A method for quickly identifying green-calyx plums mixed with other plum blossoms

Abstract

The invention discloses a method for quickly identifying green-calyx plum mixed with other plum blossoms. The method is designed based on the fact that the invention finds that green-calyx plum does not contain 4,6(E),8(E)-column triene, while big red plum and white green plum that are easily confused with green-calyx plum both contain 4,6(E),8(E)-column triene. By detecting the presence or absence of 4,6(E),8(E)-column triene in plum blossom samples, it is quickly identified whether green-calyx plum is mixed with big red plum and / or white green plum, and the detection result is accurate.

Description

Technical Field

[0001] The invention relates to the field of plum blossom identification, and in particular to a method for rapidly identifying green-calyx plum blossoms mixed with other plum blossoms. Background Art

[0002] Plum blossoms are the flowers or buds of the Rosaceae plant Prunus mume, and are one of my country's top ten famous flowers. There are many varieties of plum blossoms, some of which can be used as medicine, while others are ornamental flowers and have no medicinal value.

[0003] Green-calyx plum (Prunus mume f.viridicalyx (Makino) TYChen) is named for its green calyx. Its petals are white or red. It can be used for both medicinal and ornamental purposes. The Compendium of Materia Medica, Grass Section, Green-calyx plum records that its smell is sour, astringent, and flat. It is mainly used to treat flank pain, abdominal distension, poor appetite, and a feeling of something blocking the throat. Red-red plum (Prunus mume f.rubriflora TYChen) has red petals and is used for ornamental purposes, but has no medicinal value. White-green plum (Prunus mume f.viridiflora TYChen) has a green calyx and white petals. It is used for ornamental purposes, but has no medicinal value.

[0004] When plum blossoms are used for medicinal purposes, the dried flower buds are smaller in shape. The flower buds of green-calyx plum, big red plum and white plum are very similar, and there is basically no difference after drying. Therefore, if green-calyx plum is mixed with big red plum and / or white plum, it is often necessary for experienced technicians to select them based on their experience. If only a small amount of fragments are mixed, it is even more difficult to identify, and the accuracy is difficult to guarantee, and it is even more difficult for non-professionals to distinguish them. Even the distinction between fresh green-calyx plum, fresh big red plum and fresh white plum is difficult due to their similar appearance. Summary of the invention

[0005] The present invention finds that green-calyx plum, which can be used as a medicinal material, does not contain 4,6(E),8(E)-columntriene compounds, while big red plum and white green plum, which are easily confused with green-calyx plum, both contain 4,6(E),8(E)-columntriene compounds. Therefore, a method for quickly identifying other plum blossoms mixed with green-calyx plum by detecting 4,6(E),8(E)-columntriene in plum blossom samples is proposed, and the method has high accuracy.

[0006] The 4,6(E),8(E)-columntriene, molecular formula: C 13 H 20 , CAS: 51468-86-1, the structural formula is as follows:

[0007]

[0008] A method for quickly identifying green-calyx plum blossoms mixed with other plum blossoms comprises the following steps:

[0009] Plum blossom samples were taken to detect 4,6(E),8(E)-big columnar triene in the plum blossom samples. The plum blossom samples with 4,6(E),8(E)-big columnar triene detected were judged to be green-calyx plum mixed with one or two of big red plum and white and blue plum. The plum blossom samples without 4,6(E),8(E)-big columnar triene detected were judged to be pure green-calyx plum.

[0010] The method for detecting 4,6(E),8(E)-columntriene in the plum blossom sample adopts the existing method for detecting 4,6(E),8(E)-columntriene.

[0011] Preferably, the method for detecting 4,6(E),8(E)-columntriene in the plum blossom sample adopts a gas chromatography-mass spectrometry (GC-MS) detection method for 4,6(E),8(E)-columntriene.

[0012] Preferably, the plum blossom sample is first placed in a sealed container, and the extraction head is placed in the sealed container for adsorption for more than 30 minutes. After the extraction is completed, the extraction head is taken out and inserted into the injection port of the gas chromatograph for gas chromatography-mass spectrometry analysis.

[0013] Preferably, the extraction head is placed in a sealed container at 30°C-100°C for adsorption for more than 30 minutes. More preferably, the extraction head is placed in a sealed container at 60°C-100°C for adsorption for 30 minutes. Most preferably, the extraction head is placed in a sealed container at 80°C for adsorption for 30 minutes.

[0014] Preferably, the extraction head is placed in a sealed container at 20° C.-30° C. for adsorption for more than 1 hour.

