CN102967682A - Ganoderma lucidum spores oil or ganoderma lucidum spore powder or quality test method of preparation thereof - Google Patents
Ganoderma lucidum spores oil or ganoderma lucidum spore powder or quality test method of preparation thereof Download PDFInfo
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- CN102967682A CN102967682A CN2012105453460A CN201210545346A CN102967682A CN 102967682 A CN102967682 A CN 102967682A CN 2012105453460 A CN2012105453460 A CN 2012105453460A CN 201210545346 A CN201210545346 A CN 201210545346A CN 102967682 A CN102967682 A CN 102967682A
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- 240000008397 Ganoderma lucidum Species 0.000 title claims abstract description 67
- 235000001637 Ganoderma lucidum Nutrition 0.000 title claims abstract description 66
- 238000002360 preparation method Methods 0.000 title claims abstract description 43
- 239000000843 powder Substances 0.000 title claims abstract description 31
- 238000010998 test method Methods 0.000 title abstract 2
- 238000012360 testing method Methods 0.000 claims abstract description 37
- 238000000034 method Methods 0.000 claims abstract description 25
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 5
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 51
- 239000013558 reference substance Substances 0.000 claims description 40
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 33
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 27
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid group Chemical group C(CCCCCCC\C=C/CCCCCCCC)(=O)O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 claims description 17
- IPCSVZSSVZVIGE-UHFFFAOYSA-N palmitic acid group Chemical class C(CCCCCCCCCCCCCCC)(=O)O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 claims description 13
- 238000000105 evaporative light scattering detection Methods 0.000 claims description 11
- 239000012467 final product Substances 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 10
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 8
- 239000000945 filler Substances 0.000 claims description 7
- 238000004811 liquid chromatography Methods 0.000 claims description 5
- 239000000047 product Substances 0.000 claims description 4
- 239000000284 extract Substances 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 3
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 2
- 239000000243 solution Substances 0.000 description 66
- 239000003921 oil Substances 0.000 description 43
- 235000019198 oils Nutrition 0.000 description 43
- 238000003556 assay Methods 0.000 description 12
- 238000005303 weighing Methods 0.000 description 7
- 241000222336 Ganoderma Species 0.000 description 6
- 238000011084 recovery Methods 0.000 description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 239000000741 silica gel Substances 0.000 description 5
- 229910002027 silica gel Inorganic materials 0.000 description 5
- 125000004079 stearyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 5
- PHYFQTYBJUILEZ-IUPFWZBJSA-N triolein Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(OC(=O)CCCCCCC\C=C/CCCCCCCC)COC(=O)CCCCCCC\C=C/CCCCCCCC PHYFQTYBJUILEZ-IUPFWZBJSA-N 0.000 description 4
- OILXMJHPFNGGTO-UHFFFAOYSA-N (22E)-(24xi)-24-methylcholesta-5,22-dien-3beta-ol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)C=CC(C)C(C)C)C1(C)CC2 OILXMJHPFNGGTO-UHFFFAOYSA-N 0.000 description 3
- RQOCXCFLRBRBCS-UHFFFAOYSA-N (22E)-cholesta-5,7,22-trien-3beta-ol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CCC(C)C)CCC33)C)C3=CC=C21 RQOCXCFLRBRBCS-UHFFFAOYSA-N 0.000 description 3
- OQMZNAMGEHIHNN-UHFFFAOYSA-N 7-Dehydrostigmasterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CC(CC)C(C)C)CCC33)C)C3=CC=C21 OQMZNAMGEHIHNN-UHFFFAOYSA-N 0.000 description 3
- DNVPQKQSNYMLRS-NXVQYWJNSA-N Ergosterol Natural products CC(C)[C@@H](C)C=C[C@H](C)[C@H]1CC[C@H]2C3=CC=C4C[C@@H](O)CC[C@]4(C)[C@@H]3CC[C@]12C DNVPQKQSNYMLRS-NXVQYWJNSA-N 0.000 description 3
- 241000207961 Sesamum Species 0.000 description 3
- 235000003434 Sesamum indicum Nutrition 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000000470 constituent Substances 0.000 description 3
- DNVPQKQSNYMLRS-SOWFXMKYSA-N ergosterol Chemical compound C1[C@@H](O)CC[C@]2(C)[C@H](CC[C@]3([C@H]([C@H](C)/C=C/[C@@H](C)C(C)C)CC[C@H]33)C)C3=CC=C21 DNVPQKQSNYMLRS-SOWFXMKYSA-N 0.000 description 3
- 150000003648 triterpenes Chemical class 0.000 description 3
- 229930182558 Sterol Natural products 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- 150000003432 sterols Chemical class 0.000 description 2
