Dataset Information

Simultaneous Determination of 34 Amino Acids in Tumor Tissues from Colorectal Cancer Patients Based on the Targeted UHPLC-MS/MS Method.

ABSTRACT: A targeted ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established and validated for the simultaneous determination of 34 amino acids in tissue samples from colorectal cancer (CRC) patients. The chromatographic separation was achieved on an Agilent ZORBAX SB-C18 column (3.0?×?150?mm, 5??m) with a binary gradient elution system (A, 0.02% heptafluorobutyric acid and 0.2% formic acid in water, v/v; B, methanol). The run time was 10?min. The multiple reaction monitoring mode was chosen with an electrospray ionization source operating in the positive ionization mode for data acquisition. The linear correlation coefficients were >0.99 for all the analytes in their corresponding calibration ranges. The sample was pretreated based on tissue homogenate and protein precipitation with a 100?mg aliquot sample. The average recovery and matrix effect for 34 amino acids and 3 internal standards were 39.00%?146.95% and 49.45%?173.63%, respectively. The intra- and interday accuracy for all the analytes ranged from -13.52% to 14.21% (RSD ?8.57%) and from -14.52% to 12.59% (RSD ?10.31%), respectively. Deviations of stability under different conditions were within ±15% for all the analytes. This method was applied to simultaneous quantification of 34 amino acids in tissue samples from 94 CRC patients.

SUBMITTER: Yang Y

PROVIDER: S-EPMC7416278 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Simultaneous Determination of 34 Amino Acids in Tumor Tissues from Colorectal Cancer Patients Based on the Targeted UHPLC-MS/MS Method.

Yang Yang Y Zhang Feng F Gao Shouhong S Wang Zhipeng Z Li Mingming M Wei Hua H Zhong Renqian R Chen Wansheng W

Journal of analytical methods in chemistry 20200801

A targeted ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established and validated for the simultaneous determination of 34 amino acids in tissue samples from colorectal cancer (CRC) patients. The chromatographic separation was achieved on an Agilent ZORBAX SB-C<sub>18</sub> column (3.0 × 150 mm, 5 <i>μ</i>m) with a binary gradient elution system (A, 0.02% heptafluorobutyric acid and 0.2% formic acid in water, v/v; B, methanol). The run time was 10 ...[more]

PMID: 32802550

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Project description:BACKGROUND:Millions of adults have been reported with hyperlipemia in the world. It is still unclear whether the plasma level of essential amino acids (AAs) will be influenced by the hyperlipemia. This study was aimed to investigate the AAs levels and the underlying metabolic relationship in hyperlipidemic subjects. METHODS:An ultra-high performance liquid chromatography-tandem mass spectrometric (UPLC-MS/MS) method was developed for the determination of phenylalanine (Phe), valine (Val), histidine (His), tryptophan (Trp), and methionine (Met). Plasma samples (100??L) were precipitated by acetonitrile (300??L) and analyzed on a BEH C18 (2.1?mm?×?100?mm, 1.7??m) column at 40?°C by gradient elution. The mobile phase composed of 0.1% formic acid and acetonitrile was used with flow rate at 0.2-0.4?ml/0-3?min. Five AAs were determined at positive electrospray ionization (ESI+) at m/z 118.1/72.1 (Val), 150.12/104.02(Met), 156.06/110.05(His), 166.1/120.1(Phe), and 205.2/188.02 (Trp). A total of 75 healthy subjects and 83 hyperlipidemic subjects, who had blood routine test and plasma lipid test were determined by developed UPLC-MS/MS. RESULTS:It was shown that there was good linearity for Val, Met, His, Phe, and Trp within 1-100??g/mL. The relative standard deviations of precision and accuracy were all within 15%. The level of Val, Phe, Trp, His, and Met were 35.34?±?15.64, 22.72?±?9.13, 17.23?±?4.94, 16.78?±?13.64, and 6.24?±?1.97??g/mL in healthy subjects, while they were 38.04?±?16.70, 22.41?±?8.45, 15.62?±?5.77, 18.35?±?14.49, and 6.21?±?1.97??g/mL in hyperlipidemic subjects respectively. The Spearman's correlations analysis showed that there were high correlations between Val, Phe, Trp, His, Met and triglyceride in healthy subjects. While, those correlations decreased in hyperlipemia cases. CONCLUSION:A convenient and sensitive method for simultaneous determination of Val, Phe, Trp, His, and Met in human plasma was developed. There was a high correlation between Val, Phe, Trp, His, Met and triglyceride. Hyperlipemia influences the metabolic balance of His, Phe, Trp, Met and Val.

Project description:A selective and sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for the determination of ziyuglycoside I (I), 3?,19?-dihydroxyurs-12-en-28-oic-acid 28-?-d-glucopyranosyl ester (II), 3?-[(?-l-arabinopyranosyl) oxy]-urs-12,18(19)-dien-28-oic acid ?-d-glucopyranosyl ester (III), rosamultin (IV), 1?-hydroxyeuscaphic acid (V) and alpinoside (VI) in rats after oral administration of Sanguisorba officinalis L. (S. officinalis) extract. The 3?,19?-dihydroxyurs-12-en-28-oic-acid 28-?-d-glucopyranosyl ester, 3?-[(?-l-arabinopyranosyl) oxy]-urs-12,18(19)-dien-28-oic acid ?-d-glucopyranosyl ester, rosamultin, 1?-hydroxyeuscaphic acid and alpinoside in rat plasma were the first report in the pharmacokinetics study in the present study. The analytes were quantified using the multiple reaction monitoring (MRM) mode with the electrospray ion source in positive electrospray ionization. Plasma was extracted with ethyl acetate via liquid?liquid extraction. Bifendate was used as internal standard (IS). The current method was validated for linearity, intra-day and inter-day precisions, accuracy, extraction recovery, matrix effect and stability. The lower limits of quantification of ziyuglycoside I, 3?,19?-dihydroxyurs-12-en-28-oic-acid 28-?-d-glucopyranosyl ester, 3?-[(?-l-arabinopyranosyl) oxy]-urs-12,18(19)-dien-28-oic acid ?-d-glucopyranosyl ester, rosamultin, 1?-hydroxyeuscaphic acid and alpinoside were 6.1, 4.9, 1.3, 3.8, 1.5 and 5.7 ng/mL, respectively. Intra-day and inter-day precision and the accuracy of the assay were in range from -9.48 to 12.74%. The extraction recoveries of analytes and bifendate (IS) from rat plasma ranged from 77.17% to 92.48%. Six compounds could be rapidly absorbed into blood (Tmax, 0.58?1.58 h), and then eliminated relatively slowly (t1/2, 6.86?11.63 h). The pharmacokinetic results might contribute to further guide the clinical application of S. officinalis.

Dataset Information

Simultaneous Determination of 34 Amino Acids in Tumor Tissues from Colorectal Cancer Patients Based on the Targeted UHPLC-MS/MS Method.

Publications

Simultaneous Determination of 34 Amino Acids in Tumor Tissues from Colorectal Cancer Patients Based on the Targeted UHPLC-MS/MS Method.

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