KR20120014736A - Method for analyzing aspirin in plasma with liquid chromatography-mass spectrometry - Google Patents
Method for analyzing aspirin in plasma with liquid chromatography-mass spectrometry Download PDFInfo
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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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
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Abstract
Description
본 발명은 액체크로마토그래피-질량분석기를 이용한 혈장 중의 아스피린을 분석하는 방법에 관한 것이다.
The present invention relates to a method for analyzing aspirin in plasma using liquid chromatography-mass spectrometry.
아세틸살리실산(ASA; 아스피린)은 진통제, 항염증제, 해열제 및 항혈전제로 널리 이용되어 왔다. 이러한 아스피린은 생체 내에서 빠르게 분해되어 생리학적 활성을 나타내는 살리실산(salicylic acid; SA)으로 전환된다. 따라서, 약효를 나타내는 아스피린의 양을 분석하기 위해서는 살리실산의 혈중 농도를 측정하여야 하며, 이러한 살리실산의 혈중 농도는 액체크로마토그래피-질량분석기(liquid chromatography-mass spectrometry; LC-MS)를 이용하여 측정된다. Acetylsalicylic acid (ASA; aspirin) has been widely used as an analgesic, anti-inflammatory, antipyretic and antithrombotic agent. Such aspirin is rapidly degraded in vivo and converted into salicylic acid (SA), which exhibits physiological activity. Therefore, in order to analyze the amount of aspirin exhibiting the effect of blood, the concentration of salicylic acid must be measured, and the concentration of such salicylic acid is measured by liquid chromatography-mass spectrometry (LC-MS).
그러나 생체액(biological fluid) 중의 아스피린 및 살리실산을 정량하기 위하여 HPLC를 이용할 경우에는 2가지 문제점이 존재한다. 첫번째는, 아스피린은 물 또는 메탄올과 같은 양성자성 용매 및 혈장 중에서 살리실산으로 가수분해되는데, 물 중의 아스피린은 pH가 2-3일때 가장 우수한 안정성을 나타내지만, pH가 7일 경우에는 아스피린의 약 20%가 하루 이내에 살리실산으로 분해되어 버린다. 또한, 아스피린의 반감기는 37℃의 혈장 중에서 약 1시간으로 매우 짧기 때문에, 시료를 정량하기 위해 HPLC 시스템에 적용하기도 전에 일부 분해될 수 있다. 두 번째는, 살리실산은 증발 단계가 포함된 시료의 처리과정 동안 승화되어 일부 유실될 수 있다. 따라서, HPLC를 이용한 아스피린 양의 분석방법은 정확한 양을 측정하는 데에 어려움이 있었다.However, there are two problems when using HPLC to quantify aspirin and salicylic acid in biological fluids. First, aspirin is hydrolyzed to salicylic acid in plasma with protic solvents such as water or methanol, while aspirin in water has the best stability at pH 2-3, but at 20 pH it is about 20% of aspirin. Will be broken down into salicylic acid within one day. In addition, the half-life of aspirin is very short, about 1 hour in plasma at 37 ° C., so it may be partially degraded before application to the HPLC system to quantify the sample. Secondly, salicylic acid may be sublimed and partially lost during the processing of the sample including the evaporation step. Therefore, the method of analyzing the amount of aspirin using HPLC had difficulty in determining the correct amount.
따라서, 상기한 단점을 보완하기 위한 다양한 연구들이 진행되어 왔다. Therefore, various studies have been conducted to compensate for the above disadvantages.
문헌[Frieder Kees et al., Journal of Chromatography B, 677, 172-177(1996)]에서는 혈장 중의 아스피린이 분해되는 것을 방지하기 위하여 시료를 pH 2-3의 범위로 산성화한 후 냉각하여 -70℃에서 보관한 후 분석하는 방법을 개시하고 있다.Frieder Kees et al. , Journal of Chromatography B , 677, 172-177 (1996)], in order to prevent degradation of aspirin in plasma, acidify the sample to a pH 2-3 range, cool it, store it at -70 ° C, and then analyze it. Is starting.
