Analysis of Cisplatin and its Variants in Water · 2020. 2. 14. · Analysis of Cisplatin and its...
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Analysis of Cisplatin and its Variants in Water COSMOSIL πNAP Technical Note 18 Data courtesy of Prof. Ryuji Kato, Ph D., Laboratory of Cardiovascular Pharmacotherapy and Toxicology, Osaka University of Pharmaceutical Sciences COSMOSIL πNAP uses a naphthalene-based bonded phase, and exhibits different retention behavior from C 18 . In this note, πNAP is used to analyze cisplatin (CDDP) and its mono-chloro, none-chloro (hydration complex), and OH-dimer forms in water. Standard C 18 columns have difficulty separating compounds of similar hydrophobicity. Perfomance may be improved by using Cholester, a column with improved molecular shape recognition. (1)Experiment Summary Cisplatin (CDDP) in water converts to its mono-chloro, hydration complex, and OH-dimer forms, and it is thought that they exist in an equilibrium state (fig. 1). Each of these forms was separated and quantified. Cl H 3 N H 3 N Cl Pt -Cl - , +H 2 O +Cl - , -H 2 O +H + -H + +H + -H + +H + -H + OH H 3 N H 3 N Cl -Cl - , +H 2 O +Cl - , -H 2 O -Cl - , +H 2 O -2H 2 O +2H 2 O +Cl - , -H 2 O OH H 3 N H 3 N OH OH 2 H 3 N H 3 N Cl Pt Pt 2 OH 2 H 3 N H 3 N OH 2 Pt OH 2 H 3 N H 3 N OH Pt Pt OH-dimer O H 3 N H 3 N O NH 3 H H NH 3 Pt Pt CDDP None-chloro form 2 Cisplatin Mono-chloro form Fig. 1. Equilibrium reactions of cisplatin (CDDP) in water In water, CDDP loses Cl - , which is replaced with water. Depending on the pH, it may be replaced by a hydroxide ion. The mono-chloro form may lose another Cl - , and, in the same way, the water molecules (one or both) may be replaced with a hydroxide ion. These forms are in equilibrium. (2)Quantification of Cisplatin (CDDP) and its Mono-Chloro, None-Chloro, and OH-Dimer Forms ■ Comparing a C 18 column, COSMOSIL Cholester, and πNAP under identical conditions When measuring the OH-dimer nitrate form, the dissociated NO 3 - peak elutes before the OH-dimer peak. C 18 -MS-II could not separate the two peaks, and Cholester achieved partial separation, but could not separate to baseline. πNAP, however, was able to separate the compounds completely. NACALAI TESQUE, INC Column: COSMOSIL ** Column size: 4.6 mmI.D.-250 mm Mobile phase: 100 mmol/l NaClO 4 /Acetonitrile/100 mmol/l HClO 4 = 290/10/3 Flow rate: 1.0 ml/min Temperature: 37 C Detection: UV225 nm Sample: Cisplatin OH-dimer nitrate (2 mmol/l) 1; Nitrate 2; Cisplatin OH-dimer Injection Vol. 20 μl Data courtesy of Ryuji Kato, Ph D., Laboratory of Cardiovascular Pharmacotherapy and Toxicology, Osaka University of Pharmaceutical Sciences COSMOSIL 5C 18 -MS-II COSMOSIL Application Data COSMOSIL Cholester COSMOSIL πNAP
Transcript of Analysis of Cisplatin and its Variants in Water · 2020. 2. 14. · Analysis of Cisplatin and its...
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Analysis of Cisplatin and its Variants in WaterCOSMOSIL πNAP
Technical Note18
Data courtesy of Prof. Ryuji Kato, Ph D., Laboratory of Cardiovascular Pharmacotherapy and Toxicology, Osaka University of Pharmaceutical Sciences
COSMOSIL πNAP uses a naphthalene-based bonded phase, and exhibits different retention behavior from C18. In this note, πNAP is used to analyze cisplatin (CDDP) and its mono-chloro, none-chloro (hydration complex), and OH-dimer forms in water.Standard C18 columns have difficulty separating compounds of similar hydrophobicity. Perfomance may be improved by using Cholester, a column with improved molecular shape recognition.
(1)Experiment SummaryCisplatin (CDDP) in water converts to its mono-chloro, hydration complex, and OH-dimer forms, and it is thought that they exist in an equilibrium state (fig. 1). Each of these forms was separated and quantified.
