气相色谱-负化学源质谱法测定蜂蜜和王浆中4种杀虫剂的残留

建立了气相色谱-负化学源质谱（GC-NCI/MS）测定蜂蜜和王浆中4种杀虫剂残留量的方法。蜂蜜样品由乙酸乙酯提取、乙二胺-N-丙基硅烷（PSA）净化,而王浆样品经乙腈-水（1：1,v/v）提取、C18固相萃取柱净化,采用GC-NCI/MS测定,外标法定量。结果表明：在50~500 μg/L范围内4种农药的线性良好;所有农药的LOD在0.12~5.0 μg/kg之间,LOQ在0.40~16.5 μg/kg之间;在10、15、20 μg/kg 3个添加水平下,4种农药的平均回收率在78.2%~110.0%之间,且RSD均小于14%。所有农药的测定均没有出现干扰峰。该方法简单、快速,准确度、精密度和选择性高,抗干扰能力强,可用于蜂蜜和王浆中这4种农药的快速检测。     

离子色谱法测定奶制品及蜂王浆中的乙酰胆碱

建立了奶制品及蜂王浆中乙酰胆碱的离子交换/电导检测离子色谱分析方法.样品经去离子水超声提取,离子交换/电导检测离子色谱法测定,外标法定量.采用IonPac CS17(250 mm x4.0 mm i.d.)分析柱,流动相为甲基磺酸,梯度洗脱,流速1.0 mL/min,柱温30℃,检测池温度35℃.实验结果表明,氯化乙酰...     

Optimized determination method for trans-10-hydroxy-2-decenoic acid content in royal jelly by high-performance liquid chromatography with an internal standard

An optimized reversed-phase high-performance liquid chromatography method was developed to detect the trans-10-hydroxy-2-decenoic acid (10-HDA) content in royal jelly cream and lyophilized powder. The sample was extracted using absolute ethanol. Chromatographic separation of 10-HDA and methyl 4-hydroxybenzoate as the internal standard was performed on a Nova-pak C18 column. The average recoveries were 95.0-99.2% (n = 5) with relative standard deviation (RSD) values of 1.3-2.1% for royal jelly cream and 98.0-100.0% (n = 5) with RSD values of 1.6-3.0% for lyophilized powder, respectively. The limits of detection and quantitation were 0.5 and 1.5 mg/kg, respectively, for both royal jelly cream and lyophilized powder. The method was validated for the determination of practical royal jelly products. The concentration of 10-HDA ranged from 1.26 to 2.21% for pure royal jelly cream samples and 3.01 to 6.19% for royal jelly lyophilized powder samples. For 30 royal jelly products, the 10-HDA content varied from not detectable to 0.98%.     

高效液相色谱柱后衍生法测定蜂王浆中的链霉素

采用高效液相色谱柱后衍生法测定蜂王浆中的链霉素。样品用庚烷磺酸钠的酸性溶液进行提取,通过LC-18柱和WCX柱两次固相萃 取达到净化的目的。该方法采用反相C8柱,以0.01 mol/L的庚烷磺酸钠溶液和乙腈为流动相,在碱性条件下以1,2-萘醌-4-磺酸钠溶液 为柱后衍生化试剂,用荧光检测器检测。该方法的线性范围为0.02～0.5 mg/L,相关系数为0.9958,方法的检出限和定量限分别为为 0.005 mg/kg和0.01 mg/kg,回收率范围为84.0%～104.0%,相对标准偏差不大于7.9%。结果表明,该方法能有效地减少蜂王浆中链霉素 检测的假阳性结果,符合目前检测工作的需要。     

Determination of chloramphenicol in royal jelly by liquid chromatography/tandem mass spectrometry

A liquid chromatographic/tandem mass spectrometric method was developed and validated for the determination of chloramphenicol (CAP) in royal jelly. Royal jelly samples were first denatured with lead acetate solution, and the CAP was extracted with solid-phase extraction before separation by liquid chromatography. A triple-quadrupole mass spectrometer operated in the negative electrospray ionization and selected-reaction monitoring mode was used for the detection of CAP. For method validation, royal jelly samples were fortified at CAP levels between 0.1 and 10.0 microg/kg; at these levels, recovery values (internal standard-corrected) ranged from 93.3 to 105.0%, and the within-laboratory reproducibility (relative standard deviation) was < or = 9.1%. The decision limit was 0.07 microg/kg, and the detection capability was 0.1 microg/kg.     

