CN105954453A - Kit for simultaneously quantifying and detecting niacin, nicotinamide and pantothenic acid - Google Patents
Kit for simultaneously quantifying and detecting niacin, nicotinamide and pantothenic acid Download PDFInfo
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Abstract
The invention provides a kit for simultaneously quantifying and detecting niacin, nicotinamide and pantothenic acid. The kit mainly comprises a dried blood spot slice (Whatman protein saver 903) required for collecting whole peripheral blood, a capillary blood collection tube, niacin, nicotinamide and pantothenic acid vitamin standard substances and isotope internal standards thereof as well as trichloroacetic acid and further comprises preparation of dried blood spots, pre-treatment of the dried blood spots and detection. According to the kit for simultaneously quantifying and detecting niacin, nicotinamide and pantothenic acid, a dried blood spot collection technique and a high-performance liquid chromatography-tandem mass spectrum technique are combined to be used for quantification of B-vitamins, one-time minimally-invasive treatment and trace collection of blood, evaluation of nutrition conditions of various B-vitamins is also realized, and the work which is about detection of quantification of multiple vitamins under the condition of a small sample size and cannot be completed with a metabolic enzyme dependence method or an immunoassay method can be completed by means of the kit.
Description
Technical field
The invention belongs to vitamin quantitative detection field, nicotinic acid, nicotiamide and the test kit of pantothenic acid in a kind of simultaneous quantitative detection whole blood dried blood spot.
Background technology
Pantothenic acid (VB5), nicotinic acid (VB3), nicotiamide is the vitamin B group of small-molecular-weight, the lowest at people's in-vivo content, but the maintenance to body normal physiological function is most important.Nicotinic acid (VB3) and nicotiamide are transmitted and hydrogen migration by the center electron forming nicotinamide adenine dinucleotide (NAD) and nicotinamide-adenine dinucleotide phosphate (NADP) participation cellular metabolism, at least participate in more than 200 carbohydrate, fatty acid and amino acid metabolism reaction.Pantothenic acid (VB5) is the essential component of coenzyme A (CoA), and the acetyl-CoA that CoA and acetic acid are formed is the key factor of three big substance metabolismes, and also has important function in the acylation of protein.Because the wide application that VB3, VB5 are in cellular metabolism, it lacks or deficiency all can cause special shortage syndrome, maintains its best nutritional state most important.
Vitamin B group Nutrition Evaluation can find the suboptimal nutritional status of vitamin vitamin deficiency (vitamin storage mobilizes the phase or before obvious deficiency symptom occurs) in early days.Because the avitaminosis stage in early days is easy to correct, and vitamin shortage crowd in early days is compared with significant vitamin deficiency syndrome crowd, and number is more, and therefore, vitamin assessment is the most valuable to population health.
Most widely used vitamin appraisal procedure is the determination of activity that vitamin relies on enzyme, such as, measure the content of erythrocyte Sodium Sulfacetamide based transferase Activity Assessment pantothenic acid.Although vitamin relies on whether vitamin in enzyme assay can estimate actual metabolic condition indirect antimer lacks, but a metabolite enzyme determination of activity, only reflect the situation of a kind of vitamin, owing to can not detect multivitamin content simultaneously, and metabolic enzyme activity is vulnerable to the impact of genetic polymorphism, limit its extensive application in crowd's vitamin is assessed, assist diagnosis avitaminosis usually used as lab testing at present.
Body fluid (whole blood, blood plasma, serum, urine) in vitamin B group horizontal quantitative can be with direct reaction body vitamin situation, wherein containing the vitamin recently absorbing and being transported to each tissue in plasma/serum substrate, the vitamin absorption level that more reaction is recent, testing result is easily by dietary effects.In whole blood substrate, the erythrocytic half-life is 120 days, and therefore compared to serum/plasma, whole blood or erythrocyte more can react long-term vitamin situation.High performance liquid chromatography-tandem mass method (LC-MS/MS) is the method that in the complex matrices being widely used in recent years, trace materials is quantitative, compared with the method that traditional vitamin relies on enzyme assay and immunoassay, the method of LC-MS/MS is possible not only to the level of direct reaction vitamin, and can realize multivitamin simultaneous quantitative.During previously in document, LC-MS/MS is used for serum, blood plasma, urine or food, VB3 or VB5 is quantitative, the most not yet has application LC-MS/MS method to detect the research report of VB3 and VB5 level in whole blood substrate simultaneously.Main cause is as follows, firstly because cell is the place of metabolism, therefore whole blood substrate is compared with the most celliferous plasma/serum substrate, and substrate background is increasingly complex, and the pre-treatment to sample requires higher.Secondly, VB3 (nicotinic acid, nicotiamide), pantothenic acid is polar compound, on traditional reversed phase chromatographic column, reserve capability is more weak, inferior separating effect, although it is qualitative that mass spectrographic multiple-reaction monitoring pattern is independent of retention time, but good separation can reduce the endogenous chaff interference flowed out altogether with targeted vitamins, reduce matrix effect to greatest extent, and then ensure the accurate of quantitative result.
