气相色谱-质谱分析血清葡萄糖的前处理方法研究

用气相色谱-同位素稀释质谱法(GC-ID/MS)测定血清葡萄糖,需要对血清样品进行前处理,为确保测定结果的准确性,比较了乙醇、丙酮、乙腈沉淀及微过滤4种去除蛋白的方法.将用上述4种方法处理的血清样品进行衍生,采用GC-ID/MS测定,得到其精密度、相对回收率和绝对回收率,加以比较.结果表明:4种方法中,乙醇去除血清中蛋白的效果最佳.     

Study on the quality control of cell therapy products. Determination of N-glycolylneuraminic acid incorporated into human cells by nano-flow liquid chromatography/Fourier transformation ion cyclotron mass spectrometry

N-Glycolylneuraminic acid (NeuGc), an acidic nine-carbon sugar, is produced in several animals, such as cattle and mice. Since human cells cannot synthesize NeuGc, it is considered to be immunogenic in humans. Recently, NeuGc contamination was reported in human embryonic stem cells cultured with xenogeneic serum and cells, suggesting that possibly NeuGc may harm the efficacy and safety of cell therapy products. Sialic acids have been determined by derivatization with 1,2-diamino-4,5-methylenedioxybenzene (DMB) followed by liquid chromatography/mass spectrometry (LC/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS); however, the limited availability of cell therapy products requires more sensitive and specific methods for the quality test. Here we studied the use of nano-flow liquid chromatography/Fourier transformation ion cyclotron resonance mass spectrometry (nanoLC/FTMS) and nanoLC/MS/MS for NeuGc-specific determination at a low femtomole level. Using our method, we found NeuGc contamination of the human cell line (HL-60RG cells) cultured with human serum. Our method needs only 2.5x10(3) cells for one injection and would be applicable to the determination of NeuGc in cell therapy products.     

Sensitive CE-MS method for monitoring of riociguat and desmethylriociguat levels in human serum

Riociguat is novel antihypertensive drug for treatment of pulmonary hypertension. As such, it is still being tested in many clinical and pharmacokinetic trials. Existing methods that determine serum riociguat and desmethylriociguat (DMR) are based solely on liquid chromatography with mass spectrometry. Therefore, we present a novel capillary electrophoresis with mass spectrometry method (CE-MS) for their determination in human serum as alternative method for ongoing trials. Complete resolution of both analytes was achieved by means of pH optimization of ammonium formate background electrolytes that are fully compatible with ESI/MS detection. Simple liquid-liquid extraction was used as sample pretreatment. The calibration dependence of the method was linear (in the range of 10–1000 ng/mL), with adequate accuracy (90.1–114.9%) and precision (13.4%). LOD and LOQ were arbitrarily set at 10 ng/mL for both analytes. Clinical applicability was validated using serum samples from patients treated with riociguat in pharmacokinetic study and the results corresponded with reference HPLC-MS/MS values. Capillary electrophoresis proved to be sensitive and selective tool for the analysis of riociguat and DMR.     

Identification and determination of fat-soluble vitamins and metabolites in human serum by liquid chromatography/triple quadrupole mass spectrometry with multiple reaction monitoring

A method for determination of fat-soluble vitamins K(1), K(3), A, D(2), D(3) and E (as alpha- and delta-tocopherol) and metabolites 25-hydroxyvitamin D(2) and D(3) and 1,25-dihydroxyvitamin D(3) in human serum by liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) in positive mode is proposed. Highly selective identification of the target compounds in serum was confirmed by the most representative transitions from precursor ion to product ion. Quantitative MS/MS analysis was carried out by multiple reaction monitoring optimizing the most sensitive transition for each analyte in order to achieve low detection limits (from 0.012 to 0.3 ng/mL estimated with serum). The analysis was performed with 1 mL of serum, which was subjected to protein precipitation, liquid-liquid extraction to an organic phase, evaporation to dryness and reconstitution with methanol. The precision of the overall method ranged from 3.17-6.76% as intra-day variability and from 5.07-11.53% as inter-day variability. The method, validated by the standard addition method, provides complete information on the fat-soluble vitamins profile, which is of interest in clinical and metabolomics studies.     

