CN103018242A - Rapid colorimetric determination method for capsaicin content of capsicum products - Google Patents
Rapid colorimetric determination method for capsaicin content of capsicum products Download PDFInfo
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Abstract
The invention relates to a rapid colorimetric determination method for the capsaicin content of capsicum products. According to the method, a sodium carbonate-phosphotungstic acid-phosphomolybdic acid coloration liquid is prepared, capsaicin methanol liquids of different concentration gradients are then mixed with the sodium carbonate-phosphotungstic acid-phosphomolybdic acid coloration liquid and subjected to coloration, a standard colorimetric plate is manufactured, a capsaicin methanol liquid to be determined is finally mixed with the sodium carbonate-phosphotungstic acid-phosphomolybdic acid coloration liquid and subjected to coloration, the concentration of capsaicin is obtained through comparing with the standard colorimetric plate, and the capsaicin content of the capsaicin methanol liquid to be determined is determined to be 0.02-2.00 mg/mL. The method disclosed by the invention is simple, convenient and easy to learn, the reaction is quick, the determination of each time needs only 50 min, the determination is more accurate, the cost is low, the requirements on environments and personnel are low, and the deficiencies of a high-performance liquid chromatography that the cost is high, the time consumed is long (3-4 hours), the device/equipment investment is large, the requirements on fields and operators are harsh, the high-performance liquid chromatography is inconvenient, and the requirements on rapid site determination of the capsaicin content of the capsicum products cannot be met can be overcome.
Description
Technical field
The invention belongs to the food analysis technical field, be specifically related to a kind of Capsaicinoid Content in Capsicum Products content rapid colorimetric determination method.
Background technology
Along with the continuous increase of popular current consumption chilli products amount, and people are to the variation of the peppery degree chilli products of difference demand, and the peppery degree of product becomes the main reference factor that the consumer selects chilli products.So the peppery degree of accurate quick and convenient detection chilli products has great importance and application prospect for the production and consumption of chilli products.The peppery degree of chilli products is mainly determined by capsaicin content, can substantially judge the peppery degree of product by measuring capsaicin content.The method that is used at present the capsaicin content detection in the national standard mainly is high performance liquid chromatography, this method accuracy is high, but cost is high, length consuming time, instrument and equipment has high input, place and operating personnel are required harshness, and inconvenience can not satisfy the demand of field quick detection Capsaicinoid Content in Capsicum Products content.Therefore, in actual production and life, be badly in need of setting up a kind of easy to learn, reaction easy and simple to handle fast, measure accurately, low price, to environment and the less demanding peppery degree assay method of personnel.Up to the present, there is no relevant research report both at home and abroad.
Summary of the invention
The object of the present invention is to provide a kind of Capsaicinoid Content in Capsicum Products content rapid colorimetric determination method, set up a kind of easy, quick, semiquantitative capsicim detection method.
A kind of Capsaicinoid Content in Capsicum Products content rapid colorimetric determination method of the present invention, realize by following steps:
(1) preparation sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion: compound concentration is the sodium carbonate liquor of 0.1mol/L and phosphomolybdic acid-Salkowski's solution of 1.0mol/L respectively, with sodium carbonate liquor and phosphomolybdic acid-Salkowski's solution by volume 30:1 mix, namely get sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion, concentration is that the compound method of phosphomolybdic acid-Salkowski's solution of 1.0mol/L is: take by weighing 100g sodium tungstate and 20g phosphomolybdic acid, adding 60mL volume by volume concentration is 85% phosphoric acid and 700mL distilled water, boil 2h, cooled and filtered is settled to 1000mL with distilled water;
(2) production standard color board: adopt concentration gradient: 0,0.02,0.05,0.08,0.10,0.20,0.40,0.60,0.80, the capsicim methanol solution of 1.00,2.00mg/mL, respectively with sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion by volume 1:3 mix colour developing, according to reacted color and corresponding capsicim concentration production standard color board;
(3) sample preparation: for the chilli products that contains protein, at first be the ethanolic solution of 65-75% and the chilli products that contains protein that takes by weighing with volume by volume concentration, ratio in 50 mL:1g is mixed, be 80-120w at ultrasonic power, process 35-45min under the temperature 45-55 ℃ of condition to remove protein, process the layering of afterreaction thing, the chilli products behind the protein is removed in separation, with methyl alcohol and tetrahydrofuran 1:1 volume ratio mixed liquor extraction capsicim 10-20min wherein, re-extract is once again for residue after filtering, merging filtrate also is settled to the ratio constant volume of 100mL in every 10-20g sample, gets capsicim solution to be measured; For the chilli products that does not contain protein, directly take by weighing chilli products, with methyl alcohol and tetrahydrofuran 1:1 volume ratio mixed liquor extraction capsicim 10-20min wherein, re-extract is once again for residue after filtering, merging filtrate also is settled to the ratio constant volume of 100mL in every 5-10g sample, gets capsicim solution to be measured;
(4) with capsicim solution to be measured and sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion by volume 1:3 mix colour developing, by the contrast standard color board, draw capsicim concentration.
