CN102426212A - Enhanced fluorescence thin layer identification method for Cyperus Rotundus L - Google Patents
Enhanced fluorescence thin layer identification method for Cyperus Rotundus L Download PDFInfo
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- CN102426212A CN102426212A CN201110246268XA CN201110246268A CN102426212A CN 102426212 A CN102426212 A CN 102426212A CN 201110246268X A CN201110246268X A CN 201110246268XA CN 201110246268 A CN201110246268 A CN 201110246268A CN 102426212 A CN102426212 A CN 102426212A
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Abstract
The invention discloses an enhanced fluorescence thin layer identification method for Chinese herbal medicines. According to the invention, an enhanced fluorescer is added on the base of ordinary thin layer identification, ideal results are obtained in identification of a plurality of Chinese herbal medicines which have no detection information, detection approaches of thin layer identification are greatly broadened, and a thin layer identification revision rate of prescribed Chinese herbal preparations can be effectively improved.
Description
Patented claim of the present invention is for dividing an application, and original bill information is following:
Application number: 2010105281179
The applying date: on November 2nd, 2010
invention and created name: a kind of Chinese crude drug increase fluoroscopic examination thin layer discrimination method.
Technical field
The present invention relates to a kind of thin layer discrimination method of Chinese crude drug, particularly, what relate to a kind of rhizoma cyperi increases the fluorescence thin layer discrimination method.
Background technology
In all kinds of Chinese medicine preparation quality standards of country, the thin layer of its medicinal material differentiates that the rate of revising and enlarging is low at present, on average is about 30% of prescription Chinese crude drug total amount, can't the conduct monitoring at all levels quality of the pharmaceutical preparations.Especially extract preparation, the microscopic features of medicinal material have not attached existence, and microscopical identification is felt simply helpless to it, and thin layer is differentiated to seek less than characteristic again and detected information, with regard to the uncontrollable quality of the pharmaceutical preparations.Cause some illegal operators in the production run of medicine, to use inferior materials and turn out substandard goods, take place with bad Dai You, the situation of passing a fake product off as a genuine one, the lighter affects patient's the state of an illness adversely, and weight person directly endangers patient's health.
Summary of the invention
Differentiate the detection approach for widening thin layer, the thin layer that improves in the Chinese traditional patent formulation preparation is differentiated the rate of revising and enlarging, and the inventor has carried out increasing FLUORESCENCE STUDY to the Chinese crude drug that multiple nothing detects information, and obtains ideal results.
What the invention provides a kind of Chinese crude drug increases fluoroscopic examination thin layer discrimination method, and this method comprises following steps:
A, respectively get test sample medicinal material and control medicinal material powder 0.01-0.5 the gram, add methyl alcohol 3-10 milliliter, sonicated 10-15 minute, get supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 2-4 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, launch, take out, dry with developping agent;
Spray is with fluorescence-enhancing agent on c, the thin layer plate, and 105 ℃ to be heated to spot colour developing clear;
D, put under the 365nm uviol lamp and inspect, the test sample chromatogram with the corresponding position of reference substance chromatogram on show the fluorescence spot of same color.
As preferred version, the Chinese crude drug of thin layer discrimination method kind of the present invention is preferably a kind of in the dried venom of toads, the bighead atractylodes rhizome, the capsule of weeping forsythia, calculus bovis factitius, the root of three-nerved spicebush, oriental wormwood, moutan bark or the rhizoma cyperi, increases fluorescer and is preferably 10% ethanol solution of sulfuric acid or 1% aluminium choride ethanolic solution.
In theory, originally do not have the Chinese crude drug of any detection information, can carry out thin layer through the method that increases fluorescence of the present invention and differentiate, the inventor is preferred especially several kinds of medicinal materials increase fluoroscopic examination thin layer discrimination method, respectively as follows:
1, the fluoroscopic examination thin layer discrimination method that increases of the dried venom of toads is preferably:
A, get dried venom of toads medicinal material and dried venom of toads control medicinal material powder 0.05 gram respectively, add 3 milliliters of methyl alcohol, sonicated 15 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 2 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-formic acid that with the volume ratio is 4:6:0.2 is that developping agent launches, and takes out, and dries;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put under the 365nm uviol lamp and inspect, dried venom of toads test sample chromatogram with the corresponding position of dried venom of toads reference substance chromatogram on show the fluorescence spot of same color.
