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1.
Moser C Gschliesser A Mattle V Wildt L Griesmacher A Seger C 《Analytical and bioanalytical chemistry》2011,400(8):2655-2662
A selective and sensitive liquid chromatography-tandem mass spectrometry method for the determination of very low levonorgestrel
(d-(−)-norgestrel) serum levels such as those found in patients using levonorgestrel-releasing intrauterine devices (IUDs) was
developed. To achieve the sub-nanomolar sensitivity needed to measure such serum levels, a diethyl ether extraction sample
preparation protocol was applied prior to the online solid-phase extraction-liquid chromatography-tandem mass spectrometry
(SPE-LC-MS/MS) assay. Analyte quantification from the selected reaction monitoring experiments relied on the use of sixfold
deuterated norgestrel as internal standard. The final method was linear up to 1.50 ng/ml with a lower limit of quantification
(LLOQ) of 0.05 ng/ml. It was found to be precise and accurate with imprecision <8% and bias <6% assessed at three control
levels. Total analyte recovery measured in patient pools at three concentration levels was found to exceed 92%. Matrix interferences
were excluded by post-column analyte infusion experiments. As a proof of concept, a set of IUD patient serum samples was screened
for their levonorgestrel content. A total of 97.5% (n = 94) of the samples did show serum levels exceeding the LLOQ, proving the applicability of the assay in relevant clinical
cohorts. This method must not be used for diagnostic or therapeutic purposes, since it did not undergo formal performance
evaluation in the sense of the in vitro diagnostic directive (98/79/EG) of the European community. 相似文献
2.
A convenient, selective and sensitive liquid chromatographic-electrospary ionization mass spectrometry (LC–ESI–MS) method
was developed and validated to determine lovastatin in human plasma. The analyte was extracted from human plasma samples by
typical liquid–liquid extraction, separated on a C18 column by using the mobile phase consisting of water–methanol (13:87, v/v). Simvastatin was used as the internal standard (IS). The method was linear within the range of 0.1–20 ng mL−1. The lower limit of quantification (LLOQ) was 0.1 ng mL−1. The intra- and inter-run precision, calculated from quality control (QC) samples was less than 10.2%. The accuracy as determined
from QC samples was in the range of 99.3–102.9% for the analyte. The mean recoveries for lovastatin and IS were 84.8 and 88.0%,
respectively. The method was successfully applied for evaluation of the pharmacokinetic of lovastatin in healthy volunteers. 相似文献
3.
A sensitive LC-MS/MS assay for quantification of total and free concentrations of R- and S-warfarin in plasma was required
to support clinical studies on warfarin enantiomers. Several ultrafiltration devices were evaluated for separation of free
warfarin from plasma proteins. The highest precision and lowest non-specific binding was obtained for Centrifree ultrafiltration
devices. R- and S-warfarin were extracted from plasma (total) and ultrafiltrate (free) by liquid–liquid extraction with methyl
tert-butyl ether using d6-warfarin as internal standard. Mean extraction recovery was 68 ± 4%. The enantiomers were separated on a Chirobiotic V column
with isocratic elution using 40% methanol and 0.03% acetic acid in water. Negative mode electrospray ionisation was used for
MS/MS detection, monitoring the ion transition m/z 307/161. Calibration curves (quadratic, weighted 1/x) were fitted over the range of 20–2,000 ng/ml (r
2 ≥ 0.995) in plasma and 0.5–20 ng/ml (r
2 ≥ 0.998) in ultrafiltrate. The lower limit of quantification for R- and S-warfarin was 0.5 ng/ml in ultrafiltrate. Intra-
and interday precision (% RSD) and bias were within 10% in all cases, and matrix effects were negligible. The assay was applied
successfully to analysis of samples from clinical studies. 相似文献
4.