蜂王浆产品中5种大环内酯类抗生素残留量的高效液相色谱-质谱/质谱检测方法

建立了高效液相色谱-质谱/质谱测定蜂王浆中5种大环内酯类抗生素(螺旋霉素、竹桃霉素、泰乐菌素、罗红霉素、交沙霉素)残留的方法。先用三氯乙酸沉淀蜂王浆中的蛋白质,上层清液再用乙腈提取、C18小柱净化。每种抗生素选择一个母离子和两个子离子进行监测。5种大环内酯类抗生素在0.002～0.05 mg/L 范围内与其峰面积具有良好的线性关系,检测低限均为20 μg/kg,3个加标水平（每种抗生素的添加水平均为20, 100 和 200 μg/kg）下的加标回收率为73.0%～90.2%,相对标准偏差为5.6%～10.5%。     

Development and validation of a simple solid‐phase extraction method coupled with liquid chromatography–triple quadrupole tandem mass spectrometry for simultaneous determination of lincomycin,tylosin A and tylosin B in royal jelly

We have developed an analytical method for the determination of lincomycin, tylosin A and tylosin B residues in royal jelly using liquid chromatography–triple quadrupole tandem mass spectrometry analysis. For extraction and purification, we employed 1% trifluoroacetic acid and 0.1 m Na₂EDTA solutions along with an Oasis HLB cartridge. The target antibiotics were well separated in a Kinetex EVO C₁₈ reversed‐phase analytical column using a combination of 0.1% formate acid in ultrapure water (A) and acetonitrile (B) as the mobile phase. Good linearity was achieved over the tested concentration range (5–50 μg/kg) in matrix‐matched standard calibration. The coefficients of determination (R²) were 0.9933, 0.9933 and 0.996, for tylosin A, tylosin B and lincomycin, respectively. Fortified royal jelly spiked with three different concentrations of the tested antibiotics (5, 10 and 20 μg/kg) yielded recoveries in the range 80.94–109.26% with relative standard deviations ≤4%. The proposed method was applied to monitor 11 brand of royal jelly collected from domestic markets and an imported brand from New Zealand; all the samples tested negative for lincomycin, tylosin A and tylosin B residues. In conclusion, 1% trifluoroacetic acid and 0.1 m Na₂EDTA aqueous solvents combined with solid‐phase extraction could effectively complete the sample preparation process for royal jelly before analysis. The developed approach can be applied for a routine analysis of lincomycin, tylosin A and tylosin B residues in royal jelly.     

高效液相色谱-串联质谱法同时测定蜂王浆中7种高风险农药残留

建立了高效液相色谱-串联质谱（HPLC-MS/MS）同时测定蜂王浆中氟胺氰菊酯、三唑醇、蝇毒磷、吡氟乙草灵、多菌灵、乙基硫菌灵和甲基硫菌灵7种高风险农药残留的分析方法。样品在碱性条件下经乙腈提取,无水硫酸钠脱水后,采用HLB固相萃取小柱富集净化。采用Venusil MP C18色谱柱分离,以0.5 mmol/L乙酸铵水溶液（含0.1%（v/v）甲酸）-甲醇（含0.1%（v/v）甲酸）为流动相,梯度洗脱。在电喷雾离子（ESI）源、正离子模式和多反应监测（MRM）模式下采集数据,内标法定量。结果表明,7种高风险农药在5~100 μg/kg范围内线性关系良好,相关系数（r²）为0.9921~0.9996;方法的检出限和定量限分别为0.5~2.0 μg/kg和1.0~5.0 μg/kg。在高、中、低3个添加水平下7种农药的加标回收率为80.5%~101.3%,相对标准偏差（RSD）为3.6%~9.4%。该法操作简单,灵敏度高,准确可靠,能够满足出口蜂王浆农残限量检测的要求。     