High throughput on-line solid phase extraction/tandem mass spectrometric determination of paclitaxel in human serum

The feasibility of high throughput on-line solid phase extraction/tandem mass spectrometry (SPE/MS/MS) is tested for target analysis of paclitaxel in human serum. The use of a dual Prospekt system, with parallel SPE and elution directly to the mass spectrometer, resulted in a cycle time of 80 seconds for the entire, fully automated assay. The assay proved to be linear from 1 to 1000 ng/mL. Cartridges packed with small sorbent particles functioned both as SPE cartridges and as short analytical columns.     

Doping control analysis for adrafinil and its major metabolites in human urine

A new and reliable two‐step liquid chromatography/tandem mass spectrometry (LC/MS/MS) method in combination with gas chromatography/mass spectrometry (GC/MS) for the screening and confirmation of adrafinil and its major metabolites, modafinil and modafinil acid, in human urine has been developed and validated. The method involved reversed‐phase C18 solid‐phase extraction (SPE) cartridge extraction and MS analysis by means of LC/MS/MS and GC/MS. The study illustrated that the ESI capillary temperature played a key role in the formation of the protonated molecule. The limits of detection (LODs) of the developed method for the three compounds were lower than the minimum required performance limit (MRPL) of the World Anti‐Doping Agency (WADA). The human urine samples obtained after the oral administration of modafinil and from the Beijing 2008 Olympic Games were analyzed by using the described method, which has also been successfully applied to routine analyses and the WADA Proficiency Test. Copyright © 2009 John Wiley & Sons, Ltd.     

High turbulence liquid chromatography online extraction and tandem mass spectrometry for the simultaneous determination of suberoylanilide hydroxamic acid and its two metabolites in human serum

A reliable and sensitive method incorporating high turbulence liquid chromatography (HTLC) online extraction with tandem mass spectrometry (MS/MS), for simultaneous determination of suberoylanilide hydroxamic acid (SAHA) and its two metabolites, SAHA-glucuronide (M1) and 4-anilino-4-oxobutanoic acid (M2), in human serum, has been developed to support clinical studies. The HTLC technology significantly reduces the time required for sample clean-up since sample extraction and analysis are performed online. Clinical samples, internal standards (IS) and buffer are transferred into 96-well plates using a robotic liquid handling system. A 20 microL aliquot of prepared sample is directly injected into the HTLC/LC-MS/MS system where the matrix is rapidly washed away to waste and the analytes are retained on the narrow-bore extraction column (0.5 x 50 mm), using an aqueous mobile phase at 1.5 mL/min. Analytes are then eluted from the extraction column and transferred to the analytical column using a gradient mobile phase prior to detection by MS/MS. Interference with determination of SAHA from in-source dissociation of M1 is eliminated by the chromatographic separation. The resolution of SAHA and M1 did not change for more than 1500 serum sample injections by applying an acid wash (15% acetic acid) on the extraction column. The linear calibration ranges for SAHA, M1, and M2 are 2-500, 5-2000, and 10-2000 ng/mL, respectively. Assay intraday validation was conducted using five calibration curves prepared in five lots of human control serum. The precision expressed as relative standard deviation (RSD) is less than 6.8% and accuracy is 94.6-102.9% of nominal values for all three analytes. Assay specificity, freeze/thaw stability, storage stability, and matrix effects were also assessed.     

On-line extraction coupled with liquid chromatography tandem mass spectrometry for quantitation of pravastatin and its metabolite in human serum

A high-throughput bioanalytical method for simultaneous quantitation of pravastatin and its metabolite (M1) in human serum was developed and validated using on-line extraction following liquid chromatography tandem mass spectrometry (LC-MS/MS). The on-line extraction was accomplished by the direct injection of a 50 microL serum sample, mixed 4:1 with an aqueous internal standard solution, into one of the extraction columns with aqueous 1 mm formic acid at flow rate of 3 mL/min. The separation and analysis were achieved by back-eluting the analytes from the extraction column and the analytical column to the mass spectrometer with an isocratic mobile phase consisting of 62% aqueous 1 mm formic acid and 38% acetonitrile at a flow rate of 0.8 mL/min. The second extraction column was being equilibrated while the first column was being used for analysis, and vice versa. The standard curve range was 0.500-100 ng/mL for pravastatin and M1. The lower limit of quantitation, 0.500 ng/mL for all the analytes, was achieved when 50 microL of human serum was used. The intra- and inter-day precisions were within 7.4%, and the accuracy was between 95 and 103%. The on-line extraction was finished in 0.5 min and total analysis time was 2.5 min per sample.     

