CN115406998A - Quantitative determination method for platinum prodrug in biological sample - Google Patents
Quantitative determination method for platinum prodrug in biological sample Download PDFInfo
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- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 title claims abstract description 111
- 229910052697 platinum Inorganic materials 0.000 title claims abstract description 51
- 239000012472 biological sample Substances 0.000 title claims abstract description 29
- 229940002612 prodrug Drugs 0.000 title claims abstract description 22
- 239000000651 prodrug Substances 0.000 title claims abstract description 22
- 238000004445 quantitative analysis Methods 0.000 title claims abstract description 8
- 238000009616 inductively coupled plasma Methods 0.000 claims abstract description 25
- 238000000034 method Methods 0.000 claims abstract description 25
- 239000000523 sample Substances 0.000 claims abstract description 13
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 12
- 238000005516 engineering process Methods 0.000 claims abstract description 6
- 230000035945 sensitivity Effects 0.000 claims abstract description 4
- 238000001927 high performance liquid chromatography-inductively coupled plasma mass spectrometry Methods 0.000 claims abstract description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 75
- 239000003814 drug Substances 0.000 claims description 26
- 229940079593 drug Drugs 0.000 claims description 22
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 21
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims description 10
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 10
- 239000007789 gas Substances 0.000 claims description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 9
- 239000012071 phase Substances 0.000 claims description 9
- DWAFYCQODLXJNR-BNTLRKBRSA-L oxaliplatin Chemical class O1C(=O)C(=O)O[Pt]11N[C@@H]2CCCC[C@H]2N1 DWAFYCQODLXJNR-BNTLRKBRSA-L 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 8
- 238000005259 measurement Methods 0.000 claims description 7
- 229960001756 oxaliplatin Drugs 0.000 claims description 7
- 230000006920 protein precipitation Effects 0.000 claims description 6
- 238000005070 sampling Methods 0.000 claims description 6
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- 229910052786 argon Inorganic materials 0.000 claims description 5
- 239000012159 carrier gas Substances 0.000 claims description 5
- 239000001307 helium Substances 0.000 claims description 5
- 229910052734 helium Inorganic materials 0.000 claims description 5
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 claims description 5
- 229910052757 nitrogen Inorganic materials 0.000 claims description 5
- 229920001223 polyethylene glycol Polymers 0.000 claims description 5
- 238000007664 blowing Methods 0.000 claims description 4
- 238000000589 high-performance liquid chromatography-mass spectrometry Methods 0.000 claims description 4
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 claims description 4
- 239000002253 acid Substances 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
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- 190000008236 carboplatin Chemical compound 0.000 claims description 3
- 229960004316 cisplatin Drugs 0.000 claims description 3
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 claims description 3
- XSMVECZRZBFTIZ-UHFFFAOYSA-M [2-(aminomethyl)cyclobutyl]methanamine;2-oxidopropanoate;platinum(4+) Chemical compound [Pt+4].CC([O-])C([O-])=O.NCC1CCC1CN XSMVECZRZBFTIZ-UHFFFAOYSA-M 0.000 claims description 2
- 210000003608 fece Anatomy 0.000 claims description 2
- 238000000703 high-speed centrifugation Methods 0.000 claims description 2
- 229950008991 lobaplatin Drugs 0.000 claims description 2
- 238000000746 purification Methods 0.000 claims description 2
- 210000002966 serum Anatomy 0.000 claims description 2
- 210000002700 urine Anatomy 0.000 claims description 2
- 238000003556 assay Methods 0.000 claims 3
- 239000007791 liquid phase Substances 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 8
- 229920000642 polymer Polymers 0.000 abstract description 6
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- 206010051779 Bone marrow toxicity Diseases 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
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- 238000002512 chemotherapy Methods 0.000 description 1
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- 210000003462 vein Anatomy 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
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- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
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Abstract
The invention relates to a quantitative determination method of a platinum prodrug in a biological sample, which can be used for the pharmacokinetics and toxicity kinetics research of the platinum prodrug. The method comprises the steps of firstly preparing a standard curve, then measuring the concentration of the platinum prodrug in a biological sample, and then calculating the content of the platinum prodrug in a sample to be detected through the standard curve. The detection method is based on a High Performance Liquid Chromatography (HPLC) and inductively coupled plasma mass spectrometry (ICP/MS) combined technology, the platinum prodrug is qualitatively separated through HPLC, and the content of the platinum prodrug in a biological sample is determined through ICP/MS. The invention has the characteristics of high sensitivity, good reproducibility and the like, and can solve the pharmacokinetic research problem caused by the large molecular weight and the non-unique molecular weight of the polymer-coupled platinum prodrug.