- 235000003702 sterols Nutrition 0.000 description 2
- 150000003626 triacylglycerols Chemical class 0.000 description 2
- DQJCDTNMLBYVAY-ZXXIYAEKSA-N (2S,5R,10R,13R)-16-{[(2R,3S,4R,5R)-3-{[(2S,3R,4R,5S,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}-5-(ethylamino)-6-hydroxy-2-(hydroxymethyl)oxan-4-yl]oxy}-5-(4-aminobutyl)-10-carbamoyl-2,13-dimethyl-4,7,12,15-tetraoxo-3,6,11,14-tetraazaheptadecan-1-oic acid Chemical class NCCCC[C@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)CC[C@H](C(N)=O)NC(=O)[C@@H](C)NC(=O)C(C)O[C@@H]1[C@@H](NCC)C(O)O[C@H](CO)[C@H]1O[C@H]1[C@H](NC(C)=O)[C@@H](O)[C@H](O)[C@@H](CO)O1 DQJCDTNMLBYVAY-ZXXIYAEKSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241001489090 Ganoderma japonicum Species 0.000 description 1
- 241000222341 Polyporaceae Species 0.000 description 1
- FPWNQPQTICPCOM-UHFFFAOYSA-N acetonitrile;propan-2-ol Chemical compound CC#N.CC(C)O FPWNQPQTICPCOM-UHFFFAOYSA-N 0.000 description 1
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- 229910001410 inorganic ion Inorganic materials 0.000 description 1
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- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses ganoderma lucidum spores oil or ganoderma lucidum spore powder or a quality test method of a preparation thereof. Contents of 1,2-dioleic acid-3-linoleic acid-triglyceride, 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride, 1,2-dilinoleic acid-3-oleic acid-triglyceride and 1,3-dipalmitic acid-2-acid-triglyceride in a product are tested by a high performance liquid chromatography. 1g of ganoderma lucidum spore powder, 1g of ganoderma lucidum spore oil or the preparation equivalent to 1g of ganoderma lucidum spore powder and 1g ganoderma lucidum spore oil respectively contain 15.0mg-60.0mg of 1,2-dioleic acid-3-linoleic acid-triglyceride and 50.0mg-200.0mg of 1,2-dioleic acid-3-linoleic acid-triglyceride, or 12.0mg-50.0mg of 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride and 40.0mg-150.0mg of 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride or 10.0mg-50.0mg, 30.0mg-140.0mg of 1,2-dilinoleic acid-3-oleic acid-triglyceride or 5..0mg-30.0mg, 15.0mg-80.0mg of 1,3-dipalmitic acid-2-acid-triglyceride. The method can effectively test the quality of the product, and is strong in operability, high in reproducibility and strong in specificity.
Description
Technical field
The present invention relates to medicine or health food mass analysis method, specifically, relate to the quality determining method of a kind of ganoderma lucidum spore oil or lucidum spore powder or its preparation.
Background technology
Glossy ganoderma is the red sesame of Basidiomycetes Polyporaceae (Polyproraceae) Ganoderma (Ganoderma) fungi [Ganoderma lucidium (Leyss.ex Fr.) Karst.] and purple sesame Ganoderma japonicum (Fr.) Lloyd] general name, have the effect such as strengthen the body resistance to consolidate the constitution, be called top grade by " herbal classic ".Reishi sporule (Ganoderma lucidiumspore) is the red sesame growth and maturity phase to eject ultrafine spore from cap, is the reproduction cell of glossy ganoderma, has the whole hereditary active substance of glossy ganoderma, and its medical value also comes into one's own just day by day.That the pharmacological action of Reishi sporule mainly contains is antitumor, immunological regulation, adjusting blood fat and neural, cardiovascular and respiratory system had the improvement of adjusting effect.Product about Reishi sporule is also a lot, such as G. lucidum spores Softgel, and Ganoderma Lucidum Spore Powder Capsule, but true and false difficulty is divided difficult minute of quality good or not, the present method that a kind of more comprehensive detection quality is not also arranged.
The complex chemical composition of Reishi sporule, bibliographical information have following a few class: sterols, triterpenes, alkaloids, fatty acid, lactone, protein and amino acids, glycopeptide class, vitamins, carrotene and inorganic ions class etc.Bibliographical information control lucidum spore powder quality mainly concentrates on the assay of water soluble ingredient ganoderan, and mainly be to measure total triterpene contents in the ganoderma lucidum spore oil with ultraviolet spectroscopy to the assay of liposoluble constituent, but the content of the triterpene in the ganoderma lucidum spore oil seldom, specificity is not strong during mensuration, can measure this constituents in other vegetable oil uses the same method yet.