Development and evaluation of a candidate reference measurement procedure for the determination of testosterone in human serum using isotope dilution liquid chromatography/tandem mass spectrometry

A candidate reference measurement procedure for total testosterone in human serum involving isotope dilution (ID) coupled with liquid chromatography/tandem mass spectrometry (LC/MS/MS) has been developed and critically evaluated. The endogenous testosterone and its internal standard (testosterone-d ₃) were extracted from the serum matrix using a combination of solid-phase extraction and liquid–liquid extraction prior to reversed-phase LC/MS/MS. Accuracy of the measurements was evaluated by a recovery study using testosterone-spiked serum. The recovery of the added testosterone ranged from 100.0 to 100.3%. This method was applied to the determination of testosterone in frozen serum samples from three individual donors (one female and two males) with the testosterone concentrations ranging from 0.3 to 8.5 ng g⁻¹. Repeatability with within-set coefficients of variation (CVs) from 0.1 to 1.0% and intermediate precision with between-set CVs from 0.1 to 0.5% for both female and male serum materials were demonstrated. Excellent linearity was obtained for all linear regression lines. The detection limit at a signal-to-noise ratio of approximately 3 was 2 pg of testosterone in serum. Structural analogs as well as testosterone metabolites were tested and found to not interfere with the measurement of testosterone. This well-characterized LC/MS/MS method for serum testosterone, which demonstrates good accuracy and precision, and low susceptibility to interferences, qualifies as a reference measurement procedure that can be used to provide an accuracy base to which routine methods for testosterone can be compared and that will serve as a standard of higher order for measurement traceability.     

High‐throughput salting‐out assisted liquid–liquid extraction with acetonitrile for the determination of anandamide in plasma of hemodialysis patients with liquid chromatography tandem mass spectrometry

Anandamide (AEA) is an endocannabinoid present in human plasma that is associated with several physiological functions and disease states. However, low AEA plasma levels pose challenges in terms of analytical characterization. Classical liquid‐based lipid extraction and solid‐phase extraction require complicated procedures and the drying down of relatively large volumes of solvents, making them unsuitable for high‐throughput analysis. Here a high‐throughput salting‐out assisted liquid–liquid extraction (SALLE) method with acetonitrile and mass spectrometry compatible salts for liquid chromatography–tandem mass spectrometry (LC‐MS/MS) analysis of AEA in human plasma has been developed and validated. The seamless interface of SALLE and LC‐MS eliminated the drying‐down step, only 100 μL of plasma is required and minimal volumes of organic solvent are used. Good reproducibility, accuracy and precision were demonstrated during the method validation. The method is linear up to 10 ng/mL with a lower limit of quantitation of 0.1 ng/mL for AEA, the accuracy for AEA was from 93.3 to 96.7% and the precision was <8.57%. This new methodology was successfully applied to analysis of clinical samples from maintenance hemodialysis patients. Copyright © 2015 John Wiley & Sons, Ltd.     

母乳中多种含卤持久性有机污染物的联合检测方法

建立了母乳中多种含卤持久性有机污染物(POPs)的联合检测方法,目标化合物主要包括六溴环十二烷(HBCDs)、多溴联苯醚(PBDEs)、多氯联苯(PCBs)和有机氯农药(OCPs)等.样品的前处理采用液液萃取、凝胶渗透色谱(GPC)净化和固相萃取(SPE)等技术,目标化合物经气相色谱-质谱联用仪(GC-MS)、液相色谱-三重四极杆串联质谱联用仪(LC-MS/MS)和气相色谱-三重四极杆串联质谱联用仪(GC-MS/MS)等进行检测.样品通过GPC除去脂肪,然后经SPE柱进一步净化并进行多组分分离,极大程度地减小了生物样品中复杂基质的干扰,适合样品量相对较小的人体样本中多种超痕量POPs的分析.应用灵敏度高、选择性更好的GC-MS/MS对样品中的PCBs和OCPs等进行分析,进一步降低基质的干扰.方法经过小牛血清加标实验验证,稳定可靠.POPs的加标回收率分别为88.7％～98.8％(PBDEs), 88.5％～92.5％(HBCDs), 67.9％～82.3％(PCBs)和81.7％～116.1％(OCPs),方法检出限分别为0.13～1.8 pg/mL(PBDEs), 0.31～1.2 pg/mL(HBCDs), 0.22～1.4 pg/mL(PCBs)和0.20～1.5 pg/mL(OCPs).采用本方法对潍坊地区20例母乳样品进行分析,结果显示,潍坊市母乳中HBCDs, PBDEs, PCBs、HCHs和DDTs的中值浓度分别为2.86, 7.76, 8.84、140和503 ng/g 脂重,此浓度水平与国内其它地区人群相当.     

