免疫亲和柱净化-柱后光化学衍生-高效液相色谱法同时检测粮谷中的黄曲霉素、玉米赤霉烯酮和赭曲霉毒素A

建立了同时检测粮谷中黄曲霉毒素(B1、B2、G1和G2)、玉米赤霉烯酮和赭曲霉毒素A的免疫亲和柱净化-柱后光化学衍生-高效液相色谱方法.样品经过甲醇-水(体积比为80∶20)提取,通过免疫亲和柱富集和净化,采用Waters Nova-Pak色谱柱(3.9 mm I.d.×150 mm,4 μm),以甲醇、乙腈和1%的磷酸溶液为流动相,梯度洗脱,柱后光化学衍生、改变波长荧光检测.黄曲霉毒素(B1、B2、G1和G2)、玉米赤霉烯酮和赭曲霉毒素A检出限分别为0.24,4.0和0.5 μg/kg,标准曲线的线性范围分别为0.24～6.0,4.0～100.0和0.5～40.0 μg/L;在小麦、玉米、黑麦样品中,平均加标回收率为70.8% ～94.0%,相对标准偏差为2.79% ～9.38%.     

柱上衍生反相高效液相色谱法测定疾藜提取物中痕量铅

提出了微波消解样品,在线衍生测定疾藜提取物中痕量铅的反相高效液相色谱方法.双水杨醛邻苯二胺为衍生剂,用DiamonsilTM C18色谱柱(4.6 mm×150 mm,5um)作为固定相,甲醇-水-衍生剂(64.5:34.0:1.5体积比)为流动相,检测波长为275 nm,柱温30℃.样品中铅在0.10～0.63 ng范围内呈线性,铅的平均回收率为99.5%,相对标准偏差(n=7)为1.8%.     

柱前衍生高效液相色谱法测定尼莫地平注射液中的甲醛和乙醛

建立柱前衍生高效液相色谱测定尼莫地平注射液中甲醛和乙醛含量的方法.以2,4-二硝基苯肼为衍生化试剂,室温反应30 min,采用高效液相色谱法检测,色谱柱为Waters XBridge C8柱(250 mm×4.6 mm,5μm),流动相为乙腈–水(体积比为50:50),等度洗脱,流量为1.0 mL/min,柱温为30℃,检测波长为360 nm,进样体积为20μL.甲醛、乙醛的色谱峰面积与质量浓度分别在0.01～1.00μg/mL、0.01～1.20μg/mL范围内线性关系良好,相关系数均为0.9999,测定结果的相对标准偏差分别为0.86％和1.02％(n=6),平均加标回收率分别为104.3％和92.0％.     

高效液相色谱光化学在线衍生技术在11种磺胺药物残留检测中的应用

建立了磺胺药物残留的高效液相色谱-光化学在线衍生-荧光检测方法,并应用于猪肉的检测。样品经过乙腈提取,色谱柱分离后,通过在线光化学衍生后,用荧光检测器进行直接检测。优化后的色谱条件:Eclipse Plus C18柱(250 mm×4.6 mm,5.0μm),流动相为均含0.2%甲酸的乙腈、甲醇和水梯度洗脱,检测激发波长为248 nm,发射波长为350和412 nm。各种磺胺在各自浓度范围内线性相关系数R2>0.999,回收率在85.7%~101.1%之间,RSD为1.9%~6.6%(n=6),各磺胺的检出限(S/N=3)为0.2~3.0μg/kg,定量限(S/N=10)为0.5~10.0μg/kg。     

UPLC-MS/MS法测定7类化妆品中4-氨基联苯

建立了超高效液相色谱-串联质谱(UPLC-MS/MS)检测化妆品中4-氨基联苯的方法。化妆品样品经溶剂超声提取、净化(液液萃取或固相萃取),氮气吹至近干并定容后,通过高效液相色谱-串联四极杆质谱仪检测。分离柱为Waters Acquity BEH C18柱(1.7μm,2.1 mm×100 mm);流动相为0.3%甲酸水溶液-乙腈;流速0.5 mL·min-1。7种不同基质样品中4-氨基联苯的检测限均小于1.0ng·g-1。7种不同基质样品中4-氨基联苯的回收率为85.3%～111.2%,相对标准偏差(RSD)为0.93%～4.11%(n=3),在2.5ng·mL-1～250ng·mL-1浓度范围内呈良好的线性关系,线性回归系数r2均大于0.999。     

柱前衍生高效液相色谱-质谱法测定水稻中尼克烟酰胺含量

建立了柱前衍生高效液相色谱-质谱联用(HPLC-MS)测定水稻中尼克烟酰胺含量的方法.样品中尼克烟酰胺经水提取后,与9-芴甲基氯甲酸酯(FMOC-Cl)衍生,采用液相色谱质谱联用仪测定.系统研究了衍生剂浓度和衍生介质等条件对衍生效率的影响.通过优化流动相酸度和梯度洗脱等条件,提高了方法灵敏度.尼克烟酰胺在0.1～5.0 mg/L范围内呈良好的线性关系(r=0.9983),对水稻的根、茎、叶及大米的标准加入实验表明,方法的添加回收率在72.0%～89.2%之间; 相对标准偏差为2.3%～9.6%; 方法检出限为0.05 mg/kg.方法简便、准确可靠,可以满足水稻中生理水平尼克烟酰胺的定性定量分析.     