Babu Rao Chandu Sreekanth Nama Kanchanamala Kanala Balasekhara Reddy Challa Rihana Parveen Shaik Mukkanti Khagga 《Analytical and bioanalytical chemistry》2010,398(3):1367-1374
A novel simple, sensitive, selective, and rapid high-performance liquid chromatography coupled with tandem mass spectrometry
method was developed and validated for quantification of riluzole in human plasma. The chromatography was performed by using
a Zorbax-SB-C18 (4.6 × 75 mm, 3.5 μm) column , isocratic mobile phase 0.1% formic acid/acetonitrile (10:90 v/v), and an isotope-labeled
internal standard (IS), [13C,15N2]riluzole. The extraction of drug and internal standard was performed by liquid–liquid extraction and analyzed by MS in the
multiple reaction monitoring (MRM) mode using the respective [M+H]+ ions, m/z 235.0/165.9 for riluzole and m/z 238.1/169.0 for the IS. The calibration curve was linear over the concentration range 0.5–500.0 ng/ml for riluzole in human
plasma. The limit of quantification (LOQ) was demonstrated at 0.5 ng/ml. The within-batch and between-batch precision were
0.6–2.3% and 1.4–5.7%, and accuracy was 97.1–101.1% and 98.8–101.2% for riluzole respectively. Drug and IS were eluted within
3.0 min. The validated method was successfully applied in a bioequivalence study of riluzole in human plasma. 相似文献
5.
León-González Z Ferreiro-Vera C Priego-Capote F Luque de Castro MD 《Analytical and bioanalytical chemistry》2011,401(3):1003-1011
The proven endocrine disruption nature of the sunscreen ingredient 2-ethylhexyl 4-(N,N-dimethylamino)benzoate (EDP) calls for research to understand its distribution and bioaccumulation in the human body. A sensitive
analytical method to determine EDP and its metabolites in human semen based on online SPE-LC-MS/MS is described. The method
has been fully validated and a standard addition calibration has been used for quantification to correct the observed matrix
effects. The on-column detection limits of the analytes are between 0.2 and 0.6 ng, depending on the analyte and the sample.
The repeatability of the method, expressed as relative standard deviation, was in the range 4.6–9.4%. The method was satisfactorily
applied to semen samples from male volunteers who were subjected to single and repeated whole-body applications of an EDP-containing
sunscreen product. EDP metabolites were found at different concentrations in semen samples from the repeated application study,
thus showing evidences of bioaccumulation in humans. 相似文献
6.
Neng Zhou Yi-Zeng Liang Ben-Mei Chen Ping Wang Xian Chen Feng-Ping Liu 《Chromatographia》2007,66(7-8):481-486
A rapid, simple and specific liquid chromatography-electrospray ionization mass spectrometry method has been developed and
validated for the determination of hydroxyzine hydrochloride in human plasma. Samples were separated using a Thermo Hypersil-HyPURITYC18
reversed-phase column (150 mm × 2.1 mm i.d., 5 μm). The mobile phase consisted of 50 mM ammonium acetate (pH 4.0)–methanol–acetonitrile
(45:36:19, v/v). Hydroxyzine and its internal standard were measured by electrospray ion source in positive selective ion monitoring mode.
The method was validated with a linear range of 1.56–200.0 ng mL−1 and the lowest limit of quantification was 1.56 ng mL−1 for hydroxyzine hydrochloride (r
2= 0.9991). The extraction efficiencies were about 70% and recoveries of the method were in the range of 93.5–104.4%. The intra-day
relative standard deviation (RSD) was less than 8.0% and inter-day RSD was within 7.4%. QC samples were stable when kept at
ambient temperature for 12 h at −20 °C for 30 days and after four freeze–thaw cycles. The method has been successfully applied
to the evaluation of pharmacokinetics and bioequivalence of two hydroxyzine hydrochloride formulations in 12 healthy Chinese
volunteers after an oral dose of 25 mg. 相似文献
7.
