825 835312 26.792 1348276  4 27.677 1568980 27.58 1618192 27.628 583983 27.592 988315  5 29.766 726264 29.726 640246 29.728 504444 29.682 63799  6 30.848 1684376 30.805 1607557 30.805 1228157 30.758 1607071  7 36.568 696776 36.65 727171 36.587 508024 36.557 574568  8 41.18 1465211 41.325 1326577 41.225 1456713 41.204 1409587  9 43.845 876450 44.023 854211 43.908 70613 43.889 812621 10 44.685 1657856 44.822 1533682 44.733 1064258 44.72 1577652 11 45.334 3527758 45.451 3560299 45.371 2957408 45.372 3193504 12 46.243 7056511 46.343 7025802 46.273 5514386 46.284 6195326 13 48.844 1598122 48.918 1558022 48.894 1029473 48.919 1427036 14 52.714 628620 52.797 629451 52.773 581626 52.828 611.838 171203 171205 171207 Retention Peak Retention Peak Retention Peak Peak time area time area time area  1 5.55 1414582 5.631 983.753 5.58 1065356  2 7.543 82738 7.687 596.382 7.566 643063  3 26.654 2455499 26.937 1916.194 26.787 1880597  4 27.465 2068538 27.74 1668.563 27.598 164387  5 29.58 90696 29.857 677.514 29.676 720028  6 30.657 2340311 30.939 1782.836 30.748 1818672  7 36.413 876814 36.744 667.308 36.562 697398  8 41.006 1719622 41.459 1345.05 41.16 1346073  9 43.646 1273978 44.21 795.843 43.899 906752 10 44.536 2093408 44.962 1650.886 44.764 1671137 11 45.216 4705229 45.564 3631.176 45.416 3933584 12 46.148 9211836 46.437 7010.378 46.299 7642561 13 48.786 205742 48.987 1573.616 48.848 1610911 14 52.693 782229 52.863 670.302 52.728 675571

(101) The foregoing descriptions of specific examples and embodiments of the invention are for the purpose of exemplifying and illustration. These descriptions are not intended to limit the invention to the exact form disclosed, and it is clear that many changes and variations can be made according to the above instruction. The purpose of selecting and describing the exemplary examples and embodiments is to explain the specific principles of the invention and practical applications thereof so that the various exemplary embodiments and the various options and variations of the invention can be realized and utilized by technicians in the field. The scope of the invention is subject to what is claimed and its equivalent thereof.