CN110903244A - Huperzia serrata extract and preparation method thereof - Google Patents
Huperzia serrata extract and preparation method thereof Download PDFInfo
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- CN110903244A CN110903244A CN201911306139.8A CN201911306139A CN110903244A CN 110903244 A CN110903244 A CN 110903244A CN 201911306139 A CN201911306139 A CN 201911306139A CN 110903244 A CN110903244 A CN 110903244A
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- 239000000284 extract Substances 0.000 title claims abstract description 42
- 241001090156 Huperzia serrata Species 0.000 title claims abstract description 38
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 238000000605 extraction Methods 0.000 title claims abstract description 17
- 239000000243 solution Substances 0.000 claims abstract description 67
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 62
- 238000000034 method Methods 0.000 claims abstract description 39
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 34
- 239000002994 raw material Substances 0.000 claims abstract description 25
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims abstract description 22
- 238000001914 filtration Methods 0.000 claims abstract description 19
- 239000002253 acid Substances 0.000 claims abstract description 18
- 238000003795 desorption Methods 0.000 claims abstract description 18
- 239000000047 product Substances 0.000 claims abstract description 17
- 239000000706 filtrate Substances 0.000 claims abstract description 12
- 229910021529 ammonia Inorganic materials 0.000 claims abstract description 11
- 238000001035 drying Methods 0.000 claims abstract description 11
- 238000005507 spraying Methods 0.000 claims abstract description 10
- 238000010790 dilution Methods 0.000 claims abstract description 9
- 239000012895 dilution Substances 0.000 claims abstract description 9
- 238000005406 washing Methods 0.000 claims abstract description 6
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 claims abstract description 3
- 239000003729 cation exchange resin Substances 0.000 claims abstract description 3
- 238000004140 cleaning Methods 0.000 claims abstract description 3
- 239000012535 impurity Substances 0.000 claims abstract description 3
- 239000011347 resin Substances 0.000 claims description 10
- 229920005989 resin Polymers 0.000 claims description 10
- ZRJBHWIHUMBLCN-SEQYCRGISA-N Huperzine A Natural products N1C(=O)C=CC2=C1C[C@H]1/C(=C/C)[C@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-SEQYCRGISA-N 0.000 abstract description 28
- ZRJBHWIHUMBLCN-UHFFFAOYSA-N Shuangyiping Natural products N1C(=O)C=CC2=C1CC1C(=CC)C2(N)CC(C)=C1 ZRJBHWIHUMBLCN-UHFFFAOYSA-N 0.000 abstract description 21
- ZRJBHWIHUMBLCN-YQEJDHNASA-N huperzine A Chemical compound N1C(=O)C=CC2=C1C[C@H]1\C(=C/C)[C@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-YQEJDHNASA-N 0.000 abstract description 21
- ZRJBHWIHUMBLCN-BMIGLBTASA-N rac-huperzine A Natural products N1C(=O)C=CC2=C1C[C@@H]1C(=CC)[C@@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-BMIGLBTASA-N 0.000 abstract description 21
- 229910001385 heavy metal Inorganic materials 0.000 abstract description 17
- ZQPQGKQTIZYFEF-WCVJEAGWSA-N Huperzine Natural products C1([C@H]2[C@H](O)C(=O)N[C@H]2[C@@H](O)C=2C=CC=CC=2)=CC=CC=C1 ZQPQGKQTIZYFEF-WCVJEAGWSA-N 0.000 abstract description 9
- 102000011842 Serrate-Jagged Proteins Human genes 0.000 abstract description 9
- 108010036039 Serrate-Jagged Proteins Proteins 0.000 abstract description 9
- 239000012467 final product Substances 0.000 abstract description 5
- 238000002481 ethanol extraction Methods 0.000 abstract description 3
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 16
- 229910017604 nitric acid Inorganic materials 0.000 description 16
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 15
- 230000029087 digestion Effects 0.000 description 14
- 239000003153 chemical reaction reagent Substances 0.000 description 11
- 239000000523 sample Substances 0.000 description 11
- RQNWIZPPADIBDY-UHFFFAOYSA-N arsenic atom Chemical compound [As] RQNWIZPPADIBDY-UHFFFAOYSA-N 0.000 description 10
- 238000001179 sorption measurement Methods 0.000 description 10
- 229910052785 arsenic Inorganic materials 0.000 description 9
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 8
- 238000002835 absorbance Methods 0.000 description 8
- 239000012490 blank solution Substances 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 8
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 8
- 238000000120 microwave digestion Methods 0.000 description 8
- 239000000843 powder Substances 0.000 description 8
- 239000012085 test solution Substances 0.000 description 8
- UMGDCJDMYOKAJW-UHFFFAOYSA-N thiourea Chemical compound NC(N)=S UMGDCJDMYOKAJW-UHFFFAOYSA-N 0.000 description 8
- 238000005303 weighing Methods 0.000 description 8
- 229910052753 mercury Inorganic materials 0.000 description 7
- 239000012488 sample solution Substances 0.000 description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 6
- 229910052793 cadmium Inorganic materials 0.000 description 6
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 description 6
- 239000004744 fabric Substances 0.000 description 6
- 239000010439 graphite Substances 0.000 description 6
- 229910002804 graphite Inorganic materials 0.000 description 6
- 239000008213 purified water Substances 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 5