Ping Wang Yi-Zeng Liang Ben-Mei Chen Neng Zhou Lun-Zhao Yi Yan Yu Zhi-Biao Yi 《Chromatographia》2007,65(3-4):209-215
A rapid, simple and specific liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) method has been developed
and validated for the simultaneous determination of enalapril and its major active metabolite enalaprilat in human plasma.
Benazepril hydrochloride was used as the internal standard. Plasma was deproteinized with acetone and centrifuged. The supernatant
was transferred and evaporated to dryness and the residue dissolved in mobile phase. Samples were separated on a C18 column
with a mobile phase of methanol–20 mM ammonium acetate (53:47, v/v) containing 0.15% trifluoracetic acid (v/v) with a pH of 3.0. Enalapril, enalaprilat and the internal standard were measured by electrospray positive selective ion
monitoring mode. The method was validated over a linear range of 1.56–400 ng mL−1 and the limits of quantification were 1.56 ng mL−1 for both enalapril and enalaprilat using 0.1 mL plasma. Extraction efficiency was more than 84% and recoveries were in range
of 93.65–101.17%. The intra-day relative standard deviations (RSD) were less than 8.16 and 7.05% and inter-day RSDs were within
8.42 and 5.72% for enalapril and enalaprilat, respectively. The storage stability of QC samples was investigated under various
conditions. The method was successfully applied for the evaluation of the pharmacokinetics and bioequivalence of enalapril
and enalaprilat in 20 healthy volunteers after an oral dose of 20 mg enalapril maleate. 相似文献
8.
Summary A reversed-phase high-performance liquid chromatographic (HPLC) method has been developed for the sensitive and highly selective
determination of histamine in plasma. This method includes selective extraction of histamine from plasma, pre-column derivatisation
in aqueous phase with o-phthaldialdehyde (OPA) and HPLC analysis. The fluorescence of the histamine-OPA-complex was monitored
at wavelengths of 350nm excitation and 450nm emission, after isocratic eluation with a mixture of 0.2 M NaCl and methanol.
The reproducibility of this method including extraction, derivatisation and detection of histamine was >95% in a range of
0.35–17.6 pmol. The HPLC precision was 99±1% at 4 pmol of histamine. The lower limit of detection was 88fmol. A significantly
increased concentration of plasma histamine was detected in patients (n=46) with various liver diseases (0.3–5.2 ng/ml). In
comparison the plasma histamine levels of healthy blood donors were in the range of 0.0–0.4 ng/ml (p<0.01). 相似文献
9.
Zhang Y Zhang G Clarke PA Huang JT Takahashi E Muirhead D Steenwyk RC Lin Z 《Journal of mass spectrometry : JMS》2011,46(7):705-711
Quantitation of urinary tetranor PGDM or tetranor PGEM (tPGDM and tPGEM) in the past was performed separately using off-line SPE LC-MS/MS methods. The manual SPE procedure is generally time-consuming and cost-ineffective. In addition, simultaneous quantitation of tPGDM and tPGEM is favorable yet very challenging because of the similar chemical structures and identical MRM transitions. This work describes the development and validation of a high-throughput online SPE-LC-MS/MS method, allowing simultaneous and high-throughput measurement of tPGDM and tPGEM in human urine. The reportable range of the assay was 0.2-40 ng/ml for tPGDM and 0.5-100 ng/ml for tPGEM. Intra- and inter-assay precision and accuracy determined using quality control samples were all within acceptable ranges (% CV and % Bias < 15%). Tetranor PGDM was stable under all tested conditions while tPGEM was stable at 4 °C and after three F/T cycles but not stable at room temperature for 24 h (recovery below 80%). The assay was applied to measure urinary tPGDM and tPGEM among healthy volunteers, smokers and COPD patients. Significantly higher urinary levels of both tPGDM and tPGEM were observed in COPD patients than those of non-smoking healthy volunteers. These results demonstrated that the high-throughput online SPE-LC-MS/MS assay provides sensitive, reproducible and accurate measurement of urinary tPGDM and tPGEM as biomarkers for assessing inflammatory diseases such as COPD. 相似文献
10.