动物源性食品中磺胺类药物增效剂残留的高效液相色谱-串联质谱法测定

利用高效液相色谱-电喷雾串联质谱测定了蜂蜜、蜂王浆、鮰鱼、鳗鱼、猪肉、猪肾、猪肝、鸡肉、牛肉和牛奶中的三甲氧苄氨嘧啶、二甲氧苄氨嘧啶和奥美普林3种磺胺类药物增效剂.除蜂王浆基质直接用10%三氯乙酸溶液提取外,其余基质均用10%三氯乙酸-乙腈(体积比7 : 3)混合溶液提取,提取溶液过阳离子交换固相萃取柱进行富集和净化.采用C18色谱柱,流动相为甲醇和0.1%甲酸溶液.选择1个母离子和2个子离子进行反应监测,对3种磺胺类药物增效剂残留进行定性,选择信号最强的子离子进行定量.在2 ～100 μg/L范围内,分析物的线性相关系数r>0.992.通过实际样品添加回收实验,所有基质定量下限为5.0 μg/kg,3个添加水平的回收率为63% ～89%,相对标准偏差为3.2% ～6.9%.     

Determination of trans-10-hydroxy-2-decenoic acid content in pure royal jelly and royal jelly products by column liquid chromatography.

In this research, several royal jellies and commercial products containing royal jelly were analysed for their trans-10-hydroxy-2-decenoic acid (10-HDA) content by using a column liquid chromatography technique. Ten samples claimed to be pure royal jelly, containing 10-HDA between 0.75 and 2.54%. Seven samples claimed to contain royal jelly as an ingredient which ranged from non-detectable to 0.054%. The technique was found to be rapid with high recovery.     

Simultaneous determination of seven fluoroquinolones in royal jelly by ultrasonic‐assisted extraction and liquid chromatography with fluorescence detection

A method for the quantitative determination of seven fluoroquinolone antibacterial agents (FQs) used in beekeeping, viz. ciprofloxacin, norfloxacin, ofloxacin, pefloxacin, danofloxacin, enrofloxacin, and difloxacin, in royal jelly samples was developed on the basis of high performance liquid chromatography with fluorescence detection. Sample preparation included deproteination, ultrasonic‐assisted extraction with a mixed inorganic solution of monopotassium phosphate (KH₂PO₄) and ethylenediaminetetraacetic acid disodium salt (Na₂EDTA), and clean‐up on a solid‐phase extraction cartridge. The extraction procedure was optimized with regard to the amount of inorganic solvent and the duration of sonication for royal jelly as a complicated matrix. Overall recoveries for FQs ranged from 85.9 to 99.1% for royal jelly with standard deviations between 2.79 and 6.27%. Limits of quantification were 2–40 ng/g for seven FQs in royal jelly. A total of 57 real royal jelly samples collected from beekeepers and supermarkets were analyzed. The three most abundant honeybee‐use FQs, i. e. ofloxacin, ciprofloxacin, and norfloxacin, were determined in some royal jelly samples in concentrations ranging from 11.9 to 55.6 ng/g. Unexpectedly, however, difloxacin was found at concentrations of about 46.8 ng/g in one sample although it is rarely used in beekeeping. The presented method was successfully applied to quantify FQs in real royal jelly samples.     

Formulation of Ocular In Situ Gels with Lithuanian Royal Jelly and Their Biopharmaceutical Evaluation In Vitro

Royal jelly is a natural substance produced by worker bees that possesses a variety of biological activities, including antioxidant, anti-inflammatory, antibacterial, and protective. Although fresh royal jelly is kept at low temperatures, to increase its stability, it needs to be incorporated into pharmaceutical formulations, such as in situ gels. The aim of this study was to formulate in situ ocular gels containing Lithuanian royal jelly for topical corneal use in order to increase the retention time of the formulation on the ocular surface and bioavailability. Gels were evaluated for physicochemical characteristics (pH, rheological properties, refractive index) and in vitro drug release measuring the amount of 10-hydroxy-2-decenoic acid (10-HDA). An ocular irritation test and cell viability tests were performed using the SIRC (Statens Seruminstitut Rabbit Cornea) cell culture line. Results indicated that all the in situ gels were within an acceptable pH and refractive index range close to corneal properties. Rheology studies have shown that the gelation temperature varies between 25 and 32 °C, depending on the amount of poloxamers. The release studies have shown that the release of 10-HDA from in situ gels is more sustained than royal jelly suspension. All gel formulations were non-irritant according to the short-time exposure test (STE) using the SIRC cell culture line, and long-term cell viability studies indicated that the formulations used in small concentrations did not induce cell death. Prepared in situ gels containing royal jelly have potential for ocular drug delivery, and they may improve the bioavailability, stability of royal jelly, and formation of non-irritant ocular formulations.     