[0015] Preferably, the extraction head is a divinylbenzene / carbon molecular sieve / polydimethylsiloxane (DVB / CAR / PDMS) solid phase microextraction extraction head or a carbon molecular sieve / polydimethylsiloxane (CAR / PDMS) solid phase microextraction extraction head, and commercially available products can be used.

[0016] Preferably, the plum blossom sample is more than 2g.

[0017] When the plum blossom sample is green-calyx plum mixed with one or two of big red plum and white green plum, the weight percentage of one or two of big red plum and white green plum is more than 0.1%, which can conveniently adsorb 4,6(E),8(E)-big column triene above the detection limit. In the doped sample, if the doping amount of big red plum and / or white green plum is too low, it may lead to adsorption saturation, and the proportion of big column triene in all volatiles is too low, and it may not be adsorbed in time. In practical applications, if green-calyx plum is doped with big red plum and / or white green plum, it will generally be doped with more, and it will not be doped with only 0.1%. The method of the present invention is particularly suitable for the rapid identification and detection of green-calyx plum medicinal materials in practical applications, and is used to identify green-calyx plum doped with big red plum and / or white green plum and pure green-calyx plum.

[0018] The plum blossom sample includes one or both of a fresh plum blossom sample and a dried plum blossom sample, both of which are applicable to the method of the present invention.

[0019] Optionally, the plum blossom sample is a plum blossom bud, which is suitable for rapid identification of non-medicinal red plum and / or white plum mixed with plum medicinal materials. The plum blossom sample can be crushed first and then used for extraction, adsorption and detection of 4,6(E),8(E)-big column triene in the plum blossom sample, which can generally be crushed into fragments of 1mm-3mm; or it can be directly extracted, adsorbed and detected without treatment.

[0020] The conditions of the gas chromatography-mass spectrometry analysis are:

[0021] Agilent J&W DB-5ms column, injection port temperature 250°C; heating program: initial temperature 50°C, hold for 1 min, increase to 160°C at 8°C / min, hold for 1 min, then increase to 300°C at 5°C / min, hold for 20 min; carrier gas is high-purity helium (purity above 99.995%), flow rate: 1.0 mL / min, non-split injection; solid phase microextraction analysis time 2 min.

[0022] Electron bombardment ion source (EI), ion source temperature 230°C, transfer line temperature 280°C; mass scanning range m / z 35-600.

[0023] The gas chromatography-mass spectrometry analysis conditions can obtain suitable peak elution time and good peak shape. Adjusting the parameters does not have a significant effect on the detection of 4,6(E),8(E)-big columnar triene, but may result in poor peak shape and delayed or advanced peak elution time.

[0024] The reagents, consumables, instruments, etc. of the present invention can all be commercially available products.

[0025] The present invention has the following beneficial technical effects:

[0026] The method of the present invention is designed based on the fact that the present invention first discovered that green-calyx plum does not contain 4,6(E),8(E)-columntriene, while big red plum and white green plum that are easily confused with green-calyx plum both contain 4,6(E),8(E)-columntriene. The method is designed to quickly identify green-calyx plum mixed with other plum blossoms by detecting 4,6(E),8(E)-columntriene in plum blossom samples, and the detection result is accurate.

[0027] The method of the invention can realize the rapid identification of whether the green-calyx plum is mixed with the big red plum and / or the white green plum by means of the existing detection method of 4,6(E),8(E)-big column triene, and can make the best use of the existing technical resources.

[0028] In the method of the present invention, the GC-MS method is adopted to detect 4,6(E),8(E)-macrotriene, and the detection limit is 2 ppb, and the accuracy is very high.

[0029] The method of the invention can be applied to the rapid identification and detection of green-calyx plum medicinal materials, and can be used to identify green-calyx plums mixed with big red plums and / or white and green plums and pure green-calyx plums. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 This is the GC-MS spectrum of green-calyx plum;

[0031] Figure 2 This is the GC-MS spectrum of Dahongmei;

[0032] Figure 3 yes Figure 2 The mass spectrum of the compound corresponding to the peak at 21 min;

[0033] Figure 4 is a graph showing the effect of adsorption temperature on the peak area of ​​4,6(E),8(E)-macrotriene detected by GC-MS in Example 4;

[0034] Figure 5 This is a graph showing the effect of adsorption temperature on the peak area of ​​4,6(E),8(E)-macrotriene detected by GC-MS in Example 5. DETAILED DESCRIPTION

[0035] The present invention is further described in detail below in conjunction with the accompanying drawings and embodiments so that those skilled in the art can better understand the technical solution of the present invention; the embodiments adopt the optimal conditions of the present invention and are not intended to limit the feasibility scope of the present invention. The technical effects of the present invention can be achieved within the scope described in the present invention.