A liquid chromatography–mass spectrometry (LC-MS) method was developed and validated for the simultaneous determination of
alisol A and alisol A 24-acetate from Alisma orientale (Sam.) Juz. in rat plasma using diazepam as an internal standard. A 200-μl plasma sample was extracted by methyl tert-butyl ether and the separation was performed on Kromasil C18 column (150 × 4.6 mm, 5 μm) with the mobile phase of acetonitrile (containing 0.1% of formic acid)–water (73:27, v/v) at a flow rate of 0.8 ml/min in a run time of 10 min. The two analytes were monitored with positive electrospray ionization
by selected ion monitoring mode. The lower limit of quantitation for both alisol A and alisol A 24-acetate were 10 ng/ml.
The calibration curves were linear in the measured range 10–1,000 ng/ml for alisol A and 10–500 ng/ml for alisol A 24-acetate.
The mean extraction recoveries were above 74.7% for alisol A and above 72.4% for alisol A 24-acetate from biological matrixes.
The intra- and inter-day precision for all concentrations of quality controls was lower than 14.1% (RSD %) for each analyte.
The accuracy ranged from −12.3% to 9.8% (RE %) for alisol A, and −8.6% to 14.2% (RE %) for alisol A 24-acetate. The method
was successfully applied to the study on the pharmacokinetics of alisol A and alisol A 24-acetate in rat plasma. 相似文献
11.
Jin F 《Analytical and bioanalytical chemistry》2011,400(9):2881-2887
A rapid, sensitive, and selective liquid chromatography–tandem mass spectrometry method for the detection of tandospirone
in human plasma is described. It was employed in a pharmacokinetic study. The analyte and internal standard diphenhydramine
were extracted from plasma using liquid–liquid extraction, then separated on a Zorbax XDB C18 column using a mobile phase of methanol–water–formic acid (80:20:0.5, v/v/v). The detection was performed with a tandem mass
spectrometer equipped with an electrospray ionization source. Linearity was established in the concentration range of 10.0-5,000 pg/ml.
The lower limit of quantification was 10.0 pg/ml. The intraday and interday relative standard deviation across three validation
runs over the entire concentration range was less than 13%. Accuracy determined at three concentrations (25.0, 200, and 4,000 pg/ml
for tandospirone) ranged from 94.4 to 102.1%. Each plasma sample was chromatographed within 3.4 min. The method proved to
be highly selective and suitable for bioequivalence evaluation of different formulations containing tandospirone and clinical
pharmacokinetic investigation of tandospirone. 相似文献
12.
Paula Guerra Ethel Eljarrat Damià Barceló 《Analytical and bioanalytical chemistry》2010,397(7):2817-2824
This paper describes the development of a methodology for the simultaneous determination and quantification of hexabromocyclododecane
(HBCD), tetrabromobisphenol A (TBBPA), and related compounds (bisphenol A, monobromobisphenol A, dibromobisphenol A, and tribromobisphenol
A) in sludge and sediment samples. The selected method is based on an extraction with dichloromethane: methanol followed by
purification via SPE C18 cartridges. Instrumental determination was carried out by liquid chromatography–quadrupole linear ion trap mass spectrometry
(LC-QqLIT-MS), with quantification based on isotopic dilution method. Analyte recoveries were in the range of 39–120% and
88–126% for spiked sewage and sediment, respectively. Repeatability of replicate extractions was better than 13% relative
standard deviation. Linearity was checked in the range of 0.05 and 25 injected nanograms. Limits of detection (LODs) and limits
of quantification (LOQs) were in the range of 0.6 and 2.7 ng/g and 1.4 and 66 ng/g for sediment and sludge samples, respectively.
