CN112345575A - Method for quantitatively determining effective components in L-carnitine health-care product through nuclear magnetic resonance hydrogen spectrum - Google Patents
Method for quantitatively determining effective components in L-carnitine health-care product through nuclear magnetic resonance hydrogen spectrum Download PDFInfo
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- PHIQHXFUZVPYII-ZCFIWIBFSA-N (R)-carnitine Chemical compound C[N+](C)(C)C[C@H](O)CC([O-])=O PHIQHXFUZVPYII-ZCFIWIBFSA-N 0.000 title claims abstract description 96
- 238000000034 method Methods 0.000 title claims abstract description 60
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- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 title claims abstract description 23
- 229910052739 hydrogen Inorganic materials 0.000 title claims abstract description 23
- 239000001257 hydrogen Substances 0.000 title claims abstract description 23
- 238000001228 spectrum Methods 0.000 title claims abstract description 19
- KYQCOXFCLRTKLS-UHFFFAOYSA-N Pyrazine Chemical compound C1=CN=CC=N1 KYQCOXFCLRTKLS-UHFFFAOYSA-N 0.000 claims abstract description 102
- 238000012360 testing method Methods 0.000 claims abstract description 56
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- RZALONVQKUWRRY-FYZOBXCZSA-N 2,3-dihydroxybutanedioic acid;(3r)-3-hydroxy-4-(trimethylazaniumyl)butanoate Chemical compound OC(=O)C(O)C(O)C(O)=O.C[N+](C)(C)C[C@H](O)CC([O-])=O RZALONVQKUWRRY-FYZOBXCZSA-N 0.000 description 24
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- BQMPVGMHZKZPBH-BYZBDTJCSA-N (3r)-3-hydroxy-4-(trimethylazaniumyl)butanoate Chemical compound C[N+](C)(C)C[C@H](O)CC([O-])=O.C[N+](C)(C)C[C@H](O)CC([O-])=O BQMPVGMHZKZPBH-BYZBDTJCSA-N 0.000 description 1
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- PHIQHXFUZVPYII-UHFFFAOYSA-N carnitine Chemical compound C[N+](C)(C)CC(O)CC([O-])=O PHIQHXFUZVPYII-UHFFFAOYSA-N 0.000 description 1
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- G—PHYSICS
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- G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
- G01N24/08—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using nuclear magnetic resonance
- G01N24/082—Measurement of solid, liquid or gas content
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Abstract
The invention discloses a method for quantitatively determining effective components in L-carnitine health-care products by nuclear magnetic resonance hydrogen spectrum, which comprises the following steps: using deuterated water (D)2O) as an internal standard and pyrazine as an internal standard. Two peaks of L-carnitine delta 3.22(9H, s) and pyrazine delta 8.57(4H, s) are selected for quantitative analysis. Testing parameters: 298.15K, zg30 pulses, Spectrum Width (SW) set to 13, pulse excitation center frequency (O1P) 5.5, relaxation time D1 30S, sampling times NS 32, idle sampling times DS 0, and other parameters default to default. The method has the advantages of good precision and accuracy, lower detection limit and quantitative limit, and good linear relation between the content of the effective components of the L-carnitine and the quantitative peak area. The method measures that the contents of the effective components of the four L-carnitine health care varieties reach the official nominal value. The method is accurate, convenient, efficient and low in test cost, and can be used as a method for quantitatively analyzing the effective components in the L-carnitine health-care products.
Description
Technical Field
The invention belongs to the field of quality detection of health care products, and particularly relates to a method for quantitatively determining effective components in an L-carnitine health care product through nuclear magnetic resonance hydrogen spectrum.
Background
L-carnitine (L-carnitine), also known as L-carnitine or translaminating carnitine, has the chemical name of beta-hydroxy-gamma-trimethylammoniobutanoic acid, formula C7H15NO3Is a kind of aid for fat metabolismThe amino acid-like substance of (1). Sufficient L-carnitine is matched with aerobic exercise to obviously improve the fat metabolism efficiency and solve a series of health problems caused by fat metabolism difficulty, such as fatty liver, hyperlipidemia, hypertension and the like, so that the L-carnitine is added into a plurality of health-care foods. In addition, due to the relatively high safety of L-carnitine, the addition of L-carnitine to infant milk powder is currently allowed by the state to prevent infants from being overweight. Because L-carnitine is rich and easy to absorb moisture, L-carnitine is generally prepared into organic acid salts, and currently, L-carnitine preparations on the market mainly comprise L-carnitine fumarate and L-carnitine tartrate, wherein the L-carnitine fumarate mainly exists in some imported L-carnitine health-care products (GNC, Purpura and the like in the United states), and the L-carnitine tartrate mainly exists in L-carnitine tea polyphenol tablets produced by enterprises in the modified pharmaceutical industry, Tangchen Beijian and the like.
The effective content of the L-carnitine health-care product is an important index for measuring the quality of the L-carnitine health-care product, and the accurate determination of the L-carnitine content in the health-care food has important significance. At present, the L-carnitine is mainly determined by a high performance liquid chromatography, an ultraviolet spectrophotometry, an ion chromatography, a high performance liquid chromatography-mass spectrometry, an enzyme reaction kit method and the like. The method for measuring the L-carnitine in the national standard is a spectrophotometry, and has the problems of difficult sample recovery, complex operation, long consumed time, low detection sensitivity and the like. Quantitative Nuclear Magnetic Resonance (QNMR) realizes quantification through strict corresponding relation between peak area and the number of atomic nuclei to be measured, and has the advantages of no need of high-purity reference substances, no damage to samples, simpler operation, small artificial operation error, accurate and reliable test results and the like, so that the United states pharmacopoeia, the British pharmacopoeia and the European pharmacopoeia include the quantitative nuclear magnetic method as an effective determination method. The 2010 Chinese pharmacopoeia contains nuclear magnetic resonance quantitative method as quantitative and qualitative test method, 2020 New edition Chinese medicineQuantitative nuclear magnetic resonance spectroscopy is continuously included as an effective measurement method. Taking L-carnitine fumarate as an example, pyrazine is taken as an internal standard, heavy water is taken as a solvent, and quantification is carried out1The method is accurate, reliable, simple, convenient and feasible, and can be used for measuring the content of the effective components in the L-carnitine health-care product.
