CN1793821A - Sample pretreatment method for detecting limonin kind compound content in orange peel and seed raw material - Google Patents
Sample pretreatment method for detecting limonin kind compound content in orange peel and seed raw material Download PDFInfo
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- CN1793821A CN1793821A CN 200510101198 CN200510101198A CN1793821A CN 1793821 A CN1793821 A CN 1793821A CN 200510101198 CN200510101198 CN 200510101198 CN 200510101198 A CN200510101198 A CN 200510101198A CN 1793821 A CN1793821 A CN 1793821A
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- sample
- raw material
- limonin
- compound content
- organic solvent
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- 150000001875 compounds Chemical class 0.000 title claims abstract description 13
- 239000002994 raw material Substances 0.000 title claims description 16
- VHLJDTBGULNCGF-UHFFFAOYSA-N Limonin Natural products CC1(C)OC2CC(=O)OCC23C4CCC5(C)C(CC(=O)C6OC56C4(C)C(=O)CC13)c7cocc7 VHLJDTBGULNCGF-UHFFFAOYSA-N 0.000 title claims description 13
- KBDSLGBFQAGHBE-MSGMIQHVSA-N limonin Chemical compound C=1([C@H]2[C@]3(C)CC[C@H]4[C@@]([C@@]53O[C@@H]5C(=O)O2)(C)C(=O)C[C@@H]2[C@]34COC(=O)C[C@@H]3OC2(C)C)C=COC=1 KBDSLGBFQAGHBE-MSGMIQHVSA-N 0.000 title claims description 13
- 238000002203 pretreatment Methods 0.000 title description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 40
- 238000000034 method Methods 0.000 claims abstract description 21
- 239000003960 organic solvent Substances 0.000 claims abstract description 12
- 239000000284 extract Substances 0.000 claims description 19
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 18
- 241000675108 Citrus tangerina Species 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 8
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 6
- 239000006166 lysate Substances 0.000 claims description 6
- 238000001514 detection method Methods 0.000 claims description 5
- 238000012360 testing method Methods 0.000 claims description 5
- 239000000463 material Substances 0.000 claims description 3
- 238000010298 pulverizing process Methods 0.000 claims description 3
- 238000005303 weighing Methods 0.000 claims description 2
- 238000001035 drying Methods 0.000 abstract description 3
- 241000207199 Citrus Species 0.000 abstract 1
- 244000248349 Citrus limon Species 0.000 abstract 1
- 235000005979 Citrus limon Nutrition 0.000 abstract 1
- 235000020971 citrus fruits Nutrition 0.000 abstract 1
- 239000007788 liquid Substances 0.000 abstract 1
- 238000007781 pre-processing Methods 0.000 abstract 1
- 238000000605 extraction Methods 0.000 description 6
- 150000002630 limonoids Chemical class 0.000 description 6
- 239000002904 solvent Substances 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 238000005238 degreasing Methods 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000002798 spectrophotometry method Methods 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- 241001093501 Rutaceae Species 0.000 description 1
- 238000003811 acetone extraction Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000005714 functional activity Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
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- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
A sample preprocessing method for detecting lemon bitter principle compound content in citrus skin and seed includes drying and grinding detected sample to be 60 - 80 mu, adding 2 - 5 ml organic solvent into ground sample of 0.2 - 0.5g, oscillating and extracting it for 2 - 3 min. then centrifuging it for 5 - 10 min. to absorb off top clear liquid; adding acetone of 2 - 5ml, extracting and oscillating it for 2 - 3 min. then centrifuging it for 5 - 10min. to collect acetone extractive; adding 1-2ml of organic solvent into extractive, oscillating it then centrifuging it for 5 - 10min to collect solution for fixing volume and detecting.
Description
Technical field
The present invention relates to a kind of sample-pretreating method that detects limonin kind compound content in oranges and tangerines skin, the seed raw material.
Background technology
The biologically active of limonoid receives increasing concern in the rutaceae (as oranges and tangerines etc.), and it in the application of food medicine and other fields also more and more widely.The separation of limonoid, purifying, detection technique are also in constantly developing.Obtain at first will detect its content behind the raw material, be beneficial to carry out the extraction purifying and the product quality Control work of back.Therefore, can the content that detect the limonoid in the raw material quickly and accurately will directly have influence on the production efficiency of limonin.