The developed method was applied to sewage sludge and sediment samples collected along the Ebro River and Cinca River, one
of its tributaries (northeast of Spain). TBBPA levels in sewage sludge ranged from not quantified to 1,329 ng/g dw, whereas
levels in sediment samples were lower, between not detected and 15 ng/g dw. As regards HBCD, concentrations were between not
detected and 375 ng/g for sludge samples and 0.8 and 1850 ng/g for sediments. 相似文献
13.
Xiao-Li Liu Hong-Fang Zhang Guang-Jun Qiao Wei Cao Jian-Bin Zheng 《Chromatographia》2008,67(1-2):147-150
A rapid, sensitive, and specific method for quantification of olmesartan, the prodrug of olmesartan medoxomil, in human plasma,
using zidovudine as internal standard, is described. Sample preparation involved a simple solid-phase extraction procedure.
The extract was analyzed by high-performance liquid chromatography coupled to electrospray tandem mass spectrometry (LC–MS–MS).
Chromatography was performed isocratically on a 5 μm C18 analytical column (50 mm × 4.6 mm i.d.) with water–acetonitrile–formic acid 20:80:0.1 (v/v) as mobile phase. The response to olmesartan was a linear function of concentration over the range 4.82–1,928 ng mL−1. The lower limit of quantification in plasma was 4.82 ng mL−1. The method was successfully applied in a bioequivalence study of an olmesartan formulation after administration as a single
oral dose. 相似文献
14.
A liquid chromatographic-mass spectrometric method for the simultaneous determination of nicotine, cotinine, trans-3'-hydroxycotinine, and norcotinine in human plasma was developed and validated. Analytes and deuterated internal standards were extracted from human plasma using solid-phase extraction and analyzed by liquid chromatography/atmospheric pressure chemical ionization-mass spectrometric detection with selected ion monitoring (SIM). Limits of detection and quantification were 1.0 and 2.5 ng/ml, respectively, for all analytes. Linearity ranged from 2.5 to 500 ng/ml of human plasma using a weighting factor of 1/x; correlation coefficients for the calibration curves were > 0.99. Intra- and inter-assay precision and accuracy were < 15.0%. Recoveries were 108.2-110.8% nicotine, 95.8-108.7% cotinine, 90.5-99.5% trans-3'-hydroxycotinine, and 99.5-109.5% norcotinine. The method was also partially validated in bovine serum, owing to the difficulty of obtaining nicotine-free human plasma for the preparation of calibrators and quality control (QC) samples. This method proved to be robust and accurate for the quantification of nicotine, cotinine, trans-3'-hydroxycotinine, and norcotinine in human plasma collected in clinical studies of acute nicotine effects on brain activity and on the development of neonates of maternal smokers. 相似文献
15.
Laser-ablation inductively coupled plasma mass spectrometry (LA–ICP–MS) has been established as a very efficient and sensitive
technique for the direct analysis of solids. In this work the capability of LA–ICP–MS was investigated for determination of
trace elements in high-purity graphite. Synthetic laboratory standards with a graphite matrix were prepared for the purpose
of quantifying the analytical results. Doped trace elements, concentration 0.5 μg g–1, in a laboratory standard were determined with an accuracy of 1% to ± 7% and a relative standard deviation (RSD) of 2–13%.
Solution-based calibration was also used for quantitative analysis of high-purity graphite. It was found that such calibration
led to analytical results for trace-element determination in graphite with accuracy similar to that obtained by use of synthetic
laboratory standards for quantification of analytical results. Results from quantitative determination of trace impurities
in a real reactor-graphite sample, using both quantification approaches, were in good agreement. Detection limits for all
elements of interest were determined in the low ng g–1 concentration range. Improvement of detection limits by a factor of 10 was achieved for analyses of high-purity graphite
with LA–ICP–MS under wet plasma conditions, because the lower background signal and increased element sensitivity.
Received: 4 January 2001 / Revised: 27 March 2001 / Accepted: 28 March 2001 相似文献
16.