[0036] Example 1

[0037] Step 1: Take 50g of dried green-calyx plum bud sample, chop into 2mm size, mix evenly, take 2g sample and add to 15mL sample bottle, put the sample bottle into the metal bath heating tank, set the heating temperature to 80℃, insert the extraction head into the upper part of the sample bottle (extraction head: DVB / CAR / PDMS 50 / 30μm), extraction adsorption time 30min. After extraction, insert the extraction head into the injection port of gas chromatography for GC-MS analysis.

[0038] Step 2: GC-MS analysis conditions

[0039] Agilent J&W DB-5ms chromatographic column (30m×0.25mm×0.25μm), injection port temperature 250℃; heating program: initial temperature 50℃, hold for 1min, increase to 160℃ at 8℃ / min, hold for 1min, then increase to 300℃ at 5℃ / min, hold for 20min; carrier gas is high-purity helium (purity above 99.995%), flow rate: 1.0mL / min, non-split injection; solid phase microextraction analysis time 2min.

[0040] The electron bombardment ion source had an ion source temperature of 230°C, a transfer line temperature of 280°C, and a mass scan range of m / z 35-600.

[0041] The GC-MS spectrum of the dried green-calyx plum bud sample is shown in Figure 1 , Figure 1 No peak appeared at the 21st minute, indicating that the dried green-calyx plum bud sample did not contain 4,6(E),8(E)-column triene compounds. The spectrum search function can also be used to compare the entire spectrum data with the 4,6(E),8(E)-column triene in the internationally used NIST database, and no 4,6(E),8(E)-column triene components were found in the sample.

[0042] Example 2

[0043] 50 g of dried green-calyx plum bud sample was replaced with 50 g of dried bright red plum bud sample, and the remaining operations were the same as in Example 1 to obtain the GC-MS spectrum of the dried bright red plum bud sample. Figure 2 , Figure 2 A peak appears at the 21st minute. The structure of the corresponding compound is identified by using ion fragments. The identification results are shown in Figure 3 , Figure 3 prove Figure 2 The peak that appears at the 21st minute is the peak of the compound "4,6(E),8(E)-big column triene", indicating that the dried big red plum bud sample contains 4,6(E),8(E)-big column triene compound components.

[0044] Example 3

[0045] Take two fresh green-calyx plum bud samples, 50 g each, and record them as fresh green-calyx plum 1 and fresh green-calyx plum 2; take three dried green-calyx plum bud samples, 50 g each, and record them as dried green-calyx plum 3, dried green-calyx plum 4, and dried green-calyx plum 5;

[0046] Two fresh red plum bud samples, 50 g each, were recorded as fresh red plum 1 and fresh red plum 2; four dried red plum bud samples, 50 g each, were recorded as dried red plum 3, dried red plum 4, dried red plum 5, and dried red plum 6;

[0047] Two fresh samples of Prunus mume flower buds, 50 g each, were recorded as fresh Prunus mume 1 and fresh Prunus mume 2; two dried Prunus mume flower buds, 50 g each, were recorded as dried Prunus mume 3 and dried Prunus mume 4;

[0048] A mixed sample consisting of 47.5 g of dried green-calyx plum buds and 2.5 g of dried red plum buds, recorded as green-calyx plum + 5% red plum;

[0049] A mixed sample consisting of 47.5 g of dried green-calyx plum buds and 2.5 g of dried white-green plum buds, recorded as green-calyx plum + 5% white plum;

[0050] A mixed sample consisting of 48g of dried green-calyx plum buds, 1g of dried big red plum buds and 1g of dried white and green plum buds, recorded as green-calyx plum + 2% red plum + 2% white plum;

[0051] The remaining operations were the same as in Example 1, and the GC-MS spectra of various samples were obtained. The peak areas of 4,6(E),8(E)-big pillartriene in the spectra of various samples were calculated. The results are shown in Table 1:

[0052] Table 1

[0053]

[0054]

[0055] Table 1 shows that fresh green-calyx plum buds and dried green-calyx plum buds do not contain 4,6(E),8(E)-column triene compounds, fresh red plum buds and dried red plum buds contain 4,6(E),8(E)-column triene compounds, and fresh white-green plum buds and dried white-green plum buds contain 4,6(E),8(E)-column triene compounds. 4,6(E),8(E)-column triene compounds can be detected in samples of green-calyx plum mixed with one or both of red plum and white-green plum, indicating that by detecting the presence of 4,6(E),8(E)-column triene components in plum samples, it is possible to quickly identify whether green-calyx plum is mixed with one or both of red plum and white-green plum, and the detection results are accurate. Therefore, plum blossom samples were taken to detect the 4,6(E),8(E)-big columnar triene component in the plum blossom samples. The plum blossom samples with detected 4,6(E),8(E)-big columnar triene component were judged to be green-calyx plum mixed with one or two of the big red plum and white green plum. The plum blossom samples without detected 4,6(E),8(E)-big columnar triene component were judged to be pure green-calyx plum (without mixed with one or two of the big red plum and white green plum).