- 125000002091 cationic group Chemical group 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 239000012046 mixed solvent Substances 0.000 description 5
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Natural products NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 238000007792 addition Methods 0.000 description 4
- 229910052786 argon Inorganic materials 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000003085 diluting agent Substances 0.000 description 4
- 238000005485 electric heating Methods 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 150000004678 hydrides Chemical class 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- -1 polytetrafluoroethylene Polymers 0.000 description 4
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 4
- 239000004810 polytetrafluoroethylene Substances 0.000 description 4
- 238000002791 soaking Methods 0.000 description 4
- 239000011550 stock solution Substances 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 150000001768 cations Chemical class 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000013558 reference substance Substances 0.000 description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 239000012445 acidic reagent Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 206010039966 Senile dementia Diseases 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 1
- 229960004373 acetylcholine Drugs 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- PYKYMHQGRFAEBM-UHFFFAOYSA-N anthraquinone Natural products CCC(=O)c1c(O)c2C(=O)C3C(C=CC=C3O)C(=O)c2cc1CC(=O)OC PYKYMHQGRFAEBM-UHFFFAOYSA-N 0.000 description 1
- 150000004056 anthraquinones Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 150000001661 cadmium Chemical class 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 229910052804 chromium Inorganic materials 0.000 description 1
- 239000011651 chromium Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000003937 effect on alzheimer disease Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000002969 morbid Effects 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 239000012088 reference solution Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D221/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00
- C07D221/02—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00 condensed with carbocyclic rings or ring systems
- C07D221/22—Bridged ring systems
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention discloses a huperzia serrata extract and a preparation method thereof, wherein the preparation method comprises the following steps: (1) raw material treatment: taking raw material huperzia serrata, cleaning with water, and drying in the shade for later use; (2) extraction: adding 65-90% ethanol, and extracting at normal temperature; (3) and (3) filtering: filtering to obtain filtrate; (4) concentration: concentrating the filtrate under reduced pressure to obtain concentrated solution a without alcohol smell, and recovering ethanol; (5) acid adjustment: adding dilute acid into the concentrated solution a to adjust the pH value to 3-4, and adding water for dilution; (6) column passing: adsorbing with 732 cation exchange resin; (7) washing with water: washing impurities with water; (8) desorbing: desorbing with ammonia alcohol, and collecting desorption solution; (9) concentration: concentrating the desorption solution under reduced pressure to obtain concentrated solution b, wherein the Baume degree is 10-20; (10) and (3) drying: and transferring the concentrated solution b to a spraying process to obtain the product of the huperzia serrata extract. The invention adopts ethanol extraction, so that the final product of the huperzine serrate extract has qualified heavy metal content, and the content of huperzine A is further improved to 10.8%.
Description
Technical Field
The invention relates to the technical field of plant extraction, and particularly relates to a huperzia serrata extract and a preparation method thereof.
Background
Huperzia serrata, also called huperzia serrata, pagoda grass and the like, belongs to perennial herb plant huperzia serrata whole plant, mainly contains alkaloid, terpenoid, flavone, anthraquinone and other components, in particular huperzine A, is proved to be a reversible, efficient and strong-selectivity acetylcholine inhibitor through research pharmacology, and has very obvious curative effect on Alzheimer's disease. With the arrival of aging society, senile dementia has become the fourth most morbid state of human beings, resulting in the shortage of huperzine A supply on the market.
At present, most of the effective components of the huperzia serrata are extracted by dilute acid, and heavy metals are easy to dissolve in the dilute acid, so that the heavy metals of a final product finally exceed the standard, and a step of removing the heavy metals is required to be specially added.
In addition, the content of huperzine A in the conventional huperzia serrata extract in the current market is about 1%, and the content of huperzine A is lower.
Disclosure of Invention
The invention aims to overcome the technical defects that the huperzine A content of the effective component is not high and the heavy metal of the huperzine A extract exceeds the standard, and provides the huperzine A extract and the preparation method thereof. The invention adopts ethanol extraction, so that the final product of the huperzine serrate extract has qualified heavy metal content, and the content of huperzine A is further improved to 10.8%.