Takeshi Kumazawa Koichi Saeki Isao Yanagisawa Seisaku Uchigasaki Chika Hasegawa Hiroshi Seno Osamu Suzuki Keizo Sato 《Analytical and bioanalytical chemistry》2009,394(4):1161-1170
This paper describes a fully automated on-line method combining in-tube solid-phase microextraction (SPME) in which sample
clean-up and enrichment are conducted through an open tubular fused-silica capillary column and high-performance liquid chromatography
(HPLC)/tandem mass spectrometry (MS/MS) detection for the determination of six butyrophenone derivatives (moperone, floropipamide,
haloperidol, spiroperidol, bromperidol, and pimozide) in human plasma samples. The six butyrophenones were extracted by repeatedly
aspirating and dispensing plasma sample solutions on a DB-17 capillary column (60 cm × 0.32 mm i.d., film thickness 0.25 μm).
The analytes retained on the inner surface of the capillary column were then eluted into an acetonitrile-rich mobile phase
using a gradient separation technique. Extraction efficiencies ranged from 12.7% to 31.8% for moperone, spiroperidol, and
pimozide, and from 1.08% to 4.86% for floropipamide, haloperidol, and bromperidol. The regression equations for all compounds
showed excellent linearity, ranging from 0.05 to 50 ng/0.1 mL of plasma, except for moperone and spiroperidol (0.01 to 50 ng/0.1 mL).
The limits of detection and quantification in plasma for each drug were 0.03–0.2 and 0.1–0.5 ng/mL, respectively. The intra-
and inter-day coefficients of variation for all compounds in plasma were not greater than 13.7%. 相似文献
17.
Kacinko SL Concheiro-Guisan M Shakleya DM Huestis MA 《Analytical and bioanalytical chemistry》2008,392(5):903-911
A liquid chromatography–tandem mass spectrometry method for the simultaneous quantification of buprenorphine (BUP), norbuprenorphine
(NBUP), buprenorphine glucuronide (BUP-Gluc), and norbuprenorphine glucuronide (NBUP-Gluc) in human urine was developed and
fully validated. Extensive endogenous and exogenous interferences were evaluated and limits of quantification were identified
empirically. Analytical ranges were 5–1,000 ng/mL for BUP and BUP-Gluc and 25–1,000 ng/mL for NBUP and NBUP-Gluc. Intra-assay
and interassay imprecision were less than 17% and recovery was 93–116%. Analytes were stable at room temperature, at 4 °C,
and for three freeze–thaw cycles. This accurate and precise assay has sufficient sensitivity and specificity for urine analysis
of specimens collected from individuals treated with BUP for opioid dependence. 相似文献
18.
Erin L. Karschner Allan J. Barnes Ross H. Lowe Karl B. Scheidweiler Marilyn A. Huestis 《Analytical and bioanalytical chemistry》2010,397(2):603-611
A sensitive analytical method for simultaneous quantification of sub-nanogram concentrations of cannabidiol (CBD), Δ9-tetrahydrocannabinol (THC), 11-hydroxy-THC (11-OH-THC), and 11-nor-9-carboxy-THC (THCCOOH) in plasma is presented for monitoring
cannabinoid pharmacotherapy and illicit cannabis use. Analytes were extracted from 1 mL plasma by solid-phase extraction,
derivatized with N,O-bis(trimethylsilyl) trifluoroacetamide with 1% trimethylchlorosilane, and analyzed by two-dimensional gas chromatography
mass spectrometry (2D-GCMS) with cryofocusing. The lower calibration curve was linear from 0.25–25 ng/mL for CBD and THC,
0.125-25 ng/mL for 11-OH-THC and 0.25-50 ng/mL for THCCOOH. A second higher linear range from 5–100 ng/mL, achieved through
modification of injection parameters, was validated for THC, 11-OH-THC, and THCCOOH and was only implemented if concentrations
exceeded the lower curve upper limit of linearity. This procedure prevented laborious re-extraction by allowing the same specimen
to be re-injected for quantification on the high calibration curve. Intra- and inter-assay imprecision, determined at four
quality control concentrations, were ≤7.8% CV. Analytical bias was within ±9.2% of target and extraction efficiencies were
≥72.9% for all analytes. Analytes were stable when stored at 22°C for 16 h, 4°C for 48 h, after three freeze–thaw cycles at
−20°C and when stored on the autosampler for 48 h. This sensitive and specific 2D-GCMS assay provides a new means of simultaneously
quantifying CBD, THC and metabolite biomarkers in clinical medicine, forensic toxicology, workplace drug testing, and driving
under the influence of drugs programs. 相似文献
19.