高效液相色谱法测定牛奶中的氯霉素残留

建立牛奶中氯霉素残留的高效液相色谱测定方法。牛奶样品用乙腈提取,超声离心后采用液相色谱柱分离,外标法定量。色谱条件:Agilent ZORBAX SB-C18柱(4.6mm×250mm,5μm),流动相:甲醇-水(40∶60,V/V),波长:278nm,流速0.8mL.min-1。线性范围为0.1～10.0 mg·L-1,相关系数R2为0.9995,最低检测限为0.009 mg·L-1。平均回收率在93.0%～97.3%之间,相对标准偏差在3.67%～5.25%之间(n=6),该方法简便、准确,适用于牛奶中氯霉素的检测。     

柱后衍生-高效液相色谱-荧光检测法测定面粉中的过氧化苯甲酰

利用过氧化苯甲酰能够在氯化血红素的催化下将底物对羟基苯乙酸氧化成具有荧光的二聚体的性质,建立了柱后衍生-高效液相色谱-荧光检测法测定面粉中过氧化苯甲酰的方法。优化了色谱分离条件和柱后衍生条件,结果表明:催化剂氯化血红素的浓度为8μmol/L,底物对羟基苯乙酸的浓度为80μmol/L,衍生化试剂的pH 10.5,衍生温度为35℃时衍生效率最高。在优化条件下,过氧化苯甲酰的线性范围为0.5～100 mg/L;样品的检出限为1 mg/kg,样品的加标回收率为98.5%～99.5%。本方法与现有的高效液相色谱法相比,检测面粉中过氧化苯甲酰具有抗干扰能力强,灵敏度高。     

固相萃取-反相高效液相色谱法测定隐形眼镜护理液中聚胺丙基双胍

建立了固相萃取-反相高效液相色谱法测定隐形眼镜护理液中聚胺丙基双胍的方法.用Spe-ed WCX固相萃取柱(500 rag/3 mL)将样品浓缩富集后,以Shiseido C18色谱柱(150 min×4.6 mm,5 pm)为分离柱,以pH 4.0的乙腈-20 mm01.L-1乙酸铵(体积比16比84)为流动相,检测波长为235 nm,7 min内测定样品中聚胺丙基双胍.方法检出限(S/N=3)为0.046 mg·L-1,相对标准偏差为3.3%(n=7),线性范围为100～800 mg·L-1,加标回收率93.5%～110.9%.     

超高效液相色谱-串联质谱法同时测定尿液与血清中23种双酚类化合物

建立了基于吡啶-3-磺酰氯衍生和超高效液相色谱-串联质谱(UPLC-MS/MS)检测尿液和血清中23种双酚类化合物(BPs)的方法。尿液样品采用乙酸乙酯提取;血清样品采用乙腈提取,上清液经PRiME HLB柱净化。提取液经吡啶-3-磺酰氯衍生后,以乙腈和0.1%甲酸水溶液为流动相,采用ACQUITY UPLC BEH C_(18)(2.1 mm×100 mm,1.7μm)色谱柱进行分离。质谱离子源为电喷雾电离源,正离子模式检测。23种目标物在0.005～100μg/L范围内线性关系良好,检出限为0.002～0.030μg/L,定量下限为0.005～0.100μg/L,回收率为76.6%～122%,相对标准偏差(RSD)为0.60%～14%。使用建立的方法分别对20份尿液和20份血清样品进行了检测。该方法样品用量少,灵敏度高,可以满足尿液和血清样品中23种BPs的同时测定要求。     

Determination of sulfite in dried garlic by reversed-phase ion-pairing liquid chromatography with post-column detection