Disclosure of Invention
The technical problem to be solved is as follows: aiming at the technical problems of difficult sample recovery, complex operation, long time consumption, need of high-purity reference substances of components to be detected and the like of the existing analysis and test method, the invention develops a method for quantitatively determining effective components in L-carnitine health-care products by nuclear magnetic resonance hydrogen spectrum.
The technical scheme is as follows:
a method for quantitatively determining effective components in L-carnitine health-care products by nuclear magnetic resonance hydrogen spectrum comprises the following steps:
first, selection of deuterated reagents: weighing 20mg L-carnitine compound, selecting D2O is used as a deuterated solvent, and the topsppin 2.1 software with Proton standard parameters is selected for respective determination1H NMR spectrum, active hydrogen is deuterated;
secondly, selecting a quantitative peak and an internal standard substance: direct determination of L-carnitine using default parameters of Topspin2.1 software1H NMR spectrum with D2Calibrating chemical shift of residual solvent peak of O, selecting pyrazine as internal standard, and obtaining quantitative peak delta 8.57(4H, s);
thirdly, testing parameter optimization: selecting zg30 pulse sampling, setting the sampling time (aq) to be 4.15S, the relaxation time D1 to be 30S, the sampling frequency NS to be 32, the idle sampling frequency DS to be 0, the spectrum width SW to be 13.00, the center frequency o1p to be 6.1, and other parameters to be defaulted;
step four, sample testing and data processing: using the optimised test parameters of the third step for the determination of the sample1H NMR spectrum; accurately integrating the peak area at delta 3.19(9H, s) based on the single peak at delta 8.57(4H, s) of pyrazine, and selecting the peak profile line of quantitative peak or internal standard peak and water to reduce errorThe interval between the intersections of the flat baselines is taken as an integration area, and the internal standard peak and the quantitative peak are integrated for 5 times, and the average value is taken as the actual integration area.
The fifth step: the methodology verification comprises linear relation verification, precision experiment, repeatability experiment and stability experiment.
Further, according to the sampling amount, the internal standard substance and the deuterated reagent, the mass of the L-carnitine fumarate in the sampled samples is calculated according to the following formula:
in the above formula, mL-carnitine，mPyrazine estersDenotes the mass of L-carnitine salt and pyrazine, ML-carnitineRepresents the molecular weight of L-carnitine, SL-carnitine、SPyrazine estersThe quantitative peak area of L-carnitine and pyrazine are respectively shown.
Further, purchasing two imported L-carnitine fumarate health-care products, respectively taking one tablet, weighing the tablet, grinding the tablet into powder by using a mortar, accurately weighing 20-30mg of sample, dissolving the sample by using 1000 mu L of standard solution, carrying out ultrasonic treatment for 10min to fully dissolve effective components, standing for 3h to clarify the upper layer, accurately transferring 600 mu L of supernatant into a nuclear magnetic tube, and measuring the upper layer by using optimized test parameters1H NMR spectrum, the content of the L-carnitine effective component in the solution is calculated according to the peak area by the formula in claim 2.
Further, in the fifth step, a verification test of the linear relationship is performed: sample preparation: accurately weighing 280.2mg pyrazine samples, dissolving the pyrazine samples with heavy water, transferring the pyrazine samples into a 10mL volumetric flask, fixing the volume to a scale mark with the heavy water, and shaking up for later use as a standard solution; weighing 35.9mg of L-carnitine fumarate, filling the L-carnitine fumarate into a disposable PE tube, adding 1.5mL of standard solution, performing ultrasonic treatment for 20min to completely dissolve a sample to obtain uniform, clear and transparent solution, sequentially adding 100 mu L, 150 mu L, 200 mu L, 250 mu L, 300 mu L and 450 mu L of the L-carnitine fumarate into a 1.5mL of disposable PE tube, diluting the solution to 600 mu L by using the standard solution, performing ultrasonic treatment for 10min to completely dissolve the solution, transferring the solution to a No. 1-6 nuclear magnetic tube to obtain No. 1-6 solution to be tested, and testing and processing data according to the fourth step; and (3) taking the ratio of the L-carnitine quantitative peak area to the pyrazine quantitative peak area as a vertical coordinate, and taking the actual mass of the L-carnitine fumarate in the sample as a horizontal coordinate to perform plotting to obtain a working curve, and performing linear fitting on the working curve to obtain a regression equation, wherein the regression equation shows that the sample mass and the peak area have a good linear relationship.
Further, the repeatability test is as follows: sample preparation: weighing 40-50mg of L-carnitine salt and 400mg of pyrazine 270- -; testing and processing data according to the fourth step method; the RSD is calculated to be less than 1% according to the ratio of the L-carnitine fumarate quantitative peak area to the pyrazine quantitative peak area, and the method has good repeatability.
Further, the precision test is as follows: and (3) taking the No. 1 sample subjected to linear test, testing and processing data according to the fourth step, carrying out parallel test for 5 times, calculating the standard deviation according to the peak area ratio of the L-carnitine fumarate quantitative peak to the internal standard quantitative peak, and calculating the RSD to be less than 0.5%, so that the sample and the internal standard substance have good precision under the room-temperature solution state.
Has the advantages that:
1. the invention establishes a method for quantitatively analyzing the effective components in the L-carnitine health-care product by nuclear magnetic resonance hydrogen spectrum with high efficiency, accuracy and convenience.
2. The method established by the invention has better universality and can be used for quantitative analysis of effective components such as L-carnitine tartrate, L-carnitine fumarate and L-carnitine prototype compound in L-carnitine health-care products.