2, the fluoroscopic examination thin layer discrimination method that increases of the bighead atractylodes rhizome is preferably:
A, get bighead atractylodes rhizome medicinal material and bighead atractylodes rhizome control medicinal material powder 0.5 gram respectively, add 5 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 4 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-formic acid that with the volume ratio is 8:2:0.2 is that developping agent launches, and takes out, and dries;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put under the 365nm uviol lamp and inspect, bighead atractylodes rhizome test sample chromatogram with the corresponding position of bighead atractylodes rhizome reference substance chromatogram on show the fluorescence spot of same color.
3, the fluoroscopic examination thin layer discrimination method that increases of the capsule of weeping forsythia is preferably:
A, get capsule of weeping forsythia medicinal material and capsule of weeping forsythia control medicinal material powder 0.4 gram respectively, add 5 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 4 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-formic acid that with the volume ratio is 10:2:0.2 is that developping agent launches, and takes out, and dries;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put under the 365nm uviol lamp and inspect, capsule of weeping forsythia test sample chromatogram with the corresponding position of capsule of weeping forsythia reference substance chromatogram on show the fluorescence spot of same color.
4, the fluoroscopic examination thin layer discrimination method that increases of calculus bovis factitius is preferably:
A, get calculus bovis factitius medicinal material and calculus bovis factitius control medicinal material powder 0.15 gram respectively, add 6 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 2 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-acetic acid-methyl alcohol that with the volume ratio is 10:25:2:3 is that developping agent launches, and takes out, and dries;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put under the 365nm uviol lamp and inspect, calculus bovis factitius test sample chromatogram with the corresponding position of calculus bovis factitius reference substance chromatogram on show the fluorescence spot of same color.
5, the fluoroscopic examination thin layer discrimination method that increases of the root of three-nerved spicebush is preferably:
A, get root of three-nerved spicebush medicinal material and root of three-nerved spicebush control medicinal material powder 0.3 gram respectively, add 4 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 4 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-formic acid that with the volume ratio is 8:2:0.2 is that developping agent launches, and takes out, and dries;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put under the 365nm uviol lamp and inspect, root of three-nerved spicebush test sample chromatogram with the corresponding position of root of three-nerved spicebush reference substance chromatogram on show the fluorescence spot of same color.
6, the fluoroscopic examination thin layer discrimination method that increases of oriental wormwood is preferably:
A, get oriental wormwood medicinal material and oriental wormwood control medicinal material powder 0.4 gram respectively, add 8 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 4 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate that with the volume ratio is 7:3 is that developping agent launches, and takes out, and dries;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put under the 365nm uviol lamp and inspect, oriental wormwood test sample chromatogram with the corresponding position of oriental wormwood reference substance chromatogram on show the fluorescence spot of same color.
7, the fluoroscopic examination thin layer discrimination method that increases of moutan bark is preferably:
A, get moutan bark medicinal material and moutan bark control medicinal material powder 0.01 gram respectively, add 10 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 3 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-acetone-formic acid that with the volume ratio is 8:2:1:0.2 is that developping agent launches, and takes out, and dries;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 1% aluminium choride ethanolic solution in spray on c, the thin layer plate;
D, put under the 365nm uviol lamp and inspect, moutan bark test sample chromatogram with the corresponding position of moutan bark reference substance chromatogram on show the fluorescence spot of same color.
8, the fluoroscopic examination thin layer discrimination method that increases of rhizoma cyperi is preferably:
A, get rhizoma cyperi medicinal material and rhizoma cyperi control medicinal material powder 0.5 gram respectively, add 4 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 4 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-formic acid that with the volume ratio is 10:2:0.2 is that developping agent launches, and takes out, and dries;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put under the 365nm uviol lamp and inspect, rhizoma cyperi test sample chromatogram with the corresponding position of rhizoma cyperi reference substance chromatogram on show the fluorescence spot of same color.