Said method step (2) capsicim methanol solution and sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion are after 1:3 mixes colour developing by volume, can utilize L*, a*, b* value after colour photometer is measured colour developing, the capsicim concentration that record is corresponding, carry it into and produce corresponding color in the Photoshop software, thereby make standard colorimetric plate, step (4) is used mutually colour photometer and is measured color, draws capsicim concentration according to the variation of Δ L* and Δ b* value.
The described chilli products that does not contain protein of step (3) comprises capsicum oleoresin, capsicim crystal and chilli oil.
The scope that the present invention measures capsaicin content in the capsicim solution to be measured is 0.02-2.00 mg/mL, and wherein concentration limit is 0.02 mg/mL.In the mark-on control test, the relative error of the present invention and high performance liquid chromatography measured result is up to 22.84% all below 23%, and average relative error is that 11.37%, RSD is 18.28%.In the practical application, measuring once needs time spent 50min, the present invention to measure the relative error of capsaicin content in 30%, and its maximal value is 29.82%, can be used as the semiquantitative determination of capsaicin content.The capsicim solution to be measured that surpasses 2mg/mL for capsaicin content then needs first by a certain percentage dilution, measures again.Judged result for peppery degree is then in full accord with the judged result of high performance liquid chromatography.
Advantage of the present invention is: the inventive method is simple and convenient and easy to study, reaction is quick, measuring once only needs time spent 50min, measure accurate, low price, less demanding to environment and personnel, can overcome high performance liquid chromatography cost height, length consuming time (3-4h), instrument and equipment and have high input, place and operating personnel are required harshness, inconvenience, can not satisfy the deficiency of fast-field evaluation Capsaicinoid Content in Capsicum Products content.
Embodiment
The invention will be further described below in conjunction with embodiment.
Compound concentration is the sodium carbonate liquor of 0.1mol/L and phosphomolybdic acid-Salkowski's solution of 1.0mol/L respectively, with sodium carbonate liquor and phosphomolybdic acid-Salkowski's solution by volume 30:1 mix, namely get sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion, concentration is that the compound method of phosphomolybdic acid-Salkowski's solution of 1.0mol/L is: take by weighing 100g sodium tungstate and 20g phosphomolybdic acid, adding 60mL volume by volume concentration is 85% phosphoric acid and 700mL distilled water, boil 2h, cooled and filtered is settled to 1000mL with distilled water;
Press the described method production standard of summary of the invention color board, 0,0.02,0.05,0.08,0.10,0.20,0.40,0.60,0.80 1.00,2.00mg/mL concentration gradient capsicim methanol solution and sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion are after 1:3 mixes colour developing by volume, with L*, a*, the b* value after the colour developing of colour photometer mensuration, the capsicim concentration that record is corresponding carries it into and produces corresponding color in the Photoshop software, thereby makes standard colorimetric plate.
Embodiment 1: the mensuration that does not contain protein Capsaicinoid Content in Capsicum Products content.
Take by weighing capsicum oleoresin, capsicim crystal and each 10g of chilli oil, proceed as follows respectively: in the 100mL beaker in tall form, add methyl alcohol and tetrahydrofuran 1:1 volume ratio mixed liquor 40mL extracting capsicum element 10min, residue re-extract once (10min) again after filtering, merging filtrate also is settled to 100mL, again with sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion by volume 1:3 mix colour developing, by the contrast standard color board, carry out the subjective appreciation of color, and the variation of mensuration color difference meter data Δ L* and Δ b* value is obtained a result.
Each sample replication 6 times, calculating mean value compares with the Capsaicinoid Content in Capsicum Products of high performance liquid chromatography Accurate Determining and to draw: capsaicin content is measured relative error and is respectively 4.45%, 5.37% and 14.19% in capsicum oleoresin, capsicim crystal and the chilli oil.