또한, M.P. Bouche 등의 포스터[Combining selectivity from chromatography and fast ion separation by MS in bioanalysis]에서는 ESI(-)-LC-MS/MS 방법을 이용하였다. 하지만, 분석 시간이 길고 꼬리끌림(tailing) 현상이 발생하였으며, 높은 pH로 인하여 안정성의 일정하지 못하였다. 또한, 정량제한(LOQ)이 아스피린은 80 ng/ml 및 살리실산은 60 ng/ml로서 감도가 높지 않아 인간의 혈장 정량에는 적합하지 못하였다. In addition, M.P. In Bouche et al. [Combining selectivity from chromatography and fast ion separation by MS in bioanalysis], the ESI (-)-LC-MS / MS method was used. However, the analysis time was long, tailing (tailing) occurred, the stability was not constant due to the high pH. In addition, the limit of quantitation (LOQ) was 80 ng / ml for aspirin and 60 ng / ml for salicylic acid, which is not suitable for quantifying plasma in humans.
문헌[배수경 등, Biomed chromatogra 22: 590-595 (2008)] 및 문헌[Xiangrong Xu et al., Biomed chromatogra 23: 973-979 (2009)]에서는 이동상으로 아세토니트릴:0.1% 포름산 함유하는 물의 혼합액을 각각 80:20(v/v(%)) 및 63:37(v/v(%))로 사용하였으나, 완전한 분리는 이루어지지 않았다.In the literature (Drainer et al., Biomed chromatogra 22: 590-595 (2008)) and Xiangrong Xu et al., Biomed chromatogra 23: 973-979 (2009), a mixture of water containing acetonitrile: 0.1% formic acid as a mobile phase Was used at 80:20 (v / v (%)) and 63:37 (v / v (%)), respectively, but no complete separation occurred.
이에, 본 발명자들은 혈장 중의 아스피린 및 살리실산의 정확한 농도를 측정하기 위하여 예의 연구한 결과, 액체크로마토그래피-질량분석기를 이용하여 혈장 중의 아스피린과 살리실산을 안정하고 간편하게 분리할 수 있는 방법을 발견함으로써 본 발명을 완성하였다.
Therefore, the present inventors made a thorough study to determine the exact concentration of aspirin and salicylic acid in the plasma, by using a liquid chromatography-mass spectrometer to find a method for stably and conveniently separating the aspirin and salicylic acid in plasma Was completed.
따라서, 본 발명의 목적은 액체크로마토그래피-질량분석기를 이용한 혈장 중의 아스피린의 분석 방법을 제공하는 것이다.
It is therefore an object of the present invention to provide a method for the analysis of aspirin in plasma using liquid chromatography-mass spectrometry.
상기 목적을 달성하기 위해 본 발명은 The present invention to achieve the above object
1) 혈장 시료에 내부표준물질 및 추출용매를 첨가한 후 원심분리하여 상등액을 회수하는 단계; 1) recovering the supernatant by adding an internal standard and an extraction solvent to the plasma sample and centrifuging;
2) 상기 상등액을 농축한 후 아세토니트릴에 용해시키는 단계; 및 2) concentrating the supernatant and dissolving in acetonitrile; And
3) 상기 단계 2)에서 수득한 용해물을 액체 크로마토그래피-질량분석기로 정량분석하는 단계를 포함하는, 액체크로마토그래피-질량분석기를 이용한 혈장 중의 아스피린의 분석 방법을 제공한다.
3) It provides a method for analyzing aspirin in plasma using a liquid chromatography-mass spectrometer comprising the step of quantitatively analyzing the lysate obtained in step 2) by a liquid chromatography-mass spectrometer.
본 발명의 아스피린 분석법을 이용하면 분석 과정 중에 아스피린이 살리실산으로 분해되는 것을 최소화할 뿐만 아니라, 생체 내에서 생성된 살리실산을 우선적으로 분리하여 분석 과정에서 생성된 살리실산과의 혼합을 방지함으로써 혈중 살리실산의 양을 보다 정확하게 측정할 수 있다.
The aspirin assay of the present invention not only minimizes the breakdown of aspirin into salicylic acid during the analysis, but also preferentially separates salicylic acid produced in vivo and prevents mixing with salicylic acid produced in the analysis, thereby increasing the amount of salicylic acid in the blood. Can be measured more accurately.
도 1은 아스피린의 MS/MS 스펙트럼 결과를 나타낸 것이다.
도 2는 살리실산의 MS/MS 스펙트럼 결과를 나타낸 것이다.
도 3은 이부프로펜의 MS/MS 스펙트럼 결과를 나타낸 것이다.