The technical scheme adopted by the invention for solving the technical problems is as follows:
a method for preparing herba Lycopodii Serrati extract comprises the following steps:
(1) raw material treatment: taking raw material huperzia serrata, cleaning with water, and drying in the shade for later use;
(2) extraction: adding 65-90% ethanol, and extracting at normal temperature;
(3) and (3) filtering: filtering to obtain filtrate;
(4) concentration: concentrating the filtrate under reduced pressure to obtain concentrated solution a without alcohol smell, and recovering ethanol;
(5) acid adjustment: adding dilute acid into the concentrated solution a to adjust the pH value to 3-4, and adding water for dilution;
(6) column passing: adsorbing with 732 cation exchange resin;
(7) washing with water: washing impurities with water;
(8) desorbing: desorbing with ammonia alcohol, and collecting desorption solution;
(9) concentration: concentrating the desorption solution under reduced pressure to obtain concentrated solution b, wherein the Baume degree is 10-20;
(10) and (3) drying: and transferring the concentrated solution b to a spraying process to obtain the product of the huperzia serrata extract.
Preferably, in the step (2), the adding amount of the ethanol is 12-16 times of that of the raw materials according to the mass-to-volume ratio; the mass volume ratio of 1 means that 1000g of raw material and 1000ml of ethanol are added.
Preferably, in the step (2), the time for normal-temperature extraction is 1-3 h, and the operation of filtering residues is repeated for 1-2 times.
Preferably, in the step (2), the normal temperature is 20-30 ℃.
Preferably, in the step (3), the filtration is performed by using a filter cloth.
More preferably, in the step (3), the mesh number of the screen during the filtration is 200 meshes.
Preferably, in the step (4), the reduced pressure concentration is carried out under the conditions of-0.06 MPa to-0.1 MPa and 40 ℃ to 60 ℃.
Preferably, in the step (5), the addition amount of the water is 1 to 3 times of the volume of the solution after the pH adjustment according to the volume ratio.
Preferably, in the step (6), the addition amount of the resin is 0.8-1 time of the raw material by mass-volume ratio.
Preferably, in the step (6), the speed of the adsorption is 2.0-3.0 BV/h.
Preferably, in the step (7), the addition amount of the water is 2-4 BV.
Preferably, in the step (8), the ammonia alcohol is 4-5% of ammonia and 70-80% of ethanol.
Preferably, in the step (8), the flow rate during desorption is 1.0-1.5 BV/h.
Preferably, in the step (9), the reduced pressure concentration condition is 40-60 ℃ and-0.06-0.1 MPa.
Preferably, in the step (10), the inlet temperature during spraying is 180-195 ℃, the outlet temperature is 80-90 ℃, and the flow rate is 1000-2000 mL/h.
A herba Lycopodii Serrati extract is prepared by the above method.
In the technical scheme, the percentage is mass percentage.
Through a large number of experiments, the invention finds that the specific technical effect of the invention can be realized only by adopting the specific technical scheme of the invention, and other technical schemes can not realize the specific technical effect of the invention.
The basic principle of the invention is as follows:
in the prior art, most of products are extracted, and one step of heavy metal removal (chloroform extraction) is specially added, but the invention directly changes dilute acid extraction into ethanol extraction, so that heavy metal residue can be qualified in one step, the cost is low, the efficiency is high, and the period is short.
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention adopts specific cation resin, so that the content of huperzine A in the product huperzine serrate extract is more than or equal to 10.5 percent;
(2) the invention uses ethanol for extraction, so that the heavy metal content of the final product is qualified, and the total heavy metal content is less than 0.3 ppm;
(3) the process has the characteristics of science and practicality, low cost, short period and high efficiency;
(4) the huperzine A content of the huperzine A extracted from the huperzia serrata obtained by the process is more than or equal to 10.5 percent, and the huperzine A yield is more than or equal to 98 percent.
Detailed Description
For a better understanding of the present invention, reference is made to the following examples. It is to be understood that these examples are for further illustration of the invention and are not intended to limit the scope of the invention. In addition, it should be understood that the invention is not limited to the above-described embodiments, but is capable of various modifications and changes within the scope of the invention.
