CN110794075A - Analysis method of oil-tea camellia seed oil saponin compounds - Google Patents

Analysis method of oil-tea camellia seed oil saponin compounds Download PDF

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CN110794075A
CN110794075A CN201911145546.5A CN201911145546A CN110794075A CN 110794075 A CN110794075 A CN 110794075A CN 201911145546 A CN201911145546 A CN 201911145546A CN 110794075 A CN110794075 A CN 110794075A
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seed oil
tea
saponin
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CN110794075B (en
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王晓琴
邢晨
方恩华
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Huaqiao University
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    • G—PHYSICS
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    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
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Abstract

The invention relates to an analysis method of oil tea seed oil saponin compounds, which comprises the steps of rapidly extracting the oil tea seed oil saponin compounds under a non-heating condition, and analyzing the extract by adopting a UHPLC-Q-TOF/MS technology. The analysis method of the camellia seed oil saponin compound provided by the invention is rapid and accurate, is a scientific separation and analysis method, can lay a foundation for standardization of the analysis method of the camellia seed oil saponin compound, and provides a reference for further deep analysis and development of camellia seed oil.

Description

Analysis method of oil-tea camellia seed oil saponin compounds
Technical Field
The invention relates to the technical field of analysis methods, in particular to an analysis method of a sasanqua seed oil saponin compound.
Background
The camellia oleifera as one of four woody oil plants in the world is the characteristic woody oil plant in China, is widely cultivated in more than ten provinces (cities) such as Hunan, Guangxi, Jiangxi and the like, does not compete for fields with grains, and belongs to important national strategic resources in China. The camellia seed oil is high-grade woody plant oil, the unsaturated fatty acid can reach about 90 percent, and the fatty acid composition is similar to that of olive oil. The camellia seed oil is also rich in phenolic compounds, vitamin E, squalene and the like. In addition, the camellia seeds also contain abundant saponin compounds, and the cake after oil extraction can be used for developing tea saponin products, but the analysis reports of the saponin compounds in the camellia seed oil are less, and the research on the extraction process is possibly limited. Most of the reported tea saponin is water-soluble, and the related extraction process is not suitable for separating the saponin compounds of the oil-tea camellia seed oil.
However, the fat-soluble saponin compounds are used as the specific active ingredients of the camellia oleosa seed oil, and research on the fat-soluble saponin compounds can provide basic data for the quality standard and application development of the camellia oleosa seed oil. At present, the basic data of the oil-tea camellia seed oil saponin compounds are almost blank.
Disclosure of Invention
The invention aims to overcome the defects of analytical data of camellia seed oil saponin compounds in the prior art, provides an analytical method of camellia seed oil saponin compounds, and lays a scientific foundation for qualitative and quantitative work of the camellia seed oil saponin compounds.
The specific scheme is as follows:
an analysis method of oil-tea camellia seed oil saponin compounds is characterized by comprising the following steps: the method comprises the following steps:
step 1: rapidly extracting oil tea seed oil saponin compounds with organic solvent under non-heating condition, filtering the extractive solution with a membrane, and collecting filtrate;
step 2: respectively sending the filtrate obtained in the step 1 into a high performance liquid chromatograph and a high resolution time-of-flight mass spectrometer, wherein:
the chromatographic conditions are as follows: a chromatographic column: agilent Poroshell 120EC-C18, mobile phase A: 0.1% formic acid water, mobile phase B: 0.1% formic acid acetonitrile, gradient elution, flow rate: 0.3mL/min, column temperature: 30 ℃, sample introduction: 4 mu L of the solution; % in chromatographic conditions are volume fractions;
the mass spectrum conditions are as follows: ionization mode: ESI-The temperature of the drying gas is 300 ℃, the flow rate of the dryer is 7 +/-2L/min, the voltage of a capillary tube is 3.5 +/-0.5 kV, the pressure of an atomizer is 35 +/-5 psig, the temperature of a sheath gas is 350 +/-10 ℃, the flow rate of the sheath gas is 10 +/-2L/min, the fragmentation voltage is 120 +/-10V, the collision voltage of a primary mass spectrum is 45 +/-5V, the collision voltage of a secondary mass spectrum is 65 +/-5V, and the m/z of an ion scanning range is 100-;
and step 3: analyzing the collected data by using UHPLC-Q-TOF/MS, wherein the analysis comprises the following steps: collecting names and molecular formulas of tea saponin compounds in camellia oleifera seeds in the prior art, accurately calculating relative molecular mass, and establishing a database containing the tea saponin compounds by using MassHunter PCDLManager software; retrieving the collected original data by MassHunter qualitative software in combination with saponin compound database to obtain a chromatographic peak contained in the extracting solution and having the same accurate relative molecular mass with the tea saponin compound database; selecting molecular ions corresponding to chromatographic peaks by utilizing a quadrupole rod of a Q-TOF/MS mass spectrometer, performing collision induced dissociation on the molecular ions in a collision cell, and scanning fragment ions obtained by cracking through TOF to obtain a secondary mass spectrogram; deducing the type and structure of the tea saponin contained in the oil-tea camellia seed oil saponin compound extracting solution according to the cracking condition of ions.