The precision test of the inventive method: getting capsicim concentration is the capsicim methanol solution of 0.06mg/mL, with the inventive method replication 6 times, contrast by the color with standard colorimetric plate, capsaicin content is respectively 0.05mg/mL, 0.05mg/mL, 0.08mg/mL, 0.08mg/mL, 0.08mg/mL 0.05mg/mL calculates its relative standard deviation RSD=0.015.
Do not contain the chilli products of disturbance of protein for capsicum oleoresin, capsicim crystal and this class of chilli oil, the inventive method can as capsaicin content fast, semiquantitative determination, its relative error is in 15%, precision RSD is 0.015.
Embodiment 2: the mensuration that contains protein Capsaicinoid Content in Capsicum Products content.
The chilli products that 13 kinds of peppery degree of difference are contained protein respectively takes by weighing 20g, proceed as follows respectively: be that 70% ethanolic solution mixes with 1000 mL volume by volume concentrations at first, be 100w at ultrasonic power, process 40min under the temperature 50 C condition to remove protein, the clearance of protein reaches 92.00% in the chilli products, process the layering of afterreaction thing, the chilli products behind the protein is removed in separation, in the 100mL beaker in tall form, add methyl alcohol and tetrahydrofuran 1:1 volume ratio mixed liquor 40mL extracting capsicum element 20min, residue re-extract once (20min) again after filtering, merging filtrate also is settled to 100mL, again with sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion by volume 1:3 mix colour developing, by the contrast standard color board, carry out the subjective appreciation of color, and the variation of mensuration color difference meter data Δ L* and Δ b* value is obtained a result; According to the capsaicin content that records, adopt Scovil index (Scoville Heat Units, SHU) method that the peppery degree of chilli products is estimated.
Each sample replication 6 times, calculating mean value compares with the Capsaicinoid Content in Capsicum Products of high performance liquid chromatography Accurate Determining, and concrete outcome is as shown in table 1:
Table 1 contains the measurement result of protein Capsaicinoid Content in Capsicum Products content
| Sample number into spectrum | The HPLC measured value (g/kg） | Actual pungent degree | The inventive method (colorimetric) measured value (g/kg） | The pungent degree of measuring | Relative error (%) |
| 1 | 0.547 | In peppery | 0.60 | In peppery | ?9.69 |
| 2 | 0.245 | In peppery | 0.20 | In peppery | ? 18.37 |
| 3 | 0.946 | High peppery | 1.00 | High peppery | ? 5.71 |
| 4 | 0.285 | In peppery | 0.20 | In peppery | ? 29.82 |
| 5 | 0.927 | High peppery | 0.80 | High peppery | 13.70 |
| 6 | 1.254 | High peppery | 1.00 | High peppery | ? 20.26 |
| 7 | 0.895 | High peppery | 1.00 | High peppery | ? 11.73 |
| 8 | 0.603 | In peppery | 0.60 | In peppery | ? 0.50 |
| 9 | 0.241 | In peppery | 0.20 | In peppery | ? 17.01 |
| 10 | 1.120 | High peppery | 1.00 | High peppery | ? 10.71 |
| 11 | 0.275 | In peppery | 0.20 | In peppery | ? 27.27 |
| 13 | 0.506 | In peppery | 0.60 | In peppery | ? 18.58 |
| 13 | 0.043 | Low peppery | 0.05 | Low peppery | ? 16.28 |
As can be seen from Table 1, chilli products carries out capsaicin content mensuration, the result that the inventive method is measured after removing disturbance of protein, the result who records with high performance liquid chromatography contrasts, all in 30%, wherein the relative error of sample 4 is maximum, is 29.82% for relative error; The relative error of sample 8 measured results is minimum, only is 0.50%.Reached the Expected Results of rapid semi-quantitative mensuration Capsaicinoid Content in Capsicum Products content.According to capsaicin content the peppery degree of chilli products is estimated, the peppery degree that the peppery degree of judging for the high effective liquid chromatography for measuring value and the inventive method measured value are judged compares rear discovery, and the result is in full accord.Show for purpose to be to judge the mensuration of the peppery degree of chilli products, the inventive method can satisfy well measures requirement.
Get sample No. 13, carry out precision test and mark-on recovery test: getting capsaicin content is No. 13 sample 5.0g of 0.043mg/g, by carrying out fast detecting according to the inventive method mensuration program behind the preceding method removal protein, duplicate detection 6 times, contrast by the color with standard colorimetric plate, determine capsaicin content, calculate relative standard deviation RSD=0.020.And result and high performance liquid chromatography compared, concrete outcome is as shown in table 2.