Example 1
A method for preparing herba Lycopodii Serrati extract comprises the following steps:
(1) raw material treatment: 1000g of raw material huperzia serrata is washed by water and dried in the shade for standby;
(2) extraction: adding 12 times of 65% ethanol, extracting at normal temperature for 1 hr, and repeating the residue for 1 time;
(3) and (3) filtering: filtering with 200 mesh filter cloth to obtain filtrate;
(4) concentration: concentrating under reduced pressure at-0.06 MPa and 60 deg.C until no alcohol smell exists, and recovering ethanol;
(5) acid adjustment: adding diluted acid into the concentrated solution obtained in the step 4 to adjust the pH value to 3, and adding 1 time of water for dilution;
(6) adsorption: adsorbing the diluent obtained in the step 5 by 800mL of 732 cationic resin at the adsorption flow rate of 2.3 BV/h;
(7) desorbing: desorbing with mixed solvent of 4% ammonia and 70% ethanol at a desorption flow rate of 1.0 BV/h;
(8) concentration: concentrating the desorption solution collected in the step 7 under reduced pressure at 60 ℃ and-0.06 MPa to obtain Baume degree of 10;
(9) and (3) drying: transferring the concentrated solution obtained in the step 8 to a spraying process, wherein the inlet temperature is 180 ℃, the outlet temperature is 80 ℃, and the flow rate is 1000mL/h, so as to obtain the product of the huperzia serrata extract.
Example 2
A method for preparing herba Lycopodii Serrati extract comprises the following steps:
(1) raw material treatment: 1000g of raw material huperzia serrata is washed by water and dried in the shade for standby;
(2) extraction: adding 90% ethanol in an amount which is 16 times that of the raw materials, extracting for 2 hours at normal temperature, and repeating the operation on filter residues for 1 time;
(3) and (3) filtering: filtering with 200 mesh filter cloth to obtain filtrate;
(4) concentration: concentrating under reduced pressure at-0.06 MPa and 60 deg.C until no alcohol smell exists, and recovering ethanol;
(5) acid adjustment: adding diluted acid into the concentrated solution obtained in the step 4 to adjust the pH value to 4, and adding 3 times of water for dilution;
(6) adsorption: adsorbing the diluent obtained in the step 5 by 1000mL of 732 cationic resin at the adsorption flow rate of 2.8 BV/h;
(7) desorbing: desorbing with 5% ammonia and 80% ethanol mixed solvent at desorption flow rate of 1.2 BV/h;
(8) concentration: concentrating the desorption solution collected in the step 7 under reduced pressure at 40 ℃ and-0.09 MPa to obtain a Baume degree of 15;
(9) and (3) drying: transferring the concentrated solution obtained in the step 8 to a spraying process, wherein the inlet temperature is 190 ℃, the outlet temperature is 90 ℃, and the flow rate is 1500mL/h, so as to obtain the product huperzia serrata extract.
Example 3
A method for preparing herba Lycopodii Serrati extract comprises the following steps:
(1) raw material treatment: 1000g of raw material huperzia serrata is washed by water and dried in the shade for standby;
(2) extraction: adding 15 times of 90% ethanol, extracting at normal temperature for 3 hr, and repeating the above steps for 2 times;
(3) and (3) filtering: filtering with 200 mesh filter cloth to obtain filtrate;
(4) concentration: concentrating under reduced pressure at 50 deg.C under-0.09 MPa until no alcohol smell exists, and recovering ethanol;
(5) acid adjustment: adding dilute acid into the concentrated solution obtained in the step 4 to adjust the pH value to 4, and adding 2 times of water for dilution;
(6) adsorption: absorbing the diluent obtained in the step 5 by 900mL of 732 cationic resin at the absorption flow rate of 2.5 BV/h;
(7) desorbing: desorbing with mixed solvent of 4.5% ammonia and 75% ethanol at a desorption flow rate of 1.5 BV/h;
(8) concentration: concentrating the desorption solution collected in the step 7 under reduced pressure at 50 ℃ and-0.09 MPa to obtain Baume degree of 18;
(9) and (3) drying: transferring the concentrated solution obtained in step 8 to spraying process at inlet temperature of 185 deg.C, outlet temperature of 85 deg.C and flow rate of 2000mL/h to obtain the product herba Lycopodii Serrati extract.
Comparative example 1
A method for preparing herba Lycopodii Serrati extract comprises the following steps:
(1) raw material treatment: 1000g of raw material huperzia serrata is washed by water and dried in the shade for standby;
(2) extraction: adding 12 times of 0.5% diluted hydrochloric acid, extracting at normal temperature for 1 hr, and repeating the residue for 1 time;
(3) and (3) filtering: filtering with 200 mesh filter cloth to obtain filtrate;
(4) adsorption: adsorbing the filtrate obtained in the step 3 by 800mL of 732 cationic resin at the adsorption flow rate of 2.3 BV/h;
(5) desorbing: desorbing with mixed solvent of 4% ammonia and 70% ethanol at a desorption flow rate of 1.0 BV/h;
(6) concentration: concentrating the desorption solution collected in the step 7 under reduced pressure at 60 ℃ and-0.06 MPa to obtain Baume degree of 10;
(7) and (3) drying: transferring the concentrated solution obtained in the step 8 to a spraying process, wherein the inlet temperature is 180 ℃, the outlet temperature is 80 ℃, and the flow rate is 1000mL/h, so as to obtain the product of the huperzia serrata extract.