高效液相色谱法测定蜜蜂蜂体王浆酸

建立了蜂体中10－HDA的高效液相色谱分析方法。对10－HDA的提取方法进行了研究。在选择的最佳色谱条件下,线性范围为10～1000ng,r=0．9998,回收率96．5％～99．2％,检测限为0.53μg／g。     

Determination of proline enantiomers in honey and royal jelly by LC-UV

The chiral separation and quantification of D-proline and L-proline in honey and royal jelly were examined by LC with UV detection. Most of the endogenous compounds existing in honey, such as sugars, were removed by using SPE cartridges containing C18 and strong cation-exchange sorbent. Other components, such as primary amino acids, were also removed by two-step derivatization with o-phthalaldehyde (OPA) and 9-fluorenylmethyl chloroformate (FMOC-CI). The components that were derivatized with OPA were separated from proline with a C18 cartridge. Proline was then converted into an FMOC derivative that could be subsequently measured by LC-UV. Sufficient chiral separation of D-proline and L-proline was achieved with an LC chiral column made of a beta-cyclodextrin phase in the polar organic-phase mode. The average recoveries of D-proline and L-proline from honey and royal jelly were in the range of 81.3-98.6% (RSD of < 1.8%). When this method was applied to commercial honey and royal jelly samples, L-proline was detected at concentrations of 369-1930 microg/g, whereas D-proline was not detected.     

Development and validation of an UPLC-DAD-MS method for the determination of leonurine in Chinese motherwort (Leonurus japonicus)

In the present study, an ultra-performance liquid chromatography method with diode array detection (UPLC-DAD) was developed for the analysis and determination of leonurine from motherwort (Leonurus japonicus), a traditional Chinese medicinal plant. This method was validated in terms of specificity, linearity (R(2) = 0.9995, linear range: 0.005 ~ 0.5 mg/mL), precision (< 5.0% RSD), and recovery (103.2%). The extracted amount of leonurine is 0.15 mg/g. Moreover, the target analyte was identified or tentatively characterized using UPLC coupled with electrospary tandem mass spectrometry (UPLC-ESI-MS).     

气相色谱法测定化妆品中二甘醇的含量及其测量不确度评定

采用气相色谱法测定了化妆品中的二甘醇,并用计量学方法对测量不确定度进行了评定。结果显示,二甘醇含量在0.01~0.1 mg/mL范围内与色谱峰面积呈线性关系,线性相关系数为0.999 9,检测限为0.41μg/mL,不同类别的3种样品中二甘醇测定结果的相对标准偏差在0.24%~1.52%范围内(n=6),平均回收率在93.65%~104.36%之间。当啫喱水样品中二甘醇的测定结果为48.1 mg/kg时,其扩展不确定度为1.4 mg/kg。该方法可用于化妆品中二甘醇含量的质量控制。     

高效液相色谱-串联质谱法测定水果中残留的烯唑醇

建立了水果中烯唑醇残留量的高效液相色谱-串联质谱（LC-MS/MS）测定方法。样品采用丙酮-正己烷液液振荡提取,经分散固相萃取（DSPE）净化后,采用ODS-C18柱为分离柱,以乙腈-0.1％甲酸溶液为流动相,采用液相色谱-串联质谱(ESI)多反应监测（MRM）正离子模式测定,外标法定量。烯唑醇在0.002～0.2 mg//L质量浓度范围内线性关系良好,方法定量下限（LOQ）为0.001 mg/kg。在0.005～0.1 mg/kg之间的3个添加浓度水平下,添加回收率为78.8 %~108.0 %,相对标     