ClH3N
H3N ClPt
-Cl-, +H2O
+Cl-, -H2O
+H+-H+ +H+-H+
+H+-H+
OHH3N
H3N Cl
-Cl-, +H2O
+Cl-, -H2O
-Cl-, +H2O
-2H2O
+2H2O
+Cl-, -H2O
OHH3N
H3N OH
OH2H3N
H3N Cl
Pt
Pt
2OH2H3N
H3N OH2
Pt
OH2
H3N
H3N
OH
Pt
Pt
OH-dimer
OH3N
H3N O
NH3H
HNH3
Pt Pt
CDDP None-chloro form
2
Cisplatin
Mono-chloro form
Fig. 1. Equilibrium reactions of cisplatin (CDDP) in waterIn water, CDDP loses Cl-, which is replaced with water. Depending on the pH, it may be replaced by a hydroxide ion. The mono-chloro form may lose another Cl-, and, in the same way, the water molecules (one or both) may be replaced with a hydroxide ion. These forms are in equilibrium.
(2)Quantification of Cisplatin (CDDP) and its Mono-Chloro, None-Chloro, and OH-Dimer Forms
■ Comparing a C18 column, COSMOSIL Cholester, and πNAP under identical conditionsWhen measuring the OH-dimer nitrate form, the dissociated NO3- peak elutes before the OH-dimer peak. C18-MS-II could not separate the two peaks, and Cholester achieved partial separation, but could not separate to baseline. πNAP, however, was able to separate the compounds completely.
NACALAI TESQUE, INC
Column: COSMOSIL **
Column size: 4.6 mmI.D.-250 mm
Mobile phase: 100 mmol/l NaClO4/Acetonitrile/100 mmol/l HClO4= 290/10/3
Flow rate: 1.0 ml/min
Temperature: 37 �CDetection: UV225 nm
Sample: Cisplatin OH-dimer nitrate (2 mmol/l)
1; Nitrate
2; Cisplatin OH-dimer
Injection Vol. 20 μl
Data courtesy of Ryuji Kato, Ph D., Laboratory of Cardiovascular Pharmacotherapy and Toxicology, Osaka University of Pharmaceutical Sciences
COSMOSIL
5C18-MS-II
COSMOSIL Application DataCOSMOSIL
Cholester
COSMOSIL
πNAP
-
For research use only, not intended for diagnostic or drug use.
NACALAI TESQUE, INC.
TELFAXWebsiteE-mail
: +81-(0)75-251-1730: +81-(0)75-251-1763: www.nacalai.com: [email protected] 1901
Nijo Karasuma, Nakagyo-ku, Kyoto 604-0855 JAPAN
■ Analysis Using COSMOSIL πNAPBelow is a typical chromatogram; the peak separation was excellent. The calibration curve for cisplatin (CDDP) was linear in the range of 0.01 to 4 mM, and the CV values for within-day and between-day variation were within 5%.
NACALAI TESQUE, INC
Column: COSMOSIL **
Column size: 4.6 mmI.D.-250 mm
Mobile phase: 100 mmol/l NaClO4/Acetonitrile/
100 mmol/l HClO4 = 290/10/3
Flow rate: 1.0 ml/min
Temperature: 37 �CDetection: UV225 nm
Sample: Cisplatin (5 mmol/l)
1; Di-chloro form
2; Mono-chloro form
3; OH-dimer form
4; None-chloro form
Injection Vol. 20 μl
Data courtesy of Ryuji Kato, Ph D., Laboratory of Cardiovascular Pharmacotherapy and Toxicology, Osaka University of Pharmaceutical Sciences
COSMOSIL
5C18-MS-II
COSMOSIL Application Data COSMOSIL πNAP
ClH3N
H3N ClPt
-Cl-, +H2O
+Cl-, -H2O
+H+-H+ +H+-H+
+H+-H+
-Cl-, +H2O
+Cl-, -H2O
-Cl-, +H2O
-2H2O
+2H2O
+Cl-, -H2O
OHH3N
H3N OH
OH2H3N
H3N Cl
Pt
OHH3N
H3N Cl
Pt
OH2H3N
H3N OH2
2
Pt
OH2
H3N
H3N
OH
Pt
Pt
OH-dimer form
OH3N
H3N O
NH3H
HNH3
Pt Pt
Di-chloro form Mono-chloro form None-chloro form
2
Cisplatin
(3)ObservationsBy using COSMOSIL πNAP, it is possible to separate and quantify cisplatin (CDDP) and its mono-chloro, none-chloro, and OH-dimer forms using HPLC.
(4)ReferencesKato R. et al. A Novel Analytical Method of Cisplatin Using the HPLC with a Naphthylethyl Group Bonded with Silica Gel (πNAP) Column. Biol Pharm Bull. 40(3), 290-296(2017).