Comparative example 2
A method for preparing herba Lycopodii Serrati extract comprises the following steps:
(1) raw material treatment: 1000g of raw material huperzia serrata is washed by water and dried in the shade for standby;
(2) extraction: adding 12 times of 65% ethanol, extracting at normal temperature for 1 hr, and repeating the residue for 1 time;
(3) and (3) filtering: filtering with 200 mesh filter cloth to obtain filtrate;
(4) concentration: concentrating under reduced pressure at-0.06 MPa and 60 deg.C until no alcohol smell exists, and recovering ethanol;
(5) acid adjustment: adding diluted acid into the concentrated solution obtained in the step 4 to adjust the pH value to 3, and adding 1 time of water for dilution;
(6) adsorption: adsorbing the diluent obtained in the step 5 by 800mL of 001X 7 cationic resin at the adsorption flow rate of 2.3 BV/h;
(7) desorbing: desorbing with mixed solvent of 4% ammonia and 70% ethanol at a desorption flow rate of 1.0 BV/h;
(8) concentration: concentrating the desorption solution collected in the step 7 under reduced pressure at 60 ℃ and-0.06 MPa to obtain Baume degree of 10;
(9) and (3) drying: transferring the concentrated solution obtained in the step 8 to a spraying process, wherein the inlet temperature is 180 ℃, the outlet temperature is 80 ℃, and the flow rate is 1000mL/h, so as to obtain the product of the huperzia serrata extract.
Effects of the embodiment
Method for detecting content of heavy metal lead
1 instrument and equipment:
analytical balance (accuracy: one in ten thousand grams);
graphite furnace atomic absorption spectrophotometer TAS-990;
argon cylinder (outlet pressure 0.5 MPa);
microsyringe: 10 mu L of the solution;
lead element hollow cathode lamps;
a microwave digestion instrument WX-600;
the intelligent temperature control electric heater DKQ-1800.
2, reagent:
purified water;
nitric acid (UP grade).
3 preparation of standard lead solution:
precisely sucking 1ml of lead standard stock solution (1000 mu g/ml) into a 100ml volumetric flask, diluting the solution to the scale with 2% nitric acid solution, and shaking up. I.e., lead standard use solution (10. mu.g/ml). Then 0 mul, 5 mul, 10 mul, 15 mul, 20 mul and 30 mul of lead standard use solution are sucked into 5 10ml volumetric flasks, diluted to the scale with 2% nitric acid solution and shaken up to prepare 0ng/ml, 5ng/ml, 10ng/ml, 15 ng/ml, 20ng/ml and 30ng/ml lead standard series respectively. A 2% nitric acid reagent was used as a standard blank. The absorbance at 283.3nm was measured and a standard curve was drawn.
4, preparation of test solution:
precisely weighing about 0.1g of powder, placing the powder into a polytetrafluoroethylene digestion tank, adding 3-5ml of nitric acid, uniformly mixing, soaking overnight, covering an inner cover, screwing a jacket, placing the tank into a proper microwave digestion furnace, carrying out digestion (according to the digestion program operation of an instrument), after complete digestion, canceling the inner tank, placing the inner tank on an electric heating plate (the temperature is 190 ℃ for 30min), slowly heating until reddish brown steam is volatilized, continuously slowly concentrating to 2-3ml, cooling, transferring the solution into a 25ml volumetric flask with purified water, fixing the volume, and uniformly shaking to obtain the finished product. The reagent blank solution is prepared simultaneously by the same method.
5, measurement:
precisely measuring 10 mu L of blank solution and test solution, injecting into a graphite furnace, measuring absorbance according to the method under the preparation term of the standard curve, reading out the content of lead in the test solution from the curve, and calculating to obtain the lead-free test solution.
Sample content results average of two replicates
Second, detection method of heavy metal arsenic content
1 instrument and equipment:
analytical balance (accuracy: one in ten thousand grams);
graphite furnace atomic absorption spectrophotometer TAS-990;
a hydride device;
argon cylinder (outlet pressure 0.5 MPa);
an arsenic element hollow cathode lamp;
a microwave digestion instrument WX-600;
the intelligent temperature control electric heater DKQ-1800.