Further, in the step 1, taking a 2.5g oil tea seed oil sample as an example, and the dosage of each reagent corresponds to 2.5g oil tea seed oil sample, the method for extracting the oil tea seed oil saponin compounds comprises the following steps: accurately weighing 2.5g of camellia oleosa seed oil sample in a centrifuge tube, adding 5mL of n-hexane, adding 20-30mL of ethanol-water mixed solution, carrying out vortex for 3-7min, centrifuging for 8-12min at 3000-4000rmp, collecting the lower layer, carrying out vortex drying, adding 8-12mL of distilled water for dissolving, transferring the extracting solution into the centrifuge tube, adding 8-12mL of n-butanol, carrying out vortex oscillation for 2-5min, carrying out centrifugal separation, repeating for 2-4 times, combining n-butanol extracting solutions, carrying out vortex evaporation till the n-butanol extracting solution is dried, and re-dissolving with methanol water to obtain the extracting solution.
Further, in the step 1, the method for extracting the sasanqua seed oil saponin compound comprises the following steps: accurately weighing 2.5g of camellia oleosa seed oil sample in a centrifuge tube, adding 5mL of n-hexane, adding 20-30mL of ethanol-water mixed solution, wherein the ethanol content is 65-85%, performing vortex for 5min, centrifuging 3500rmp for 10min, collecting the lower layer, performing vortex drying, adding 10mL of distilled water for dissolving, transferring the extract into the centrifuge tube, adding 10mL of n-butanol, performing vortex oscillation for 3min, performing centrifugal separation, repeating for 3 times, mixing n-butanol extract, performing rotary evaporation to dryness, re-dissolving with 250 μ l of 50% methanol water to obtain extract, passing the extract through a filter membrane with the aperture of 0.2-0.3 μm, and collecting filtrate for later use; the above% are volume fractions.
Further, in the step 1, the method for extracting the sasanqua seed oil saponin compound comprises the following steps: accurately weighing 2.5g of camellia oleosa seed oil sample in a centrifuge tube, adding 5mL of n-hexane, adding 25mL of ethanol-water mixed solution, wherein the ethanol content is 85%, performing vortex for 5min, centrifuging at 3500rmp for 10min, collecting the lower layer, performing vortex drying, adding 10mL of distilled water for dissolving, transferring the extract into the centrifuge tube, adding 10mL of n-butanol, performing vortex oscillation for 3min, performing centrifugal separation, repeating for 3 times, mixing n-butanol extract, performing rotary evaporation to dryness, re-dissolving with 250 μ l of 50% methanol-water to obtain extract, passing the extract through a filter membrane with the aperture of 0.22 μm, and collecting filtrate for later use; the above% are volume fractions.
Further, the chromatographic conditions in the step 2 are as follows: a chromatographic column: agilent Poroshell 120EC-C18(3.0 mm. times.100 mm, 2.7 μm), mobile phase A: 0.1% formic acid water, mobile phase B: 0.1% formic acid acetonitrile, gradient elution (0-2min, 90% A, 10% B; 2-6min, 90% A → 65% A, 10% B → 35% B; 6-16min, 65% A → 35% A, 35% B → 65% B; 16-18min, 35% A → 5% A, 65% B → 95% B; 18-19.9min, 5% A, 95% B; 19.9-20min, 5% A → 90A, 95% B → 10% B), flow rate: 0.3mL/min, column temperature: 30 ℃, sample introduction: 4 mu L of the solution; the% in chromatographic conditions are both volume fractions.
Further, the mass spectrum conditions in the step 2 are as follows: ionization mode: ESI-, drying gas temperature 300 ℃, dryer flow rate 7L/min, capillary voltage 3.5kV, atomizer pressure 35psig, sheath gas temperature 350 ℃, sheath gas flow rate 10L/min, fragmentation voltage 120V, primary mass spectrum collision voltage 45V, secondary mass spectrum collision voltage 65V, and ion scanning range m/z of 100-1500.
Further, in the step 3, the types and structures of the tea saponins contained in the oil-tea camellia seed oil saponin compound extracting solution are deduced according to the ion cracking condition, and the method comprises the following steps: and reasonably speculating the structure of the tea saponin according to the identified structure of the tea saponin, the m/z difference and the functional group site of the existing tea saponin.
Further, comparing the unknown compound with the known tea saponin, if the difference between the fragments of the two tea saponins at each cracking stage is 2Da except the 451Da fragment which represents glycosyl at last, supposing that a certain functional group on the sapogenin of the unknown compound A is two hydrogens more than that of the existing tea saponin, and then reasonably replacing the known tea saponin according to the distribution characteristics of the functional groups on the tea sapogenin skeleton; if the fragments of glycosyl in the cracking process are the same after glycosyl is gradually lost, and the difference between the fragments of glycosyl in the rest cracking processes is 30Da, the outermost glycosyl on the glycosyl chain of the unknown compound B is presumedIs a six carbon sugar; if the fragment difference in the R2 group cleavage process is 14Da, the fragments in the rest cleavage processes are the same, and the unknown compound C is presumed to have more CH than the R2 group2Possibly a butyryl group; no reported compound is found, and reasonable speculation is carried out based on the reported theasaponin structure and the cracking rule of the theasaponin structure.
Has the advantages that:
the invention adopts the ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry technology to effectively eliminate the interference of the tea saponin isotope peak and the compounds with similar mass-to-charge ratio on the qualitative determination. Under the influence of factors such as optical rotation of sugar, cis-trans isomerization of vinyl, position isomerization of substituent and the like, a plurality of peaks appear in the tea saponin compound with the same mass-to-charge ratio. Analyzing the tea saponin in the oil-tea camellia seed oil in an anion mode, and identifying 21 main tea saponin compounds according to the accurate relative molecular mass of each chromatographic peak and the secondary fragment ion information by referring to related documents and combining with a self-built saponin compound database. 15 of the above-mentioned plants are reported in the literatures of camellia seeds, camellia flowers and the like, and the rest 6 are not reported. Because the tea saponin compounds have strong cracking regularity in mass spectrum, the structure can be reasonably speculated according to the identified tea saponin structure, the m/z difference value and the functional group site of the existing tea saponin. The tea saponin compounds only exist in the crude oil of the camellia seeds, all tea saponins are decomposed after the crude oil is subjected to alkali refining, a large amount of tea sapogenins are possibly generated, the technology cannot provide enough sapogenin cracking information, and the subsequent exploration can be carried out by adopting ion trap mass spectrometry.