Dried blood spot Sampling techniques (DBS), will be collected on filter paper by Peripheral whole blood, compared with tradition venous blood collection, there is Wicresoft, store convenient transportation and Nurses level professional technology is required the multiple advantages such as low, it is highly suitable for blood specimen collection during crowd's vitamin condition evaluation on a large scale, is particularly suited for locality and does not possess vitamin detection by quantitative equipment, need to transport the outlying district of specimen.The combination of DBS Yu LC-MS/MS technology has become as an important method in little molecular method quantification analysis, but in the quantitative field of vitamin, does not the most apply DBS to combine LC-MS/MS technology and carry out the report that vitamin B group is quantitative.Except liquid whole blood needs the sample pre-treatments and the separation problem that solve, whole blood dried blood spot due to the botal blood volume seldom (50 μ) gathered, be also easy to occur vitamin to be measured can not meet detection quantitative limit so that can not be quantitative situation.
Summary of the invention
The present invention provides a kind of simultaneous quantitative detection nicotinic acid, nicotiamide and the test kit of pantothenic acid, this test kit application DBS combines isotopic dilution LC-MS/MS technology, use the pre-treating method of trichloroacetic acid solution ultrasonic extraction, can the vitamin B group such as nicotinic acid, nicotiamide and pantothenic acid in simultaneous quantitative denier whole blood sample.
The summary of the invention of the present invention is a kind of simultaneous quantitative detection nicotinic acid, nicotiamide and the test kit of pantothenic acid, mainly gathered required dried blood spot (Whatman protein saver 903) by Peripheral whole blood, capillary blood taking tube, nicotinic acid, nicotiamide and pantothenic acid vitamin standard substance and Isotopic Internal Standard thereof, trichloroacetic acid forms, also include dried blood spot prepare, dried blood spot pre-treatment and detecting step, wherein: prepared by dried blood spot, gather 50 μ L blood with capillary blood taking tube and gather on required dried blood spot in Peripheral whole blood, dark place stands to dry in the shade makes dried blood spot, seals and preserves；Dried blood spot pre-treatment, takes dried blood spot, adds the inner mark solution configured by Isotopic Internal Standard and the solution having trichloroacetic acid to configure, and vortex also makes liquid level soak dried blood spot, lucifuge supersound extraction, and vortex shakes, and centrifugal treating makes solution to be detected；Detecting step, takes part band detection solution, carries out LC-MS/MS analysis.Described vitamin standard substance are: deuterated nicotinic acid (Nicotinic-d4, No. CAS: 66148-15-0, purity: 98%), deuterated nicotiamide (Nicotinamide-d4),13C,15N for pantothenic acid (B5-[13C6,15N2], No. CAS: 137-08-6, purity: > 97%).Described Isotopic Internal Standard is: deuterated nicotinic acid, No. CAS: 66148-15-0, and purity 98%；Deuterated nicotiamide；13C, 15N for pantothenic acid, No. CAS: 137-08-6, purity > 97%.The configuration process of standard solution is as follows: weigh appropriate nicotinic acid, nicotiamide, pantothenic acid standard substance, adding 50g/L trichloroacetic acid to dissolve, preparation obtains concentration respectively is 150 μ g/mL, 700 μ g/mL, the nicotinic acid of 400 μ g/mL, nicotiamide, pantothenic acid standard solution storing solution, be stored in-20 DEG C of refrigerators；To be configured to, with above-mentioned three kinds of solution storing solutions, two grades of solution that concentration is 5 μ g/mL respectively with 50g/L trichloroacetic acid, then re-use the preparation of 3 kinds of two grades of solution to obtain Nicotinic Acid/Nicotinamide/pantothenic acid concentration and be respectively 0.5/2/1,1/4/2,2.5/10/5,5/20/10,25/100/50,50/200/100,80/320/160, the standard solution working solution mixed liquor of 100/400/200ng/mL and 1.5/6/3, the Quality Control solution mixed liquor of 30/120/60,75/300/150ng/mL, is stored in-20 DEG C.The configuration process of inner mark solution is as follows: weigh appropriate deuterated nicotinic acid, deuterated nicotiamide, 13C, 15N are for pantothenic acid, add 50g/L trichloroacetic acid to dissolve, preparation obtains concentration respectively is 160 μ g/mL, 90 μ g/mL, the deuterated nicotinic acid of 135 μ g/mL, deuterated nicotiamide, 13C, 15N, for pantothenic acid standard solution storing solution, are stored in-20 DEG C of refrigerators；Taking above-mentioned internal standard standard solution storing solution and be configured to deuterated nicotinic acid, deuterated nicotiamide, 13C, 15N, for the internal standard working solution that pantothenic acid concentration is 50/100/100ng/mL, are stored in-20 DEG C of refrigerators.Described dried blood spot preparation is specially, with 75% ethanol cotton piece sterilization finger pulp loss, after ethanol volatilizes, puncture skin with disposable blood taking needle, gathering 50 μ L droplets of whole blood on DBS card with capillary blood taking tube, dark place stands within 1-2 hour, to dry in the shade to be placed in lucifuge seals bag and preserves.Described dried blood spot pre-treatment and detecting step are specially, take dried blood spot, it is separately added into inner mark solution 50 μ L, 200g/L trichloroacetic acid solution 450 μ L, vortex also makes liquid level soak dried blood spot, lucifuge supersound extraction 30min, vortex concussion 5min, 13000rpm is centrifuged 5min, takes 40 μ L and carries out LC-MS/MS analysis.
Including chromatographically pure rank formic acid, analytical pure trichloroacetic acid, C8 reversed phase chromatographic column (ACE, 4.6*100mm).
This test kit is applicable to liquid chromatography-tandem mass spectrometry instrument (LC-MS/MS), electron spray (ESI) source cation many reflections monitoring (MRM) pattern, parent ion, daughter ion, goes the mass spectrometry parameters such as a bunch voltage (DP), collision voltage (CE), collision cell injection voltage (CXP) to be shown in Table 1.Used in chromatograph mobile phase A is 0.3% aqueous formic acid, and Mobile phase B is 0.3% formic acid methanol solution, and flow velocity is 0.8mL/min, and flowing phase condition is shown in Table 2.
The nicotinic acid of present invention offer, nicotiamide, pantothenic acid dried blood spot quantification kit, DBS with LC-MS/MS technology is combined, Wicresoft, the blood collection of trace realizes the assessment to multiple vitamin B group nutriture simultaneously, it is capable of detecting multivitamin paced work under the conditions of metabolic enzyme relies on the low sample size that method, immunoassay cannot complete simultaneously, reaches following Detection results:
(1) plurality of target vitamin is detected simultaneously
Trichloroacetic acid solution ultrasonic extraction can be by VB3 (nicotinic acid, nicotiamide), VB5 (pantothenic acid) dissolution from whole blood dried blood spot, and the at utmost protein in precipitation system, reach the purpose of enrichment and purification vitamin to be measured, in conjunction with the type of elution in table 2, three kinds of vitamin separating degrees to be measured are good, make detection method that the present invention set up can in single injected sampling the VB3 (nicotinic acid in simultaneous quantitative whole blood, nicotiamide), VB5 (pantothenic acid), waits 3 vitamin level.Quickly obtain testing result, time-consuming, man power and material's cost.
(2) highly sensitive, the quantitation curves range of linearity wide,
In the present invention, the lower limit of quantitation (LLOQ) of nicotinic acid, nicotiamide and pantothenic acid is respectively 0.5ng/mL, 2ng/mL, 1ng/mL, it is significantly better than the quantitative nicotinic acid of LC-MS/MS method in recent literature, nicotiamide and the lower limit of quantitation of pantothenic acid, the quantitative sensitivity that three kinds of vitamin are good makes in dried blood spot targeted vitamins in micro whole blood quantitatively be possibly realized.Exemplary operation curve is as it is shown in figure 1,3 kinds of vitamin B group standard curves all comprise 8 concentration levels and linearly good, and concrete lower limit of quantitation (LLOQ) and curvilinear equation, the range of linearity, correlation coefficient (R) are shown in Table 3.