2, reagent:
purified water;
nitric acid (UP grade).
3 preparation of standard arsenic solution:
precisely sucking 0.1ml of arsenic standard stock solution (1000 mug/ml) into a 100ml volumetric flask, diluting the solution to the scale with 2% hydrochloric acid solution, and shaking up to obtain the concentration (1 mug/ml) of arsenic standard use solution. 0. mu.l, 50. mu.l, 100. mu.l, 150. mu.l, 200. mu.l and 250. mu.l of the standard arsenic solutions were pipetted into 5 100ml volumetric flasks, diluted to the scale with 0.5% thiourea and 2% hydrochloric acid solutions, shaken up to prepare standard arsenic series of 0ng/ml, 0.5ng/ml, 1.0ng/ml, 1.5ng/ml, 2.0ng/ml and 2.5ng/ml, respectively. The same method as used for the reagents is used as a standard blank. Absorbance at 193.7nm was measured and a standard curve was drawn.
4, preparation of test solution:
precisely weighing about 0.1g of powder, placing the powder into a polytetrafluoroethylene digestion tank, adding 3-5ml of nitric acid, uniformly mixing, soaking overnight, covering an inner cover, screwing an outer cover, placing the outer cover into a proper microwave digestion furnace, digesting (operating according to a digestion program of an instrument), after complete digestion, canceling the inner tank, placing the inner tank on an electric heating plate, slowly heating until reddish brown steam is volatilized completely, continuously slowly concentrating to 2-3ml, cooling, transferring 0.5% thiourea and water into a 25ml volumetric flask, fixing the volume, and shaking uniformly to obtain the finished product. The reagent blank solution is prepared simultaneously by the same method.
5, measurement:
precisely measuring a proper amount of blank solution and sample solution, injecting into a hydride device, measuring absorbance according to the method under the preparation item of a standard curve, reading out the content of arsenic in the sample solution from the curve, and calculating to obtain the product.
Sample content results average of two replicates
Third, detection method of heavy metal mercury content
1 instrument and equipment:
analytical balance (accuracy: one in ten thousand grams);
graphite furnace atomic absorption spectrophotometer TAS-990;
a hydride device;
argon cylinder (outlet pressure 0.5 MPa);
mercury element hollow cathode lamps;
a microwave digestion instrument WX-600;
the intelligent temperature control electric heater DKQ-1800.
2 reagent
Purified water;
nitric acid (UP grade).
3 preparation of standard mercury solution:
1.0ml of mercury standard stock solution (1000. mu.g/ml) is taken into a 100ml volumetric flask, diluted to the mark with 2% nitric acid solution and shaken up. I.e., mercury standard use solution (10. mu.g/ml). Then 0. mu.l, 50. mu.l, 100. mu.l, 150. mu.l, 200. mu.l and 300. mu.l of the mercury standard use solution are sucked into 5 100ml volumetric flasks, diluted to the scale with 0.5% thiourea and 4% nitric acid solution, shaken up and prepared into 0ng/ml, 5ng/ml, 10ng/ml, 15 ng/ml, 20ng/ml and 30ng/ml mercury standard series respectively. The same method as used for the reagents is used as a standard blank. Absorbance was measured at 253.6nm and a standard curve was drawn.
4, preparation of test solution:
precisely weighing about 0.1g of powder, placing the powder into a polytetrafluoroethylene digestion tank, adding 3-5ml of nitric acid, uniformly mixing, soaking overnight, covering an inner cover, screwing an outer cover, placing the outer cover into a proper microwave digestion furnace, digesting (operating according to a digestion program of an instrument), after complete digestion, canceling the inner tank, placing the inner tank on an electric heating plate, slowly heating until reddish brown steam is volatilized completely, continuously slowly concentrating to 2-3ml, cooling, transferring 0.5% thiourea and water into a 25ml volumetric flask, fixing the volume, and shaking uniformly to obtain the finished product. The reagent blank solution is prepared simultaneously by the same method.
5, measurement:
precisely measuring a proper amount of blank solution and sample solution, injecting into a hydride device, measuring absorbance according to the method under the preparation item of a standard curve, reading out the content of arsenic in the sample solution from the curve, and calculating to obtain the product.
Sample content results average of two replicates
Fourth, detection method of heavy metal cadmium content
1 instrument and equipment:
analytical balance (accuracy: one in ten thousand grams);
graphite furnace atomic absorption spectrophotometer TAS-990;
argon cylinder (outlet pressure 0.5 MPa);
microsyringe: 10 mu L of the solution;
a cadmium element hollow cathode lamp;
a microwave digestion instrument WX-600;
the intelligent temperature control electric heater DKQ-1800.