Description
Technical Field
The invention relates to a quantitative determination method of a platinum prodrug in a biological sample, in particular to a quantitative determination method of a polymer coupled platinum prodrug in a biological sample.
Background
The platinum drugs including cisplatin, carboplatin, oxaliplatin and the like are developed and marketed in the 60 th century in the 20 th century, have unique anticancer mechanism and wide anticancer spectrum, and become one of the most widely used chemotherapeutic drugs in clinic at present. Although the curative effect is remarkable, the platinum drugs have poor targeting property, are easy to cause serious gastrointestinal tract reaction and nerve and bone marrow toxicity, have strong dose limiting toxicity and seriously limit the clinical use effect and the administration dose of the platinum drugs. In recent years, a drug delivery strategy based on nanotechnology and a polymer coupling technology provide a new solution for oxaliplatin chemotherapy. For example, in CN106074379B reported by Shanghai medicine of Chinese academy of sciences, oxaliplatin is oxidized, platinum is changed from a high-bioactivity divalent state into a low-bioactivity tetravalent state, and the platinum can be self-assembled into micelles after being coupled by polyethylene glycol, so that the in-vivo metabolism behavior of the medicine can be improved, the systemic, bone marrow and immune toxicity of the platinum medicine are reduced, and the anti-tumor activity is improved.
The in vivo metabolism behavior of the drug has important guiding significance for the research and development of new drugs, but the existing determination method has certain limitation on the in vivo metabolism research of platinum drugs. After the platinum drugs are modified by polymers, the molecular weight of the drugs is in a normal distribution form, the conventional HPLC-MS/MS is large in challenge, and effective tracking of active ingredients of the platinum drugs is lacked. Traditional platinum drugs usually adopt ICP/MS to measure total platinum in biological samples to study pharmacokinetic behaviors in vivo, but the method for measuring total platinum by using polymer modified platinum prodrugs cannot effectively distinguish pharmacokinetics behaviors of bivalent platinum and tetravalent platinum in vivo, so that clinical transformation of the polymer modified platinum prodrugs is limited. The high performance liquid chromatography is a commonly used detection method for detecting the platinum drugs, but the detection limit is 1 mug/mL by adopting an HPLC-UV method, and the detection method is not suitable for detecting the content of the platinum prodrugs in a biological sample.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a quantitative determination method of a platinum prodrug in a biological sample, which has the advantages of simple operation, reliable result, specific number, high sensitivity, short analysis time and good linear relation. The method adopts an HPLC-ICP/MS combined technology, wherein HPLC can well separate platinum metabolites with different valence states, and the ICP/MS method can measure the content of platinum drugs with different valence states.
In the combined method, the liquid chromatography mobile phase selects methanol, and compared with acetonitrile and other organic solvents, the methanol has higher theoretical plate number of chromatographic peak when measuring platinum metabolites and has better chromatographic behavior. The addition of acetic acid in the water phase can make the chromatographic peak width narrower, thus being suitable for detection.
The specific technical scheme is as follows:
a quantitative determination method of a platinum prodrug in a biological sample adopts a High Performance Liquid Chromatography (HPLC) and inductively coupled plasma mass spectrometry (ICP/MS) combined technology, and the determination steps comprise:
a. extracting the obtained biological sample;
b. establishing a drug determination standard curve;
c. and (c) measuring the sample to be detected by using HPLC-ICP/MS, and calculating the drug concentration of the platinum prodrug in the sample to be detected by using the standard curve obtained in the step b.
Further, biological samples include plasma, serum, cells, tissue homogenates, urine, and feces.
Further, the biological sample processing method in the step a comprises a methanol protein precipitation method, an acetonitrile protein precipitation method and a column purification method.