(3) speed is analyzed fast
Under the conditions of the flowing shown in table 2 mutually, 3 target determinands realized good chromatographic isolation (Fig. 2) in 4 minutes, and the high analyte speed of 4 minutes detection by quantitative nicotinic acid, nicotiamide and pantothenic acid makes the present invention be applicable to crowd's sample analysis extensive, high-throughout.
(4) precision is good, accuracy is high
After the whole blood dried blood spot Quality Control sample of basic, normal, high concentration is carried out Sample pretreatment according to identical pre-treating method, the working curve criticized with same analysis calculates the concentration of quality-control sample, the veracity and precision of the measurement result computational methods according to Quality Control sample, the results are shown in Table 4, relative standard deviation (RSD%) in batch, between Pi, RSD% is respectively less than 10%, and accuracy is between 91.38%-103.80%.
(5) recovery of standard addition, matrix effect meet biological specimen LC-MS/MS standard measure requirement
After sample pre-treatments, the response rate of three concentration of pantothenic acid is respectively 124.41%, 97.65%, 82.17%, and average recovery rate value is 101.41%；The response rate of three concentration of nicotinic acid is respectively 69.98%, 80.56%, 81.15%, average recovery rate value is 77.23%, and the response rate of three concentration of nicotiamide is respectively 56.47%, 68.90%, 63.16%, and average recovery rate value is 62.84%, it is satisfied by corresponding requirements, the results are shown in Table 5.The investigation of matrix effect is that the solution after deriving from dried blood spot process prepared by 6 people's whole bloods is divided into two parts, a part adds pure standard solution (A), a part tests (B) as background, together enter LC-MS/MS with the pure standard solution of the equivalent (C) added to be analyzed, calculate corresponding vitamin matrix effect with (A-B)/C*100%.
Test result indicate that the matrix effect factor of nicotinic acid between 78.63% to 98.30%, meansigma methods is 91.14%；The matrix effect factor of nicotiamide is between 75.25% to 112.71%, and meansigma methods is 89.36%；The matrix effect factor of pantothenic acid is between 80.45% to 96.90%, and meansigma methods is 88.47% (table 6)；Between between the sample in 6 different substrates sources matrix effect after internal standard is corrected, relative standard deviation (RSD%) is less than 15%, illustrate under the experiment condition of the present invention, different substrates is little and stable on the impact of targeted vitamins peak area, and this result complies fully with the U.S. FDA requirement to analysis method confirmation mesostroma effect.
(6) specificity is high
The MRM pattern of tandem mass spectrum realizes qualitative to determinand by the daughter ion after gathering the parent ion of target determinand and fragmentation, having the different compounds of same molecular amount, still to produce the probability of identical fragmentation fragment after fragmentation minimum, and therefore mass spectrum MRM pattern possesses high specific.As seen in Figure 2, in bare substrate sample, there is no the response of targeted vitamins.
(7) store convenient transportation, can be used for basic unit crowd vitamin condition evaluation
The sample mode of dried blood spot is simple, blood sampling professional skill is required low, there is vitamin condition evaluation demand crowd to be in and prepare dried blood spot sample voluntarily,, the feature of solid-state little plus its volume and deposit the advantage with convenient transport so that the present invention is especially suitable for crowd's vitamin level examination.
Accompanying drawing explanation
Fig. 1 is the canonical plotting of nicotinic acid
Fig. 2 is the canonical plotting of nicotiamide
Fig. 3 is the canonical plotting of pantothenic acid
Fig. 4 is the MRM chromatogram of nicotinic acid
Fig. 5 is the MRM chromatogram of nicotiamide
Fig. 6 is the MRM chromatogram of pantothenic acid
Fig. 7 a, 7b are specificity figures
Detailed description of the invention
Below in conjunction with embodiment, the present invention will be further described.