도 4는 아스피린 250 ng/ml, 살리실산 78.125 ng/ml 및 내부 표준물질로서 이부프로펜(ISPD) 500 ng/mL인 경우의 액체크로마토그래피 측정 결과를 나타낸 것이다.Figure 1 shows the MS / MS spectral results of aspirin.
Figure 2 shows the MS / MS spectral results of salicylic acid.
Figure 3 shows the MS / MS spectral results of ibuprofen.
Figure 4 shows the results of liquid chromatography measurement of
본 발명에서는 1) 혈장 시료에 내부표준물질 및 추출용매를 첨가한 후 원심분리하여 상등액을 회수하는 단계; 2) 상기 상등액을 농축한 후 아세토니트릴에 용해시키는 단계; 및 3) 상기 단계 2)에서 수득한 용해물을 액체 크로마토그래피-질량분석기로 정량분석하는 단계를 포함하는, 액체크로마토그래피-질량분석기를 이용한 혈장 중의 아스피린의 분석 방법을 제공한다.1) recovering the supernatant by adding the internal standard and the extraction solvent to the plasma sample and then centrifuging; 2) concentrating the supernatant and dissolving in acetonitrile; And 3) quantifying the lysate obtained in step 2) with a liquid chromatography-mass spectrometer, which provides a method for analyzing aspirin in plasma using a liquid chromatography-mass spectrometer.
본 발명의 단계 1)에서는 혈장 시료에 내부표준물질 및 추출용매를 첨가한 후 원심분리하여 상등액을 회수한다. 이때, 상기 내부표준물질은 이부프로펜이 바람직하며, 추출 용매는 0.5% 아세트산을 함유하는 메틸 터트-부틸에테르(methyl tert-butylether)가 바람직하다. 혈장 중의 아스피린은 pH가 2 내지 3일 경우 가장 우수한 안정성을 나타내는데, 추출 용매로 0.5% 아세트산을 함유하는 메틸 터트-부틸에테르를 사용함으로써 pH를 일정하게 유지함으로써 안정성에 도움을 줄 수 있다.In step 1) of the present invention, the supernatant is recovered by adding an internal standard and an extraction solvent to the plasma sample, followed by centrifugation. At this time, the internal standard is preferably ibuprofen, the extraction solvent is preferably methyl tert-butylether containing 0.5% acetic acid. Aspirin in plasma shows the best stability when the pH is 2-3, which can help stability by keeping the pH constant by using methyl tert-butyl ether containing 0.5% acetic acid as the extraction solvent.
단계 2)에서는 상기 상등액을 건조 및 농축시켜 제조한 건고물에 아세토니트릴, 바람직하게는 50% 아세토니트릴을 첨가하여 재용해시킨다. In step 2), the supernatant is redissolved by adding acetonitrile, preferably 50% acetonitrile, to the dried product prepared by drying and concentrating.
단계 3)에서는 상기에서 제조된 용해물을 액체크로마토그래피-질량분석기에 투입한 후 액체크로마토그래피의 분석 컬럼을 통과하여 질량분석기로 도입한 시료를 이온화시켜 정량분석함으로써 혈장 중의 아스피린의 양을 분석할 수 있다.In step 3), the amount of aspirin in the plasma is analyzed by inputting the lysate prepared above to the liquid chromatography-mass spectrometer and ionizing and quantitatively analyzing the sample introduced into the mass spectrometer through the analysis column of the liquid chromatography. Can be.
이때, 상기 액체크로마토그래피의 이동상으로는 아세토니트릴 및 포름산의 혼합용매를 사용할 수 있으며, 바람직하게는, 70% 아세토니트릴 및 0.01% 포름산을 70:30(v/v%)으로 혼합한 혼합용매를 첨가하는 것을 특징으로 한다. In this case, a mixed solvent of acetonitrile and formic acid may be used as the mobile phase of the liquid chromatography, and preferably, a mixed solvent obtained by mixing 70% acetonitrile and 0.01% formic acid at 70:30 (v / v%) is added. Characterized in that.
또한 본 발명의 방법에서 상기 질량분석기로 분석시에는 네가티브 이온화 모드(negative ionization mode)로 이온화하고, 다중반응탐색(Multiple Reaction Monitoring; MRM) 모드에서 정량화하는 것을 특징으로 한다.In the method of the present invention, the mass spectrometer may be ionized in a negative ionization mode and quantified in a multiple reaction monitoring (MRM) mode.