In a word, the analysis method of the camellia seed oil saponin compound provided by the invention is rapid and accurate, is a relatively good qualitative determination method, can be used as an analysis method of the camellia seed oil saponin compound, and provides a reference for further deep analysis and development of camellia seed oil.
Drawings
In order to illustrate the technical solution of the present invention more clearly, the drawings will be briefly described below, and it is apparent that the drawings in the following description relate only to some embodiments of the present invention and are not intended to limit the present invention.
Fig. 1 is a schematic diagram of the cracking of camellia oleosa seed oil theasaponin provided by an embodiment 6 of the invention;
fig. 2 is a structural diagram of camellia oil saponin functional groups provided in an embodiment 6 of the present invention;
fig. 3 is a second-order mass spectrum of theasaponin provided in example 6 of the present invention.
Detailed Description
Preferred embodiments of the present invention will be described in more detail below. While the following describes preferred embodiments of the present invention, it should be understood that the present invention may be embodied in various forms and should not be limited by the embodiments set forth herein. The examples do not specify particular techniques or conditions, and are performed according to the techniques or conditions described in the literature in the art or according to the product specifications. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available. In the following examples, "%" refers to volume percent, unless otherwise specified.
The apparatus and equipment used below included:
a high performance liquid chromatograph (Agilent 1290), a high-resolution time-of-flight mass spectrometer (Agilent 6540), a quaternary pump, an online degassing system, a column oven, an automatic sample feeding system, a dual JetStream ESI electrospray ion source, a MassHunterData acquirement (B.07.00) real-time Acquisition workstation and a Qualitative Analysis (B.07.00) offline Analysis software are all Agilent corporation of America; analytical balance (BSM220.4) was purchased from shanghai zhuojing electronics, ltd; a rotary evaporator (R215), a constant temperature water bath (B491), a vacuum pump (V700) purchased from BUCHI, Switzerland; the electric heating air blowing drying box (101) is purchased from Shanghai leaf development instrument and meter company Limited; centrifuge (TGL-16M) was purchased from Hunan instruments, Inc.; vortex mixer (QL-861) was purchased from Onychoma Linbel instruments manufacturing; ultrapure water systems (Milli-Q) are available from Millipore corporation of America.
The following main reagents were used:
the crude oil of the camellia seeds is collected from Fujian oil-saving production enterprises.
Acetonitrile (4L HPLC), formic acid (100ml HPLC) from Sigma, USA; ethanol (500ml AR), methanol (500ml AR), n-hexane (500ml AR), n-butanol (500ml AR) were purchased from national pharmaceutical group chemical Co., Ltd; the laboratory water is self-made ultrapure water; sample bottles (2ml) were purchased from agilent technologies, inc; organic filters (0.22 μm) were purchased from Whatman, UK.
Example 1
Extracting oil tea seed oil saponin compounds: accurately weighing 2.5g of oil tea seed oil sample into a centrifuge tube, adding 5mL of n-hexane, adding 20mL of 85% ethanol, carrying out vortex for 5min, carrying out 3500rmp centrifugation for 10min, collecting the lower layer, carrying out vortex concentration, adding 10mL of distilled water for dissolving, transferring the extract into the centrifuge tube, adding 10mL of n-butanol, carrying out vortex oscillation for 3min, carrying out centrifugal separation, repeating for three times, combining the extracts, carrying out vortex evaporation till the extracts are dried, re-dissolving with 10mL of pure ethanol, determining the content of saponin compounds by adopting a vanillin-sulfuric acid colorimetric method, and finally determining the content of tea saponin to be 3.18mg/g of oil tea seed oil.
Example 2
Extracting oil tea seed oil saponin compounds: accurately weighing 2.5g of oil tea seed oil sample into a centrifuge tube, adding 5mL of n-hexane, adding 25mL of 85% ethanol, carrying out vortex for 5min, carrying out 3500rmp centrifugation for 10min, collecting the lower layer, carrying out vortex concentration, adding 10mL of distilled water for dissolving, transferring the extract into the centrifuge tube, adding 10mL of n-butanol, carrying out vortex oscillation for 3min, carrying out centrifugal separation, repeating for three times, combining the extracts, carrying out vortex evaporation till the extract is dried, re-dissolving with 10mL of pure ethanol, determining the content of saponin compounds by adopting a vanillin-sulfuric acid colorimetric method, and finally determining the content of tea soap to be 3.71mg/g of oil tea seed oil.
Example 3
Extracting oil tea seed oil saponin compounds: accurately weighing 2.5g of oil tea seed oil sample into a centrifuge tube, adding 5mL of n-hexane, adding 30mL of 85% ethanol, carrying out vortex for 5min, carrying out 3500rmp centrifugation for 10min, collecting the lower layer, carrying out vortex concentration, adding 10mL of distilled water for dissolving, transferring the extract into the centrifuge tube, adding 10mL of n-butanol, carrying out vortex oscillation for 3min, carrying out centrifugal separation, repeating for three times, combining the extracts, carrying out vortex evaporation till the extracts are dried, re-dissolving with 10mL of pure ethanol, determining the content of saponin compounds by adopting a vanillin-sulfuric acid colorimetric method, and finally determining the content of tea saponin to be 3.52mg/g of oil tea seed oil.