Embodiment 1
Select the whole blood dried blood spot of less than 10 years old child of 15 example, take an ecchymosis, be equivalent to whole blood 50 μ L, being sequentially added into Isotopic Internal Standard solution 50 μ L, 200g/L trichloroacetic acid solution 450 μ L, vortex also makes liquid level soak dried blood spot, lucifuge supersound extraction 30min, vortex concussion 5min, 13000rpm are centrifuged 5min, take 40 μ L and carry out LC-MS/MS analysis.After same batch standard curve calculates, quantitatively draw the content (being shown in Table 7) of 3 kinds of vitamin B group in whole blood.
Fig. 1-3 nicotinic acid, nicotiamide, pantothenic acid standard working curve, abscissa is the ratio of vitamin and corresponding Isotopic Internal Standard concentration, and vertical coordinate is the ratio of vitamin and corresponding Isotopic Internal Standard response, and weight mode is 1/X2.
Nicotinic acid in Fig. 4-6 whole blood substrate, nicotiamide and pantothenic acid MRM chromatographic fractionation figure under the conditions of the flowing mutually of this test kit, wherein nicotinic acid, nicotiamide, the retention time of pantothenic acid is respectively 3.19min, 3.05min 3.58min, Isotopic Internal Standard retention time corresponding to three is respectively 3.14min, 3.01min, 3.56min, in figure before nicotinic acid retention time (at 3.05min), (3.28min after (at 3.52min) and nicotiamide-d4 retention time after nicotinic acid-d4 retention time, at 3.50min) all there is endogenous chaff interference to occur, good separating effect can effectively reduce substrate and flow out altogether, reduce matrix effect.
Fig. 7 blank dried blood spot is after pre-treatment sample introduction, and under MRM pattern, corresponding retention time goes out driftlessness vitamin and Isotopic Internal Standard response thereof.
Table 1:3 kind vitamin B group and the mass spectrometry parameters of Isotopic Internal Standard thereof
The flowing phase condition of table 2:3 kind vitamin B group chromatographic isolation
| Time (min) | A (%)/water (0.3% formic acid) | B (%)/methanol (0.3% formic acid) |
| 0-0.50 | 95 | 5 |
| 0.51-3.00 | 25 | 75 |
| 3.01-4.00 | 25 | 75 |
| 4.01-4.60 | 0 | 100 |
| 4.61-6.50 | 0 | 100 |
| 6.51-6.60 | 95 | 5 |
| 6.61-8.00 | 95 | 5 |
Table 3:3 kind vitamin B group standard curve equation, the range of linearity, R, weight mode and sensitivity (lower limit of quantitation)
Table 4: precision and accuracy
Table 5:3 kind vitamin B group response rate under experiment condition of the present invention
Table 6: the matrix effect (IS-ME%) of Internal standard correction methods and the relative standard deviation of the sample substrate effect in 6 different substrates sources
6 kinds of vitamin testing results in less than 10 years old children whole blood dried blood spot of table 7:15 example, wherein nicotiamide and individual one pantothenic acid concentration have exceeded the curvilinear range limit of mark, are therefore sample introduction quantitative result after diluted sample
Above content is structure and the work process further description made thereto combining the present invention, it is impossible to assert the present invention be embodied as be confined to these explanations.For general technical staff of the technical field of the invention, without departing from the inventive concept of the premise, it is also possible to make some simple deduction or replace, all should be considered as belonging to protection scope of the present invention.
Claims (7)
1. simultaneous quantitative detection nicotinic acid, nicotiamide and a test kit for pantothenic acid, mainly gathered by Peripheral whole blood
Required dried blood spot, capillary blood taking tube, nicotinic acid, nicotiamide and pantothenic acid vitamin standard substance and Isotopic Internal Standard thereof,
Trichloroacetic acid forms, it is characterised in that: also include dried blood spot prepare, dried blood spot pre-treatment and detecting step,
Wherein:
Prepared by dried blood spot, gather 50 μ L Peripheral whole bloods on dried blood spot with capillary blood taking tube, and dark place stands dries in the shade
Make dried blood spot, seal and preserve；
Dried blood spot pre-treatment, takes dried blood spot, adds the inner mark solution configured by Isotopic Internal Standard and has three chloroethenes
The solution of acid configuration, vortex also makes liquid level soak dried blood spot, lucifuge supersound extraction, and vortex shakes, centrifugal
Process, make solution to be detected；
Detecting step, takes part solution to be detected, carries out LC-MS/MS analysis.