3. The method has the testing accuracy similar to that of a High Performance Liquid Chromatography (HPLC) method, and compared with the HPLC method, the method does not need a high-purity reference substance of a component to be tested, does not need complex sample pretreatment processes such as acidification, extraction, filtration and the like, and greatly saves the testing cost.
4. The method of the invention has lower detection limit and quantification limit (less than 10 mu g/mL level),
description of the drawings:
FIG. 1 is the NMR spectrum of the mixture of L-carnitine fumarate and pyrazine.
FIG. 2 is the NMR spectrum of the mixture of L-carnitine tartrate and pyrazine.
FIG.3 is a plot of the linear relationship between mass and peak area for L-carnitine fumarate according to the present application.
FIG. 4 is a graph of the linear relationship between the mass and peak area of L-carnitine tartrate according to the present application.
Detailed Description
The following examples illustrate specific steps of the present invention, but are not intended to limit the invention.
Terms used in the present invention generally have meanings commonly understood by those of ordinary skill in the art, unless otherwise specified. The invention is described in further detail below with reference to specific examples and with reference to data. It will be understood that these examples are intended to illustrate the invention and are not intended to limit the scope of the invention in any way.
In the following examples, various procedures and methods not described in detail are conventional methods well known in the art.
The present invention can be better understood from the following examples and comparative examples. However, it is easily understood by those skilled in the art that the descriptions of the examples and the comparative examples are only for illustrating the present invention and should not be construed as limiting the present invention described in detail in the claims.
Example 1
The effective ingredients in the L-carnitine fumarate health-care product are quantitatively determined by a nuclear magnetic resonance hydrogen spectrum through a nuclear magnetic resonance hydrogen spectrum, and the method comprises the following specific steps:
first, selection of deuterated reagents: the L-carnitine fumarate is organic salt, can be dissolved in polar solvent such as methanol and water, and is prepared by weighing 20mg L-carnitine compound such as L-carnitine fumarate and 600 μ L deuterization reagent (D)2O，CD3OD, DMSO-d6), and respectively determining with topspin2.1 software with Proton standard parameters1H NMR spectrum, which was analyzed to find that: by using D2When O is used as a solvent, all active hydrogen is deuterated, and the peak shape is relatively sharp, so that the method is suitable for quantitative tests; sample in CD3The solubility in OD is poor, the residual peak of a deuterated reagent solvent is strong, the quantitative analysis accuracy is influenced, when DMSO-D6 is selected as the solvent, the quality of a map is high, but the residual peak of active hydrogen is wide, degeneracy exists between the residual peak and the quantitative peak of a common internal standard substance, quantitative analysis cannot be carried out, and the price of DMSO-D6 is high, so that the test cost is greatly increased, comprehensive consideration is carried out, and D is selected2O is most suitable as a deuterated solvent;
secondly, selecting a quantitative peak and an internal standard substance: direct determination of L-carnitine using default parameters of Topspin2.1 software1H NMR spectrum with D2The chemical shift of the residual solvent peak of O is calibrated, and the internal standard peak is generally required to be a sharp single peak and cannot be coincided with other peaks. Selection of N (CH) by comprehensive analysis3) The corresponding peak delta 3.21(9H, s) is a quantitative peak, common internal standard substances suitable for heavy water comprise maleic acid and pyrazine, but the quantitative peak of the maleic acid is overlapped with a hydrogen peak of fumaric acid in a substance to be detected, so that the internal standard substance is not suitable for the experiment. Comprehensively considering and selecting pyrazine as an internal standard, wherein the quantitative peak of the pyrazine is delta 8.57(4H, s);
thirdly, testing parameter optimization: considering that the relaxation waiting time can be greatly shortened by zg30 pulse sampling, the relaxation waiting time has better accumulated acceleration, the signal to noise ratio in unit time is better, and zg30 pulse sampling is selected from the time economy aspect; sampling time (aq) atlas quality and quantitative effect have important effects: the sampling time is too short, namely the signal stops sampling without being completely attenuated, the signal is truncated, false information can be obtained, the effective nuclear magnetic resonance signal is mainly concentrated at the front end of an FID curve, the noise is mainly used at the back, the signal-to-noise ratio is reduced and the testing time is increased instead of excessively increasing the sampling time, the quantitative nuclear magnetic resonance experiment is unfavorable, the FID curve obtained by observing the sampling time for 4.15s shows that the signal is just attenuated and has little margin, and the sampling time (aq) is set to be 4.15 s. For quantitative nuclear magnetic experiments, relaxation waiting time D1 and sampling times NS are the most important two parameters, NS determines the signal-to-noise ratio of a nuclear magnetic resonance spectrogram, and enough sampling times can enable the spectrogram to have enough signal-to-noise ratio, so that a more accurate quantitative result is obtained; sufficient time for D1 is sufficient to allow all corresponding nuclei sufficient time to return to the ground state to avoid disproportionate numbers of nuclei and peak areas due to relaxation saturation. The experiment adopts the fixed sampling times, the influence of D1 on the quantitative peak-to-peak area is inspected by prolonging D1, the result shows that D1 is increased after D1 is prolonged to 30S, the peak area is not changed greatly, so D1 is selected to be 30S, D1 is fixed, the sampling times NS is increased, and the result shows that the influence of the increased sampling times on the experiment result can be ignored after NS is increased to 32 times. Comprehensively considering, the modified sampling parameters are as follows: zg30 pulse, sampling time (AQ) is 4.15S, relaxation time D1 is 30S, sampling times NS is 32, idle sampling times DS is 0, spectrum width SW is 13.00, center frequency o1p is 6.1, and other parameters are defaulted;