Example 4
Extracting oil tea seed oil saponin compounds: accurately weighing 2.5g of camellia oleosa seed oil sample into a centrifuge tube, adding 5mL of n-hexane, adding 25mL of 65% ethanol, performing vortex for 5min, centrifuging 3500rmp for 10min, collecting the lower layer, performing vortex concentration, adding 10mL of distilled water for dissolving, transferring the extract into the centrifuge tube, adding 10mL of n-butanol, performing vortex oscillation for 3min, performing centrifugal separation, repeating for three times, mixing the extracts, performing vortex evaporation till the extract is dry, and re-dissolving with 10mL of pure ethanol for later use. And finally, measuring the content of the tea saponin to be 2.79mg/g of the oil-tea camellia seed oil.
Example 5
Extracting oil tea seed oil saponin compounds: accurately weighing 2.5g of camellia oleosa seed oil sample into a centrifuge tube, adding 5mL of n-hexane, adding 25mL of 75% ethanol, performing vortex for 5min, centrifuging 3500rmp for 10min, collecting the lower layer, performing vortex concentration, adding 10mL of distilled water for dissolving, transferring the extract into the centrifuge tube, adding 10mL of n-butanol, performing vortex oscillation for 3min, performing centrifugal separation, repeating for three times, mixing the extracts, performing vortex evaporation till the extract is dry, and re-dissolving with 10mL of pure ethanol for later use. And finally, determining the content of the tea saponin to be 3.03mg/g of the oil-tea camellia seed oil.
Comparative example 1
The method for extracting the camellia seed oil saponin compounds in the prior art comprises the following steps: accurately weighing 2.5g of camellia oleosa seed oil sample into a centrifuge tube, adding 27.5mL of pure ethanol, stirring for 1.25h in a water bath at 60 ℃, centrifuging for 10min at 3500rmp, collecting the lower layer, carrying out rotary evaporation and concentration, adding 10mL of distilled water for dissolving, transferring the extracting solution into the centrifuge tube, adding 10mL of n-butyl alcohol, carrying out vortex oscillation for 3min, carrying out centrifugal separation, repeating for 3 times, combining n-butyl alcohol extracting solutions, carrying out rotary evaporation until the n-butyl alcohol extracting solutions are dried, and re-dissolving with 10mL of pure ethanol for later use. And finally, determining the content of the tea saponin to be 3.46mg/g of the oil-tea camellia seed oil.
The extraction methods provided in examples 1-5 can avoid heating treatment, reduce energy consumption, avoid saponins from degradation after heating, and maintain activity; in addition, the method greatly shortens the treatment time, replaces the traditional water bath heating treatment with vortex oscillation for 5 minutes, and improves the working efficiency. In addition, the extraction method provided in examples 1-5 adopts a method of n-butanol elution to enrich saponin compounds, does not relate to a chemical method, and avoids the influence of the extraction process on the chemical structure of the saponin compounds.
In the experiment, the extracts obtained in examples 1-5 were found to collect more comprehensive and more abundant test data than other pretreatment methods.
Example 6
Analyzing the oil-tea camellia seed oil saponin compounds, which comprises the following steps:
step 1:
accurately weighing 2.5g of camellia oleosa seed oil sample into a centrifuge tube, adding 5ml of n-hexane, uniformly mixing, adding 25ml of 80% ethanol, carrying out vortex for 5min, carrying out 3500rmp centrifugation for 10min, collecting an ethanol layer, carrying out vortex concentration, adding 10ml of distilled water for dissolving, transferring an extracting solution into the centrifuge tube, adding 10ml of n-butanol, carrying out vortex oscillation for 3min, carrying out centrifugal separation, repeating for three times, combining n-butanol layers, carrying out vortex evaporation till the n-butanol layers are dried, carrying out re-dissolution by using 250 mu l of 50% methanol water, and passing through a 0.22 mu m filter membrane for later use.
Step 2: respectively sending the filtrate obtained in the step 1 into a high performance liquid chromatograph and a high resolution time-of-flight mass spectrometer, wherein:
chromatographic conditions are as follows:
a chromatographic column: agilent Poroshell 120EC-C18(3.0 mm. times.100 mm, 2.7 μm), mobile phase A: 0.1% formic acid water, mobile phase B: 0.1% formic acid acetonitrile, gradient elution (0-2min, 90% A, 10% B; 2-6min, 90% A → 65% A, 10% B → 35% B; 6-16min, 65% A → 35% A, 35% B → 65% B; 16-18min, 35% A → 5% A, 65% B → 95% B; 18-19.9min, 5% A, 95% B; 19.9-20min, 5% A → 90A, 95% B → 10% B), flow rate: 0.3mL/min, column temperature: 30 ℃, sample introduction: 4 μ L.
Mass spectrum conditions:
ionization mode: ESI-, drying gas temperature 300 ℃, dryer flow rate 7L/min, capillary voltage 3.5kV, atomizer pressure 35psig, sheath gas temperature 350 ℃, sheath gas flow rate 10L/min, fragmentation voltage 120V, primary mass spectrum collision voltage 45V, secondary mass spectrum collision voltage 65V, and ion scanning range m/z of 100-1500.