And a further step a, adding 5-10 times of methanol into the biological sample by adopting a methanol protein precipitation method, uniformly mixing by vortex, carrying out high-speed centrifugation to extract supernatant, putting the supernatant into a nitrogen blowing instrument for volatilizing, adding methanol for redissolving, and then using the obtained product for measuring the content of the platinum drugs.
Still further, the processing method comprises the following specific steps: adding 10 times of methanol into a certain amount of biological sample, vortex mixing, centrifuging at 12000rpm for 10min in a centrifuge, collecting supernatant, volatilizing in a nitrogen blower, adding 200 μ L methanol, and vortex for 10min to obtain the final product.
Further, the operation procedure of step b is:
1) Preparing bivalent platinum standard solutions with different diluted concentrations;
2) Adding a bivalent platinum standard solution and a blank plasma sample into an EP tube, uniformly mixing by vortex, adding methanol, mixing by vortex, centrifuging to obtain a supernatant, volatilizing the methanol under a nitrogen blowing instrument, adding the methanol, concentrating and redissolving, centrifuging, and injecting the supernatant into HPLC-ICP/MS for analysis;
3) And (3) repeating the operation 2) on the platinum standard solutions with different concentrations, recording a chromatogram, taking the divalent platinum concentration as a horizontal coordinate and the chromatographic peak area as a vertical coordinate, and performing regression operation by using a weighted least square method to obtain a linear equation, namely the standard curve.
Furthermore, by adopting the high performance liquid chromatography and inductively coupled plasma mass spectrometry combined technology,
wherein, the high performance liquid chromatograph adopts the following chromatographic conditions:
mobile phase: organic phase-aqueous phase
And (3) chromatographic column: reversed C18 column, preferably Xbridge column (waters, 250mm 4.6mm,5 μm)
Column temperature: 20-40 deg.C, preferably 30 deg.C
Flow rate: 1mL/min
Sample introduction amount: 10 μ L-100 μ L, preferably 20 μ L
The organic phase in the mobile phase adopts methanol, the water phase is an aqueous solution system containing acetic acid, the percentage content of the acetic acid is 1-0.05 percent, and the preference is 0.1 percent, namely the mobile phase of the high performance liquid chromatography adopts a methanol A-aqueous solution B system containing acid to carry out isocratic elution.
The organic phase-aqueous phase volume ratio is a: B =70, preferably 75-95, further preferably 90.
Wherein, the ICP/MS measuring conditions are as follows: argon is used as ionization gas, helium is used as carrier gas, the flow rate of an atomizer is 1L/min, the flow rate of a cooler is 10-14L/min, auxiliary gas is 0.8L/min, the sampling depth is 3-8mm, the plasma power is 1200-1800W, and the measurement is carried out according to the molecular weight of platinum atoms.
Further, ICP/MS measurement conditions are as follows: a kinetic energy discrimination mode (KED mode) is adopted, argon is used as ionized gas, helium is used as carrier gas, the flow rate of an atomizer is 1L/min, the flow rate of a cooler is 14L/min, auxiliary gas is 0.8L/min, the sampling depth is 5mm, the plasma power is 1500W, the pump speed is 40rpm, and the measurement is carried out according to the molecular weight of platinum atoms.
The HPLC can separate the platinum drugs with different valence states, the ICP/MS can measure the platinum content with high sensitivity, and the HPLC-ICP/MS combination can be applied to the separation and detection of the platinum drugs with different valence states in a biological sample.
Further, the platinum prodrug comprises oxaliplatin modified by polyethylene glycol, cisplatin, carboplatin and lobaplatin, and preferably the oxaliplatin modified by polyethylene glycol.
Drawings
FIG. 1 is a limit of quantitation (5 ng/mL) chromatogram of HPLC-ICP/MS in the present application;
FIG. 2 is a graph of PEG-OXA total platinum versus tetravalent platinum in vivo drug delivery in the present application.
Detailed Description
The principles and features of this invention are described below in conjunction with examples, which are set forth to illustrate, but are not to be construed to limit the scope of the invention.
Materials and equipment: pegylated oxaliplatin (PEG-OXA), provided by the cigarette desk drug research institute according to the synthetic process of CN109988297B, with the purity of more than 98.5%. Reagents such as chromatographic methanol and acetic acid are purchased from the national medicine group, the high performance liquid is Agilent 1200 type, and the inductively coupled plasma mass spectrometry (ICP/MS) is Agilent 7500 type.