and step four, testing and data processing: all samples were tested using the test conditions modified in the third step1H NMR spectrum. Accurately integrating the peak area (not including a satellite peak) at delta 3.19(9H, s) by taking the single peak at the position of delta 8.57(4H, s) of pyrazine as a reference (the peak area is determined to be 1, and no satellite peak is included), selecting the intersection between the peak profile contour line of a quantitative peak or an internal standard peak and a horizontal base line as an integration area for reducing errors, integrating the internal standard peak and the quantitative peak for 5 times, and taking the average value of the internal standard peak and the quantitative peak as an actual integration area;
the fifth step: and (3) verification of methodology:
and (3) linear testing: sample preparation: accurately weighing 280.2mg pyrazine samples, dissolving the pyrazine samples with heavy water, transferring the pyrazine samples to a 10mL volumetric flask, fixing the volume to a scale mark with the heavy water, and shaking up for later use as a standard solution. Weighing 35.9mg of L-carnitine fumarate, filling the L-carnitine fumarate into a disposable PE tube, adding 1.5mL of standard solution, performing ultrasonic treatment for 20min to completely dissolve a sample to obtain a uniform, clear and transparent solution, sequentially adding 100 mu L, 150 mu L, 200 mu L, 250 mu L, 300 mu L and 450 mu L of the L-carnitine fumarate into a 1.5mL of disposable PE tube, diluting the solution to 600 mu L with the standard solution, performing ultrasonic treatment for 10min to completely dissolve the solution, transferring the solution to a No. 1-6 nuclear magnetic tube to obtain a No. 1-6 solution to be tested, and testing and processing data according to the fourth step. The ratio of the L-carnitine quantitative peak area to the pyrazine quantitative peak area is used as a vertical coordinate, the actual mass of L-carnitine fumarate in the sample is used as a horizontal coordinate to perform drawing to obtain a working curve (FIG.3), and the working curve is subjected to linear fitting to obtain a regression equation Y of 0.03944X-0.00131(R of 0.99986, n of 6), which shows that when the mass of the internal standard is 16.81mg and the mass of the L-carnitine fumarate is 2.39mg-10.77mg, the sample mass and the peak area have a good linear relationship
And (3) repeatability test: sample preparation: weighing 46.4mg of L-carnitine fumarate and 280.2mg of pyrazine, dissolving the L-carnitine fumarate and the pyrazine in heavy water, transferring the solution into a 10mL volumetric flask, metering the volume to a scale mark with the heavy water, shaking up, adding 6 parts of 0.6mL sample into a No. 1 '-6' nuclear magnetic tube to obtain a No. 1 '-6' solution to be detected; the data is tested and processed according to the fourth step method. RSD was calculated to be 0.197% from the ratio (0.1084, 0.1085, 0.1084, 0.1085, 0.1087, 0.1081) of the l-carnitine fumarate quantitative peak area to the pyrazine quantitative peak area. Shows that the method has good repeatability
And (3) testing precision: taking the sample No. 1 of the linear test, testing and processing data according to the fourth step, testing for 5 times in parallel, calculating the standard deviation according to the peak area ratio of the L-carnitine fumarate quantitative peak to the internal standard quantitative peak, and calculating the RSD to be 0.173% (the peak area ratio is 0.0920, 0.0921, 0.0920, 0.0920 and 0.0924), which indicates that the sample and the internal standard have good precision under the state of room-temperature solution.
The method for calculating the mass of the L-carnitine fumarate in the solution comprises the following steps:
the l-carnitine fumarate in the test solution can be calculated according to the following formula:
in the above formula, mL-carnitine，mPyrazine estersDenotes the mass of L-carnitine and pyrazine, SL-carnitine、SPyrazine estersThe quantitative peak area of L-carnitine and pyrazine are respectively shown.
The seventh step, apply: purchasing two imported L-carnitine fumarate health-care products, taking one tablet respectively, weighing mass, grinding into powder by using a mortar, accurately weighing a certain mass sample, dissolving by using 1000 mu L of standard solution, carrying out ultrasonic treatment for 10min to fully dissolve effective components, standing for 3hr to clarify the upper layer, accurately transferring 600 mu L of supernatant into a nuclear magnetic tube, testing and processing data according to a fourth step method, and calculating the L-carnitine fumarate content in the solution by using a sixth step formula according to the peak area.
TABLE 1 actual L-carnitine content of two commercially available health products
Through detection, the contents of the effective components of two imported L-carnitine fumarate health-care products (GNC, pleplenile) exceed the nominal value, particularly the GNC product contains 500mg of L-carnitine fumarate per tablet nominally, but the actual measurement value is 917.01mg, and the deviation from the nominal value is large. Interestingly, if we converted the measured l-carnitine fumarate to the amount of l-carnitine active ingredient, each tablet containing 536.45mg of l-carnitine was closer to its nominal value, so we speculated that the product description might be mislabeled because of the clear labeling of the body in its product: each tablet contains 500mg of free form L-carnitine fumarate. It should be noted that although L-carnitine is a high-safety dietary supplement, excessive intake of L-carnitine causes adverse reactions, and the safety range of L-carnitine is currently considered to be 1000mg/d-5000 mg/d. The consumption of L-carnitine is regulated in various countries: the daily limit of L-carnitine is 20mg/kg as specified by the FDA in the United states; the european union stipulates a daily limit of 1-2g for l-carnitine. The maximum usage amount of the L-carnitine of the beverage specified in the national food additive sanitary standard is 3 g/L. In order to ensure the edible safety of the product, in the evaluation of health food, the dosage of the L-carnitine is mainly referred to the regulations of European Union, and the daily edible amount of the adult L-carnitine should not exceed 2 g. The nominal value of the GNC product body is inconsistent with the actual content, the daily dosage is not easy to control, and adverse reactions can be caused by excessive administration. In contrast, the nominal effective component content of the plenilla product is closer to the actual value, so that the daily dosage can be more conveniently controlled.