固相萃取-超高效液相色谱-串联质谱法测定乳粉和果冻中的氯化胆碱

建立了固相萃取-超高效液相色谱-串联质谱测定乳粉和果冻中氯化胆碱的分析方法。样品在10 mL 0.02mol/L乙酸铵溶液(冰乙酸调节pH至3.0)中水解3 h后离心,上清液经DIKMA ProElut PLS固相萃取柱(60 mg/3 mL)净化后,用Agilent ZORBAX 300 SCX色谱柱(150 mm×2.1 mm,5μm)进行分离,通过电喷雾正离子(ESI~+)模式电离,多反应监测(MRM)模式进行定性和定量分析。方法的检出限(LOD,S/N=3)和定量限(LOQ,S/N=10)分别为0.15 mg/kg和0.50 mg/kg,加标回收率为70.8%~100.2%,相对标准偏差(RSD)均不大于6.83%(n=6)。目标化合物在0.05~8.0 mg/L范围内线性关系良好,线性方程为Y=2.05×10~5X+3.24×10~4,相关系数(r)为0.996。对市售乳粉和果冻中的氯化胆碱进行检测,结果表明,乳粉和果冻中氯化胆碱的含量分别为251.0~2 448 mg/kg和0.261~0.314 mg/kg。该法准确可靠、灵敏度高,适用于乳粉和果冻中氯化胆碱的测定。     

Tetracycline residues in royal jelly and honey by liquid chromatography tandem mass spectrometry: validation study according to Commission Decision 2002/657/EC

A specific, sensitive and robust liquid chromatography tandem mass spectrometry method for determining oxytetracycline, tetracycline, chlortetracycline and doxycycline in royal jelly and honey samples is presented. Extraction of drug residues was performed by ammonium acetate buffer as extractant followed by a clean-up with metal chelate affinity chromatography and solid-phase extraction. Tetracycline analysis was performed using liquid chromatography–electrospray ionisation–tandem mass spectrometry. The presented method is the first validated for royal jelly and in accordance with the requirements set by Commission Decision 2002/657/EC. Recoveries of the methods, calculated spiking the samples at 5.0, 10.0, 20.0 and 30.0 μg kg⁻¹, were 79% to 90% for honey and 77% to 90% for royal jelly. The intra-day precision (RSD) ranged between 8.1% and 15.0% for honey and from 9.1% to 16.3% for royal jelly, while inter-day precision values were from 10.2% to 17.6% and from 10.6% to 18.4% respectively for honey and royal jelly. Linearity for the four analytes was calculated from 5.0 to 50.0 μg kg⁻¹. The decision limits (CCα) ranged from 6.2 to 6.4 μg kg⁻¹ and from 6.1 to 6.5 μg kg⁻¹ for honey and royal jelly, respectively. Detection capabilities values (CCβ) ranged between 7.2 and 7.7 μg kg⁻¹ and from 7.3 to 7.9 μg kg⁻¹ respectively for honey and royal jelly. The developed method is currently in use for confirmation of the official control analysis of honey and royal jelly samples.     

Rapid gas chromatographic method for the determination of famoxadone, trifloxystrobin and fenhexamid residues in tomato, grape and wine samples

Trifloxystrobin, fenhexamid and famoxadone belong to the generation of fungicides acting against a broad spectrum of fungi and widely used in Integrated Pest Management strategies in different agricultural crops but mainly in viticulture. In the present work, a gas chromatographic (GC) method for their determination was developed and validated on tomato, grape and wine matrices. The method was based on a simple one step liquid-liquid microextraction with cyclohexane/dichloromethane (9+1, v/v) and determination of fungicides by gas chromatography with nitrogen phosphorous (NP-) and electron capture (EC-) detection, and ion trap mass spectrometry (ITMS) for confirmation. The method was validated by recovery experiments, assessment of matrix effect and calculation of the associated uncertainty. Recoveries for GC-NPD and GC-ECD were found in the range of 81-102% with RSD <12%, while matrix-matched calibration solutions were imposed for quantification. LOQs ranged from 0.005 to 0.05 mg/kg and 0.01 to 0.10 mg/kg for the GC-ECD and GC-NPD, respectively, depending on the sensitivity of each compound with trifloxystrobin being the most sensitive. The expanded uncertainty, calculated for a sample concentration of 0.10 mg/kg, ranged from 4.8 to 13% for the GC-ECD and from 5.4 to 29% for the GC-NPD. The concentration levels for famoxadone residues found in tomato and grape samples from field experiments were clearly below the EU established MRL values, thus causing no problems in terms of food safety.