EXAMPLE 1 creation of Standard Curve
Preparing a series of standard solutions:
precisely weighing 25mg of PEG-OXA standard substance, placing the PEG-OXA standard substance in a 25mL volumetric flask, adding methanol to dissolve and shake the PEG-OXA standard substance evenly, diluting the PEG-OXA standard substance to a scale mark, and preparing a standard stock solution of 1 mg/mL. Taking a proper amount of the stock solution, respectively diluting the stock solution to 10, 20, 200, 1000 and 2000ng/mL by using methanol, and uniformly shaking the stock solution for later use.
Preparing a standard curve in a body:
and (3) taking 10 mu L of blank plasma, respectively adding the same amount of the prepared series of standard solutions, and performing vortex for 30s to prepare series of standard solutions with the in vivo concentration of 5, 10, 100, 500 and 1000 ng/mL. Adding 1mL of methanol, uniformly mixing by vortex, placing in a centrifuge at 12000rpm for 10min, taking supernatant, placing under a nitrogen blowing instrument to volatilize the methanol, and adding 100 mu L of methanol to carry out vortex redissolution.
High performance liquid chromatography conditions:
mobile phase methanol: 0.1% aqueous acetic acid =90:10
Column temperature: 30 deg.C
Flow rate: 1mL/min
Sample injection amount: 20 μ L
A chromatographic column: XBridge C18 (Waters, 250mm 4.6mm,5 μm)
The high performance liquid chromatography system comprises a binary infusion pump, an automatic sample injector and a column incubator.
ICP/MS conditions:
the measurement is carried out according to the molecular weight of the platinum element by adopting a KED mode, taking argon as an ionized gas, taking helium as a carrier gas, taking the flow rate of an atomizer as 1L/min, the flow rate of a cooler as 14L/min, taking auxiliary gas as 0.8L/min, taking the sampling depth as 5mm, taking the plasma power as 1500W and taking the pump speed as 40 rpm.
When the standard substance is detected under the conditions, the signal-to-noise ratio of the sample with the concentration of 5ng/mL is more than 10, and the quantitative requirement is met, which is detailed in figure 1. Standard curve: y =0.047x +0.117R 2 ＝0.998。
Example 2 detection of Pt content of PEG-OXA in biological samples by HPLC-ICP/MS
After reconstitution of PEG-OXA for injection, blood was collected from the orbital venous plexus of rats at the following time points, respectively, after administration to the tail vein of rats at a dose of 5 mg/kg: 0,0.05,0.083,0.5,1,2,4,8, 12, 24h, using the biological sample treatment method of example 1, preparing a sample to be measured, sequentially entering an HPLC-ICP/MS system, carrying out quantitative determination according to the molecular weight of platinum element, substituting the measured result into the standard curve of example 1, determining the concentration of the sample, and drawing a pharmaceutical time curve, wherein the result is shown in figure 2.
Example 3 detection of Total Pt content of PEG-OXA in biological samples by ICP/MS
On the basis of example 2, 100. Mu.L of the plasma sample of the same group was digested overnight at 40 degrees by adding 5mL of nitric acid. After cooling, adding purified water to a constant volume of 50mL, sampling, quantifying the platinum element according to the following ICP/MS conditions, and determining the total Pt content of the biological sample PEG-OXA.
ICP/MS conditions: the measurement is carried out according to the molecular weight of the platinum element by adopting a KED mode, taking argon as ionized gas and helium as carrier gas, taking the flow rate of an atomizer as 1L/min, the flow rate of a cooler as 14L/min, auxiliary gas as 0.8L/min, sampling depth as 5mm, plasma power as 1500W and pump speed as 40 rpm.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (9)
1. A quantitative determination method of a platinum prodrug in a biological sample is characterized in that a High Performance Liquid Chromatography (HPLC) and inductively coupled plasma mass spectrometry (ICP/MS) combined technology is adopted, and the determination steps comprise:
a. extracting the obtained biological sample;
b. establishing a drug determination standard curve;
c. and (c) measuring the sample to be detected by using HPLC-ICP/MS, and calculating the drug concentration of the platinum prodrug in the sample to be detected by using the standard curve obtained in the step b.
