免疫亲和净化-光化学衍生-高效液相色谱法 检测动物肝脏中六种黄曲霉毒素

以动物肝脏为研究对象,二氯甲烷为提取溶剂,建立免疫亲和柱净化、高效液相色谱-光化学衍生-荧光检测器检测六种黄曲霉毒素的分析方法。该方法样品经提取、过滤、浓缩、复溶后经免疫亲和柱净化,进样后采用光化学衍生器在线对黄曲霉毒素G1、B1衍生,荧光检测器分析B1、B2、G1、G2、M1、M2六种黄曲霉毒素。结果表明,在不同加标水平,不同动物肝脏回收率在70.2%-94.1%,变异系数为2.67%-12.9%,准确度与精密度较高。方法检出限为最低为0.16μg/kg,满足痕量分析要求。     

多功能净化柱-高效液相色谱法检测玉米油中呕吐毒素

研究建立了多功能净化柱-高效液相色谱法检测玉米油中呕吐毒素(DON)的方法。样品经乙腈-水(体积比80∶20)混合溶剂振荡提取、超声波辅助萃取、多功能净化柱净化处理后,以乙腈-水(体积比16∶84)为流动相进行高效液相色谱检测。结果表明:DON在质量浓度为0.01~5μg/m L范围内具有良好的线性关系;线性方程为y=63 626.6x+390.1,相关系数为0.999 97,检出限(S/N)为3μg/kg;3个不同水平平均加标回收率为87.6%~104.8%,相对标准偏差为1.8%~5.3%。该方法可在8 min内完成检测、重现性好、灵敏度高,对取自不同工厂的13个成品玉米油样品进行检测,DON含量均低于国标限量(1 000μg/kg)。     

Survey of aflatoxins in rice from Iran using immunoaffinity column clean-up and HPLC with fluorescence detection

A study was undertaken to determine levels of aflatoxins in rice. A total of 261 rice samples were analyzed by HPLC using a method was based on the extraction of 50?g of finely ground rice plus 5?g?NaCl with 200?ml of 80% methanol. After filtration and immunoaffinity clean-up, 20?µl was injected onto the HPLC. HPLC analysis was carried out using a Genesis RP C₁₈ column (250?×?4.6, 4?µm I.D.) and a mobile phase with a linear gradient of water/methanol/acetonitrile (6?:?2?:?2?v/v) over 16?min. Aflatoxins were determined after post-column derivatisation with iodine by fluorescence detection at excitation and emission wavelengths of 365 and 445?nm, respectively. It was found that 68.9% of the rice samples contained aflatoxin B₁ at levels greater than 0.2?ng?g^?1.     

多功能柱净化-高效液相色谱法检测粮谷 及其制品中的呕吐毒素

建立了呕吐毒素的应用多功能柱净化-高效液相色谱法检测粮谷及其制品中的呕吐毒素方法、并建立了液相色谱-采用串联质谱进行确证的方法。样品经86%乙腈-水溶液(84：16,v/v)提取,再以、多功能柱净化、吹干、定容、离心后,最后以液相色谱-紫外检测器法进行定量分析,阳性检出以液相色谱-串联质谱法确证。该方法定量限为0.3 μg/g,在0.5、1.0、1.5 μg/g的添加水平下, 样品回收率为66.0 %-~97.3 %,标准偏差(SD)为1.4 %~-11.6 %,室内相对标准偏差(RSDr)为1.9 %~-12.7 %,室内HorRat值为0.1-~0.7。该方法操作简便,灵敏度高,选择性好,适用于粮谷及其制品中呕吐毒素的检测。     

Determination of gossypol in feeds by HPLC

An HPLC method for determination of goosypol in feed was developed. Gossypol in food was extracted with acetic acid-water-phosphoric acid (85 : 15 : 1) for 20 min in a water bath at 100 degrees C. The extract was diluted with acetone-water (1 : 1), and injected into the HPLC. HPLC was performed with a Shodex C18M4E (4.6 mm i.d.x250 mm) column at a flow-rate of 1.0 mL/min, using a mobile phase of methanol-water (9 : 1) adjusted to pH 2.6 with phosphoric acid, and gossypol was detected with a UV detector (254 nm). A recovery test was conducted with cottonseed spiked with gossypol at 1,000 and 5,000 mg/kg, and with 2 kinds of formula feed spiked with gossypol at 58 and 580 mg/kg. The mean recoveries of gossypol were 90.8-105.0% and the relative standard deviations (RSD) were within 3.0%. A collaborative study was conducted with cottonseed and formula feed spiked with gossypol at 305 mg/kg in 8 laboratories. The average content of gossypol in cottonseed was 6,090 mg/kg, and the repeatability and reproducibility as the relative standard deviation (RSD(r) and RSD(R)) were 3.3% and 4.4%, while the mean recovery, RSD(r) and RSD(R) of gossypol in formula feed were 87.0%, 2.7% and 5.5%, respectively.     