The fat oil that contains high level in the Reishi sporule, about 20～30%, can obtain ganoderma lucidum spore oil by extracting method.Ganoderma lucidum spore oil mainly contains fatty acid ester and sterols, and existing patent and bibliographical information cross in the ganoderma lucidum spore oil 1, the content assaying method of 2-two oleic acid-3-palmitic acid-triglyceride, olein and ergosterol.As in report a kind of mensuration lucidum spore powder or ganoderma lucidum spore oil and the preparation thereof among the patent No. CN 100473389C 1, the method of 2-two oleic acid-3-palmitic acid-triglyceride or olein content, CN100515434C reports a kind of method of measuring Quantitative Determination of Ergosterol in lucidum spore powder or ganoderma lucidum spore oil and the preparation thereof.CN 101518551A reports in a kind of mensuration Reishi sporule oil emu 1,2-two oleic acid-3-palmitic acid-triglyceride, the method for olein content and Quantitative Determination of Ergosterol.But 1,2-two oleic acid-3-palmitic acid-triglyceride or olein are the wherein two kinds of triglycerides in the ganoderma lucidum spore oil, the ratio that these two kinds of triglycerides account in the ganoderma lucidum spore oil is 30～45%, and 1,2-two oleic acid-3-linoleic acid-triglyceride, 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride, 1,2-dilinoleic acid-3-oleic acid-triglyceride, 1,3-, two palmitic acids-2-oleic acid-triglyceride also is the triglyceride constituents in the ganoderma lucidum spore oil, therefore sets up lucidum spore powder, in ganoderma lucidum spore oil and the preparation thereof 1,2-two oleic acid-3-linoleic acid-triglyceride, 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride, 1,2-dilinoleic acid-3-oleic acid-triglyceride, the content assaying method of 1,3-, two palmitic acids-2-oleic acid-triglyceride is a kind of effective ways that further detect its quality.Up to the present, also there is not bibliographical information with 1,2-two oleic acid-3-linoleic acid-triglyceride, 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride, 1,2-dilinoleic acid-3-oleic acid-triglyceride, 1,3-, the two palmitic acids-content of 2-oleic acid-triglyceride is estimated the quality of lucidum spore powder or ganoderma lucidum spore oil and preparation thereof.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, provide a kind of and can effectively detect lucidum spore powder, the method for the quality of ganoderma lucidum spore oil and preparation thereof.
Purpose of the present invention is achieved through the following technical solutions:
The quality determining method of a kind of ganoderma lucidum spore oil or lucidum spore powder or its preparation, with containing 1 in its product of high effective liquid chromatography for measuring, 2-two oleic acid-3-linoleic acid-triglyceride (being called for short OOL), 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride (being called for short POL), 1,3-two palmitic acids-2-oleic acid-triglyceride (being called for short POP), 1,2-dilinoleic acid-3-oleic acid-triglyceride (being called for short LLO), every 1g lucidum spore powder, ganoderma lucidum spore oil or be equivalent to the 1g lucidum spore powder, contain 1 in the preparation of ganoderma lucidum spore oil, the amount of 2-two oleic acid-3-linoleic acid-triglyceride is respectively 15.0mg～60.0mg, 50.0mg～200.0mg, or contain 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride 12.0mg～50.0mg, 40.0mg～150.0mg, or contain 1,2-dilinoleic acid-3-oleic acid-triglyceride 10.0mg～50.0mg, 30.0mg～150.0mg, or contain 1,3-, two palmitic acids-2-oleic acid-triglyceride 5.0mg～30.0mg, 15.0mg～80.0mg.