본 발명에서는 MRM 모드에서 정량화된 아스피린, 살리실산 및 이부프로펜의 MS/MS 스펙트럼 결과를 각각 도 1 내지 3에 나타내었다.In the present invention, the MS / MS spectral results of aspirin, salicylic acid and ibuprofen quantified in MRM mode are shown in FIGS.
도 1에 나타난 바와 같이, 본 발명의 MRM 모드에서 확인된 아스피린의 MS/MS 스펙트럼은 Q1이 약 179(m/z), Q3이 약 137 및 93(m/z)에서 확인되었다.As shown in FIG. 1, the MS / MS spectrum of aspirin confirmed in the MRM mode of the present invention was found at about 179 (m / z) Q1 and about 137 and 93 (m / z) Q3.
도 2에 나타난 바와 같이, 본 발명의 MRM 모드에서 확인된 살리실산의 MS/MS 스펙트럼은 Q1이 약 137(m/z), Q3이 약 93 및 65(m/z)에서 확인되었다.As shown in FIG. 2, the MS / MS spectrum of salicylic acid identified in the MRM mode of the present invention was found at about 137 (m / z) for Q1 and at about 93 and 65 (m / z) for Q3.
도 3에 나타난 바와 같이, 본 발명의 MRM 모드에서 확인된 아스피린의 MS/MS 스펙트럼은 Q1이 약 205(m/z), Q3이 약 161(m/z)에서 확인되었다.
As shown in FIG. 3, the MS / MS spectrum of the aspirin confirmed in the MRM mode of the present invention was found at about 205 (m / z) for Q1 and about 161 (m / z) for Q3.
[실시예][Example]
이하, 본 발명을 하기 실시예에 의하여 더욱 상세하게 설명한다. 단, 하기 실시예는 본 발명을 예시하기 위한 것일 뿐, 본 발명의 범위가 이들만으로 한정되는 것은 아니다.
Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.
실시예 1: 검량선의 작성Example 1: Preparation of calibration curve
아스피린과 살리실산의 각 표준용액 100 ug/mL 및 2 mg/ml을 개의 공혈장으로 경시적으로 희석하여 각각 1000 ng/mL 및 10000 ng/ml을 최고 농도로 희석하여 표준시료(아스피린: 2 내지 1000 ng/mL 및 살리실산 20 내지 10000 ng/ml)로서 사용하였다. 이어, 농도별 표준시료를 20 uL씩 정확하게 취하여 뚜껑 달린 시험관에 옮겼다. 이어, 내부표준물질(이부프로펜 500 ng/mL) 50 uL를 가하고, 0.5% 아세트산 함유하는 메틸 터트-부틸에테르 1 mL을 첨가하여 10분간 1400rpm에서 혼합하고 원심분리하였다. 상등액을 취하여 35℃에서 진공건조기를 이용하여 농축시켰다. 농축된 건고물에 50% 아세토니트릴 500 uL를 첨가하여 재용해시킨 후 하기와 같이 액체크로마토그래피-질량분석을 수행하였다. 100 ug / mL and 2 mg / ml of each standard solution of aspirin and salicylic acid were diluted with dog co-plasma over time, diluting 1000 ng / mL and 10000 ng / ml to the highest concentration, respectively. ng / mL and salicylic acid 20-10000 ng / ml). Subsequently, 20 uL of the concentration-specific sample was accurately taken and transferred to a test tube with a lid. Then, 50 uL of internal standard (500 ng / mL of ibuprofen) was added, 1 mL of methyl tert-butylether containing 0.5% acetic acid was added, mixed at 1400 rpm for 10 minutes, and centrifuged. The supernatant was taken and concentrated at 35 ° C. using a vacuum dryer. After redissolved by adding 500 uL of 50% acetonitrile to the concentrated dry matter, liquid chromatography-mass spectrometry was performed as follows.
먼저 상기 시료를 하기 조건의 액체크로마토그래피(LC)에 적용시켰다.First, the sample was subjected to liquid chromatography (LC) under the following conditions.
<LC 조건><LC condition>
기기: HP1100 시리즈 (에질런트사(Agillent Co.), 미국);Instrument: HP1100 series (Agillent Co., USA);
컬럼: xterra MS C18(내경 2.1mm ⅹ 길이 50㎜, 입자크기 3.5㎛, 워터스사);Column: xterra MS C18 (inner diameter 2.1 mm ⅹ
컬럼 온도: 30℃;Column temperature: 30 ° C .;
이동상: 70% 아세토니트릴/0.01% 포름산 (70/30 (v/v%));Mobile phase: 70% acetonitrile / 0.01% formic acid (70/30 (v / v%));
유속: 200 uL/분;Flow rate: 200 uL / min;
주입량: 5 uL; 및Injection volume: 5 uL; And
분석시간: 5분.