Example 2
The method comprises the following steps of quantitatively determining the content of L-carnitine tartrate in the health care product by nuclear magnetic resonance hydrogen spectrum:
first, selection of deuterated reagents: the L-carnitine tartrate belongs to organic salts, can be dissolved in large polar solvent such as methanol and water, 30mg of L-carnitine tartrate is weighed, and 600 mu L of deuterization reagent (D) is added2O，CD3OD, DMSO-d6), and respectively determining with topspin2.1 software with Proton standard parameters1H NMR spectrum, which was analyzed to find that: by using D2When O is used as a solvent, all active hydrogen is deuterated, and the peak shape is relatively sharp, so that the method is suitable for quantitative tests;
secondly, selecting a quantitative peak and an internal standard substance: direct determination of L-carnitine tartrate using default parameters of Topspin2.1 software1H NMR spectrum with D2O residual solvent peak calibration chemical shift. It is generally required that the internal standard peak be a sharp single peak and not coincide with other peaks. Selection of N (CH) by comprehensive analysis3) The corresponding peak delta 3.21(9H, s) is a quantitative peak, the common internal standard substance suitable for the heavy water comprises maleic acid and pyrazine, both meet the requirements of the internal standard substances, and considering that the heavy water-pyrazine system has wider application range in the quantitative determination of the L-carnitine in the health-care product and can be used for determining the content of various forms of the L-carnitine (L-carnitine, L-carnitine fumarate and L-carnitine tartrate) in the health-care product, the pyrazine is selected as the internal standard, the quantitative peak is delta 8.59(4H, s), and the quantitative peak is delta 8.59(4H, s)1The results of H NMR measurement are shown in FIG. 3.
Thirdly, testing parameter optimization: considering that the relaxation waiting time can be greatly shortened by zg30 pulse sampling, the relaxation waiting time has better accumulated acceleration, the signal to noise ratio in unit time is better, and zg30 pulse sampling is selected from the time economy aspect; sampling time (aq) atlas quality and quantitative effect have important effects: the sampling time is too short, namely the signal stops sampling without being completely attenuated, the signal is truncated, false information can be obtained, the effective nuclear magnetic resonance signal is mainly concentrated at the front end of an FID curve, the noise is mainly used at the back, the signal-to-noise ratio is reduced and the testing time is increased instead of excessively increasing the sampling time, the quantitative nuclear magnetic resonance experiment is unfavorable, the FID curve obtained by observing the sampling time for 5.15s shows that the signal is just attenuated and has little margin, and the sampling time (aq) is set to be 5.15 s. For quantitative nuclear magnetic experiments, relaxation waiting time D1 and sampling times NS are the most important two parameters, NS determines the signal-to-noise ratio of a nuclear magnetic resonance spectrogram, and enough sampling times can enable the spectrogram to have enough signal-to-noise ratio, so that a more accurate quantitative result is obtained; sufficient time for D1 is sufficient to allow all corresponding nuclei sufficient time to return to the ground state to avoid disproportionate numbers of nuclei and peak areas due to relaxation saturation. In the experiment, the influence of D1 on the quantitative peak area is inspected by fixing the sampling times and prolonging D1, and the result shows that D1 is increased after D1 is prolonged to 25s, the peak area is not changed greatly, so that D1 is selected to be 25s, D1 is fixed, the sampling times NS is increased, and the result shows that the influence of increasing the sampling times to 32 times on the experiment result can be ignored. Comprehensively considering, the modified sampling parameters are as follows: zg30 pulse, sampling time (AQ) is 5.15s, relaxation time D1 is 25s, sampling times NS is 32, idle sampling times DS is 0, spectrum width SW is 13.01, center frequency o1p is 5.88, and other parameters are defaulted;
and step four, sample testing and data processing: sequentially adopting the modified test conditions of the third step to measure each sample1H NMR spectrum. The peak area (not including satellite peak) at delta 3.22(9H, s) is accurately integrated by taking a single peak at pyrazine delta 8.59(4H, s) as a reference (the peak area is determined to be 1, no satellite peak is included), in order to reduce errors, an integration area is selected between the intersection of a peak profile line of a quantitative peak or an internal standard peak and a horizontal base line, the internal standard peak and the quantitative peak are integrated for 5 times, and an average value is taken as an actual integration area.
Fifth step methodology validation:
and (3) linear verification: 311.4mg pyrazine sample is accurately weighed, dissolved in heavy water and transferred to a 10mL volumetric flask, and the volume is determined to the scale mark by the heavy water, and the sample is shaken up for standby use as a standard solution. Weighing 42.3mg of L-carnitine tartrate, filling the L-carnitine tartrate into a disposable PE tube, adding 1.5mL of standard solution, performing ultrasonic treatment for 20min to completely dissolve a sample to obtain a uniform, clear and transparent solution, sequentially adding 100 mu L, 150 mu L, 200 mu L, 250 mu L, 300 mu L and 450 mu L of the L-carnitine tartrate into a 1.5mL of disposable PE tube, diluting the solution to 600 mu L by using the standard solution, performing ultrasonic treatment for 10min to completely dissolve the solution, and transferring the solution to a No. 1-6 nuclear magnetic tube to obtain a No. 1-6 solution to be detected. And completing the test and data processing according to the fourth step. Taking the ratio (table 1) of the quantitative peak area of the L-carnitine tartrate to the quantitative peak area of the pyrazine as a vertical coordinate, and drawing the actual mass of the L-carnitine tartrate in the sample as a horizontal coordinate to obtain a working curve (figure 3), and performing linear fitting on the working curve to obtain a regression equation Y (0.04188X-0.00608) (R (0.99992, n (6)), which shows that when the mass of the internal standard is 18.68mg and the mass of the L-carnitine tartrate is 2.82mg-12.69mg, the sample mass and the peak area have a good linear relationship
And (3) repeatability test: weighing 52.9mg of L-carnitine tartrate and 311.5mg of pyrazine in heavy water, dissolving, transferring to a 10mL volumetric flask, metering to a scale mark with deuterated water, shaking up, adding 6 parts of 0.6mL sample into a No. 1 '-6' nuclear magnetic tube to obtain No. 1 '-6' to-be-detected liquid. And completing the test and data processing according to the fourth step. RSD was calculated to be 0.0505% from ratios (0.1419, 0.1424, 0.1415, 0.1425, 0.1428, 0.1418) of the l-carnitine tartrate quantitative peak area to the pyrazine quantitative peak area.