Said high performance liquid chromatography is to carry out in the steps below:
One, the preparation of need testing solution:
(1) preparation of ganoderma lucidum spore oil and preparation thereof: get 5mg～100mg ganoderma lucidum spore oil and contain the preparation of ganoderma lucidum spore oil 5mg～100mg, add the mobile phase of 0.2～1ml with every 1mg, dissolving is also fixed molten, shakes up, and get final product, used mobile phase is identical with mobile phase in the chromatographic condition; (2) preparation of lucidum spore powder and preparation test sample thereof: get lucidum spore powder 0.1～10g or contain the preparation of lucidum spore powder 0.1～10g, adopt extracted by ether, reclaim solvent and get the Reishi sporule powder extracts, the mobile phase that adds 0.2～1ml with every 1mg, dissolving is also fixed molten, shake up, and get final product, used mobile phase is identical with mobile phase in the chromatographic condition;
Two, the preparation of reference substance solution: get 1,2-two oleic acid-3-linoleic acid-triglyceride, 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride, 1,2-dilinoleic acid-3-oleic acid-triglyceride, 1,3-, two palmitic acids-2-oleic acid-triglyceride reference substance add with the mobile phase of need testing solution dissolving and fixed molten, shake up, be made into every 1ml and contain 1,2-, two oleic acid-3-linoleic acid-triglyceride 0.35～0.55mg, or 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride 0.15～0.35mg, or 1,2-dilinoleic acid-3-oleic acid-triglyceride 0.15～0.35mg, or 1,3-, two palmitic acids-2-oleic acid-triglyceride 0.15～0.35mg.
Three, chromatographic condition: be filling agent with octadecyl silane; Form mixed liquor as mobile phase with acetonitrile, isopropyl alcohol, methylene chloride take binary or the ternary of arbitrary proportion, the flow velocity of mobile phase is 0.5～2.0ml/min; Detect with evaporative light-scattering detector or differential refractive index detector; Number of theoretical plate is pressed 1,2-, two oleic acid-3-linoleic acid-triglyceride, 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride, and 1,2-dilinoleic acid-3-oleic acid-triglyceride, 1,3-, two palmitic acids-2-oleic acid-triglyceride calculates, and all should be not less than 2000; Chromatographic column temperature is 10～50 ℃;
Four, measure: precision is drawn reference substance solution and each 10～20ul of need testing solution respectively, injects high performance liquid chromatograph, measure, and the calculating of taking the logarithm, and get final product.
The concrete operation method of high performance liquid chromatography can be undertaken by the routine techniques of Chinese Pharmacopoeia appendix regulation.
In order effectively and correctly to obtain determination data, adopt evaporative light-scattering detector to detect as good in the chromatographic condition of high effective liquid chromatography for measuring method step 3.
Be to adopt acetonitrile in the chromatographic condition of described step 3: the volume ratio of isopropyl alcohol is 40～60:60～40 or acetonitrile: the volume ratio of methylene chloride is that 40～60:60～40 are better mobile phase as mobile phase, and take acetonitrile: the volume ratio of isopropyl alcohol is as 53:47 or acetonitrile: the volume ratio of methylene chloride as 59:41 as mobile phase as the optimal flow phase.Chromatographic column optimum column temperature in the chromatographic condition of said step 3: 30 ℃; The flow velocity 1.0ml/min of optimal flow phase.
Compared with prior art, the present invention has following beneficial effect: this detection method can accurately be measured the content or one of four of OOL, POL, LLO and POP in lucidum spore powder, ganoderma lucidum spore oil and the preparation, can be used as method and foundation that this product quality detects, method is easy, workable, reappearance is high, is the effective ways that detect lucidum spore powder and ganoderma lucidum spore oil and the quality of the pharmaceutical preparations thereof.
Embodiment
Further specify technical scheme of the present invention below by embodiment.
The research of the method for content in the embodiment 1 high effective liquid chromatography for measuring ganoderma lucidum spore oil
(1) instrument and reagent
Instrument: Waters2695 high performance liquid chromatograph, chromatographic column: KromasilC
18Post (4.6mm * 250mm, 5 μ m), the Sedex75 evaporative light-scattering detector.
Reagent: reference substance OOL, POL, LLO and POP reference substance are provided by Sigma-Aldrich company; Ganoderma lucidum spore oil is provided by Guangzhou company, and acetonitrile and isopropyl alcohol are chromatographically pure; It is pure that other reagent is analysis.(2) preparation of need testing solution: take by weighing ganoderma lucidum spore oil 100.34mg, accurately weighed (removing), and to 25ml, filtering with microporous membrane namely gets need testing solution with the mobile phase constant volume, and adopt acetonitrile: the volume ratio of isopropyl alcohol is that the solution of 53:47 is made mobile phase;
The preparation of reference substance solution: a, take by weighing OOL reference substance 10.31mg, add the mobile phase constant volume to 25ml, namely get the reference substance solution that OOL concentration is 0.4124mg/ml;
B, get POL reference substance 7.06mg, add the mobile phase constant volume to 25ml, namely get the reference substance solution that POL concentration is 0.2824mg/ml;
C, get LLO reference substance 6.48mg, add the mobile phase constant volume to 25ml, namely get the reference substance solution that LLO concentration is 0.2592mg/ml;
D, get POP reference substance 4.95mg, add the mobile phase constant volume to 25ml, namely get the reference substance solution that POP concentration is 0.198mg/ml.