Increase the thin-layer chromatogram comparative analysis before and after the fluorescence from 8 kinds of medicinal materials, increase before the fluorescence, no matter under 254nm or the 365nm; All not having detection information clearly appears; Can't make judgement to the result, but increase after the fluorescence, the amplitude of containing much information increases; Sensitivity improves, and demonstrates 8~9 indigo plants, green fluorescence spot like the root of three-nerved spicebush; The bighead atractylodes rhizome demonstrates 5~6 indigo plants, green fluorescence spot; The capsule of weeping forsythia demonstrates 3 Huangs and sapphirine fluorescence spot etc., clear spot, and intensity is very high, can be used for qualitative even quantitative examination, sees each thin-layer chromatogram of embodiment testing result for details.
In a word; The present invention increases fluoroscopic examination thin layer discrimination method and has sensitivity, quick, characteristics of high efficiency; Solved not only do not have uv absorption, but also the discriminating problem of non-blooming medicinal material; Make the discriminating of these medicinal materials can high sensitivity, carry out exactly, the thin layer that improves Chinese crude drug and Chinese traditional patent formulation preparation is differentiated the rate of revising and enlarging, one has use value to invent very much beyond doubt.
Description of drawings
Fig. 1 dried venom of toads thin layer is differentiated collection of illustrative plates; A inspects photo for increasing before the fluorescence under the 254nm uviol lamp, and B inspects photo for increasing under the preceding 365nm uviol lamp of fluorescence, and C inspects photo for increasing behind the fluorescence under the 365nm uviol lamp; From left to right first three is a sample to spot, and back three is control medicinal material.
Fig. 2 bighead atractylodes rhizome thin layer is differentiated collection of illustrative plates; A inspects photo for increasing before the fluorescence under the 254nm uviol lamp, and B inspects photo for increasing under the preceding 365nm uviol lamp of fluorescence, and C inspects photo for increasing behind the fluorescence under the 365nm uviol lamp; From left to right first three is a sample to spot, and back three is control medicinal material.
Fig. 3 capsule of weeping forsythia thin layer is differentiated collection of illustrative plates; A inspects photo for increasing before the fluorescence under the 254nm uviol lamp, and B inspects photo for increasing under the preceding 365nm uviol lamp of fluorescence, and C inspects photo for increasing behind the fluorescence under the 365nm uviol lamp; From left to right first three is a sample to spot, and back three is control medicinal material.
Fig. 4 calculus bovis factitius thin layer is differentiated collection of illustrative plates; A inspects photo for increasing before the fluorescence under the 254nm uviol lamp, and B inspects photo for increasing under the preceding 365nm uviol lamp of fluorescence, and C inspects photo for increasing behind the fluorescence under the 365nm uviol lamp; From left to right first three is a sample to spot, and latter two is a control medicinal material.
Fig. 5 root of three-nerved spicebush thin layer is differentiated collection of illustrative plates; A inspects photo for increasing before the fluorescence under the 254nm uviol lamp, and B inspects photo for increasing under the preceding 365nm uviol lamp of fluorescence, and C inspects photo for increasing behind the fluorescence under the 365nm uviol lamp; From left to right first three is a sample to spot, and latter two is a control medicinal material.
Fig. 6 oriental wormwood thin layer is differentiated collection of illustrative plates; A inspects photo for increasing before the fluorescence under the 254nm uviol lamp, and B inspects photo for increasing under the preceding 365nm uviol lamp of fluorescence, and C inspects photo for increasing behind the fluorescence under the 365nm uviol lamp; From left to right first three is a sample to spot, and latter two is a control medicinal material.
Fig. 7 moutan bark thin layer is differentiated collection of illustrative plates; A inspects photo for increasing before the fluorescence under the 254nm uviol lamp, and B inspects photo for increasing under the preceding 365nm uviol lamp of fluorescence, and C inspects photo for increasing behind the fluorescence under the 365nm uviol lamp; From left to right first three is a sample to spot, and back three is control medicinal material.
Fig. 8 rhizoma cyperi thin layer is differentiated collection of illustrative plates; A inspects photo for increasing under the preceding 254nm uviol lamp of fluorescence; B inspects photo for increasing before the fluorescence under the 365nm uviol lamp, and C inspects photo for increasing behind the fluorescence under the 365nm uviol lamp, spot from left to right first three individual be sample; Next three is control medicinal material, and latter two is a Rhizoma Cyperi (vinegar processed).