Automated method for targeting analysis of prostanoids in human serum by on-line solid-phase extraction and liquid chromatography-mass spectrometry in selected reaction monitoring

Prostanoids are potent biologically active lipid molecules demanding for analysis methods combining precision, sensitivity and high-throughput for pharmacological and clinical applications. The present research describes the development and validation of an on-line automated method based on solid-phase extraction liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) for the quantification of prostanoids in human serum. This approach overcomes the main limitation of previous methods involving manual protocols, such as analyte losses, metabolites degradation and time-consuming protocols, are minimized. Human serum (100 μL) was directly injected into an automatic solid-phase extraction workstation for cleanup and preconcentration of the target metabolites. The eluate was on-line transferred to a reversed-phase analytical column for chromatographic separation prior to mass spectrometry detection in selected reaction monitoring mode. The detection limits for the target analytes ranged from 2.3 to 63.3 pg on column. The precision (expressed as relative standard deviation) was within 3.30 and 6.15% for repeatability and from 4.16 to 11.11% for within-laboratory reproducibility. Accuracy was evaluated with spiked and non-spiked serum samples to estimate concentration differences that could be affected by matrix effects or inefficient SPE performance. Accuracy, estimated as recovery factor, was from 87.7 to 100% for the target compounds. The proposed method is reliable and has an excellent potential for high-throughput use in both clinical and research laboratories by minimizing analyst intervention.     

Determination of antimigraine compounds rizatriptan, zolmitriptan, naratriptan and sumatriptan in human serum by liquid chromatography/electrospray tandem mass spectrometry

Development of a rapid, sensitive and selective method for the determination of antimigraine drugs from human serum is essential for understanding the pharmacokinetics of these drugs when administered concurrently. Solid phase extraction (SPE) using Oasis HLB was used to extract the drugs (sumatriptan, naratriptan, zolmitriptan and rizatriptan) and the internal standard bufotenine from serum. A method based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) was developed and validated to simultaneously quantitate these antimigraine drugs from human serum. The precursor and major product ions of the analytes were monitored on a triple quadrupole mass spectrometer with positive ion electrospray ionization (ESI) in the multiple reaction monitoring (MRM) mode. The base peak in all the analytes is formed by alpha cleavage associated with protonation of the secondary amine. Mechanisms for the formation of the collision-induced dissociation products of these antimigraine compounds are proposed. Linear calibration curves were generated from 1-100 ng/mL with all coefficients of determination greater than 0.99. The inter- and intraday precision (%RSD) were less than 9.3% and accuracy (%error) was less than 9.8% for all components. The limits of detection (LOD) for the method were 250 pg/mL for sumatriptan and 100 pg/mL for the remaining analytes based on a signal-to-noise ratio of 3.     

Rapid bioanalysis of vancomycin in serum and urine by high-performance liquid chromatography tandem mass spectrometry using on-line sample extraction and parallel analytical columns

A novel high-performance liquid chromatography tandem mass spectrometry (LC/MS/MS) method is described for the determination of vancomycin in serum and urine. After the addition of internal standard (teicoplanin), serum and urine samples were directly injected onto an HPLC system consisting of an extraction column and dual analytical columns. The columns are plumbed through two switching valves. A six-port valve directs extraction column effluent either to waste or to an analytical column. A ten-port valve simultaneously permits equilibration of one analytical column while the other is used for sample analysis. Thus, off-line analytical column equilibration time does not require mass spectrometer time, freeing the detector for increased sample throughput. The on-line sample extraction step takes 15 seconds followed by gradient chromatography taking another 90 seconds. Having minimal sample pretreatment the method is both simple and fast. This system has been used to successfully develop a validated positive-ion electrospray bioanalytical method for the quantitation of vancomycin. Detection of vancomycin was accurate and precise, with a limit of detection of 1 ng/mL in serum and urine. The calibration curves for vancomycin in rat, dog and primate were linear in a concentration range of 0.001-10 microg/mL for serum and urine. This method has been successfully applied to determine the concentration of vancomycin in rat, dog and primate serum and urine samples from pharmacokinetic and urinary excretion studies.     