免疫亲和层析荧光光度法测定枸杞中黄曲霉毒素

利用免疫亲和层析荧光光度法测定枸杞中的黄曲霉毒素(AFT),样品由甲醇-水提取,提取液经过滤、稀释,用黄曲霉毒素免疫亲和柱净化。以甲醇洗脱,溴溶液显色,荧光光度计测定黄曲霉毒素(B1+B2+G1+G2)含量。枸杞的最低检出限为1μg/kg,5μg/kg的加标回收率为99.6%,10μg/kg的加标回收率为114.0%。该方法准确、快速、安全。     

A simple quantitative HPLC method for determination of aflatoxins in cereals and animal feedstuffs using gel permeation chromatography clean-up

A simple and sensitive procedure is described for the analysis of aflatoxins B1, B2, G1 and G2 in cereal and animal feedstuff samples. Aflatoxins are extracted with dichloromethane:water (10:1). Gel permeation chromatography (GPC) using a column packed with Bio-beads S-X3 and dichloromethane:hexane (3:1) as the eluent is used for clean-up of extracts prior to separation and quantification of aflatoxins by HPLC. The eluent fraction containing the aflatoxins is concentrated by evaporation under reduced pressure and the aflatoxins separated by reverse phase HPLC on an ODS column. Quantitative results have been obtained at levels down to 1 microgram/kg, and average recoveries from samples of wheat, maize and compound feed spiked at levels of 10 and 40 micrograms/kg were greater than 80%, 70% and 65% respectively.     

New method of citrinin determination by HPLC after polyamide column clean-up

In the past, various analytical methods of citrinin determination have been published; their application is not unproblematic. In this article, a HPLC method for sensitive fluorescence detection, based on solid phase extraction in combination with a HPLC gradient, is described. The samples are extracted with dichloromethane with the addition of phosphoric acid, and the extract is cleaned up on polyamide columns. According to this method, citrinin is detectable in cereals and milling products up to a detection limit of ~1–2 g/kg (limit of quantification: 3–5 g/kg). The mean recovery rates amounted to 74–90%. Analyses of some samples containing ochratoxin A (OTA) show that citrinin is especially detectable in cereal products, such as brans, wheatings and shorts, which contain increased amounts of the outer layers of the kernel. Citrinin, as well as OTA, was also detected in cocoa shells and raisins. 14 OTA-containing samples had citrinin contents of 1–8 g/kg. Further, it was demonstrated that the new method is also applicable for citrinin determination in red-fermented rice.     

The determination of aflatoxins in spices by immunoaffinity column extraction using HPLC

Seventy‐five samples of different spices marketed in Turkey were purchased from bazaars, herbal shops and supermarkets. Equal amounts of paprika, chilli, black peppers and cumin were purchased and used to test and compare the amount of aflatoxin contamination. Two different analytical methods were examined for their efficacy by adding a known amount of aflatoxin to the blank samples of paprika. Twenty‐seven paprika, all the chilli powder and four ground black pepper samples were contaminated with aflatoxin B₁ in the range of 0.5–116.4, 1.6–80.4 and 0.3–1.2 μg kg^?1 respectively. Twenty‐three (30%) paprika and chilli powder samples were above the regulatory limits used in the European Union. No aflatoxin contamination was detected in the cumin samples at a detection limit of 0.2 μg kg^?1.     