A kind of simultaneous quantitative the most according to claim 1 detection nicotinic acid, nicotiamide and the test kit of pantothenic acid,
It is characterized in that: described vitamin standard substance are: nicotinic acid, No. CAS: 59-67-8, purity >=99.5%；
Nicotiamide, No. CAS: 98-92-0, purity >=99.5%)；Pantothenic acid, No. CAS: P5155.
A kind of simultaneous quantitative the most according to claim 1 detection nicotinic acid, nicotiamide and the test kit of pantothenic acid,
It is characterized in that: described Isotopic Internal Standard is: deuterated nicotinic acid, No. CAS: 66148-15-0, purity 98%；
Deuterated nicotiamide；13C, 15N for pantothenic acid, No. CAS: 137-08-6, purity > 97%.
A kind of simultaneous quantitative the most according to claim 2 detection nicotinic acid, nicotiamide and the test kit of pantothenic acid,
It is characterized in that: also including being configured to standard solution by vitamin standard substance, the configuration process of standard solution is such as
Under: weigh appropriate nicotinic acid, nicotiamide, pantothenic acid standard substance, add 50g/L trichloroacetic acid and dissolve, join respectively
Preparing concentration is 150 μ g/mL, and 700 μ g/mL, the nicotinic acid of 400 μ g/mL, nicotiamide, pantothenic acid standard are molten
Liquid storing solution, is stored in-20 DEG C of refrigerators；Respectively will be with above-mentioned three kinds of solution storing solutions with 50g/L trichloroacetic acid
It is configured to two grades of solution that concentration is 5 μ g/mL, then re-uses 3 kinds of two grades of solution preparations and obtain nicotinic acid/cigarette
Amide/pantothenic acid concentration is respectively 0.5/2/1, and 1/4/2,2.5/10/5,5/20/10,25/100/50,50/200/100,
The standard solution working solution mixed liquor of 80/320/160,100/400/200ng/mL and 1.5/6/3,30/120/60,
The Quality Control solution mixed liquor of 75/300/150ng/mL, is stored in-20 DEG C.
A kind of simultaneous quantitative the most according to claim 3 detection nicotinic acid, nicotiamide and the test kit of pantothenic acid,
It is characterized in that: also including configuring internal standard working solution, the configuration process of inner mark solution is as follows: weigh appropriate
Deuterated nicotinic acid, deuterated nicotiamide, 13C, 15N for pantothenic acid, add 50g/L trichloroacetic acid and dissolve, join respectively
Preparing concentration is 160 μ g/mL, 90 μ g/mL, the deuterated nicotinic acid of 135 μ g/mL, deuterated nicotiamide, 13C,
15N, for pantothenic acid standard solution storing solution, is stored in-20 DEG C of refrigerators；Take above-mentioned internal standard standard solution storing solution
Being configured to deuterated nicotinic acid, deuterated nicotiamide, 13C, 15N are the internal standard of 50/100/100ng/mL for pantothenic acid concentration
Working solution, is stored in-20 DEG C of refrigerators.
A kind of simultaneous quantitative the most according to claim 1 detection nicotinic acid, nicotiamide and the test kit of pantothenic acid,
It is characterized in that: described dried blood spot preparation is specifically, with 75% ethanol cotton piece sterilization finger pulp loss, treat ethanol
After volatilizing, puncture skin with disposable blood taking needle, gather 50 μ L droplets of whole blood in DBS card with capillary blood taking tube
On, dark place stands within 1-2 hour, to dry in the shade to be placed in lucifuge seals bag and preserves.