And step 3:
analyzing the camellia oil saponin extract by UHPLC-Q-TOF/MS under the conditions of the chromatogram and the mass spectrum, in order to extract effective compound information related to saponin from the extract, compounds of formula I are collected by reference to the references (see 1, Guo, N.; Tong, T.T.; Ren, N.; Tu Y.Y.; Li, B., Saponin from seeds of Genus Camellia: Phytochemistry and bioactivity.Phytocohemistry 2018, 149, 42-55; 2, Cui, C.J.; Zong, J.F.; Sun, Y.; Zhang, L.; Ho, C.T.; Wan, X.C.; Hou R.Y., Triterpen from the Genus Camllia: structures, biological activities, and molecular engineering for structure-activity reaction, 3091, FolAb 3069, and 3069), and the relative molecular masses were calculated accurately and a database containing these theasaponin compounds was built using MassHunterPCDL Manager software. The acquired original data is retrieved by MassHunter qualitative software combined with a self-built saponin compound database to obtain a chromatographic peak (the chromatogram of the extracted ion under each m/z is shown in appendix) contained in the extracting solution and consistent with the accurate relative molecular mass in the tea saponin compound database. And then, selecting molecular ions corresponding to the chromatographic peaks by utilizing a quadrupole of a Q-TOF/MS mass spectrometer, performing collision induced dissociation on the molecular ions in a collision cell, and performing TOF scanning on fragment ions obtained by cracking to obtain a secondary mass spectrogram. Then, according to the ion cleavage situation and further comparison in combination with the literature, the 21 tea saponins in the camellia oleosa seed oil n-butanol extract are preliminarily deduced (see table 1), the cleavage schematic diagram is shown in fig. 1, and the structure of the cleavage functional group is shown in fig. 2.
The cleavage rules of theasaponin are referred to and improved in the related reports (see: Feng, M.X.; Zhu, Z.L.; Zuo, L.M.; Chen, L.; Yuan, Q.P.; Shan, G.Z.; Luo, S.Z., A stream for Rapid structural characterization of saponin and vitamins from the term of Camellia oleifera Abel seeds by ultra-high-precision synthesis 2015, 7, 5942-. Among the 21 tea saponins identified qualitatively, 15 tea saponins (1, 2, 3, 5, 6, 7, 12, 13, 14, 15, 16, 17, 18, 19, 20) have been reported in the literature, and these 15 tea saponins conform to the lysis scheme of fig. 1, while the tea saponin No. 3 (Rt: 10.345) has the highest response and the most abundant secondary information, so the lysis pathways of these 15 tea saponin compounds in the mass spectrum are disclosed by taking the tea saponin No. 3 (Rt: 10.345) as an example.
The second-order mass spectrum of the No. 3 theasaponin (Rt: 10.345) is shown in figure 2, and the peak corresponds to the quasi-molecular ion peak [ M-H [ ]]1201.5673, the elemental composition C of which was analyzed by qualitative software58H90O26In secondary mass spectrometry, there are three cleavage routes for the parent ion, ① dehydration of the parent ion after collision-induced dissociation to lose Ang or Tig groups ([ C ]58H90O26-H-C5H7O-O]-) Yield m/z 1101.5114([ C ]53H80O24]-) ② parent ion lost a pentose group ([ C)58H90O26-H-C5H9O5]-or[C58H90O26-H-C5H7O4]-) Yield m/z 1051.5097([ C ]53H80021]-) Or m/z is 1069.5222([ C ]53H82O22]-) Fragment ions; [ C ]53H82O22]-Further ion collision induced dissociation lost the six carbon sugar group ([ C)53H82O22-H-C6H9O5]-or[C53H82O22-H-C6H11O6]-) Yield m/z 907.4665([ C ]47H72O17]-) Or m/z is 889.4606([ C ]47H70O16]-) Fragment ions; [ C ]47H70O16]-The ion continues to lose the six carbon sugar group and the carboxyl group ([ C ]47H70O16-H-C6H11O6-CO2]-) Yield m/z 665.4027([ C ]40H58O8]-) Fragment ion, [ C ]40H58O8]-The ions further lose residual sugar groups ([ C ]40H58O8-H-C5H5O3]-or[C40H58O8-H-C5H3O2]-) Yield m/z 551.3746([ C ]35H52O5]-) Or m/z is 569.3847([ C ]35H54O6]-) Fragment ion ③ parent ion lost a six carbon sugar group ([ C ] upon collision-induced dissociation58H90O26-H-C6H9O5]-or[C58H90O26-C6H11O5-HO]-) Yield m/z 1039.5129([ C ]52H80O21]-) Or m/z is 1021.5019([ C ]52H78O20]-) Fragment ions; [ C ]52H78O20]-Ion-removal of aglycone yields 451.1090([ C ]17H24O14]-) Fragment ions. The above-mentioned mass spectrometric information and the literature reported Isomer of calolliaseaponin B1The information of the secondary mass spectrum is consistent, so that the compound is identified as Isomer of calolliasesonin B1。
The structures of the rest 6 (No. 4, 8, 9, 10, 11 and 21) theasaponin are not reported in the literature. Because the cracking of the theasaponin compounds in the mass spectrum has a certain rule, the 6 theasaponin structures can be reasonably speculated based on the reported theasaponin structures. The method comprises the following specific steps:
mixing the fragment information of No. 4 tea saponin (m/z 1205) with No. 6 tea saponin Isomer of camelliaseaponin C1(m/z 1203) the two teasaponins differ by 2Da in the fragment at each cleavage stage, except for the 451Da fragment which finally represents the glycosyl group, presumably by two hydrogens on one of the functional groups on its sapogenin over theasaponin No. 6, and R2The group and the group G are the same as those of theasaponin No. 6. Furthermore, Isomer of camelliapasonin C1The R1, R3 and R4 groups of the theasaponin are all hydrogen and can not meet the condition of adding two hydrogen, so that only R5There is a possibility that the group satisfies the condition. And theasaponin R5The radical may be CH2OH just satisfying the conditions, so that it is presumed that theasaponin R No. 45The radical being CH2And (5) OH. Due to the remaining structure and Isomer of calomelasonin C1All of which are the same, can be regarded as Isomer of calolliaseponin C1And the theasaponin No. 4 is named as Isomer of camelliaponin C1derivative。
Comparing the No. 10 tea saponin fragments with the No. 7 tea saponin, the difference between the glycosyl fragments and the No. 7 tea saponin is 30Da, which shows that the No. 10 tea saponin substitutes penta-carbon sugar for hexa-carbon sugar of a G group on the basis of the No. 7 tea saponin structure. Comparing the fragments of other cracking paths, when the G group is lost, the fragment information of the two is consistent, which shows that only the G group of the two is different, so the No. 10 theasaponin is named as Camellia oilsaponin 7 derivative.