[0056] Example 4 Effect of adsorption temperature on the adsorption amount of 4,6(E),8(E)-columnar triene:

[0057] Take 50g of dried red plum buds, chop them into 2mm size, mix them evenly, take 2g of sample and add them to a 15mL sample bottle, set up multiple sample bottles with samples, put the sample bottles into a metal bath heating tank, set the heating temperature at 30℃, 40℃, 50℃, 60℃, 70℃, 80℃, 90℃, 100℃, insert the extraction head into the upper part of the sample bottle (extraction head: DVB / CAR / PDMS 50 / 30μm), and the extraction adsorption time is 30min. After the extraction is completed, insert the extraction head into the injection port of the gas chromatograph for GC-MS analysis.

[0058] The GC-MS analysis conditions were the same as in Example 1, and the GC-MS spectra of each group of samples were obtained. The peak area of ​​4,6(E),8(E)-macrotriene in the spectra of each group of samples was calculated. The results are shown in Figure 4 .from Figure 4It can be seen that for the same extraction head, at 30°C-100°C, the adsorption amount of 4,6(E),8(E)-large column triene gradually increases with the increase of temperature, and the adsorption amount of 4,6(E),8(E)-large column triene presents a maximum value at 80°C. As the temperature further increases, the adsorption amount of 4,6(E),8(E)-large column triene decreases, but the decrease is not large. Therefore, the extraction head is placed in a sealed container at 30°C-100°C for adsorption for more than 30 minutes, and the adsorption amount of 4,6(E),8(E)-large column triene is relatively high. More preferably, the extraction head is placed in a sealed container at 60°C-100°C for adsorption for 30 minutes. From the perspective of saving energy and reducing experimental difficulty, it is most preferred to place the extraction head in a sealed container at 80°C for adsorption for 30 minutes.

[0059] Example 5 Effect of adsorption temperature on the adsorption amount of 4,6(E),8(E)-columnar triene:

[0060] Take a mixed sample consisting of 48g of dried green-calyx plum buds, 1g of dried big red plum buds and 1g of dried white and blue plum buds, record it as green-calyx plum + 2% red plum + 2% white plum; chop it into 2mm size, mix it evenly, take 2g of sample and add it to a 15mL sample bottle, set a plurality of sample bottles filled with samples, put the sample bottle into a metal bath heating tank, set the heating temperature at 30℃, 40℃, 50℃, 60℃, 70℃, 80℃, 90℃, 100℃ respectively, insert the extraction head into the upper part of the sample bottle (the extraction head is: DVB / CAR / PDMS 50 / 30μm), and the extraction adsorption time is 30min. After the extraction is completed, insert the extraction head into the injection port of the gas chromatograph for GC-MS analysis.

[0061] The GC-MS analysis conditions were the same as in Example 1, and the GC-MS spectra of each group of samples were obtained. The peak area of ​​4,6(E),8(E)-macrotriene in the spectra of each group of samples was calculated. The results are shown in Figure 5 .from Figure 5 It can be seen that for the same extraction head, at 30℃-100℃, the adsorption amount of 4,6(E),8(E)-big columnar triene gradually increases with the increase of temperature. At 80℃, the adsorption amount of 4,6(E),8(E)-big columnar triene presents a maximum value. As the temperature further increases, the adsorption amount of 4,6(E),8(E)-big columnar triene decreases, but the decrease is not large, which is consistent with the Figure 4The trend presented in the experiment is consistent; therefore, the extraction head is placed in a sealed container at 30℃-100℃ for adsorption for more than 30 minutes, and a higher amount of 4,6(E),8(E)-big column triene is adsorbed. It is further preferred that the extraction head is placed in a sealed container at 60℃-100℃ for adsorption for 30 minutes. From the perspective of energy saving and reducing experimental difficulty, it is most preferred that the extraction head is placed in a sealed container at 80℃ for adsorption for 30 minutes.

