A method based on ultra-high performance liquid chromatography coupled with triple quadrupole linear ion trap-tandem mass spectrometry(UHPLC-QTRAP-MS/MS) was developed for the simultaneous determination of 41 bioactive constituents of flavonoids, organic acids, nucleosides, and amino acids in Lysimachiae Herba. The content of multiple bioactive constituents was compared among the samples from different habitats. The chromatographic separation was performed in a Waters XBridge®C_(18) column(4.6 mm×100 mm, 3.5 μm) at 30 ℃. The gradient elution was performed with 0.4% methanol(A)-formic acid water(B) as the mobile phase at a flow rate of 0.8 mL·min~(-1), and the multiple-reaction monitoring(MRM) mode was adopted. According to the content of 41 constituents, hierarchical cluster analysis(HCA), orthogonal partial least squares discriminant analysis(OPLS-DA), and gray relational analysis(GRA) were perfomed to comprehensively evaluate the samples from different habitats. The results showed that the 41 constituents exhibited good linear relationship within the tested concentration ranges, with the correlation coefficients(r) greater than 0.999 4. The method featured good precision, repeatability, and stability with the relative standard deviations(RSDs) less than 5.0%. The average recoveries of the 41 constituents ranged from 98.06% to 101.9%, with the RSDs of 0.62%-4.6%. HCA and OPLS-DA separated 48 batches of Lysimachiae Herba samples from different habitats into three categories: the producing areas in Sichuan and Chongqing, the producing areas in Jiangsu, Zhejiang, and Jiangxi, and the producing areas in Guizhou. The content of 41 constituents varied among the Lysimachiae Herba samples from different habitats. The GRA results revealed that the Lysimachiae Herba sample from Nanchong City, Sichuan Province had the best comprehensive quality. The method developed in this study was accurate and reliable and thus can be used for comprehensive evaluation of Lysimachiae Herba quality and provide basic information for the selection of habitats.
Tandem Mass Spectrometry/methods* ; Multivariate Analysis ; Chromatography, High Pressure Liquid/methods* ; Drugs, Chinese Herbal/chemistry* ; Amino Acids/analysis*

This study aimed to provide data support for resource utilization of the stems and leaves of Astragalus membranaceus var. mongholicus(SLAM) by analyzing and evaluating the chemical constituents. The crude protein, crude fiber, and soluble saccharide of SLAM were analyzed by Kjeldahl method, filtration method, and UV-Vis spectrophotometry, respectively. The nucleosides, amino acids, flavonoids, and saponins of SLAM were analyzed by ultraperformance liquid chromatography-triple quadrupole mass spectrometry(UPLC-TQ-MS). Combined with principal component analysis(PCA), the quality difference of resource components of SLAM was comprehensively evaluated. The results showed that the average content of crude protein, crude fiber, total polysaccharide, and redu-cing sugar in SLAM was 5.11%, 30.33%, 11.03 mg·g~(-1), and 31.90 mg·g~(-1), respectively. Six nucleosides, 15 amino acids, 22 flavonoids, and one saponin were detected, with an average content of 1.49 mg·g~(-1), 6.00 mg·g~(-1), 1.86 mg·g~(-1), and 35.67 μg·g~(-1), respectively. The content of various types of chemical components in SLAM differed greatly in different harvesting periods and growing years. The results of PCA showed that the quality of SLAM produced in Ningxia was superior. The results can provide references for the utilization of SLAM.
Astragalus propinquus/chemistry* ; Gas Chromatography-Mass Spectrometry ; Flavonoids/analysis* ; Plant Leaves/chemistry* ; Amino Acids ; Saponins/analysis*

In this study, we analyzed the composition and content of 25 free amino acids in 32 batches of different forms of Cervi Cornu Pantotrichum(CCP; one-branched, two-branched, and three-branched) from 15 producing areas. The clustering analysis and orthogonal partial least squares discriminant analysis(OPLS-DA) were performed based on the content of 25 free amino acids. Potential differential metabolites were identified based on VIP value. The results showed that there were 25 free amino acids in CCP, and the average content of essential, non-essential, and total amino acids was 6.13, 32.99, and 39.12 mg·g~(-1), respectively. The clustering analysis and OPLS-DA demonstrated that 25 free amino acids had different content among the three forms of CCP, of which two-branched CCP samples were separately gathered into a group. Five differential components, including glutamic acid, tryptophan, ornithine, γ-aminobutyric acid, and hydroxylysine, were screened out as potential quality markers for the identification of different forms of CCP. This study provides a theoretical basis for the quality evaluation, processing, and utilization of different forms of CCP.
Amino Acids/analysis* ; Animals ; Cornus ; Deer ; Gastropoda ; Glutamic Acid