At present, the research work of limonoid mostly around its structure of matter and functional activity research, improves very few to the limonoid detection technique.Detecting the sample-pretreating method of limonin kind compound content in oranges and tangerines skin, the seed raw material, generally is that 100g～500g raw material is pulverized the back with sherwood oil or extracted by ether repeatedly, afterwards again with acetone extraction repeatedly.Acetone extract is dissolved in methylene chloride after concentrating, and collects soluble that a part of constant volume, measures target compound content.Mainly there is following problem in this class sample-pretreating method:
1) pulverizing of sample
General compound test, sample all will be made break process to a certain degree.In theory, break process can increase the specific surface area of raw material.In fact, in leaching process, broken raw material is not to be evenly dispersed in the solvent, but deposits to the bottom of extraction vessel, and resistance to mass tranfer is bigger, equally also can cause extraction time long, and solvent load is big, extracts problems such as not thorough.
2) environmental issue
The general used solvent of pre-treating method all is that boiling point is lower, and the organic solvent of certain toxic action is arranged.Heating for multiple times is extracted, concentrates, and the leakage that can bring partial solvent, long-term use will produce harm to operator's health.
3) operating process and cost
Conventional method sample pretreatment process complexity, the time-consuming reagent that takes, cost is higher.Because the concentrated solvent amount is bigger, still available Rotary Evaporators is finished in laboratory with good conditionsi, but most of basic units plant laboratory is difficult to reach.At present, the price of internal reagent acetone, methylene chloride, methenyl choloride and sherwood oil is not low, therefore, does some improvement in the pre-treatment of sample, it is developed to low toxicity, low cost, efficient direction, is the developing direction of limonin kind compound content detection method in the raw material.
Summary of the invention
The object of the present invention is to provide a kind of sample-pretreating method that detects limonin kind compound content in oranges and tangerines skin, the seed raw material, the raw extract after this method is handled can be stablized the contents level that reflects limonoid exactly after testing.
A kind of sample-pretreating method that detects limonin kind compound content in oranges and tangerines skin, the seed raw material provided by the invention comprises the following steps:
(1) with testing sample crushed after being dried to 60～80 order;
(2) take by weighing pulverizing sample 0.200～0.500g, add organic solvent 2～5ml then, 2～3min is extracted in vibration on the high speed vortex mixer, centrifugal 5～the 10min of 4000r/min, siphon away supernatant, material repeats above operation 3～5 times, and residue carries out next step operation;
(3) add acetone 2～5ml, 2～3min is extracted in vibration, and the centrifugal 5～10min of 4000r/min collects acetone extract, repeats this operation 3～5 times, and extract merges and dried up, and obtains acetone extract;
(4) add organic solvent 1～2ml in the acetone extract, ultrasonic or vibration dissolving, the centrifugal 5～10min of 4000r/min collects lysate, repeats this operation 3～5 times, and lysate is merged, and constant volume detects.
The described organic solvent of step in the technique scheme (2) is sherwood oil or ether; The described organic solvent of step (4) is methylene chloride or methenyl choloride.
Compared with prior art, the present invention has following characteristics:
1. vibration is extracted and is pulverized sample at a high speed, and sample is contacted with extracting solvent, and the sample effective ratio area strengthens, and extraction efficiency is improved.
On a small quantity repeatedly extraction and application concentration difference, make extraction more thorough, the result is more reliable.
3. the pre-treatment behavior is to carry out at normal temperatures, can protect measured object to exempt from destruction effectively.
4. sample pretreatment process is simple to operate, and temperature is low, and consumption of organic solvent is few, makes that operating process toxicity is low, and experiment material and reagent expend and lacks simultaneously, and experimental cost is also low.
5. sample pre-treatments is simple to operate, and workload is little, and occupation of land is few, once can handle a plurality of samples, has improved work efficiency greatly.
Embodiment
The present invention all can realize purpose of the present invention by above relevant technological parameter of putting down in writing and reagent, and not only is confined to following examples.