Embodiment
Following embodiment is used to illustrate the embodiment of the inventive method; But it can not constitute any restriction to scope of the present invention; For confirming the technique effect of the inventive method, all after launching to dry with developping agent, spray increases before the fluorescence each embodiment; Under 254nm and 365nm uviol lamp, inspect and take pictures, compare with chromatogram after doing to increase fluorescence.
Embodiment 1
The fluoroscopic examination thin layer that increases of the dried venom of toads is differentiated:
A, get dried venom of toads medicinal material and dried venom of toads control medicinal material powder 0.05 gram respectively, add 3 milliliters of methyl alcohol, sonicated 15 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 2 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-formic acid that with the volume ratio is 4:6:0.2 is that developping agent launches, and takes out, and dries, and under 254nm and 365nm uviol lamp, inspects respectively and takes pictures;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put to inspect under the 365nm uviol lamp and take pictures;
The result sees Fig. 1, visible increase after the fluorescence dried venom of toads test sample chromatogram with the corresponding position of dried venom of toads reference substance chromatogram on show the fluorescence spot of 4 same colors, and do not increase in the thin-layer chromatography of fluorescence, under 254nm and 365nm uviol lamp, all do not show obvious spot.
Embodiment 2
The fluoroscopic examination thin layer that increases of the bighead atractylodes rhizome is differentiated:
A, get bighead atractylodes rhizome medicinal material and bighead atractylodes rhizome control medicinal material powder 0.5 gram respectively, add 5 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 4 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-formic acid that with the volume ratio is 8:2:0.2 is that developping agent launches, and takes out, and dries, and under 254nm and 365nm uviol lamp, inspects respectively and takes pictures;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put to inspect under the 365nm uviol lamp and take pictures;
The result sees Fig. 2, visible increase after the fluorescence bighead atractylodes rhizome test sample chromatogram with the corresponding position of dried venom of toads reference substance chromatogram on show the fluorescence spot of a plurality of same colors, and do not increase in the thin-layer chromatography of fluorescence, under 254nm and 365nm uviol lamp, all do not show obvious spot.
Embodiment 3
The fluoroscopic examination thin layer that increases of the capsule of weeping forsythia is differentiated:
A, get the capsule of weeping forsythia medicinal material and capsule of weeping forsythia contrast medicine inborn ability end 0.4 gram respectively, add 5 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 4 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-formic acid that with the volume ratio is 10:2:0.2 is that developping agent launches, and takes out, and dries, and under 254nm and 365nm uviol lamp, inspects respectively and takes pictures;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put to inspect under the 365nm uviol lamp and take pictures;
The result sees Fig. 3, visible increase after the fluorescence capsule of weeping forsythia test sample chromatogram with the corresponding position of capsule of weeping forsythia reference substance chromatogram on show the fluorescence spot of 3 same colors, and do not increase in the thin-layer chromatography of fluorescence, under 254nm and 365nm uviol lamp, all do not show obvious spot.
Embodiment 4
The fluoroscopic examination thin layer that increases of calculus bovis factitius is differentiated:
A, get calculus bovis factitius medicinal material and calculus bovis factitius control medicinal material powder 0.15 gram respectively, add 6 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 2 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-acetic acid-methyl alcohol that with the volume ratio is 10:25:2:3 is that developping agent launches, and takes out, and dries, and under 254nm and 365nm uviol lamp, inspects respectively and takes pictures;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put to inspect under the 365nm uviol lamp and take pictures;
The result sees Fig. 4; It is thus clear that increase calculus bovis factitius test sample chromatogram after the fluorescence with the corresponding position of calculus bovis factitius reference substance chromatogram on show the fluorescence spot of 4 same colors; And do not increase in the thin-layer chromatography of fluorescence, under 254nm and 365nm uviol lamp, all do not show obvious spot.