A kind of simultaneous quantitative the most according to claim 5 detection nicotinic acid, nicotiamide and the test kit of pantothenic acid,
It is characterized in that: described dried blood spot pre-treatment and detecting step are specifically, take dried blood spot, in being separately added into
Mark solution 50 μ L, 200g/L trichloroacetic acid solution 450 μ L, vortex also makes liquid level soak dried blood spot, keeps away
Light supersound extraction 30min, vortex concussion 5min, 13000rpm are centrifuged 5min, take 40 μ L and carry out
LC-MS/MS analyzes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610304169.5A CN105954453A (en) | 2016-05-10 | 2016-05-10 | Kit for simultaneously quantifying and detecting niacin, nicotinamide and pantothenic acid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610304169.5A CN105954453A (en) | 2016-05-10 | 2016-05-10 | Kit for simultaneously quantifying and detecting niacin, nicotinamide and pantothenic acid |
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Cited By (9)
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| CN106442790A (en) * | 2016-10-09 | 2017-02-22 | 汤臣倍健股份有限公司 | Method for detection of B vitamins |
| CN106770802A (en) * | 2017-02-23 | 2017-05-31 | 广州市丰华生物工程有限公司 | It is a kind of to detect multivitamin method and kit in dry blood spot simultaneously |
| CN106814147A (en) * | 2016-12-30 | 2017-06-09 | 广州市宜健医学技术发展有限公司 | The detection method of B family vitamin in food |
| CN108802213A (en) * | 2018-04-20 | 2018-11-13 | 首都儿科研究所 | Vitamin A detection dried blood spot reagent preparation box and detection method |
| CN110234974A (en) * | 2016-12-19 | 2019-09-13 | 生物梅里埃公司 | Blood sample is deposited on waterleaf paper and the method for subsequent mechanical extraction blood culture |
| US10656059B2 (en) | 2018-03-07 | 2020-05-19 | Alcala Pharmaceutical, Inc. | Method for qualitative and quantitative multiplexing of drug analytes from biological samples |
| CN112710767A (en) * | 2020-12-07 | 2021-04-27 | 浙江大学 | Tandem mass spectrometry kit for measuring NNMT catalytic substrate and NNMT catalytic product |
| CN114047267A (en) * | 2021-11-15 | 2022-02-15 | 贵州省烟草科学研究院 | Method for analyzing nicotinic acid in tobacco root system by derivatization-gas chromatography-mass spectrometry |
| CN117099774A (en) * | 2023-10-24 | 2023-11-24 | 西湖维泰(杭州)诊断技术有限公司 | Preservation and detection methods of nicotinic acid metabolite blood sample, kit and application of nicotinic acid metabolite blood sample |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106442790A (en) * | 2016-10-09 | 2017-02-22 | 汤臣倍健股份有限公司 | Method for detection of B vitamins |
| CN110234974B (en) * | 2016-12-19 | 2021-08-17 | 生物梅里埃公司 | Method for depositing blood samples on absorbent paper and subsequent mechanical extraction of blood cultures |
| CN110234974A (en) * | 2016-12-19 | 2019-09-13 | 生物梅里埃公司 | Blood sample is deposited on waterleaf paper and the method for subsequent mechanical extraction blood culture |
| CN106814147A (en) * | 2016-12-30 | 2017-06-09 | 广州市宜健医学技术发展有限公司 | The detection method of B family vitamin in food |
| CN106770802A (en) * | 2017-02-23 | 2017-05-31 | 广州市丰华生物工程有限公司 | It is a kind of to detect multivitamin method and kit in dry blood spot simultaneously |
| US10656059B2 (en) | 2018-03-07 | 2020-05-19 | Alcala Pharmaceutical, Inc. | Method for qualitative and quantitative multiplexing of drug analytes from biological samples |
| US11054349B2 (en) | 2018-03-07 | 2021-07-06 | Alcala Pharmaceutical, Inc. | Method for preparation of dried blood sample for multiplexing of analytes |
| CN108802213A (en) * | 2018-04-20 | 2018-11-13 | 首都儿科研究所 | Vitamin A detection dried blood spot reagent preparation box and detection method |
| CN112710767A (en) * | 2020-12-07 | 2021-04-27 | 浙江大学 | Tandem mass spectrometry kit for measuring NNMT catalytic substrate and NNMT catalytic product |
| CN114047267A (en) * | 2021-11-15 | 2022-02-15 | 贵州省烟草科学研究院 | Method for analyzing nicotinic acid in tobacco root system by derivatization-gas chromatography-mass spectrometry |
| CN114047267B (en) * | 2021-11-15 | 2023-11-21 | 贵州省烟草科学研究院 | Method for analyzing nicotinic acid in tobacco root system by derivatization-gas chromatography-mass spectrometry |
| CN117099774A (en) * | 2023-10-24 | 2023-11-24 | 西湖维泰(杭州)诊断技术有限公司 | Preservation and detection methods of nicotinic acid metabolite blood sample, kit and application of nicotinic acid metabolite blood sample |
| CN117099774B (en) * | 2023-10-24 | 2024-03-15 | 西湖维泰(杭州)诊断技术有限公司 | Preservation and detection methods of nicotinic acid metabolite blood sample, kit and application of nicotinic acid metabolite blood sample |
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