Analysis time: 5 minutes.
이어, 상기 분석컬럼을 통과한 시료를 하기 조건의 질량분석기(MS, Mass Spectrophotometer)에 적용시켜 네가티브 이온화 모드(negative ionization mode)에서 이온화한 후, 하기 표 1에 기재된 MRM 모드에서 정량화하였다.Subsequently, the sample that passed the analysis column was subjected to a mass spectrometer (MS) under the following conditions, ionized in a negative ionization mode, and then quantified in the MRM mode described in Table 1 below.
<MASS 조건><MASS condition>
기기: API 4000 Qtrap(어플라이드 바이오시스템스사(Applied Biosystems)/MDS SCIEX, 캐나다);Instrument: API 4000 Qtrap (Applied Biosystems / MDS SCIEX, Canada);
이온화 모드: 터보 이온 스프레이 이온화 모드(네가티브);Ionization mode: turbo ion spray ionization mode (negative);
차단 가스(Curtain Gas; CUR): 15psi;CURtain Gas (CUR): 15 psi;
충돌 가스(Collision Gas; CAD): Medium;Collision Gas (CAD): Medium;
이온 전압: -4500V;Ion voltage: -4500 V;
GS 1: 50 psi;GS 1: 50 psi;
GS 2: 40 psi;GS 2: 40 psi;
터보가스 온도: 450℃; 및Turbogas temperature: 450 ° C .; And
CUR, CAD, GS(1, 2): 질소. CUR, CAD, GS (1, 2): nitrogen.
[표 1][Table 1]
DP: 탈군락화 전위(Declustering Potential), EP: 입구 전위(Entrance Potential), CE: 충돌 에너지(Collision Energy) 및 CXP: 충돌 셀 출구 전위(Collision Cell Exit potential).
DP: Declustering Potential, EP: Entrance Potential, CE: Collision Energy and CXP: Collision Cell Exit Potential.
이어, 측정된 내부표준물질의 피크면적에 대한 아스피린과 살리실산의 피크 면적비를 이용하여 검량선을 작성하였다. 검량선은 직선회귀방식(weighed (1/x) least squares)을 이용하여 구하였다. 도 4에 아스피린을 250 ng/ml 및 살리실산을 78.125 ng/ml 사용하였을 경우의 액체 크로마토그래피를 나타내었다.
Then, a calibration curve was prepared using the peak area ratio of aspirin and salicylic acid to the peak area of the measured internal standard. The calibration curve was obtained by using linear regression (weighed (1 / x) least squares). 4 shows liquid chromatography when 250 ng / ml of aspirin and 78.125 ng / ml of salicylic acid were used.
실시예 2: 시료의 분석Example 2: Analysis of Sample
아스피린을 투여한 개로부터 혈액을 채취하여 -70℃에서 보관하였다. 상기 시료를 실온에 방치하여 녹인 후 0.5분간 탁상용 혼합기에서 잘 섞은 다음 20 uL를 취하여 뚜껑 달린 시험관에 옮겼다. 이어, 내부표준물질(이부프로펜 500 ng/mL) 50 uL를 가하고, 0.5% 아세트산을 함유하는 메틸 터트-부틸에테르 1 mL을 첨가하여 10분간 1400rpm에서 혼합하고 원심분리하였다. 상등액을 취하여 35℃에서 진공건조기를 이용하여 농축시켰다. 농축된 건고물에 50% 아세토니트릴 500 uL를 첨가하여 재용해시킨 후, 상기 실시예 1에서 기재된 것과 동일한 방식으로 액체크로마토그래피-질량분석을 수행하였다. 상기 질량분석기로 분석한 아스피린 및 살리실산의 농도를 정량하기 위하여 실시예 1에서 작성한 검량선을 통해 혈장 중의 아스피린 및 살리실산의 농도를 정량화하였다. Blood was collected from dogs given aspirin and stored at -70 ° C. The sample was left at room temperature to dissolve, mixed well in a tabletop mixer for 0.5 minutes, and then taken to 20 μL and transferred to a test tube with a lid. Then, 50 uL of internal standard (Ibuprofen 500 ng / mL) was added, 1 mL of methyl tert-butylether containing 0.5% acetic acid was added, mixed at 1400 rpm for 10 minutes, and centrifuged. The supernatant was taken and concentrated at 35 ° C. using a vacuum dryer. After redissolution by adding 500 uL of 50% acetonitrile to the concentrated dry matter, liquid chromatography-mass spectrometry was performed in the same manner as described in Example 1 above. In order to quantify the concentrations of aspirin and salicylic acid analyzed by the mass spectrometer, the concentrations of aspirin and salicylic acid in plasma were quantified through the calibration curve prepared in Example 1.