免疫亲和层析净化荧光光度法快速测定酱油及醋中黄曲霉毒素

为检测酱油和醋中黄由霉毒素的含量，建立了免疫亲和层析净化荧光光度法。试样由甲醇一水提取，提取液被过滤、稀释后，上交联着黄曲霉毒素B1、B2、G1、G2特异抗体的免疫亲和层析柱净化。以甲醇通过免疫亲和层析柱洗脱，溴溶液衍生，荧光光度计测定衍生物的黄曲霉毒素(B1 B2 G1 G2)含量。酱油和醋中检出限分别为2．5μgkg和1μg／kg。添加回收率在85％以上。该方法准确、简单、快速、安全。     

免疫亲和柱净化-光化学柱后衍生高效液相色谱 荧光法测定粮食中黄曲霉毒素

建立了粮食中黄曲霉毒素B1、B2、G1、G2的免疫亲和柱净化-光化学柱后衍生高效液相色谱荧光检测法。样品经甲醇-水提取,免疫亲和柱净化,高效液相色谱分离,光化学柱后衍生,荧光检测器测定。结果表明,黄曲霉毒素B1、B2、G1、G2的检出限分别为0.50、0.25、0.50、0.25μg/kg,回收率为67.2%～91.7%,RSD小于10%。该方法快速、准确、灵敏度高、重现性好,能满足我国对粮食中黄曲霉毒素限量的检测要求。     

HPLC column switching technique for the clean-up of steroid samples

Summary A simple column-switching system was used for the clean-up of steroid samples. Fatty tissues were extracted as described in a previous paper [1]. The filtered and concentrated extract is injected on to a HPLC system consisting of three columns. After a front-cut between column 2 and column 3, the fraction of interest is separated on column 3. Gradient elution offers the possibility to isolate a single or multi- component fraction, which can be examined by HPTLC or another detection technique.

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HPLC-Kolonne-Umkehrtechnik fur die Reinigung von Anabolica-Proben

Zusammenfassung Ein einfaches Kolonnen-Umkehrsystem wurde fur die Reinigung von Steroid-Proben angewandt. Fettgewebe wurde, wie vorher beschrieben [1], extrahiert. Der filtrierte and konzentrierte Extrakt wurde in eine aus drei Kolonnen bestehende HPLC-Anlage injiziert. Nach Abschneiden der Front zwischen Kolonne 2 and 3 wurde die interessierende Fraktion durch die Kolonne 3 chromatographiert. Die Gradiente-Elution bietet die Möglichkeit, eine ein- oder multikomponente Fraktion zu isolieren, die durch HPTLC oder eine andere Nachweistechnik geprüft werden kann.

     

免疫亲和柱结合液相色谱串联质谱法测定蜂花粉中黄曲霉毒素

建立了一种测定蜂花粉中黄曲霉毒素(B1、B2、G1、G2)的液相色谱串联质谱方法。样品经V(乙腈)∶V(水)=60∶40提取,通过低温脂肪沉淀,免疫亲和柱净化,之后用甲醇洗脱4种毒素,氮吹后复溶,采用液相色谱-串联质谱(LC-MS/MS)定量分析。方法在3个浓度添加水平的回收率为74.3%⁹6.5%,精密度低于10.0%。相关系数(r2)均大于0.997。B1、B2、G1、G2的定量限分别为0.05,0.1,0.013,0.025μg/kg。测定30个实际样品,未检出4种毒素。     

液相色谱法自制净化柱测定饲料中5种黄曲霉毒素

通过用自制净化柱净化,利用高效液相色谱仪对饲料中的黄曲霉毒素B1、B2、G1、G2、M1同时进行检测,样品经体积分数为84%的乙腈溶液提取,提取液通过自制净化柱净化、浓缩,三氟乙酸(TFA)柱前衍生,C18色谱柱分离,荧光检测器检测,外标法定量。5种黄曲霉毒素经过衍生后线性良好,对添加黄曲霉饲料样品进行加标回收,回收率在85%~102%,效果良好。     

免疫亲和柱-高效液相色谱法同时检测谷物中T-2毒素和HT-2毒素含量

建立了免疫亲和柱净化-柱前化学衍生-高效液相色谱荧光检测器同时检测谷物中T-2毒素和HT-2毒素的方法。样品经溶剂[V(甲醇)∶V(水)=80∶20]提取,通过免疫亲和柱净化(IAC),以氰酸蒽(1-AN)为衍生化试剂、4-二甲基氨基吡啶(DMAP)为催化剂进行衍生,以ZORBAX Eclipse XDB苯基柱为分离柱,乙腈-水为流动相进行高效液相色谱分离和检测。在0.005⁰.5μg/g内呈良好线性,检出限为0.005μg/g,添加回收率为82.0%0.5μg/g内呈良好线性,检出限为0.005μg/g,添加回收率为82.0%¹08.0%,RSD<15.5%。     