The No. 8, 9, 11 tea saponin can be selected from No. 12 tea saponin Theasaponin F3Presumably their structure. Since the number 11 sapogenin is two more hydrogens than the number 8 sapogenin, the number 8 sapogenin is more than Theasaponin F3Two more hydrogens, while the fragments generated after the first cleavage route for theasaponin Nos. 11 and 8 are the same, indicating that No. 11 and No. 8 are only in R2With differences in radicals, R No. 112The radical is R of MB (Isov), No. 82The group should be ang (tig). No. 8 and Theasaponin F3The fragments differed by 2Da after the first cleavage route, indicating that the long alkyl chain group at number 12 should be Ang (Tig). Considering the possibility of hydrogenation of the rest groups on theasapogenin No. 12, R is first hydrogenated5The group was presumed. Due to theasaponin R5The groups have 4 possibilities and are respectively CH3、CHO、CH2OH and COOCH3. To satisfy the condition of hydrogenation, R85The group hydrogen should be as much as possible, thus excluding CHO. If it is CH3And adding two oxygens from other R positions, and comparing with the existing theasaponin literature, a proper functional group cannot be found. If R is number 85The radical being CH2OH, one oxygen, one carbon and two hydrogen are added from other R positions, and the functional group meeting the condition is CH2OH, reported in the literature, theasaponin R1The radical may be CH2OH, just satisfiesConditions, therefore theasaponin R No. 8 was presumed1The radical being CH2OH，R2The radical is Ang (Tig), R5The radical being CH2OH, the remaining groups with Therasaponin F3The tea saponin is the same, and the No. 8 tea saponin is named as Theasaponin F3derivitive a. According to the fragment information of No. 8 and No. 11 theasaponin, No. 11 theasaponin R can be easily estimated2The group is changed into MB (Isov), the rest is the same as No. 8, and the No. 11 theasaponin is named as TheasaponINF3deritive B. The No. 9 theasaponin is G on the basis of No. 111Become G2The No. 9 Theasaponin is named as Theasaponin F3derivative C。
The comparison of the No. 21 tea saponin fragment and the No. 20 tea saponin Isomer of teadsaponin D shows that the difference between the glycosyl fragment and Isomer of teadsaponin D is 30Da, which indicates that the No. 21 tea saponin substitutes hexose for pentose of G group on the basis of Isomer of teadsaponin D structure, and the information of the two fragments is consistent after the G group is lost, which indicates that only the G group is different, so the No. 21 tea saponin is named Isomer of teadsaponin D derivative.
Figure BDA0002281946830000151
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the various features described in the above embodiments may be combined in any suitable manner without departing from the scope of the invention. The invention is not described in detail in order to avoid unnecessary repetition.
In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.

Claims (8)

1. An analysis method of oil-tea camellia seed oil saponin compounds is characterized by comprising the following steps: the method comprises the following steps:
step 1: rapidly extracting oil tea seed oil saponin compounds with organic solvent under non-heating condition, filtering the extractive solution with a membrane, and collecting filtrate;
step 2: sending the filtrate obtained in the step 1 into a high performance liquid chromatograph and a high resolution time-of-flight mass spectrometer, wherein:
the chromatographic conditions are as follows: a chromatographic column: agilent Poroshell 120EC-C18, mobile phase A: 0.1% formic acid water, mobile phase B: 0.1% formic acid acetonitrile, gradient elution, flow rate: 0.3mL/min, column temperature: 30 ℃, sample introduction: 4 mu L of the solution; % in chromatographic conditions are volume fractions;
the mass spectrum conditions are as follows: ionization mode: ESI-The temperature of the drying gas is 300 ℃, the flow rate of the dryer is 7 +/-2L/min, the voltage of a capillary tube is 3.5 +/-0.5 kV, the pressure of an atomizer is 35 +/-5 psig, the temperature of a sheath gas is 350 +/-10 ℃, the flow rate of the sheath gas is 10 +/-2L/min, the fragmentation voltage is 120 +/-10V, the collision voltage of a primary mass spectrum is 45 +/-5V, the collision voltage of a secondary mass spectrum is 65 +/-5V, and the m/z of an ion scanning range is 100-;
and step 3: analyzing the collected data by using UHPLC-Q-TOF/MS, wherein the analysis comprises the following steps: collecting names and molecular formulas of tea saponin compounds in camellia oleifera seeds in the prior art, accurately calculating relative molecular mass, and establishing a database containing the tea saponin compounds by using MassHunter PCDL Manager software; retrieving the collected original data by MassHunter qualitative software in combination with saponin compound database to obtain a chromatographic peak contained in the extracting solution and having the same accurate relative molecular mass with the tea saponin compound database; selecting molecular ions corresponding to chromatographic peaks by utilizing a quadrupole rod of a Q-TOF/MS mass spectrometer, performing collision induced dissociation on the molecular ions in a collision cell, and scanning fragment ions obtained by cracking through TOF to obtain a secondary mass spectrogram; deducing the type and structure of the tea saponin contained in the oil-tea camellia seed oil saponin compound extracting solution according to the cracking condition of ions.