The method is proved to have good repeatability;
and (3) testing precision: taking sample No. 2, carrying out parallel test for 5 times by adopting optimized test parameters, calculating the standard deviation by the peak area ratio of the L-carnitine tartrate quantitative peak to the internal standard quantitative peak, and calculating the RSD (the peak area ratio is 0.1699, 0.1701, 0.1697, 0.1698, 0.1705 and 0.1698) to be 0.0294 percent, thereby indicating that the sample and the internal standard substance have good precision under the state of room-temperature solution.
Sixth step method for calculating mass of L-carnitine fumarate in solution
The mass of l-carnitine in the solution to be tested can be calculated according to the following formula:
in the above formula, mL-carnitine，mPyrazine estersDenotes the mass of L-carnitine and pyrazine, SL-carnitine、SPyrazine estersThe quantitative peak areas of L-carnitine tartrate and pyrazine are respectively shown.
The seventh step, apply: according to the instruction of two L-carnitine products, the effective component L-carnitine tea polyphenol tablet per 100g of the decoction ministerial double-health L-carnitine tea polyphenol tablet contains 22g of the effective component L-carnitine tartrate, and the effective component 15g per 100g of the modified pharmaceutical product. Taking one tablet, carefully grinding the tablet by using a mortar, respectively weighing 20-30mg of powder after calculating according to the nominal active ingredient content, adding the powder into a disposable PE tube, adding 1.0mL of standard solution, carrying out ultrasonic treatment at 40 ℃ for 10min, standing overnight, taking 0.6mL of supernatant, adding the supernatant into a nuclear magnetic tube, completing the test and data processing according to the fourth step, calculating the mass of the L-carnitine tartrate in each sample according to the sixth step, reversely deducing the L-carnitine content in the original tablet, and showing the results in table 3 that the active ingredient content in the two L-carnitine tartrate health-care products exceeds the nominal value.
TABLE 3 actual L-Carnitine content in two commercially available L-Carnitine tea polyphenols health products
Through detection, the contents of the effective components (L-carnitine tartrate) of two L-carnitine tea polyphenol health-care products (decoction, minister, health-care and correction pharmaceutical industry) exceed the nominal value and are closer to the nominal value, and the difference is that the water-soluble components in the L-carnitine tea polyphenol tablet of the correction pharmaceutical industry are more complex, the atlas is relatively complex, but the quantitative determination is not influenced, and the components in the aqueous solution of the product of the decoction, minister and health-care are simpler and more convenient to analyze. Experiments show that the tea polyphenol in the domestic L-carnitine tea polyphenol health-care product is analyzed by the method without influence on quantitative determination. In addition, the marking of the effective components in the domestic L-carnitine health care product is more standard: the content of the effective components is marked to be closer to the measured value, and the marking name is more standard.
Comparative example:
the conventional methods for determining L-carnitine in health-care products are multiple, wherein a High Performance Liquid Chromatography (HPLC) method is the simplest and practical method, so that the method applied by the invention is used for determining the L-carnitine content in the health-care products by a High Performance Liquid Chromatography (HPLC) method [ Mozimei, Wiverman, high performance liquid chromatography package method, food safety quality detection bulletin (J),2016,7(5), 2093-: the contents of the effective components in the two L-carnitine health products in the examples are respectively measured by a nuclear magnetic resonance hydrogen spectrometry and a high performance liquid chromatography, and the results of the two test methods are compared, as shown in Table 4: the method has no obvious deviation in the determination result compared with the high performance liquid chromatography.
TABLE 4 determination of actual content of effective components of two commercially available L-carnitine health products by high performance liquid chromatography
Compared with the high performance liquid chromatography, the quantitative nuclear magnetic resonance method has similar accuracy, realizes the quantification by adopting the corresponding relation between the peak area and the number of atomic nuclei, does not need a high-purity reference substance of the components to be detected, and can obviously save the test cost. In addition, the method only needs to crush the sample to be tested, extract with deuterium-substituted water, stand and then directly take the supernatant for testing, complex sample pretreatment processes such as acidification, extraction, filtration and the like are not needed, the testing process can be greatly simplified, in addition, the method does not damage the sample, and the sample is convenient to recover.
Claims (6)
1. A method for quantitatively determining effective components in L-carnitine health-care products by nuclear magnetic resonance hydrogen spectrum is characterized by comprising the following steps:
first, selection of deuterated reagents: weighing 20mgL-Carnitine Compound, selection D2O is used as a deuterated solvent, and the topsppin 2.1 software with Proton standard parameters is selected for respective determination1H NMR spectrum, active hydrogen is deuterated;
secondly, selecting a quantitative peak and an internal standard substance: direct determination of L-carnitine using default parameters of Topspin2.1 software1H NMR spectrum with D2Calibrating chemical shift of residual solvent peak O, wherein the L-carnitine quantitative peak is delta 3.22(9H, s), pyrazine is selected as an internal standard, and the quantitative peak is delta 8.57(4H, s);
thirdly, testing parameter optimization: selecting zg30 pulse sampling, setting the sampling time (aq) to be 4.15S, the relaxation time D1 to be 30S, the sampling frequency NS to be 32, the idle sampling frequency DS to be 0, the spectrum width SW to be 13.00, the center frequency o1p to be 6.1, and other parameters to be defaulted;
step four, sample testing and data processing: using the optimised test parameters of the third step for the determination of the sample1H NMR spectrum; the peak area at delta 3.19(9H, s) is accurately integrated by taking the single peak at the position of delta 8.57(4H, s) of pyrazine as a reference, in order to reduce errors, the intersection between the peak profile line of a quantitative peak or an internal standard peak and a horizontal base line is selected as an integration area, the internal standard peak and the quantitative peak are integrated for 5 times, and the average value is taken as the actual integration area.
The fifth step: the methodology verification comprises linear relation verification, precision experiment, repeatability experiment and stability experiment.