2 reagent
Purified water;
nitric acid (UP grade).
3, preparation of standard cadmium solution:
precisely sucking 0.1ml of cadmium standard stock solution (1000 mug/ml) into a 100ml volumetric flask, diluting the solution to the scale with 2% nitric acid solution, and shaking up to obtain the cadmium standard use solution (1 mug/ml). Then 0 mul, 50 mul, 100 mul, 150 mul, 200 mul and 300 mul of the standard cadmium use solution are sucked into 5 100ml volumetric flasks, diluted to the scale with 2 percent nitric acid solution and shaken up to prepare 0ng/ml, 0.5ng/ml, 1.0ng/ml, 1.5ng/ml, 2.0ng/ml and 3.0ng/ml standard cadmium series respectively. A 2% nitric acid reagent was used as a standard blank. Measuring the absorbance at 228.8nm, and drawing a standard curve
4, preparation of test solution:
precisely weighing about 0.1g of powder, placing the powder into a polytetrafluoroethylene digestion tank, adding 3-5ml of nitric acid, uniformly mixing, soaking overnight, covering an inner cover, screwing a jacket, placing the tank into a proper microwave digestion furnace, carrying out digestion (according to the digestion program operation of an instrument), after complete digestion, canceling the inner tank, placing the inner tank on an electric heating plate (the temperature is 190 ℃ for 30min), slowly heating until reddish brown steam is volatilized, continuously slowly concentrating to 2-3ml, cooling, transferring the solution into a 25ml volumetric flask with purified water, fixing the volume, and uniformly shaking to obtain the finished product. The reagent blank solution is prepared simultaneously by the same method.
5, measurement:
precisely measuring 10 μ L of blank solution and sample solution, injecting into graphite furnace, measuring absorbance according to the method under the preparation of standard curve, reading out the content of lead in the sample solution from the curve, and calculating.
Sample content results average of two replicates
Fifthly, measuring the content of huperzine A
1, calculating formula:
1.1 chromatographic conditions:
1.1.1 chromatography column: Gemini-C18，4.6×250mm
1.1.2 detection wavelength 310nm
1.1.3 detector: VWD
1.1.4 column temperature: 30 deg.C
1.1.5 flow rate: 1.0ml/min
1.1.6 mobile phase: phosphate buffer acetonitrile (86: 14)
2 preparation of control solutions:
precisely weighing huperzine A reference substance, adding 0.01moL/L hydrochloric acid solution, dissolving, and making into 40 μ g/ml solution as reference substance solution.
3, preparation of a test solution:
taking a sample of about 10mg, precisely weighing, placing in a 25ml measuring flask, adding 0.01moL/L hydrochloric acid solution for dissolving, placing at room temperature, adding 0.01moL/L hydrochloric acid solution to scale, and shaking up to obtain the final product.
4, determination method:
precisely transferring 20 μ l of each of the reference solution and the sample solution, and measuring with liquid chromatograph.
Calculated as peak area by external standard method.
5 calculation of
ASample (A): the peak area of the test sample;
Wsign board: weighing (g) a reference substance;
Vsample (A): dilution factor (ml) of the test sample;
Asign board: peak area of control;
Wsample (A): weighing (g) a sample;
Vsign board: control dilution fold (ml).
6, reagent preparation:
(1) phosphate buffer solution: taking 2.72g of monopotassium phosphate, adding 1000ml of water for dissolving, and adjusting the pH value to 2.5 by using phosphoric acid;
(2)0.01moL/L hydrochloric acid solution: 0.821ml of hydrochloric acid is taken and added with water until 1000ml of water is added to the volume to be calibrated.
Measurement of yield of huperzine A
Huperzine a yield (effective component yield): w1X 100% (product net/raw material net).
The huperzine A content in the raw material huperzine serrate in the raw materials of the huperzine serrate in the embodiments 1-3 and the comparative examples 1 and 2 is 0.031%, the heavy metal lead is 15ppm, the arsenic is 8.6ppm, the chromium is 12ppm, and the mercury is 10 ppm.
The huperzia serrata extracts prepared in examples 1-3 and comparative examples 1-2 were tested by the above method, wherein the huperzia serrata extract prepared in example 1 contains huperzine a 10.5%, and the yield of huperzine a as the active ingredient is 98%.
The huperzine A content of the huperzine serrate extract prepared in example 2 is 10.8%, and the yield of the huperzine A serving as an effective component is 99%.
The huperzine A content of the huperzine serrate extract prepared in example 3 is 10.5%, and the yield of the huperzine A serving as an effective component is 98%.