免疫亲和柱净化-在线柱后光化学衍生-高效液相色谱-荧光检测法快速测定食品中的黄曲霉毒素含量

目的 通过建立免疫亲和柱净化-在线柱后光化学衍生-高效液相色谱-荧光检测法探索快速测定食品中的黄曲霉毒素含量的方法。方法 样品经甲醇-水(V/V, 7︰3)提取, 免疫亲和柱净化、富集、淋洗后, 收集洗脱液, 洗脱液经Acclaim?120 C18 柱分离, 在254 nm紫外光下进行光化学衍生, 用荧光检测器进行测定。结果 黄曲霉毒素G2, G1, B2, B1检测限均小于2 μg/kg, 加标回收率为94.3%~108.8%。 结论 该方法能连续操作, 节省人力, 可用于食品中黄曲霉毒素含量的测定。     

Determination of nitrofurans residues in animal feeds by flow injection chemiluminescence procedure

A simple flow injection chemiluminescence (FI-CL) method was developed for the rapid and sensitive determination of nitrofurans, including furazolidone, nitrofurantoin and nitrofurazone, in animal feeds based on its chemiluminescence induced by potassium permanganate in sulphuric acid medium. The method involves the injection of nitrofuran samples or standards into H₂SO₄ carrier stream, which then merges at a T-piece with a reagent stream consisting of KMnO₄ in the H₂SO₄ carrier solution. The elicited chemiluminescence intensity of the resulting reaction mixture was measured by photomultiplier tube operated at a voltage of 950 V. Optimum CL signals were given using 2.5 × 10⁻⁵ mol L⁻¹ potassium permanganate in 0.1 mol L⁻¹ sulphuric acid as an oxidant stream and a carrier stream of 0.1 mol L⁻¹ sulphuric acid with a total flow rate of 7.0 mL min⁻¹. Results detailing the optimisation of the analytical signal, calibration, and common interferences of animal feeds were also discussed. The proposed FI-CL method was successfully applied to the determination of nitrofurans in animal feeds, with excellent recoveries, as the determination is free from interference. The method validation has been compared versus HPLC method for animal feed samples.     

Analysis of sterigmatocystin in cereals,animal feed,seeds, beer and cheese by immunoaffinity column clean-up and HPLC and LC-MS/MS quantification

A method is reported for the analysis of sterigmatocystin in various food and feed matrices using a commercial sterigmatocystin immunoaffinity column (IAC) for sample clean-up prior to HPLC analysis by UV with mass spectrometric detection (LC-MS/MS). Cereals (wheat, oats, rye, maize and rice), sunflower seeds and animal feed were spiked with sterigmatocystin at levels from 0.75 to 50 µg kg^?1 to establish method performance. Using acetonitrile/water extraction followed by IAC clean-up, and analysis by HPLC with detection at 325 nm, recoveries ranged from 68% to 106%, with repeatability from 4.2% to 17.5%. The limit of quantification with UV detection in these matrices was 1.5 µg kg^?1. For the analysis of beer and cheese the sample preparation prior to IAC clean-up was changed to accommodate the different properties of the matrix, prior to analysis by LC-MS/MS. For beer and cheese spiked at 5.0 µg kg^?1 the recoveries were 94% and 104%, and precision (RSDs) were 1.9% and 2.9% respectively. The limits of quantification by LC-MS/MS in beer and cheese were 0.02 and 0.6 µg kg^?1 respectively. The sterigmatocystin IAC was demonstrated to provide an efficient clean-up of various matrices to enable this mycotoxin to be determined by either HPLC with UV detection or LC-MS/MS.     

HPLC法柱后衍生测定乳制品中牛磺酸的含量

建立了高效液相色谱法测定乳制品中牛磺酸含量的方法。采用荧光检测器，柱后OPA衍生剂衍生，色谱柱为Waerts氨基酸分析柱，柱温为25℃，流动相流速为0．4mL／min，衍生剂流速为0．4mL／min。标准曲线相关系数为0．9749；回收率为95．6％；8次样品测定SD为0．3832，变异系数为1．2136％。