2. The method for quantitatively determining the active ingredients in the L-carnitine health-care product through the nuclear magnetic resonance hydrogen spectrum according to the claim 1, which is characterized in that: calculating the mass of the L-carnitine fumarate in the samples according to the sampling amount of the samples, the internal standard substance and the deuterated reagent and the following formula:
in the above formula, mL-carnitine，mPyrazine estersDenotes the mass of L-carnitine salt and pyrazine, ML-carnitineRepresents the molecular weight of L-carnitine, SL-carnitine、SPyrazine estersThe quantitative peak area of L-carnitine and pyrazine are respectively shown.
3. The method for quantitatively determining the active ingredients in the L-carnitine health-care product through the nuclear magnetic resonance hydrogen spectrum according to claim 1, which is characterized in that: purchasing two imported L-carnitine fumarate health-care products, taking one tablet respectively, weighing the tablet, grinding the tablet into powder by using a mortar, accurately weighing 20-30mg of sample, dissolving the sample by using 1000 mu L of standard solution, fully dissolving effective components by using ultrasound for 10min, standing the sample for 3h to clarify the upper layer, accurately transferring 600 mu L of supernatant into a nuclear magnetic tube, and measuring the supernatant by using optimized test parameters1H NMR spectrum, the content of the L-carnitine effective component in the solution is calculated according to the peak area by the formula in claim 2.
4. The method for quantitatively determining the active ingredients in the L-carnitine health-care product by means of nuclear magnetic resonance hydrogen spectroscopy as claimed in claim 1, wherein in the fifth step, a linear relation verification test comprises the following steps: sample preparation: accurately weighing 280.2mg pyrazine samples, dissolving the pyrazine samples with heavy water, transferring the pyrazine samples into a 10mL volumetric flask, fixing the volume to a scale mark with the heavy water, and shaking up for later use as a standard solution; weighing 35.9mg of L-carnitine fumarate, filling the L-carnitine fumarate into a disposable PE tube, adding 1.5mL of standard solution, performing ultrasonic treatment for 20min to completely dissolve a sample to obtain uniform, clear and transparent solution, sequentially adding 100 mu L, 150 mu L, 200 mu L, 250 mu L, 300 mu L and 450 mu L of the L-carnitine fumarate into a 1.5mL of disposable PE tube, diluting the solution to 600 mu L by using the standard solution, performing ultrasonic treatment for 10min to completely dissolve the solution, transferring the solution to a No. 1-6 nuclear magnetic tube to obtain No. 1-6 solution to be tested, and testing and processing data according to the fourth step; and (3) taking the ratio of the L-carnitine quantitative peak area to the pyrazine quantitative peak area as a vertical coordinate, and taking the actual mass of the L-carnitine fumarate in the sample as a horizontal coordinate to perform plotting to obtain a working curve, and performing linear fitting on the working curve to obtain a regression equation, wherein the regression equation shows that the sample mass and the peak area have a good linear relationship.
5. The method for quantitative determination of effective ingredients in L-carnitine health products by means of nuclear magnetic resonance hydrogen spectrometry as claimed in claim 1, wherein said repeatability tests: sample preparation: weighing 40-50mg of L-carnitine salt and 400mg of pyrazine 270- -; testing and processing data according to the fourth step method; the RSD is calculated to be less than 1% according to the ratio of the L-carnitine fumarate quantitative peak area to the pyrazine quantitative peak area, and the method has good repeatability.
6. The method for quantitatively determining the active ingredients in the L-carnitine health-care product by means of nuclear magnetic resonance hydrogen spectrum according to claim 1, wherein the precision test comprises the following steps: and (3) taking the No. 1 sample subjected to linear test, testing and processing data according to the fourth step, carrying out parallel test for 5 times, calculating the standard deviation according to the peak area ratio of the L-carnitine fumarate quantitative peak to the internal standard quantitative peak, and calculating the RSD to be less than 0.5%, so that the sample and the internal standard substance have good precision under the room-temperature solution state.
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Citations (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010043112A1 (en) * | 2008-10-13 | 2010-04-22 | 常州高新技术产业开发区三维工业技术研究所有限公司 | Preparation and its use of derivatisation reagent for detecting l-carnitine or d-carnitine |
| JP2011085511A (en) * | 2009-10-16 | 2011-04-28 | Mitsubishi Rayon Co Ltd | Method of detecting infinitesimal impurity included in carnitine mixture |
| JP2012177643A (en) * | 2011-02-28 | 2012-09-13 | Kyorin Pharmaceutical Co Ltd | Method for measuring partition coefficient using quantitative analysis based on nmr |
| CN105004808A (en) * | 2015-07-10 | 2015-10-28 | 云南中烟工业有限责任公司 | Method for accurately characterizing and evaluating medicinal herb fragrance characteristics of cigarette fragrance |
| CN106018456A (en) * | 2016-05-16 | 2016-10-12 | 山东省分析测试中心 | Method for determining purity of daptomycin on basis of hydrogen nuclear magnetic resonance |
| CN106053510A (en) * | 2016-05-16 | 2016-10-26 | 山东省分析测试中心 | Method for rapidly measuring purity of pregabalin bulk drug based on H-nuclear magnetic resonance (HNMR) |