이와 같이, 본 발명의 액체크로마토그래피-질량분석기를 이용한 혈장 중의 아스피린 분석 방법은 샘플링한 후에도 혈장 중의 아스피린과 살리실산의 함량 소실 없이 안정하고 간편하게 정량분석할 수 있다.As described above, the method for analyzing aspirin in plasma using the liquid chromatography-mass spectrometer of the present invention can be quantitatively and simply analyzed without loss of the content of aspirin and salicylic acid in plasma even after sampling.
Claims (4)
2) 상기 상등액을 농축한 후 아세토니트릴에 용해시키는 단계; 및
3) 상기 단계 2)에서 수득한 용해물을 액체 크로마토그래피-질량분석기로 정량분석하는 단계를 포함하는, 혈장 중의 아스피린 분석방법.1) recovering the supernatant by adding an internal standard and an extraction solvent to the plasma sample and centrifuging;
2) concentrating the supernatant and dissolving in acetonitrile; And
3) quantifying the lysate obtained in step 2) with a liquid chromatography-mass spectrometer.
상기 내부표준물질이 이부프로펜인 것을 특징으로 하는 분석방법.The method of claim 1,
The internal standard is ibuprofen.
상기 추출용매가 0.5% 아세트산을 함유하는 메틸 터트-부틸에테르(methyl tert-butylether; MTBE)인 것을 특징으로 하는 분석방법.The method of claim 1,
The extraction solvent is methyl tert-butylether (MTBE) containing 0.5% acetic acid.
상기 액체 크로마토그래피의 이동상이 70% 아세토니트릴과 0.01% 포름산의 70:30(v/v%) 혼합용매인 것을 특징으로 하는 분석방법.
The method of claim 1,
The mobile phase of the liquid chromatography is a 70:30 (v / v%) mixed solvent of 70% acetonitrile and 0.01% formic acid.
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| KR101495450B1 (en) * | 2013-11-22 | 2015-03-02 | 한국마사회 | A method for analysis of acidic drugs in equine blood sample using on-column methylation |
| KR102033857B1 (en) | 2018-05-30 | 2019-10-17 | 경상대학교산학협력단 | Method for increasing the amino acid recovery of serum or plasma and Analytical method for determination of free amino acid in serum or plasma using LC-MS/MS |
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| CN109085262A (en) * | 2018-08-03 | 2018-12-25 | 杭州佰勤医疗器械有限公司 | Serum plasma pharmaceutical extraction composition and application thereof |
| CN112485340A (en) * | 2019-11-27 | 2021-03-12 | 南京品生医学检验实验室有限公司 | Method for detecting 1, 5-sorbitan in plasma by ultra-high performance liquid chromatography tandem mass spectrometry |
| CN112345683A (en) * | 2020-11-24 | 2021-02-09 | 辽宁药联制药有限公司 | LC-MS simultaneous determination of ligustrazine and aspirin in blood plasma |
| CN114814018A (en) * | 2022-04-18 | 2022-07-29 | 合肥创新医药技术有限公司 | Method for determining doxylamine in human plasma through LC-MS/MS |
| CN114778737A (en) * | 2022-04-27 | 2022-07-22 | 天津国科医工科技发展有限公司 | Liquid chromatography detection sample pretreatment method capable of shortening time |
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| KR101495450B1 (en) * | 2013-11-22 | 2015-03-02 | 한국마사회 | A method for analysis of acidic drugs in equine blood sample using on-column methylation |
| KR102033857B1 (en) | 2018-05-30 | 2019-10-17 | 경상대학교산학협력단 | Method for increasing the amino acid recovery of serum or plasma and Analytical method for determination of free amino acid in serum or plasma using LC-MS/MS |
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