Embodiment 1
Get tangerine seed 100g, drying and crushing to 80 order, get 9 parallel sample, each sample 0.2g (being accurate to 0.001g) adds the limonin titer that 0.5ml concentration is 100ug/ml respectively in 3 samples therein, carries out sample pretreatment by following method: sample is earlier with the 3ml sherwood oil degreasing 2min that vibrates at a high speed, centrifugal 10min under 4000r/min speed then, siphon away supernatant, in sample, add the 3ml sherwood oil again, repeat this operation 2 times; Sample adding 3ml acetone after the degreasing vibrates at a high speed and extracts 2min, and centrifugal 10min under the 4000r/min speed collects supernatant, repeats to extract 3 times, merges acetone extract, under 40 ℃ of conditions acetone extract is dried up; Add the 1ml methylene chloride in the acetone extract, ultrasonic dissolution, centrifugal 5min under the 4000r/min speed collects supernatant, redissolve 3 times, the combined dichloromethane lysate, and constant volume is to be measured to 10ml.It (is that a certain amount of solution to be measured adds absolute ethyl alcohol to 2ml that the spectrophotometric method of solution to be measured under the same terms detects, add concentration respectively and be right-dimethylaminobenzaldehyde-ethanol solution of sulfuric acid 5ml of 0.125%, shake up and leave standstill, survey light absorption value down in the 500nm wavelength behind the 30min, according to the content of limonin substances in the typical curve Equation for Calculating sample).The result is that the content of 6 parallel samples is respectively 13.12mg/g, 13.09mg/g, 13.15mg/g, 13.07mg/g, 13.11mg/g, 13.16mg/g, mean value 13.12mg/g; 3 recovery of standard addition are respectively 87.6%, 89.1%, 88.9%.
Embodiment 2
Get tangerine seed raw material 500g, drying and crushing to 60 order, parallelly get 3 samples, each sample 0.2g (being accurate to 0.001g), earlier with the 3ml sherwood oil degreasing 2min that vibrates at a high speed, centrifugal 10min under 4000r/min speed siphons away supernatant then, in sample, add the 3ml sherwood oil again, repeat this operation 2 times; Sample adding 3ml acetone after the degreasing vibrates at a high speed and extracts 2min, and centrifugal 10min under the 4000r/min speed collects supernatant, repeats to extract 3 times, merges acetone extract, under 40 ℃ of conditions acetone extract is dried up; Add the 1ml methenyl choloride in the acetone extract, vibration dissolving at a high speed, centrifugal 5min under the 4000r/min speed collects supernatant, and redissolve 3 times merge the methenyl choloride lysate, and constant volume is to be measured to 10ml.Spectrophotometric method testing result as embodiment 1 is respectively 13.02%, 12.95%, 13.07%, average out to 13.01%.
Claims (3)
1, a kind of sample-pretreating method that detects limonin kind compound content in oranges and tangerines skin, the seed raw material is characterized in that comprising the following steps:
(1) with testing sample crushed after being dried to 60～80 order;
(2) take by weighing pulverizing sample 0.200～0.500g, add organic solvent 2～5ml then, 2～3min is extracted in vibration on the high speed vortex mixer, centrifugal 5～the 10min of 4000r/min, siphon away supernatant, material repeats above operation 3～5 times, and residue carries out next step operation;
(3) add acetone 2～5ml, 2～3min is extracted in vibration, and the centrifugal 5～10min of 4000r/min collects acetone extract, repeats this operation 3～5 times, and extract merges and dried up, and obtains acetone extract;
(4) add organic solvent 1～2ml in the acetone extract, ultrasonic or vibration dissolving, the centrifugal 5～10min of 4000r/min collects lysate, repeats this operation 3～5 times, and lysate is merged, and constant volume detects.
2, the sample-pretreating method of limonin kind compound content in detection oranges and tangerines skin according to claim 1, the seed raw material is characterized in that the described organic solvent of step (2) is sherwood oil or ether.