A method is described for determining sulfite in dried garlic. Garlic is extracted with an HCl solution to inhibit the formation of allicin, which interferes with the determination of sulfite. After cleanup of the extract on a C18 solid-phase extraction column, sulfite is converted to hydroxymethylsulfonate (HMS) by adding formaldehyde and heating to 50 degrees C. HMS is determined by reversed-phase ion-pairing liquid chromatography with post-column detection. The post-column reaction system consists of the addition of KOH to convert HMS to sulfite ion, followed by the addition of 5,5'-dithiobis(2-nitrobenzoic acid) to produce 5-mercapto-2-nitrobenzoic acid which is detected spectrophotometrically at 450 nm. Background levels in unsulfited dried garlic equivalent to < 20 ppm SO2 were found. Recoveries of HMS from spiked garlic averaged 94.8% with a coefficient of variation of 3.8%. Sulfite was found in 13 of 21 samples of dried garlic produced in China, with sulfite ranging from 114 to 445 ppm. Sulfite was found in 60% of commercial dried garlic products purchased locally. The suitability of the Monier-Williams method for determining sulfite in garlic is discussed.     

高效液相色谱柱后衍生测定脱水蔬菜中的亚硫酸盐

建立了检测脱水蔬菜中亚硫酸盐的反相硅胶柱净化-柱后衍生-高效液相色谱方法.样品经甲醛提取,通过反相硅胶固相萃取小柱净化,采用Discover ODS-C18柱,流动相为0.005 mol/L氢氧化四丁基铵和乙腈,在碱性条件下以5,5’-二硫代双(2-硝基苯甲酸)为柱后衍生化试剂,445 nm检测.实验结果表明,在0.050～50.00 mg/L浓度范围内,相关系数为0.9987,方法的检出限和定量限分别为1 mg/kg和2 mg/kg,添加浓度在2～900mg/kg范围内,平均添加回收率为62％～88％,相对标准偏差不大于7.8％.该法能有效地避免脱水蔬菜中亚硫酸盐测定的假阳性结果,可满足实际检测工作的需要.     

Simultaneous determination of sulfite, sulfate, and hydroxymethanesulfonate in atmospheric waters by ion-pair HPLC technique

A reversed-phase ion-pair HPLC with indirect photometric detection for the simultaneous determination of sulfite, sulfate, hydroxymethanesulfonate (HMS), and other inorganic anions in atmospheric water has been developed. Separations were accomplished in less than 10 min in a cetylpyridinium-coated C₁₈ column with 0.5 mM potassium hydrogen phthalate-0.015% triethanolamine-3% methanol at pH 7.9 as mobile phase. Quantitation was carried out by the peak area method, with detection limits in the pmol range. UV light absorption responses were linear over a wide concentration range from several hundred μmoles to the detection limits of each anion. The application of the method provides a rapid and efficient technique for the quantitative determination of sulfur and other inorganic species in atmospheric liquids.     

Validated liquid chromatographic method for quantitative determination of allicin in garlic powder and tablets

In the present study, an RP high performance liquid chromatographic method was developed and validated for the determination of allicin in garlic powder and tablets. Chromatographic separation was carried out on an RP-18(e )column (125 mm x 4 mm), using a mobile phase, consisting of methanol-water (50:50 v/v), at a flow rate of 0.5 mL/min and UV detection at 220 nm. Ethylparaben was used as the internal standard. The assay was linear for allicin concentrations of 5.0-60.0 microg/mL. The RSD for precision was <6.14%. The accuracy was above 89.11%. The detection and quantification limits were 0.27 and 0.81 microg/mL, respectively. This method was used to quantify allicin in garlic powder samples. The results showed that the method described here is useful for the determination of allicin in garlic powder and tablets.     

Ultraviolet derivatization of steroidal saponin in garlic and commercial garlic products as p-nitrobenzoate for liquid chromatographic determination

A method is described for determination of the steroidal saponin, eruboside B, originating in garlic and garlic products as the p-nitrobenzoyl chloride (PNBC) derivative by reversed-phase liquid chromatography (with ultraviolet detection at 260 nm. Proto-eruboside B was extracted from garlic (Allium sativum L.); subjected to solid-phase extraction (SPE) with a C18 cartridge, Florisil column chromatography, and silica gel column chromatography; and then enzymatically converted to eruboside B, which was applied as an external standard. Steroidal saponins in garlic and commercial garlic products were extracted with methanol and purified by SPE cartridges, followed by enzymatic treatment. A frostanol saponin such as proto-eruboside B is enzymatically transformed to a spirostanol saponin, eruboside B. After the derivatization with PNBC, the saponin derivative was chromatographed on a C8 column with a gradient elution of (A) 80% aqueous acetonitrile and (B) 100% acetonitrile. The detection limit of the developed method was 1 microg/g for the samples. The method was applied to the analysis of garlic and garlic health food products available in Japan.     