2. The method as claimed in claim 1, wherein HPLC is used to separate platinum drugs with different valence states, ICP/MS is used to measure platinum content with high sensitivity, and HPLC-ICP/MS can be used to separate and detect platinum drugs with different valence states in biological samples.
3. The assay of claim 1, wherein the biological sample comprises plasma, serum, cells, tissue homogenates, urine, and feces.
4. The method according to claim 1, wherein the biological sample treatment method of step a comprises methanol protein precipitation, acetonitrile protein precipitation, and column purification.
5. The determination method according to claim 4, wherein the biological sample treatment method in the step a adopts a methanol protein precipitation method, 5-10 times of methanol is added into the biological sample, the biological sample is uniformly mixed by vortex, the supernatant is extracted by high-speed centrifugation, the supernatant is placed in a nitrogen blowing instrument to be volatilized, and the platinum drug content determination is performed after the methanol is added for redissolution.
6. The determination method according to claim 1, wherein a high performance liquid chromatography and inductively coupled plasma mass spectrometry technique is adopted, wherein a mobile phase of the high performance liquid chromatography is eluted at equal speed by adopting a methanol A-acid-containing aqueous solution B system, the acid is acetic acid, the percentage content of the acetic acid is 1% -0.05%, a chromatographic column under liquid phase conditions is a reversed C18 column, the column temperature is 20-40 ℃, and the mobile phase ratio is A: B = 70.
7. The method according to claim 1, wherein a high performance liquid chromatography and inductively coupled plasma mass spectrometry technique is used, wherein the ICP/MS measurement conditions are as follows: argon is used as ionized gas, helium is used as carrier gas, the flow rate of the atomizer is 1L/min, the flow rate of the cooler is 10-14L/min, the auxiliary gas is 0.8L/min, the sampling depth is 3-8mm, the plasma power is 1200-1800W, and the measurement is carried out according to the molecular weight of platinum atoms.
8. An assay method as claimed in claim 1 wherein said platinum prodrug comprises polyethylene glycol modified oxaliplatin, cisplatin, carboplatin, lobaplatin.
9. The assay of claim 8, wherein the platinum prodrug is oxaliplatin modified with polyethylene glycol.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202211160560.4A CN115406998A (en) | 2022-09-22 | 2022-09-22 | Quantitative determination method for platinum prodrug in biological sample |
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| CN202211160560.4A CN115406998A (en) | 2022-09-22 | 2022-09-22 | Quantitative determination method for platinum prodrug in biological sample |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140329272A1 (en) * | 2013-05-02 | 2014-11-06 | The Board Of Trustees Of The Leland Stanford Junior University | Metal-Based Covalent Viability Reagent for Single Cell Analysis |
| CN108169369A (en) * | 2017-12-19 | 2018-06-15 | 岛津企业管理（中国）有限公司 | The method that LC-ICP-MS measures Pt constituent contents in blood plasma |
| CN114441671A (en) * | 2021-12-23 | 2022-05-06 | 北京悦康科创医药科技股份有限公司 | Method for determining content of platinum impurity cyclohexanediamine dihydrate in oxaliplatin by combination of HPLC-ICP-MS |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140329272A1 (en) * | 2013-05-02 | 2014-11-06 | The Board Of Trustees Of The Leland Stanford Junior University | Metal-Based Covalent Viability Reagent for Single Cell Analysis |
| CN108169369A (en) * | 2017-12-19 | 2018-06-15 | 岛津企业管理（中国）有限公司 | The method that LC-ICP-MS measures Pt constituent contents in blood plasma |
| CN114441671A (en) * | 2021-12-23 | 2022-05-06 | 北京悦康科创医药科技股份有限公司 | Method for determining content of platinum impurity cyclohexanediamine dihydrate in oxaliplatin by combination of HPLC-ICP-MS |
Non-Patent Citations (1)
| Title |
|---|
| 曹鹏 等: "LC-MS/MS法联合ICP-MS法测定非小细胞肺癌患者血浆培美曲塞及顺铂的浓度", 中国医院药学杂志, vol. 42, no. 12, 30 June 2022 (2022-06-30), pages 1181 - 1185 * |
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