| US20160318972A1 (en) * | 2015-04-30 | 2016-11-03 | Yang-Chang Wu | Preparation method and analytic method for the extract of antrodia cinnamomea |
| CN106124550A (en) * | 2016-05-16 | 2016-11-16 | 山东省分析测试中心 | A kind of method quickly measuring vildagliptin purity based on hydrogen nuclear magnetic resonance |
| CN107421975A (en) * | 2017-07-17 | 2017-12-01 | 天津中医药大学 | The detection method of glycerol content in a kind of injection |
| CN108152316A (en) * | 2017-12-14 | 2018-06-12 | 中国日用化学工业研究院 | It is a kind of using maleic anhydride content and sulphonation rate as the quality inspection method of the glucosides sulfosuccinate ester salt product of index |
| JP2018090571A (en) * | 2016-11-29 | 2018-06-14 | 国立研究開発法人産業技術総合研究所 | 2,5-bis(aminomethyl)furan dihydrohalide and production method of the same, and production method of 2,5-bis(aminomethyl)furan |
| CN109374667A (en) * | 2018-11-05 | 2019-02-22 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of NMR spectrum method identifying ultra-high-temperature sterilized milk and reconstituted milk |
| CN109632784A (en) * | 2019-01-21 | 2019-04-16 | 淮阴师范学院 | A kind of method of quantitative detection ascorbic acid |
| CN109696449A (en) * | 2018-12-13 | 2019-04-30 | 北京市食品安全监控和风险评估中心（北京市食品检验所） | The method for detecting a variety of small molecule water soluble substances in milk powder product simultaneously |
| CN109991259A (en) * | 2018-12-11 | 2019-07-09 | 北京市食品安全监控和风险评估中心（北京市食品检验所） | The method for detecting a variety of small molecule water soluble substances in milk powder product simultaneously |
| US20190227011A1 (en) * | 2016-06-28 | 2019-07-25 | Ocean University Of China | Method for measuring sterol content by utilizing quantitative nuclear magnetic resonance technology |
| CA3047665A1 (en) * | 2018-06-21 | 2019-12-21 | Oregon Institute of Science and Medicine | Metabolic profiling with magnetic resonance mass spectrometry (mrms) |
| CN111855730A (en) * | 2020-08-12 | 2020-10-30 | 黄河三角洲京博化工研究院有限公司 | Method for testing water content in butyl rubber glue solution |
-
2020
- 2020-11-10 CN CN202011249090.XA patent/CN112345575B/en active Active
Patent Citations (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010043112A1 (en) * | 2008-10-13 | 2010-04-22 | 常州高新技术产业开发区三维工业技术研究所有限公司 | Preparation and its use of derivatisation reagent for detecting l-carnitine or d-carnitine |
| JP2011085511A (en) * | 2009-10-16 | 2011-04-28 | Mitsubishi Rayon Co Ltd | Method of detecting infinitesimal impurity included in carnitine mixture |
| JP2012177643A (en) * | 2011-02-28 | 2012-09-13 | Kyorin Pharmaceutical Co Ltd | Method for measuring partition coefficient using quantitative analysis based on nmr |
| US20160318972A1 (en) * | 2015-04-30 | 2016-11-03 | Yang-Chang Wu | Preparation method and analytic method for the extract of antrodia cinnamomea |
| CN105004808A (en) * | 2015-07-10 | 2015-10-28 | 云南中烟工业有限责任公司 | Method for accurately characterizing and evaluating medicinal herb fragrance characteristics of cigarette fragrance |
| CN106018456A (en) * | 2016-05-16 | 2016-10-12 | 山东省分析测试中心 | Method for determining purity of daptomycin on basis of hydrogen nuclear magnetic resonance |
| CN106053510A (en) * | 2016-05-16 | 2016-10-26 | 山东省分析测试中心 | Method for rapidly measuring purity of pregabalin bulk drug based on H-nuclear magnetic resonance (HNMR) |
| CN106124550A (en) * | 2016-05-16 | 2016-11-16 | 山东省分析测试中心 | A kind of method quickly measuring vildagliptin purity based on hydrogen nuclear magnetic resonance |
| US20190227011A1 (en) * | 2016-06-28 | 2019-07-25 | Ocean University Of China | Method for measuring sterol content by utilizing quantitative nuclear magnetic resonance technology |
| JP2018090571A (en) * | 2016-11-29 | 2018-06-14 | 国立研究開発法人産業技術総合研究所 | 2,5-bis(aminomethyl)furan dihydrohalide and production method of the same, and production method of 2,5-bis(aminomethyl)furan |
| CN107421975A (en) * | 2017-07-17 | 2017-12-01 | 天津中医药大学 | The detection method of glycerol content in a kind of injection |
| CN108152316A (en) * | 2017-12-14 | 2018-06-12 | 中国日用化学工业研究院 | It is a kind of using maleic anhydride content and sulphonation rate as the quality inspection method of the glucosides sulfosuccinate ester salt product of index |
| CA3047665A1 (en) * | 2018-06-21 | 2019-12-21 | Oregon Institute of Science and Medicine | Metabolic profiling with magnetic resonance mass spectrometry (mrms) |
| CN109374667A (en) * | 2018-11-05 | 2019-02-22 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of NMR spectrum method identifying ultra-high-temperature sterilized milk and reconstituted milk |
| CN109991259A (en) * | 2018-12-11 | 2019-07-09 | 北京市食品安全监控和风险评估中心（北京市食品检验所） | The method for detecting a variety of small molecule water soluble substances in milk powder product simultaneously |
| CN109696449A (en) * | 2018-12-13 | 2019-04-30 | 北京市食品安全监控和风险评估中心（北京市食品检验所） | The method for detecting a variety of small molecule water soluble substances in milk powder product simultaneously |
| CN109632784A (en) * | 2019-01-21 | 2019-04-16 | 淮阴师范学院 | A kind of method of quantitative detection ascorbic acid |
| CN111855730A (en) * | 2020-08-12 | 2020-10-30 | 黄河三角洲京博化工研究院有限公司 | Method for testing water content in butyl rubber glue solution |
Non-Patent Citations (5)
| Title |
|---|
| HO CS,: "Rapid liquid chromatography electrospray tandem ma-ss spectrometry method for serum free and total carnitine", CLINICAL CHEMISTRY, 31 March 2003 (2003-03-31), pages 1189 - 1191 * |
| 张鑫鑫;杜利君;杨燕强;杨蒲晨;张琪;: "离子迁移谱在减肥类保健食品检测中的应用", 食品研究与开发, no. 13 * |
| 李先娜;董辉;孙晖;: "代谢组学在中医药现代化研究中的应用", 中医药信息, no. 05 * |
| 杨晓惠, 检测与标准, 28 February 2018 (2018-02-28), pages 40 - 43 * |
| 高泽岳: "左旋肉碱在食品检测中的研究新进展", 食品安全, 31 May 2019 (2019-05-31), pages 83 * |
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