光化学柱后衍生–高效液相色谱法测定婴幼儿营养米粉中的黄曲霉毒素B1

建立婴幼儿营养米粉中黄曲霉毒素B1的高效液相色谱荧光检测器测定方法。样品以甲醇–水(体积比70∶30)溶液匀质提取,过黄曲霉毒素B1免疫层析亲和柱净化,经CNW Athena C18色谱柱分离和光化学柱后衍生反应器衍生后,用带有荧光检测器的高效液相色谱仪测定。采用峰面积外标法定量黄曲霉毒素B1含量。黄曲霉毒素B1在0～10μg/L的浓度范围内线性关系良好,相关系数为0.999 8,检出限为0.25μg/kg。在3个添加水平下加标回收率为97.7%～106.9%,测定结果的相对标准偏差为1.7%(n=6)。该方法的灵敏度、准确度、精密度均符合黄曲霉毒素B1的检测技术要求,适用于婴幼儿营养米粉中黄曲霉毒素B1的日常检测。     

Automated pre-column derivatization of amines in biological samples with dansyl chloride and with or without post-column chemiluminescence formation by using TCPO-H2O2.

On-line automation of two different liquid chromatographic procedures, a pre-column derivatization system and a pre- and post-column system, in order to generate chemiluminescence is reported. Dansyl chloride (Dns-Cl) was used as a pre-column reagent to form fluorophores and bis(2,4,6-trichlorophenyl) oxalate (TCPO) and hydrogen peroxide (H2O2) as a post-column reagent to generate chemiluminescence. This procedure is based on the employment of a primary column packed with C18 material inserted in a multi-dimensional assembly for sample clean-up and derivatization with Dns-Cl. The dansyl derivatives formed are transferred and separated in a LiChrospher 100 RP18 analytical column (125 x 4 mm id, 5 microns film thickness) using acetonitrile-imidazole buffer (pH 6.8) (70 + 30) as eluent. The separated derivatives were transferred to the detector for fluorescence detection or to the post-column system where the chemiluminescence response was generated by using TCPO-H2O2 and the products were detected by chemiluminescence. The procedure was optimised for amphetamine and related compounds. A comparison between the on-line pre-column and pre- and post-column systems was performed. The results show that the sensitivity of chemiluminescence detection can be higher than that of fluorescence detection. The recoveries obtained ranged from 98 +/- 8 up to 108 +/- 8% for amphetamine and methamphetamine, respectively. The accuracy and precision of these methods were evaluated.     

高效液相色谱法测定动物组织样品中黄曲霉毒素的残留量

采用免疫亲合技术,对动物肝脏和肌肉中黄曲霉毒素残留进行了提取和纯化处理,建立了一种快速、灵敏、简便的测定动物组织中黄曲霉毒素B1、B2、G1、G2的高效液相色谱方法。用反相HPLC分离,柱后光化学衍生,荧光检测器测定黄曲霉毒素含量,保留时间定性,峰面积定量,测定了样品和标准样。结果表明,4种毒素可在10min内完成分离;线性范围为15～1500pg;线性回归系数大于0．9998。用本方法对动物肝脏和肌肉样品进行了加标回收实验,4种黄曲霉毒素的平均回收率为68．71％～83．42％;相对标准偏差为3．51％～7．40％;检出限均达到2．65pg。     

Determination of retinoids in galenicals by column liquid chromatography with fluorescence and diode-array detection

Simple and rapid reversed-phase gradient column liquid chromatography (LC) with fluorescence detection at different wavelengths was developed for the simultaneous analysis of all-trans, 13-cis, 9-cis retinoic acids, vitamin A palmitate and beta-carotene in galenicals. The assay results agreed with those obtained by an LC method with diode-array UV detection. A post-column on-line photochemical reactor (irradiation at 254 and 366 nm) was inserted between the LC column and the fluorescence detector to enhance the performance of the method. Two fluorescence spectra (photoreactor on and off) were obtained for each analyte which proved useful for the unambiguous identification of the various analytes.     

Determination of metal complexes of ethylenediaminetetraacetate in the presence of organic matter by high-performance liquid chromatography

A high-performance liquid chromatography method is described here for the determination of the Cd(II), Co(II), Cu(II), Pb(II), and Zn(II) complexes of ethylenediaminetetraacetate (EDTA) in municipal wastewaters and surface waters. The method involves separation by ion-exchange chromatography on a reversed-phase C18 column coated with ion-pair reagent, followed by post-column conversion to FeEDTA and subsequent detection by UV absorbance. Although Co(II) and Cu(II) coelute, they can be quantified by analyzing absorbance by CuEDTA2- prior to post-column conversion. The method detection limit of 6-8 x 10(-8) M (5-7 ng) is an order of magnitude improvement over previous UV absorbance post-column reaction methods. The technique can be used in the presence of organic matter encountered in matrices such as untreated wastewater without pre-concentration or sample cleanup.