2. The analysis method of the camellia seed oil saponin compound according to claim 1, characterized by comprising the following steps: in the step 1, taking a 2.5g oil tea seed oil sample as an example, and the dosage of each reagent corresponds to 2.5g oil tea seed oil sample, the method for extracting the oil tea seed oil saponin compounds comprises the following steps: accurately weighing 2.5g of camellia oleosa seed oil sample in a centrifuge tube, adding 5mL of n-hexane, adding 20-30mL of ethanol-water mixed solution, carrying out vortex for 3-7min, centrifuging for 8-12min at 3000-4000rmp, collecting the lower layer, carrying out vortex drying, adding 8-12mL of distilled water for dissolving, transferring the extracting solution into the centrifuge tube, adding 8-12mL of n-butanol, carrying out vortex oscillation for 2-5min, carrying out centrifugal separation, repeating for 2-4 times, combining n-butanol extracting solutions, carrying out vortex evaporation till the n-butanol extracting solution is dried, and re-dissolving with methanol water to obtain the extracting solution.
3. The analysis method of the camellia seed oil saponin compound according to claim 2, characterized by comprising the following steps: in the step 1, the method for extracting the sasanqua seed oil saponin compounds comprises the following steps: accurately weighing 2.5g of camellia oleosa seed oil sample in a centrifuge tube, adding 5mL of n-hexane, adding 20-30mL of ethanol-water mixed solution, wherein the ethanol content is 65-85%, performing vortex for 5min, centrifuging 3500rmp for 10min, collecting the lower layer, performing vortex drying, adding 10mL of distilled water for dissolving, transferring the extract into the centrifuge tube, adding 10mL of n-butanol, performing vortex oscillation for 3min, performing centrifugal separation, repeating for 3 times, mixing n-butanol extract, performing rotary evaporation to dryness, re-dissolving with 250 μ l of 50% methanol water to obtain extract, passing the extract through a filter membrane with the aperture of 0.2-0.3 μm, and collecting filtrate for later use; the above% are volume fractions.
4. The analysis method of the camellia seed oil saponin compound according to claim 3, characterized by comprising the following steps: in the step 1, the method for extracting the sasanqua seed oil saponin compounds comprises the following steps: accurately weighing 2.5g of camellia oleosa seed oil sample in a centrifuge tube, adding 5mL of n-hexane, adding 25mL of ethanol-water mixed solution, wherein the ethanol content is 85%, performing vortex for 5min, centrifuging at 3500rmp for 10min, collecting the lower layer, performing vortex drying, adding 10mL of distilled water for dissolving, transferring the extract into the centrifuge tube, adding 10mL of n-butanol, performing vortex oscillation for 3min, performing centrifugal separation, repeating for 3 times, mixing n-butanol extract, performing rotary evaporation to dryness, re-dissolving with 250 μ l of 50% methanol-water to obtain extract, passing the extract through a filter membrane with the aperture of 0.22 μm, and collecting filtrate for later use; the above% are volume fractions.
5. The analysis method of the camellia seed oil saponin compounds according to any one of claims 1 to 4, characterized by comprising the following steps: the chromatographic conditions in the step 2 are as follows: a chromatographic column: agilent Poroshell 120EC-C18(3.0 mm. times.100 mm, 2.7 μm), mobile phase A: 0.1% formic acid water, mobile phase B: 0.1% formic acid acetonitrile, gradient elution (0-2min, 90% A, 10% B; 2-6min, 90% A → 65% A, 10% B → 35% B; 6-16min, 65% A → 35% A, 35% B → 65% B; 16-18min, 35% A → 5% A, 65% B → 95% B; 18-19.9min, 5% A, 95% B; 19.9-20min, 5% A → 90A, 95% B → 10% B), flow rate: 0.3mL/min, column temperature: 30 ℃, sample introduction: 4 mu L of the solution; the% in chromatographic conditions are both volume fractions.
6. The analysis method of the camellia seed oil saponin compounds according to any one of claims 1 to 4, characterized by comprising the following steps: the mass spectrum conditions in the step 2 are as follows: ionization mode: ESI-, drying gas temperature 300 ℃, dryer flow rate 7L/min, capillary voltage 3.5kV, atomizer pressure 35psig, sheath gas temperature 350 ℃, sheath gas flow rate 10L/min, fragmentation voltage 120V, primary mass spectrum collision voltage 45V, secondary mass spectrum collision voltage 65V, and ion scanning range m/z of 100-1500.
7. The analysis method of the camellia seed oil saponin compounds according to any one of claims 1 to 4, characterized by comprising the following steps: in the step 3, the types and structures of the tea saponins contained in the oil-tea camellia seed oil saponin compound extracting solution are deduced according to the cracking condition of ions, and the method comprises the following steps: and reasonably speculating the structure of the tea saponin according to the identified structure of the tea saponin, the m/z difference and the functional group site of the existing tea saponin.