(3) chromatographic condition
Chromatographic column: KromasilC
18Post (4.6mm * 250mm, 5 μ m); Column temperature: 30 ℃; Mobile phase is the solution of 53:47 with the volume ratio of acetonitrile-isopropyl alcohol; Flow velocity is 1.0ml/min; Use the Sedex75 evaporative light-scattering detector.
(4) linear relationship is investigated
Precision is measured reference substance solution (mg/ml) OOL, POL, LLO, and POP is with 5,10, and 15,20,25,30 μ L injection liquid chromatographies are measured by above-mentioned chromatographic condition.Take sample size (μ g) as horizontal ordinate, take peak area as ordinate, the drawing standard curve the results are shown in Table 1:
Table 1 linear relationship experimental data (n=6)
OOL：lg A＝1.41lgm+5.44，R
2=0.9997
POL：lg A＝1.56lgm+5.47，R
2=0.9991
LLO：lg A＝1.71lgm+4.59，R
2=0.9991
POP:lg A＝1.81lgm+4.83，R
2=0.9997
Above result shows: OOL in 2.061～12.372ug scope, POL in 1.412～8.472ug scope, LLO in 1.296～7.776ug scope, POP in 0.990～5.940ug scope, be good linear relationship.
(5) precision test: get each 10 μ L of reference substance solution, repeat sample introduction 6 times, the RSD of OOL, POP, LLO and POP peak area＜2% the results are shown in Table 2.
Table 2 precision test data (n=6)
Evidence, precision is good.
(6) stability test: the sample thief test liquid is respectively at 0,2, and 4,6,9,12 hours, sample introduction 10 μ L respectively recorded the RSD of OOL in the sample, POL, LLO, POP peak area＜2%, the results are shown in Table 3.
Table 3 stability test data (n=6)
Test findings shows that the sample test liquid is good at 12 hours internal stabilities.
(7) reappearance test
According to operating under the content assaying method item, 6 parts of same batch samples to be measured, the RSD of OOL, POL, LLO, POP content＜2% the results are shown in Table 4.
Table 4 reappearance test figure
(8) application of sample recovery test
Get with 6 parts of batch Reishi sporule oil samples, each about 50mg, accurately weighed after, add a certain amount of OOL, POL, LLO, POP reference substance, operate under the photograph assay item, the results are shown in Table 5.
Table 5 application of sample recovery test result
Test findings shows: the OOL recovery between 98.23～102.36%, the POL recovery between 97.39～102.34%, the LLO recovery between 98.28～101.31%, the POP recovery between 98.46～102.69%, application of sample reclaims good.
(9) assay
OOL, POL, LLO, POP assay in the ganoderma lucidum spore oil according to the operation of the method for above-mentioned assay, have been measured the content of OOL, POL, LLO, POP in 3 sample ganoderma lucidum spore oils, the results are shown in Table 6.
The assay result of OOL, POL, LLO, POP in table 6 sample
| Sample | 1 | 2 | 3 |
| OOL content (mg/g) | 110.15 | 135.84 | 105.63 |
| POL content (mg/g) | 75.64 | 87.67 | 73.61 |
| LLO content (mg/g) | 60.19 | 74.29 | 58.87 |
| POP content (mg/g) | 34.82 | 41.67 | 27.64 |
Record that every 1g contains the content of OOL, POL, LLO, POP respectively between 105.63～135.84mg, 73.61～87.67mg, 58.87～74.29mg, 27.64～41.67mg in three batches of ganoderma lucidum spore oils.
The assay of OOL, POL, LLO, POP in the embodiment 2 Reishi sporule oil emu
Strong to close silica gel be filling agent to chromatographic condition with octadecyl; Chromatographic column: KromasilC
18Post (4.6mm * 250mm, 5 μ m); Column temperature: 30 ℃; The mobile phase acetonitrile: the volume ratio of isopropyl alcohol is the solution of 40:60; Evaporative light-scattering detector detects; Flow velocity is 0.5ml/min; Number of theoretical plate is pressed OOL or POL or LLO or the calculating of POP peak, all greater than 2000.
