A rapid monitoring method of paraquat and diquat in serum and urine using ion-pairing bare-silica stationary phase HPLC following a single acidification step of sample pretreatment
A rapid monitoring method of paraquat and diquat in serum and urine using ion-pairing bare-silica stationary phase HPLC following a single acidification step of sample pretreatment
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DOI：
10.1080/10826079708005835
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发表时间：
1997-01-01
影响因子：
1.3
通讯作者：
Binder, SR
中科院分区：
文献类型：
--
作者：
Itagaki, T;Lai, SJ;Binder, SR
亚洲和东欧国家经常报道百草枯（一种农业化学品）中毒的情况，因为普通民众都可以使用百草枯。由于百草枯在人体中不会发生显着的生物转化，因此在抢救和康复阶段都需要密切监测百草枯水平。设计一种有效的技术在治疗的早期阶段消除患者体内的百草枯至关重要，并且需要一种快速的方法来监测百草枯浓度。敌草快是百草枯毒性较小的类似物，通常与百草枯混合用于农业用途。最近，在所有开发用于监测血清和尿液样本中百草枯和敌草快的药物中，敌草快和百草枯混合物的致死率很高。总分析时间不到6分钟。它只需要最少的样品预处理，包括样品酸化和 9500 x g 离心 1 分钟。该方法采用离子对 HPLC，以裸硅胶 (150 mm X 4.6 mm) 作为固定相。用2M磷酸酸化后，样品上清液可直接注入HPLC系统。流动相为25/75乙腈/水(VN)；含有 10 mM 1-庚磺酸钠、4 mM 磷酸钾、10 mM 氯化钾，pH 3.0。分析在室温下以 1.0 mL/min 的流速进行。百草枯和敌草快的最佳紫外线波长分别为 257 nm 和 310 nm；测得敌草快相应的检测限为 63 ng/mL（柱上 1.25 ng），百草枯相应的检测限为 125 ng/mL（柱上 2.5 ng）。为了同时监测两种化合物，可以使用 290 mn 的共同波长；进样量为20μL时，尿液和血清中的敌草快浓度为500ng/mL（柱上10ng），百草枯浓度为125ng/mL（柱上2.5ng）。通过使用更大的进样量可以实现 290 nm 处的较低检测限。两种化合物的保留数据的 CV% 均小于 1.2%。还对常见药物和代谢物进行了干扰研究。对阳离子交换方法强烈保留的药物进行了额外的测试：氯喹、士的宁和尼古丁。所有这三种化合物均不干扰该方法。
Intoxication by paraquat, an agricultral chemical, is frequently reported in Asian and eastern European countries because of its availablity by general population. Because paraquat is not significantly biotransformed in man, it requires an intensive monitoring of paraquat level during both stages of rescue and recuperation. It is critical to design an effective technique to eliminate the paraquat from the patient at the earlier stage of the therapy, and a rapid method is required for monitoring the paraquat concentration. Diquat is a less toxic analog of paraquat, and is often mixed with paraquat for agricultural usage. Recently, a high rate of fatal cases were reported on the mixture of diquat and paraquat among all been developed for monitoring paraquat and diquat in serum and urine samples.The total analysis is less than 6 minutes. It only requires a minimal sample pretreatment, including acidification and centrifugation of sample at 9500 x g for 1 minute. This method utilizes ion-pairing HPLC with bare silica (150 mm X 4.6 mm) as stationary phase. After acidification with 2M phosphoric acid, the supernatant of samples can be injected directly into the HPLC system. The mobile phase was 25/75 acetonitrile/water (VN); containing 10 mM 1- Heptanosulphonic Sodium Salt, 4 mM potassium phosphate, 10 mM potassium chloride, pH 3.0. The analysis was performed at room temperature with a flow rate of 1.0 mL/min. The-optimum UV wavelength for paraquat and diquat were 257 nm and 310 nm, respectively; the corresponding detection limits were measured at 63 ng/mL (1.25 ng on column) for diaquat and 125 ng/mL (2.5 ng on column) for paraquat, respectively. For simultaneous monitoring of both compounds, a common wavelength of 290 mn can be used; with an injection volume of 20 mu L, paraquat could be detected at 500 ng/mL(10 ng on column) and 125 ng/mL (2.5 ng on column) for diquat in both urine and serum. A lower detection limit at 290 nm can be achieved by using a larger injection volume. The CV% of retention data are less than 1.2% for both compounds. Interference studies were also conducted for common drugs and metabolites. Additional tests were conducted on drugs which are strongly retained on cation exchange methods: chloroquine, strychnine and nicotine. All these three compounds did not interfere with the method.