8. The oil tea of claim 7The analysis method of the seed oil saponin compound is characterized by comprising the following steps: comparing the unknown compound with known tea saponin, if the difference between the fragments of the two tea saponins at each cracking stage is 2Da except the 451Da fragment which represents glycosyl at last, presuming that a certain functional group on the sapogenin of the unknown compound A is two hydrogens more than that of the existing tea saponin, and then reasonably replacing the known tea saponin according to the distribution characteristics of the functional groups on the tea sapogenin skeleton; if the fragments of glycosyl in the cracking process are the same after glycosyl is gradually lost, the difference between the fragments of glycosyl in the other cracking processes is 30Da, and the outermost glycosyl on the glycosyl chain of the unknown compound B is presumed to be hexose; if the fragment difference in the R2 group cleavage process is 14Da, the fragments in the rest cleavage processes are the same, and the unknown compound C is presumed to have more CH than the R2 group2Possibly a butyryl group; no reported compound is found, and reasonable speculation is carried out based on the reported theasaponin structure and the cracking rule of the theasaponin structure.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111855860A (en) * 2020-07-31 2020-10-30 中南林业科技大学 Preparation of detection standard substance for sasanquasaponin and quantitative detection method for sasanquasaponin

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101863953A (en) * 2010-06-30 2010-10-20 浙江工商大学 Method for separating and preparing Liriope graminifolia saponin A from Liriope graminifolia
CN102925292A (en) * 2012-10-19 2013-02-13 伊国群 Method for comprehensively extracting tea oil, flavonoids compounds, tea saponin and tea polysaccharide
CN103472165A (en) * 2013-06-17 2013-12-25 云南出入境检验检疫局检验检疫技术中心 Detection method used for rapid identification of paris polyphylla var. yunnanensis saponin composition, and applications thereof
CN106083985A (en) * 2016-06-29 2016-11-09 南昌大学 A kind of method that membrance separation is combined isolated and purified tea saponin with macroporous resin
CN108659092A (en) * 2018-06-25 2018-10-16 广东药科大学 A kind of anti-inflammatory triterpene saponin componds and its extracting method and application
CN109212088A (en) * 2018-10-22 2019-01-15 嘉兴迈维代谢生物科技有限公司 A method of 23 kinds of ginsenosides in sample are quickly detected based on UPLC- QTRAP technology simultaneously
CN109596766A (en) * 2018-12-27 2019-04-09 中国科学院上海药物研究所 Method based on offline 2D LC-MS combination metabolin prediction saponin of Radix Notoginseng leaf ingredient
CN109765308A (en) * 2018-12-25 2019-05-17 浙江大学 A kind of UPLC-UV-QTOF-MS/MS method of quantitative detection Seed of Camellia Sinensis triterpenoid saponin
CN110095524A (en) * 2019-05-15 2019-08-06 山东省分析测试中心 A kind of triterpenoid saponin mass spectroscopy structural analytic method

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101863953A (en) * 2010-06-30 2010-10-20 浙江工商大学 Method for separating and preparing Liriope graminifolia saponin A from Liriope graminifolia
CN102925292A (en) * 2012-10-19 2013-02-13 伊国群 Method for comprehensively extracting tea oil, flavonoids compounds, tea saponin and tea polysaccharide
CN103472165A (en) * 2013-06-17 2013-12-25 云南出入境检验检疫局检验检疫技术中心 Detection method used for rapid identification of paris polyphylla var. yunnanensis saponin composition, and applications thereof
CN106083985A (en) * 2016-06-29 2016-11-09 南昌大学 A kind of method that membrance separation is combined isolated and purified tea saponin with macroporous resin
CN108659092A (en) * 2018-06-25 2018-10-16 广东药科大学 A kind of anti-inflammatory triterpene saponin componds and its extracting method and application
CN109212088A (en) * 2018-10-22 2019-01-15 嘉兴迈维代谢生物科技有限公司 A method of 23 kinds of ginsenosides in sample are quickly detected based on UPLC- QTRAP technology simultaneously
CN109765308A (en) * 2018-12-25 2019-05-17 浙江大学 A kind of UPLC-UV-QTOF-MS/MS method of quantitative detection Seed of Camellia Sinensis triterpenoid saponin
CN109596766A (en) * 2018-12-27 2019-04-09 中国科学院上海药物研究所 Method based on offline 2D LC-MS combination metabolin prediction saponin of Radix Notoginseng leaf ingredient
CN110095524A (en) * 2019-05-15 2019-08-06 山东省分析测试中心 A kind of triterpenoid saponin mass spectroscopy structural analytic method

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
JIE-LUN HU ET AL: "Extraction of saponin from Camellia oleifera cake and evaluation of its antioxidant activity", 《INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY》 *
MENGXUE FENG ET AL: "A strategy for rapid structural characterization of saponins and fl avonoids from the testa of Camellia oleifera Abel seeds by ultra-high-pressure liquid chromatography combined with electrospray ionization linear ion trap-orbitrap mass spectrometry", 《ANALYTICAL METHODS》 *
XIN-FU ZHANG ET AL: "Qualitative and Quantitative Analysis of Triterpene Saponins from Tea Seed Pomace (Camellia oleifera Abel) and Their Activities against Bacteria and Fungi", 《MOLECULES》 *
XUEJIN WU ET AL: "Simultaneous Determination and Quantification of Triterpene Saponins from Camellia sinensis Seeds Using UPLC-PDA-QTOF-MS/MS", 《MOLECULES》 *
李迅: "茶皂素的提取、成分鉴定及在草莓保鲜中的应用", 《中国优秀硕士学位论文全文数据库（工程科技Ⅰ辑）》 *
石珊珊 等: "乙醇—正己烷混合溶剂同时提取茶油和茶皂素技术的研究", 《粮食与食品工业》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111855860A (en) * 2020-07-31 2020-10-30 中南林业科技大学 Preparation of detection standard substance for sasanquasaponin and quantitative detection method for sasanquasaponin

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