We present here a high-performance liquid chromatography−tandem mass spectrometry (LC-MS/MS) method for quantifying phytoestrogenic
isoflavones (daidzein, equol, genistein, and O-desmethylangolensin) and lignans (enterodiol and enterolactone) in urine without the use of extraction or the preconcentration
techniques inherent in existing methods. The development of this concept was made possible by use of atmospheric pressure
photoionization (APPI); an ionization technique that we found to improve analyte sensitivity relative to electrospray ionization
and atmospheric pressure chemical ionization for this particular group of compounds. The analytical performance of this method
was equal to or exceeded that of comparable methods. Between-run coefficients of variation (CVs) across three quality control
(QC) pool levels analyzed in duplicate over 20 days were 3.1–5.8% CV; within-run CVs were 2.3–6.0%. Accuracy, as determined
by average spike recovery in QC pools, was generally within ±10% of being quantitative (100%). Relative limits of detection
were 0.04–0.4 ng/mL urine, with absolute detection limits as low as 0.1 pg. This method was applied to the analysis of >2,500
urine specimens for the 2005–2006 Centers for Disease Control and Prevention’s National Health and Nutrition Examination Survey
(NHANES). The method was capable of quantifying these compounds in 95–100% of study samples. This work is the first ever report
of using APPI for the LC-MS/MS determination of these compounds in urine. It is also the first method of its kind to do so
without any need for analyte extraction or preconcentration prior to analysis. 相似文献
20.
Clavijo CF Hoffman KL Thomas JJ Carvalho B Chu LF Drover DR Hammer GB Christians U Galinkin JL 《Analytical and bioanalytical chemistry》2011,400(3):715-728
Opioids such as morphine are the cornerstone of pain treatment. The challenge of measuring the concentrations of morphine
and its active metabolites in order to assess human pharmacokinetics and monitor therapeutic drugs in children requires assays
with high sensitivity in small blood volumes. We developed and validated a semi-automated LC-MS/MS assay for the simultaneous
quantification of morphine and its active metabolites morphine 3β-glucuronide (M3G) and morphine 6β-glucuronide (M6G) in human
plasma and in dried blood spots (DBS). Reconstitution in water (DBS only) and addition of a protein precipitation solution
containing the internal standards were the only manual steps. Morphine and its metabolites were separated on a Kinetex 2.6-μm
PFP analytical column using an acetonitrile/0.1% formic acid gradient. The analytes were detected in the positive multiple
reaction mode. In plasma, the assay had the following performance characteristics: range of reliable response of 0.25–1000 ng/mL
(r
2 > 0.99) for morphine, 1–1,000 ng/mL (r
2 > 0.99) for M3G, and 2.5–1,000 ng/mL for M6G. In DBS, the assay had a range of reliable response of 1–1,000 ng/mL (r
2 > 0.99) for morphine and M3G, and of 2.5–1,000 ng/mL for M6G. For inter-day accuracy and precision for morphine, M3G and
M6G were within 15% of the nominal values in both plasma and DBS. There was no carryover, ion suppression, or matrix interferences.
The assay fulfilled all predefined acceptance criteria, and its sensitivity using DBS samples was adequate for the measurement
of pediatric pharmacokinetic samples using a small blood of only 20–50 μL. 相似文献