3, the sample-pretreating method of limonin kind compound content in detection oranges and tangerines skin according to claim 1, the seed raw material is characterized in that the described organic solvent of step (4) is methylene chloride or methenyl choloride.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005101011983A CN100494963C (en) | 2005-11-14 | 2005-11-14 | Sample pretreatment method for detecting limonin kind compound content in orange peel and seed raw material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005101011983A CN100494963C (en) | 2005-11-14 | 2005-11-14 | Sample pretreatment method for detecting limonin kind compound content in orange peel and seed raw material |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1793821A true CN1793821A (en) | 2006-06-28 |
| CN100494963C CN100494963C (en) | 2009-06-03 |
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|---|---|---|---|
| CNB2005101011983A Expired - Fee Related CN100494963C (en) | 2005-11-14 | 2005-11-14 | Sample pretreatment method for detecting limonin kind compound content in orange peel and seed raw material |
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| CN (1) | CN100494963C (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102116725A (en) * | 2011-03-17 | 2011-07-06 | 湖北富邦科技股份有限公司 | Method for accurately analyzing content of tartrazine |
| CN102680293A (en) * | 2012-05-03 | 2012-09-19 | 沈阳药科大学 | Toxic effect position of morning glory seed caused renal toxicity effects and preparing method thereof |
| CN102940687A (en) * | 2011-12-12 | 2013-02-27 | 苏州大学 | Szechwan Chinaberry fruit extract and application thereof |
| CN102944623A (en) * | 2012-11-21 | 2013-02-27 | 福建农林大学 | Determination method of limonin and nomilin in kumquat |
| CN103399090A (en) * | 2013-01-23 | 2013-11-20 | 苏州大学 | Detection method for limonins components in szechwan Chinaberry fruit extractive |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4305923A (en) * | 1980-05-08 | 1981-12-15 | Weiler Elmar W | Method for quantitative analysis for limonin |
| CN1279242A (en) * | 2000-05-08 | 2001-01-10 | 玉环县惠丰文旦有限公司 | Process for extracting limonin from rue fruit |
| US20020061339A1 (en) * | 2000-09-28 | 2002-05-23 | Martin Stogniew | Compositions and methods for use of extracts of rutaceae plants |
| CN1100553C (en) * | 2000-09-30 | 2003-02-05 | 傅敏恭 | Process for extracting active medicine comonents from orange seed |
-
2005
- 2005-11-14 CN CNB2005101011983A patent/CN100494963C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102116725A (en) * | 2011-03-17 | 2011-07-06 | 湖北富邦科技股份有限公司 | Method for accurately analyzing content of tartrazine |
| CN102940687A (en) * | 2011-12-12 | 2013-02-27 | 苏州大学 | Szechwan Chinaberry fruit extract and application thereof |
| CN102940687B (en) * | 2011-12-12 | 2016-01-13 | 苏州大学 | A kind of Fructus Toosendan extract and uses thereof |
| CN102680293A (en) * | 2012-05-03 | 2012-09-19 | 沈阳药科大学 | Toxic effect position of morning glory seed caused renal toxicity effects and preparing method thereof |
| CN102944623A (en) * | 2012-11-21 | 2013-02-27 | 福建农林大学 | Determination method of limonin and nomilin in kumquat |
| CN103399090A (en) * | 2013-01-23 | 2013-11-20 | 苏州大学 | Detection method for limonins components in szechwan Chinaberry fruit extractive |
| CN103399090B (en) * | 2013-01-23 | 2015-06-24 | 苏州大学 | Detection method for limonins components in szechwan Chinaberry fruit extractive |
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| Publication number | Publication date |
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| CN100494963C (en) | 2009-06-03 |
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Address after: 510610 Guangdong, Guangzhou, Tianhe District province Dongguan Zhuang Road, No. 133 Patentee after: SERICULTURE & AGRI FOOD Research Institute GAAS Patentee after: GUANGDONG BOSUN HEALTHY FOOD Co.,Ltd. Address before: 510610 Guangdong, Guangzhou, Tianhe District province Dongguan Zhuang Road, No. 133 Patentee before: SERICULTURE & AGRI FOOD Research Institute GAAS Patentee before: GUANGDONG BOSUN HEALTH FOOD RESEARCH DEVELOPMENT CENTER |
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