The preparation of reference substance solution: get OOL, POL, LLO, POP reference substance 4.53mg, 2.85mg, 2.39mg, 2.16mg place the 10ml measuring bottle, add the dissolving of mobile phase solution and be diluted to scale, making every 1ml, to contain respectively OOL, POL, LLO and POP be respectively 0.453mg/ml, and the solution of 0.285mg/ml, 0.239mg/ml, 0.216mg/ml namely gets reference substance solution;
The preparation of need testing solution: take by weighing and contain Reishi sporule oil emu (Guangzhou company provides) 1.15g, add the dissolving of mobile phase solution and be settled to 25ml, namely get need testing solution.
Measurement result: precision is drawn reference substance solution and each 10ul of need testing solution respectively, injects high performance liquid chromatograph, measure, and the calculating of taking the logarithm, and get final product.Every 1g emulsion contains the content of OOL, POL, LLO, POP respectively at 11.15mg, 7.432mg, 5.949mg, 3.505mg.
OOL assay in embodiment 3 ganoderma lucidum spore oils
Strong to close silica gel be filling agent to chromatographic condition with octadecyl; Chromatographic column: KromasilC
18Post (4.6mm * 250mm, 5 μ m); Column temperature: 30 ℃; The mobile phase acetonitrile: the volume ratio of isopropyl alcohol is the solution of 60:40; Evaporative light-scattering detector detects; Flow velocity is 0.8ml/min; Number of theoretical plate is pressed OOL and is calculated, greater than 2000.
The preparation of reference substance solution: get OOL reference substance 4.87mg, place the 10ml measuring bottle, add the dissolving of mobile phase solution and be diluted to scale, make the solution that every 1ml contains 0.487mg, namely get reference substance solution;
The preparation of need testing solution: take by weighing ganoderma lucidum spore oil 102.31mg, add the dissolving of mobile phase solution and be settled to 25ml, namely get need testing solution.
Measurement result: precision is drawn reference substance solution and each 10ul of need testing solution respectively, injects high performance liquid chromatograph, measure, and the calculating of taking the logarithm, and get final product.Containing OOL in every 1g ganoderma lucidum spore oil is 112.63mg.
POL assay in embodiment 4 ganoderma lucidum spore oils
Strong to close silica gel be filling agent to chromatographic condition with octadecyl; Chromatographic column: KromasilC
18Post (4.6mm * 250mm, 5 μ m); Column temperature: 30 ℃; The mobile phase acetonitrile: the volume ratio of methylene chloride is the solution of 59:41; Evaporative light-scattering detector detects; Flow velocity is 0.8ml/min; Number of theoretical plate is pressed OOL and is calculated, greater than 2000.
The preparation of reference substance solution: get POL reference substance 2.54mg, place the 10ml measuring bottle, add the dissolving of mobile phase solution and be diluted to scale, make the 0.254mg solution that every 1ml contains POL, namely get reference substance solution;
The preparation of need testing solution: take by weighing ganoderma lucidum spore oil 102.32mg, add the dissolving of mobile phase solution and be settled to 25ml, namely get need testing solution.
Measurement result: precision is drawn reference substance solution and each 10ul of need testing solution respectively, injects high performance liquid chromatograph, measure, and the calculating of taking the logarithm, and get final product.Containing POL in every 1g ganoderma lucidum spore oil is 75.54mg.
LLO assay in embodiment 5 ganoderma lucidum spore oils
Strong to close silica gel be filling agent to chromatographic condition with octadecyl; Chromatographic column: KromasilC
18Post (4.6mm * 250mm, 5 μ m); Column temperature: 30 ℃; Mobile phase acetonitrile: isopropyl alcohol: the volume ratio of methylene chloride is the solution of 50:35:15; Evaporative light-scattering detector detects; Flow velocity is 1.0ml/min; Number of theoretical plate is pressed OOL and is calculated, greater than 2000.
The preparation of reference substance solution: get LLO reference substance 2.65mg, place the 10ml measuring bottle, add mobile phase solution dissolving and be diluted to scale, making every 1ml, to contain LLO be 0.265mg solution, namely gets reference substance solution;
The preparation of need testing solution: take by weighing ganoderma lucidum spore oil 101.24mg, add the dissolving of mobile phase solution and be settled to 25ml, namely get need testing solution.
Measurement result: precision is drawn reference substance solution and each 10ul of need testing solution respectively, injects high performance liquid chromatograph, measure, and the calculating of taking the logarithm, and get final product.LLO contains and is 68.42mg in every 1g ganoderma lucidum spore oil.
OOL, POL, LLO, POP assay in embodiment 6 lucidum spore powders
Strong to close silica gel be filling agent to chromatographic condition with octadecyl; Chromatographic column: KromasilC18 post (4.6mm * 250mm, 5 μ m); Column temperature: 30 ℃; The mobile phase acetonitrile: the volume ratio of methylene chloride is the solution of 40:60; Evaporative light-scattering detector detects; Flow velocity is 1.0ml/min; Number of theoretical plate is pressed OOL or POL or LLO or POP calculating, all greater than 2000.