Enrichment of low molecular weight serum proteins using acetonitrile precipitation for mass spectrometry based proteomic analysis

A rapid acetonitrile (ACN)-based extraction method has been developed that reproducibly depletes high abundance and high molecular weight proteins from serum prior to mass spectrometric analysis. A nanoflow liquid chromatography/tandem mass spectrometry (nano-LC/MS/MS) multiple reaction monitoring (MRM) method for 57 high to medium abundance serum proteins was used to characterise the ACN-depleted fraction after tryptic digestion. Of the 57 targeted proteins 29 were detected and albumin, the most abundant protein in serum and plasma, was identified as the 20th most abundant protein in the extract. The combination of ACN depletion and one-dimensional nano-LC/MS/MS enabled the detection of the low abundance serum protein, insulin-like growth factor-I (IGF-I), which has a serum concentration in the region of 100 ng/mL. One-dimensional sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the depleted serum showed no bands corresponding to proteins of molecular mass over 75 kDa after extraction, demonstrating the efficiency of the method for the depletion of high molecular weight proteins. Total protein analysis of the ACN extracts showed that approximately 99.6% of all protein is removed from the serum. The ACN-depletion strategy offers a viable alternative to the immunochemistry-based protein-depletion techniques commonly used for removing high abundance proteins from serum prior to MS-based proteomic analyses.     

Simultaneous determination of benzodiazepines and their metabolites in human serum by liquid chromatography-tandem mass spectrometry using a high-resolution octadecyl silica column compatible with aqueous compounds

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method using a high-resolution octadecyl silica column compatible with aqueous compounds was developed for the simultaneous determination of benzodiazepines and their metabolites in human serum. This method enabled us to determine multiple benzodiazepines, including flurazepam, bromazepam, chlordiazepoxide, nitrazepam, clonazepam, flunitrazepam, estazolam, clobazam, lorazepam, alprazolam, triazolam, brotizolam, fludiazepam, diazepam, quazepam, prazepam and their metabolites such as 7-aminonitrazepam, 7-aminoclonazepam, 7-acetamidonitrazepam, N-desmethylclobazam and N-desmethyldiazepam. The analytes spiked into human serum were subjected to solid-phase extraction followed by liquid chromatography coupled with electrospray ionization tandem mass spectrometry. The running time was within 25 min for the measurement of 22 benzodiazepines and their metabolites. The recovery rates exceeded 58.1% for those compounds except for quazepam, which showed a recovery of 45.8%. The limit of detection ranged from 0.3 to 11.4 ng/mL. Linearity was satisfactory for all compounds. These data suggest that the present method can be applicable to routine assay for benzodiazepines in the clinical setting.     

Sensitive measurement of serum 1α,25-dihydroxyvitamin D by liquid chromatography/tandem mass spectrometry after removing interference with immunoaffinity extraction

Vitamin D plays important roles in bone health and a variety of other pathophysiological conditions. 1α,25‐Dihydroxyvitamin D is the active form of vitamin D. Quantification of serum 1α,25‐dihydroxyvitamin D is useful for evaluation of several diseases including chronic renal failure, hypoparathyroidism, sarcoidosis, and rickets. Measurement of 1α,25‐dihydroxyvitamin D is very challenging due to its low circulating concentration and presence of interfering substances in serum. In this report, a liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for quantifying serum 1α,25‐dihydroxyvitamin D is described. Lithium adducts of 1α,25‐dihydroxyvitamin D were formed prior to mass spectrometry analysis to improve ionization efficiency. We tested a number of different sample preparation procedures and found that immunoaffinity extraction was the method of choice because it completely removed isobaric interferences and matrix effects present in patient serum. Extraction efficiency, expressed as absolute recovery, was greater than 60% in both patient serum and charcoal‐stripped serum. This method was linear from 3.4 to 206.2 pg/mL for 1α,25‐dihydroxyvitamin D₃ and 3.9 to 212.6 pg/mL for 1α,25‐dihydroxyvitamin D₂ with an accuracy of 89.8–98.4% and 97.5–115.7%, respectively. Inter‐assay and intra‐assay coefficients of variance (CVs) for both analytes at two different concentration levels ranged from 2.5–7.0%. Comparison with a radioimmunoassay for measuring total 1α,25‐dihydroxyvitamin D concentration using 40 patient samples showed a Deming regression slope of 0.751, a y‐intercept of 0.84 pg/mL, an r value of 0.7909, and a mean percentage difference of –27.1%. Comparison with a reference LC/MS/MS method (n = 20) showed a Deming regression slope of 1.020, a y‐intercept of 1.32 pg/mL, an r value of 0.9797, and a mean percentage difference of –2.9%. In conclusion, usage of immunoaffinity extraction enabled a sensitive LC/MS/MS method for quantification of 1α,25‐dihydroxyvitamin D in serum. Copyright © 2011 John Wiley & Sons, Ltd.     