In the huperzine serrate extract prepared by the technical scheme of acid extraction in the comparative example 1, the content of huperzine a is 8.5 percent, and the yield of the effective component huperzine a is 90 percent.
Comparative example 2 the huperzine a content of 9.6% and the yield of the effective component huperzine a was 85% in the huperzine serrate extract prepared by other cation resin, such as 001 x 7 cation resin.
The heavy metal content of the huperzia serrata extracts prepared in examples 1-3 and comparative examples 1-2 is shown in table 1.
TABLE 1 heavy metal content of the Huperzia serrata extracts prepared in examples 1-3 and comparative examples 1-2
The above description is not intended to limit the present invention, and the present invention is not limited to the above examples. Those skilled in the art should also realize that changes, modifications, additions and substitutions can be made without departing from the true spirit and scope of the invention.
Claims (10)
1. A preparation method of a huperzia serrata extract is characterized by comprising the following steps:
(1) raw material treatment: taking raw material huperzia serrata, cleaning with water, and drying in the shade for later use;
(2) extraction: adding 65-90% ethanol, and extracting at normal temperature;
(3) and (3) filtering: filtering to obtain filtrate;
(4) concentration: concentrating the filtrate under reduced pressure to obtain concentrated solution a without alcohol smell, and recovering ethanol;
(5) acid adjustment: adding dilute acid into the concentrated solution a to adjust the pH value to 3-4, and adding water for dilution;
(6) column passing: adsorbing with 732 cation exchange resin;
(7) washing with water: washing impurities with water;
(8) desorbing: desorbing with ammonia alcohol, and collecting desorption solution;
(9) concentration: concentrating the desorption solution under reduced pressure to obtain concentrated solution b, wherein the Baume degree is 10-20;
(10) and (3) drying: and transferring the concentrated solution b to a spraying process to obtain the product of the huperzia serrata extract.
2. The method for preparing the huperzia serrata extract according to claim 1, wherein the ethanol is added in an amount 12-16 times the raw material in the step (2) by mass-to-volume ratio.
3. The method for preparing the huperzia serrata extract as claimed in claim 1, wherein in the step (2), the normal-temperature extraction time is 1-3 hours, and the filter residue is repeatedly processed for 1-2 times.
4. The method of claim 1, wherein the concentration under reduced pressure in step (4) is in the range of-0.06 MPa to-0.1 MPa, and 40 ℃ to 60 ℃.
5. The method for preparing the huperzia serrata extract according to claim 1, wherein in the step (6), the resin is added in an amount of 0.8 to 1 time of the raw materials by mass to volume ratio.
6. The method of claim 1, wherein the alcohol in step (8) is 4-5% ammonia and 70-80% ethanol.
7. The method for preparing the huperzia serrata extract as claimed in claim 1, wherein the desorption in step (8) is performed at a flow rate of 1.0 to 1.5 BV/h.
8. The method for preparing the huperzia serrata extract as claimed in claim 1, wherein in the step (9), the concentration under reduced pressure is performed at 40-60 ℃ and-0.06-0.1 MPa.
9. The method for preparing the huperzia serrata extract according to claim 1, wherein in the step (10), the inlet temperature during spraying is 180-195 ℃, the outlet temperature is 80-90 ℃, and the flow rate is 1000-2000 mL/h.
10. A huperzia serrata extract, which is prepared by the method for preparing the huperzia serrata extract as claimed in any one of claims 1 to 9.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101134743A (en) * | 2007-08-21 | 2008-03-05 | 陕西嘉禾植物化工有限责任公司 | Method for extracting and separating Huperzine from huperzine serrate |
| CN101697992A (en) * | 2007-05-31 | 2010-04-28 | 北京和润创新医药科技发展有限公司 | Method for separating huperzia serrata total alkaloids by ion exchange resin |
| CN108191762A (en) * | 2018-01-24 | 2018-06-22 | 湖南华诚生物资源股份有限公司 | A kind of extracting method of pure natural heavy metal free Huperzine Serrate P.E |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101697992A (en) * | 2007-05-31 | 2010-04-28 | 北京和润创新医药科技发展有限公司 | Method for separating huperzia serrata total alkaloids by ion exchange resin |
| CN101134743A (en) * | 2007-08-21 | 2008-03-05 | 陕西嘉禾植物化工有限责任公司 | Method for extracting and separating Huperzine from huperzine serrate |
| CN108191762A (en) * | 2018-01-24 | 2018-06-22 | 湖南华诚生物资源股份有限公司 | A kind of extracting method of pure natural heavy metal free Huperzine Serrate P.E |
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