The preparation of reference substance solution: get OOL, POL, LLO, POP reference substance 5.42mg, 2.38mg, 2.93mg, 2.75mg place the 10ml measuring bottle, add the dissolving of mobile phase solution and be diluted to scale, making every 1ml, to contain respectively OOL, POL, LLO, POP be respectively 0.542mg/ml, and the solution of 0.238mg/ml, 0.293mg/ml, 0.275mg/ml namely gets reference substance solution;
The preparation of need testing solution: take by weighing lucidum spore powder 650.32mg, put in the apparatus,Soxhlet's, add ether 50ml, extracted 8 hours in heating water bath, extract reclaims ether to doing, and residue dissolves with mobile phase, is settled to 50ml, namely gets need testing solution.
Measurement result: precision is drawn reference substance solution and each 10ul of need testing solution respectively, injects high performance liquid chromatograph, measure, and the calculating of taking the logarithm, and get final product.OOL, POL, LLO, POP are respectively 36.15mg, 22.85mg, 19.52mg, 10.84mg in every 1g lucidum spore powder.
Claims (4)
1. the quality determining method of a ganoderma lucidum spore oil or lucidum spore powder or its preparation, it is characterized in that, with containing 1 in its product of high effective liquid chromatography for measuring, 2-two oleic acid-3-linoleic acid-triglyceride, 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride, 1,2-dilinoleic acid-3-oleic acid-triglyceride, 1,3-two palmitic acids-2-oleic acid-triglyceride, every 1g lucidum spore powder, ganoderma lucidum spore oil or be equivalent to the 1g lucidum spore powder, contain 1 in the preparation of ganoderma lucidum spore oil, the amount of 2-two oleic acid-3-linoleic acid-triglyceride is respectively 15.0mg～60.0mg, 50.0mg～200.0mg, or contain 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride 12.0mg～50.0mg, 40.0mg～150.0mg, or contain 1,2-dilinoleic acid-3-oleic acid-triglyceride 10.0mg～50.0mg, 30.0mg～140.0mg, or contain 1,3-, two palmitic acids-2-oleic acid-triglyceride 5.0mg～30.0mg, 15.0mg～80.0mg;
Said high performance liquid chromatography is to carry out in the steps below:
One, the preparation of need testing solution: the preparation of (1) ganoderma lucidum spore oil and preparation test sample thereof: get 5mg～100mg ganoderma lucidum spore oil and contain the preparation of ganoderma lucidum spore oil 5mg～100mg, the mobile phase that adds 0.2～1ml with every 1mg, dissolving is also fixed molten, shake up, and get final product, used mobile phase is identical with mobile phase in the chromatographic condition; (2) preparation of lucidum spore powder and preparation test sample thereof: get lucidum spore powder 0.1～10g or contain the preparation of Reishi sporule 0.1～10g, adopt extracted by ether, reclaim solvent and get the Reishi sporule powder extracts, the mobile phase that adds 0.2～1ml with every 1mg, dissolving is also fixed molten, shake up, and get final product, used mobile phase is identical with mobile phase in the chromatographic condition;
Two, the preparation of reference substance solution: get 1,2-two oleic acid-3-linoleic acid-triglyceride, 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride, 1,2-dilinoleic acid-3-oleic acid-triglyceride, 1,3-, two palmitic acids-2-oleic acid-triglyceride reference substance add with the mobile phase of need testing solution dissolving and fixed molten, shake up, be made into every 1ml and contain 1,2-, two oleic acid-3-linoleic acid-triglyceride 0.35～0.55mg, or 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride 0.15～0.35mg, or 1,2-dilinoleic acid-3-oleic acid-triglyceride 0.15～0.35mg, or the solution of 1,3-, two palmitic acids-2-oleic acid-triglyceride 0.10～0.30mg;
Three, chromatographic condition: be filling agent with octadecyl silane; Take acetonitrile or isopropyl alcohol or three kinds of solvents of methylene chloride or acetonitrile: the volume ratio of isopropyl alcohol is as 40～60:60～40 or acetonitrile: the volume ratio of methylene chloride is as 40～60:60～40 as mobile phase, and the flow velocity of mobile phase is 0.5～2.0ml/min; Detect with evaporative light-scattering detector or differential refractive index detector; Number of theoretical plate is pressed 1,2-, two oleic acid-3-linoleic acid-triglyceride, 1-palmitic acid-2-oleic acid-3-linoleic acid-triglyceride, and 1,2-dilinoleic acid-3-oleic acid-triglyceride, 1,3-, two palmitic acids-2-oleic acid-triglyceride calculates, and all should be not less than 2000; Chromatographic column temperature is 10～50 ℃;
Four, measure: precision is drawn reference substance solution and each 10～20ul of need testing solution respectively, injects high performance liquid chromatograph, measure, and the calculating of taking the logarithm, and get final product.