Measurement of bisphenol A in human serum by gas chromatography/mass spectrometry

The purpose of this study is to establish an easy and accurate method for the determination of bisphenol A (BPA) in the human serum. The samples were applied to the C₁₈ solid phase extraction (SPE) column for clean up of samples. The BPA is conjugated with tetrabutylammonium hydrogen sulfate as the counter ion in alkali solution. The ion paired BPA is moves from the aqueous phase to the organic phase as an ion paired extraction. BPA extracted from human serum were derivatized with pentafluorobenzyl bromide (PFBBr). The derivative was analyzed by gas chromatography (GC)/mass spectrometry (MS) using negative chemical ionization (NCI). The instrumental detection limit of BPA was 5 pg/ml (10 fg). The instrumental response between 0.01 and 100 pg/ml of BPA standards was linear (r²=0.998). The recovery of BPA spiked into human serum was 101.0±4.63 (1 pg/ml) and 100.9±3.75 (10 pg/ml), respectively. The concentration of BPA in the human serum from 20 individuals was 0.54 pg/ml.     

固相萃取-超高效液相色谱/串联质谱法检测人体血清中邻苯二甲酸二(2-乙基已基)酯代谢物

建立了固相萃取-超高效液相色谱/串联质谱法检测人体血清中邻苯二甲酸二(2-乙基已基)酯(DEHP)代谢物.血清样品中加入乙酸钠溶液及内标13C4邻苯二甲酸单乙基已基酯(13C4MEHP)后,在37℃条件下用β-葡萄糖醛苷酶酶解提取12h,提取液经MAX固相萃取小柱净化并浓缩到纯水中.以甲醇5 mmol/L乙酸铵溶液为流动相,经Waters UPLC(R)HSS T3色谱柱分离后,在电喷雾离子源负离子模式下,以多反应监测(MRM)方式进行扫描,内标法定量.DEHP代谢物在1.0～100.0 μg/L范围内线性关系良好,检出限均为0.1 μg/kg,回收率89.6％～103.2％.该方法灵敏度高,准确度好,可作为监测人群内暴露的重要技术支撑.     

固相萃取结合超高效液相色谱-串联质谱法及气相色谱-负化学源质谱法测定人血清中的四溴双酚A、六溴环十二烷和多溴联苯醚

建立了固相萃取同时提取、净化血清中四溴双酚A(TBBPA)、α, β, γ-六溴环十二烷(HBCD)和8种多溴联苯醚(PBDEs)同系物的样本前处理方法,并结合色谱-质谱分离分析技术检测人血清样本中该类化合物的含量。试样在加入各自的同位素内标物后以甲基叔丁基醚/正己烷(1:1, v/v)混合溶剂进行萃取,再经浓硫酸去除脂肪后,以LC-Si固相萃取柱分离HBCD/TBBPA和PBDEs。采用分步检测的方式,在50 mm长BEH C18反相色谱柱上以超高效液相色谱-串联质谱(UPLC-MS/MS)的多反应监测模式(MRM)检测HBCD和TBBPA,在15 m长的毛细管柱上以气相色谱-负化学源质谱(GC-NCI/MS)的选择离子监测模式(SIM)检测PBDEs。以胎牛血清为空白基质,当HBCD、TBBPA和BDE-209的加标水平为0.5 ng/g和5 ng/g、三溴至七溴联苯醚的加标水平为0.05 ng/g和0.5 ng/g时,它们的平均加标回收率为80.3%～108.8%,相对标准偏差为1.02%～11.42%(n=5);以信噪比(S/N)为3计算,方法的检出限(LOD)为1.81～42.16 pg/g。采用该方法对实际样品进行测定,结果表明,本方法快速、准确、灵敏度高,能够满足血清中HBCD、TBBPA和PBDEs残留的同时提取及测定的要求。