Table 2 measurement result contrast table: X(%) be11.37,
RSD(%) be18.28
| The sample sequence number | Sampling amount (g) | Capsicim addition (mg) | HPLC method testing result (mg/g) | Colorimetric detection result (mg/g) | Relative error (%) |
| 1 | 5.0 | 0.0 | 0.043 | 0.05±0.03 | 16.28 |
| 2 | 5.0 | 0.2 | 0.087 | 0.075±0.04 | 13.79 |
| 3 | 5.0 | 0.3 | 0.102 | 0.125±0.075 | 22.55 |
| 4 | 5.0 | 0.4 | 0.126 | 0.125±0.075 | 0.79 |
| 5 | 5.0 | 0.5 | 0.145 | 0.125±0.075 | 13.79 |
| 6 | 5.0 | 1.0 | 0.246 | 0.25±0.15 | 1.63 |
| 7 | 5.0 | 2.0 | 0.442 | 0.40±0.20 | 9.50 |
| 8 | 5.0 | 3.0 | 0.648 | 0.50±0.10 | 22.84 |
| 9 | 5.0 | 4.0 | 0.841 | 0.80±0.20 | 4.88 |
| 10 | 5.0 | 5.0 | 1.028 | 0.90±0.10 | 7.00 |
| 11 | 5.0 | 6.0 | 1.246 | 1.50±0.50 | 20.39 |
| 12 | 5.0 | 7.0 | 1.457 | 1.50±0.50 | 2.95 |
| 13 | 5.0 | 10.0 | 2.053 | -- | -- |
Annotate: HPLC represents high performance liquid chromatography,--expression can't be calculated.
As can be seen from Table 2, when the capsicim concentration in the solution to be measured during greater than 2.00 mg/mL, because colour developing is too dark, color range is difficult to distinguish, and does not have the respective standard color board to do reference, and suitably dilution is measured again.The mark-on control test is the result show, the relative error of the inventive method and high performance liquid chromatography measured result is up to 22.84% all below 23%, and average relative error is that 11.37%, RSD is 18.28%.The content range that the inventive method is used for the mensuration Capsaicinoid Content in Capsicum Products is 0.02-2.00 mg/mL, can be used for quick, the semiquantitative determination of proteinaceous Capsaicinoid Content in Capsicum Products content.
Claims (3)
1. Capsaicinoid Content in Capsicum Products content rapid colorimetric determination method is characterized in that realizing by following steps:
(1) preparation sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion: compound concentration is the sodium carbonate liquor of 0.1mol/L and phosphomolybdic acid-Salkowski's solution of 1.0mol/L respectively, with sodium carbonate liquor and phosphomolybdic acid-Salkowski's solution by volume 30:1 mix, namely get sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion, concentration is that the compound method of phosphomolybdic acid-Salkowski's solution of 1.0mol/L is: take by weighing 100g sodium tungstate and 20g phosphomolybdic acid, adding 60mL volume by volume concentration is 85% phosphoric acid and 700mL distilled water, boil 2h, cooled and filtered is settled to 1000mL with distilled water;
(2) production standard color board: adopt concentration gradient: 0,0.02,0.05,0.08,0.10,0.20,0.40,0.60,0.80, the capsicim methanol solution of 1.00,2.00mg/mL, respectively with sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion by volume 1:3 mix colour developing, according to reacted color and corresponding capsicim concentration production standard color board;
(3) sample preparation: for the chilli products that contains protein, at first be the ethanolic solution of 65-75% and the chilli products that contains protein that takes by weighing with volume by volume concentration, ratio in 50 mL:1g is mixed, be 80-120w at ultrasonic power, process 35-45min under the temperature 45-55 ℃ of condition to remove protein, process the layering of afterreaction thing, the chilli products behind the protein is removed in separation, with methyl alcohol and tetrahydrofuran 1:1 volume ratio mixed liquor extraction capsicim 10-20min wherein, re-extract is once again for residue after filtering, merging filtrate also is settled to the ratio constant volume of 100mL in every 10-20g sample, gets capsicim solution to be measured; For the chilli products that does not contain protein, directly take by weighing chilli products, with methyl alcohol and tetrahydrofuran 1:1 volume ratio mixed liquor extraction capsicim 10-20min wherein, re-extract is once again for residue after filtering, merging filtrate also is settled to the ratio constant volume of 100mL in every 5-10g sample, gets capsicim solution to be measured;
(4) with capsicim solution to be measured and sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion by volume 1:3 mix colour developing, by the contrast standard color board, draw capsicim concentration.
