HPLC/PDA determination of carminic acid and 4-aminocarminic acid using relative molar sensitivities with respect to caffeine
HPLC/PDA determination of carminic acid and 4-aminocarminic acid using relative molar sensitivities with respect to caffeine
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DOI：
10.1080/19440049.2018.1440642
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发表时间：
2018-01-01
影响因子：
2.9
通讯作者：
Sato, Kyoko
中科院分区：
文献类型：
--
作者：
Nishizaki, Yuzo;Sato-Masumoto, Naoko;Sato, Kyoko
为了准确测定胭脂红酸（CA）及其衍生物4-氨基胭脂红酸（4-ACA），建立了一种新的高效液相色谱-光电二极管阵列检测器（HPLC/PDA）相对摩尔灵敏度（RMS）测定方法。该方法不需要CA和4-ACA的分析标准品;相反，它使用相对于咖啡因（CAF）的RMS值，其用作内标。H-1-qNMR与HPLC/PDA联用，能够准确测定CA（274 nm）/CAF（274 nm）和4-ACA（274 nm）/CAF（274 nm）的RMS。为了确认使用RMS的HPLC/PDA方法的性能，使用四种不同的HPLC-PDA仪器和一种HPLC-UV检测供试品中的CA和4-ACA含量。CA（274 nm）/CAF（274 nm）和4-ACA（274 nm）/CAF（274 nm）在5台色谱仪和2根色谱柱上测定结果的相对标准偏差均小于2.7%和1.1%。采用1H-1-qNMR方法直接分析了供试品中CA和4-ACA的含量。从两种方法获得的定量值之间的差异小于5%的CA和3%的4-ACA。这些结果表明，使用RMS至CAF的HPLC/PDA方法是一种简单可靠的定量方法，不需要CA和4-ACA认证的参比物质。
To accurately determine carminic acid (CA) and its derivative 4-aminocarminic acid (4-ACA), a novel, high-performance liquid chromatography with photodiode array detector (HPLC/PDA) method using relative molar sensitivity (RMS) was developed. The method requires no analytical standards of CA and 4-ACA; instead it uses the RMS values with respect to caffeine (CAF), which is used as an internal standard. An off-line combination of H-1-quantitative nuclear magnetic resonance spectroscopy (H-1-qNMR) and HPLC/PDA was able to precisely determine the RMSs of CA(274nm)/CAF(274nm) and 4-ACA(274nm)/CAF(274nm). To confirm the performance of the HPLC/PDA method using RMSs, the CA and 4-ACA contents in test samples were tested using four different HPLC-PDA instruments and one HPLC-UV. The relative standard deviations of the results obtained from five chromatographs and two columns were less than 2.7% for CA(274nm)/CAF(274nm) and 1.1% for 4-ACA(274nm)/CAF(274nm). The H-1-qNMR method was directly employed to analyse the CA and 4-ACA contents in test samples. The differences between the quantitative values obtained from both methods were less than 5% for CA and 3% for 4-ACA. These results demonstrate that the HPLC/PDA method using RMSs to CAF is a simple and reliable quantification method that does not require CA and 4-ACA certified reference materials.