[0062] Example 6

[0063] The samples can also be directly extracted, adsorbed and tested without crushing or heating:

[0064] A mixed sample consisting of 48 g of dried green-calyx plum buds, 1 g of dried big red plum buds and 1 g of dried white plum buds was taken, recorded as green-calyx plum + 2% red plum + 2% white plum; the mixed sample was put into a polyethylene (PE) bag, and an extraction head was directly placed in the middle of the sample in the PE bag containing the sample, and allowed to stand for adsorption for more than 1 hour at room temperature (20°C). After the extraction was completed, the extraction head was inserted into the injection port of a gas chromatograph for GC-MS analysis.

[0065] The GC-MS analysis conditions were the same as those in Example 1. The GC-MS spectrum of the sample was obtained, and the peak area of ​​4,6(E),8(E)-macrocolumnatriene in the sample spectrum was calculated to be 154405 mAu·min.

[0066] Example 7

[0067] A mixed sample consisting of 48 g of dried green-calyx plum buds, 1 g of dried big red plum buds and 1 g of dried white plum buds was taken, recorded as green-calyx plum + 2% red plum + 2% white plum. The solid phase extraction head used was CAR / PDMS. The remaining operations were the same as in Example 1. A GC-MS spectrum of the sample was obtained, and the peak area of ​​4,6(E),8(E)-big columnar triene in the sample spectrum was calculated to be 110372 mAu·min.

[0068] Although the adsorption effect of the CAR / PDMS solid phase microextraction head is slightly worse than that of the DVB / CAR / PDMS solid phase microextraction head, and the adsorption time may need to be further extended to increase the adsorption amount, it does not affect the detection of 4,6(E),8(E)-big columnar triene in the sample, indicating that the DVB / CAR / PDMS solid phase microextraction head, the CAR / PDMS solid phase microextraction head and other solid phase microextraction heads can be used for the extraction and adsorption of 4,6(E),8(E)-big columnar triene in the plum blossom sample, and are suitable for the GC-MS detection method of the present invention.

[0069] The change of parameters in the method of the present invention, such as the change of parameters in the extraction head and the GC-MS detection method, has little effect on the detection results. Therefore, any combination of parameters in the GC-MS detection method of the present invention can be used to quickly identify whether the green-calyx plum is mixed with one or two of the big red plum and the white green water plum by detecting the presence of 4,6(E),8(E)-big column triene components in the plum blossom sample. No further details will be given here.

Claims

1. A method for rapidly identifying green-calyx plum blossoms mixed with other plum blossoms, characterized in that: Includes steps: Take plum blossom samples and detect 4,6(E),8(E)-big column triene in the plum blossom samples. The plum blossom samples with 4,6(E),8(E)-big column triene components are judged as green-calyx plums mixed with one or two of big red plums and white and green water plums. The plum blossom samples without 4,6(E),8(E)-big column triene components are judged as pure green-calyx plums. The plum blossom samples are green-calyx plum samples mixed with one or two of big red plum and white and green plum and pure green-calyx plum samples.

2. The method according to claim 1, characterized in that The method for detecting 4,6(E),8(E)-columntriene in the plum blossom sample adopts the existing method for detecting 4,6(E),8(E)-columntriene.

3. The method according to claim 1, characterized in that The method for detecting 4,6(E),8(E)-columntriene in a plum blossom sample adopts a gas chromatography-mass spectrometry detection method for 4,6(E),8(E)-columntriene.

4. The method according to claim 3, characterized in that The plum blossom sample is first placed in a sealed container, and the extraction head is placed in the sealed container for static adsorption for more than 30 minutes. After the extraction is completed, the extraction head is taken out and inserted into the injection port of the gas chromatograph for gas chromatography-mass spectrometry analysis.

5. The method according to claim 4, characterized in that Place the extraction head in a sealed container at 30℃-100℃ for adsorption for more than 30 minutes.

6. The method according to claim 4, characterized in that Place the extraction head in a sealed container at 20℃-30℃ and let it adsorb for more than 1 hour.

7. The method according to claim 4, 5 or 6, characterized in that: The extraction head is selected from a divinylbenzene / carbon molecular sieve / polydimethylsiloxane solid phase microextraction extraction head or a carbon molecular sieve / polydimethylsiloxane solid phase microextraction extraction head.

8. The method according to claim 3 or 4, characterized in that: The plum blossom sample is more than 2g.

9. The method according to claim 1, 2 or 3, characterized in that: When the plum blossom sample is green-calyx plum mixed with one or both of big red plum and white green plum, the weight percentage of one or both of big red plum and white green plum is above 0.1%.

10. The method according to claim 1, 2 or 3, characterized in that: The plum blossom sample includes one or both of a fresh plum blossom sample and a dried plum blossom sample.

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