2. quality determining method according to claim 1 is characterized in that, adopts evaporative light-scattering detector to detect in the chromatographic condition of said high performance liquid chromatography step 3.
3. quality determining method according to claim 1 is characterized in that, in the chromatographic condition of described step 3 is to adopt take acetonitrile: the volume ratio of isopropyl alcohol is as 53:47 or acetonitrile: the volume ratio of methylene chloride is made mobile phase as 59:41.
4. quality determining method according to claim 1 is characterized in that, chromatographic column temperature is 30 ℃ in the chromatographic condition of said step 3.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106770822A (en) * | 2016-12-15 | 2017-05-31 | 南京财经大学 | A kind of extracting method of triglycerides |
| CN109212085A (en) * | 2018-10-22 | 2019-01-15 | 广州白云山汉方现代药业有限公司 | The finger-print and its construction method of a kind of ganoderma lucidum spore oil and application |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0599913A (en) * | 1991-10-11 | 1993-04-23 | Kao Corp | Analyzing method for phospholipid containing minute amount of amino group |
| CN100362345C (en) * | 2005-07-21 | 2008-01-16 | 广州汉方现代中药研究开发有限公司 | Establishment of lucid ganderma spore and lucid ganderma spore oil finger print atlas and standard finger print atlas |
| US20090013769A1 (en) * | 2004-08-19 | 2009-01-15 | Tony Larson | Assay for oils |
| CN100473389C (en) * | 2006-02-21 | 2009-04-01 | 广州汉方现代中药研究开发有限公司 | Method for controlling quality of ganoderma lucidum spore and ganoderma lucidium spore oil and preparation thereof |
| WO2009105912A1 (en) * | 2008-02-26 | 2009-09-03 | 广州汉方现代中药研究开发有限公司 | A fat emulsion containing ganoderma spore oil, its quality control methods and its use |
| CN102175800A (en) * | 2011-01-07 | 2011-09-07 | 南京工业大学 | Method for measuring content of 1,3-dioleoyl-2-palmitoyl triglyceride in dairy products |
-
2012
- 2012-12-14 CN CN201210545346.0A patent/CN102967682B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0599913A (en) * | 1991-10-11 | 1993-04-23 | Kao Corp | Analyzing method for phospholipid containing minute amount of amino group |
| US20090013769A1 (en) * | 2004-08-19 | 2009-01-15 | Tony Larson | Assay for oils |
| CN100362345C (en) * | 2005-07-21 | 2008-01-16 | 广州汉方现代中药研究开发有限公司 | Establishment of lucid ganderma spore and lucid ganderma spore oil finger print atlas and standard finger print atlas |
| CN100473389C (en) * | 2006-02-21 | 2009-04-01 | 广州汉方现代中药研究开发有限公司 | Method for controlling quality of ganoderma lucidum spore and ganoderma lucidium spore oil and preparation thereof |
| WO2009105912A1 (en) * | 2008-02-26 | 2009-09-03 | 广州汉方现代中药研究开发有限公司 | A fat emulsion containing ganoderma spore oil, its quality control methods and its use |
| CN102175800A (en) * | 2011-01-07 | 2011-09-07 | 南京工业大学 | Method for measuring content of 1,3-dioleoyl-2-palmitoyl triglyceride in dairy products |
Non-Patent Citations (8)
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106770822A (en) * | 2016-12-15 | 2017-05-31 | 南京财经大学 | A kind of extracting method of triglycerides |
| CN106770822B (en) * | 2016-12-15 | 2019-05-03 | 南京财经大学 | A kind of extracting method of triglycerides |
| CN109212085A (en) * | 2018-10-22 | 2019-01-15 | 广州白云山汉方现代药业有限公司 | The finger-print and its construction method of a kind of ganoderma lucidum spore oil and application |
| CN109212085B (en) * | 2018-10-22 | 2021-05-25 | 广州白云山汉方现代药业有限公司 | Fingerprint of ganoderma lucidum spore oil and construction method and application thereof |
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