2. a kind of Capsaicinoid Content in Capsicum Products content rapid colorimetric determination method according to claim 1, after it is characterized in that step (2) capsicim methanol solution and sodium carbonate-phosphotungstic acid-phosphomolybdic acid nitrite ion 1:3 mixing colour developing by volume, utilize L*, a*, b* value after colour photometer is measured colour developing, the capsicim concentration that record is corresponding, carry it into and produce corresponding color in the Photoshop software, thereby make standard colorimetric plate, step (4) is used mutually colour photometer and is measured color, draws capsicim concentration according to the variation of Δ L* and Δ b* value.
3. a kind of Capsaicinoid Content in Capsicum Products content rapid colorimetric determination method according to claim 1 and 2 is characterized in that the described chilli products that does not contain protein of step (3) refers to capsicum oleoresin, capsicim crystal or chilli oil.
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| CN103411966A (en) * | 2013-08-14 | 2013-11-27 | 湖南农业大学 | Capsaicin tracking and identification method for grain with excess heavy metals and application of capsaicin |
| CN108634239A (en) * | 2018-04-17 | 2018-10-12 | 文山学院 | A kind of sea sedge flavor chili biscuits and preparation method thereof |
| CN110915649A (en) * | 2019-12-03 | 2020-03-27 | 西藏自治区农牧科学院蔬菜研究所 | Rapid screening method of high-pungency pepper breeding resources |
| CN111264215A (en) * | 2020-01-15 | 2020-06-12 | 四川省丹丹郫县豆瓣集团股份有限公司 | Chili variety breeding method for Pi county bean cotyledon |
| CN112461776A (en) * | 2020-11-30 | 2021-03-09 | 湖南省蔬菜研究所 | Method for judging hot degree of hot pepper in field |
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| CN113403363A (en) * | 2021-07-26 | 2021-09-17 | 浙江省农业科学院 | Method for detecting capsaicin content by using botrytis cinerea |
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| CN113848204A (en) * | 2021-11-16 | 2021-12-28 | 河南羚锐制药股份有限公司 | Method for judging end point of capsaicin extracted from hot pepper |
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| CN103411966A (en) * | 2013-08-14 | 2013-11-27 | 湖南农业大学 | Capsaicin tracking and identification method for grain with excess heavy metals and application of capsaicin |
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| CN108634239B (en) * | 2018-04-17 | 2021-11-09 | 文山学院 | Seaweed-flavored chili crisp and preparation method thereof |
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| CN110915649A (en) * | 2019-12-03 | 2020-03-27 | 西藏自治区农牧科学院蔬菜研究所 | Rapid screening method of high-pungency pepper breeding resources |
| CN111264215A (en) * | 2020-01-15 | 2020-06-12 | 四川省丹丹郫县豆瓣集团股份有限公司 | Chili variety breeding method for Pi county bean cotyledon |
| CN112461776A (en) * | 2020-11-30 | 2021-03-09 | 湖南省蔬菜研究所 | Method for judging hot degree of hot pepper in field |
| CN112461776B (en) * | 2020-11-30 | 2023-04-28 | 湖南省蔬菜研究所 | Method for judging peppery degree of peppers in fields |
| CN113588569A (en) * | 2021-07-23 | 2021-11-02 | 湖南省药品检验研究院(湖南药用辅料检验检测中心) | Preparation method of colorimetric block for identifying dye intermediate and colorimetric method thereof |
| CN113588569B (en) * | 2021-07-23 | 2023-07-18 | 湖南省药品检验研究院(湖南药用辅料检验检测中心) | Preparation method of colorimetric block for identifying dye intermediate and colorimetric method thereof |
| CN113403363A (en) * | 2021-07-26 | 2021-09-17 | 浙江省农业科学院 | Method for detecting capsaicin content by using botrytis cinerea |
| NL2032572A (en) * | 2021-07-26 | 2023-01-31 | Zhejiang Acad Agricultural Sci | Method for determining capsaicin content by botrytis cinerea |
| CN113848204A (en) * | 2021-11-16 | 2021-12-28 | 河南羚锐制药股份有限公司 | Method for judging end point of capsaicin extracted from hot pepper |
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Application publication date: 20130403 |