Embodiment 5
The fluoroscopic examination thin layer that increases of the root of three-nerved spicebush is differentiated:
A, get root of three-nerved spicebush medicinal material and root of three-nerved spicebush control medicinal material powder 0.3 gram respectively, add 4 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 4 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-formic acid that with the volume ratio is 8:2:0.2 is that developping agent launches, and takes out, and dries, and under 254nm and 365nm uviol lamp, inspects respectively and takes pictures;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put to inspect under the 365nm uviol lamp and take pictures;
The result sees Fig. 5, visible increase after the fluorescence root of three-nerved spicebush test sample chromatogram with the corresponding position of root of three-nerved spicebush reference substance chromatogram on show the fluorescence spot of a plurality of same colors, and do not increase in the thin-layer chromatography of fluorescence, under 254nm and 365nm uviol lamp, all do not show obvious spot.
Embodiment 6
The fluoroscopic examination thin layer that increases of oriental wormwood is differentiated:
A, get oriental wormwood medicinal material and oriental wormwood control medicinal material powder 0.4 gram respectively, add 8 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 4 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate that with the volume ratio is 7:3 is that developping agent launches, and takes out, and dries, and under 254nm and 365nm uviol lamp, inspects respectively and takes pictures;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put to inspect under the 365nm uviol lamp and take pictures;
The result sees Fig. 6, visible increase after the fluorescence oriental wormwood test sample chromatogram with the corresponding position of oriental wormwood reference substance chromatogram on show the fluorescence spot of a plurality of same colors, and do not increase in the thin-layer chromatography of fluorescence, under 254nm and 365nm uviol lamp, all do not show obvious spot.
Embodiment 7
The fluoroscopic examination thin layer that increases of moutan bark is differentiated:
A, get moutan bark medicinal material and moutan bark control medicinal material powder 0.01 gram respectively, add 10 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 3 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-acetone-formic acid that with the volume ratio is 8:2:1:0.2 is that developping agent launches, and takes out, and dries, and under 254nm and 365nm uviol lamp, inspects respectively and takes pictures;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 1% aluminium choride ethanolic solution in spray on c, the thin layer plate;
D, put to inspect under the 365nm uviol lamp and take pictures;
The result sees Fig. 7, visible increase after the fluorescence moutan bark test sample chromatogram with the corresponding position of moutan bark reference substance chromatogram on show the fluorescence spot of 1 same color, and do not increase in the thin-layer chromatography of fluorescence, under 254nm and 365nm uviol lamp, all do not show obvious spot.
Embodiment 8
The fluoroscopic examination thin layer that increases of rhizoma cyperi is differentiated:
A, get rhizoma cyperi medicinal material and rhizoma cyperi control medicinal material powder 0.5 gram respectively, add 4 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;
B, absorption need testing solution and each 4 microlitre of reference substance solution are put in same GF respectively
254On the thin layer plate, the cyclohexane-ethyl acetate-formic acid that with the volume ratio is 10:2:0.2 is that developping agent launches, and takes out, and dries, and under 254nm and 365nm uviol lamp, inspects respectively and takes pictures;
As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;
D, put to inspect under the 365nm uviol lamp and take pictures;
The result sees Fig. 8, visible increase after the fluorescence rhizoma cyperi test sample chromatogram with the corresponding position of rhizoma cyperi reference substance chromatogram on show the fluorescence spot of a plurality of same colors, and do not increase in the thin-layer chromatography of fluorescence, under 254nm and 365nm uviol lamp, all do not show obvious spot.
Claims (1)
- A rhizoma cyperi increase the fluorescence thin layer discrimination method, it is characterized in that this method comprises following steps:A, get rhizoma cyperi medicinal material and rhizoma cyperi control medicinal material powder 0.5 gram respectively, add 4 milliliters of methyl alcohol, sonicated 10 minutes is got supernatant as need testing solution and reference substance solution;B, absorption need testing solution and each 4 microlitre of reference substance solution are put in same GF respectively 254On the thin layer plate, the cyclohexane-ethyl acetate-formic acid that with the volume ratio is 10:2:0.2 is that developping agent launches, and takes out, and dries;As fluorescence-enhancing agent, 105 ℃ to be heated to spot colour developing clear with 10% ethanol solution of sulfuric acid in spray on c, the thin layer plate;D, put under the 365nm uviol lamp and inspect, rhizoma cyperi test sample chromatogram with the corresponding position of rhizoma cyperi reference substance chromatogram on show the fluorescence spot of same color.
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