Galli Gigerii Endothelium Corneum(GGEC) is commonly used for the clinical treatment of indigestion, vomiting, diarrhea, and infantile malnutrition with accumulation. In recent decades, omnivorous domestic chickens, the original source of GGEC, has been replaced by broilers, which may lead to significant changes in the quality of the yielding GGEC. Through subjective and objective sensory evaluation, biological evaluation, and chemical analysis, this study compared the odor and quality between GGEC derived from domestic chickens and that from broilers. The odor intensity between them was compared by odor profile analysis and it was found that the fishy odor of GGEC derived from domestic chickens was significantly weaker than that of GGEC from broilers. Headspace-solid phase microextraction-gas chromatography-triple quadrupole tandem mass spectrometry(HS-SPME/GC-QQQ-MS/MS) suggested that the overall odor-causing chemicals were consistent with the fishy odor-causing chemicals. According to the odor activity va-lue and the orthogonal partial least squares discriminant analysis(OPLS-DA) result, dimethyl trisulfide, 2-methoxy-3-isobutylpyrazine, and 2-methylisoborneol were responsible for the fishy odor(OAV≥1) and the content of fishy odor-causing chemicals in GGEC derived from broilers was 1.12-2.13 folds that in GGEC from domestic chickens. The average pepsin potency in GGEC derived from broilers was 15.679 U·mg~(-1), and the corresponding figure for the medicinal from domestic chickens was 26.529 U·mg~(-1). The results of pre-column derivatization reverse-phase high-performance liquid chromatography(RP-HPLC) assay showed that the content of total amino acids and digestion-promoting amino acids in domestic chickens-derived GGEC was 1.12 times and 1.15 times that in GGEC from broilers, and the bitter amino acid content was 1.21 times folds that of the latter. In conclusion, GGEC derived from domestic chickens had weaker fishy odor, stronger enzyme activity, higher content of digestion-promoting amino acids, and stronger bitter taste than GGEC from broilers. This study lays a scientific basis for studying the quality variation of GGEC and provides a method for identifying high-quality GGEC. Therefore, it is of great significance for the development and cultivation of GGEC as both food and medicine and breeding of corresponding varieties.
Animals ; Odorants/analysis* ; Chickens ; Gas Chromatography-Mass Spectrometry/methods* ; Tandem Mass Spectrometry ; Solid Phase Microextraction ; Amino Acids ; Endothelium/chemistry* ; Volatile Organic Compounds/analysis*

¹³C metabolic flux analysis (¹³C-MFA) enables the precise quantification of intracellular metabolic reaction rates by analyzing the distribution of mass isotopomers of proteinogenic amino acids or intracellular metabolites through ¹³C labeling experiments. ¹³C-MFA has received much attention as it can help systematically understand cellular metabolic characteristics, guide metabolic engineering design and gain mechanistic insights into pathophysiology. This article reviews the advances of ¹³C-MFA in the past 30 years and discusses its potential and future perspective, with a focus on its application in industrial biotechnology and biomedicine.
Amino Acids ; Carbon Isotopes ; Isotope Labeling ; Metabolic Engineering ; Metabolic Flux Analysis ; Models, Biological

OBJECTIVE: To explore effects of different delivery and storage conditions on concentrations of amino acids and carnitines in neonatal dried blood spots (DBS), so as to provide evidence for improving accurate and reliable detection by tandem mass spectrometry. METHODS: A total of 1 254 616 newborn DBS samples in Newborn Screening Center of Zhejiang Province were delivered and stored at room temperature (group A, RESULTS: The concentrations of amino acids and carnitines in the three groups were skewed, and the differences in amino acid and carnitine concentrations among groups were statistically significant (all CONCLUSIONS Cold-chain logistics system and storage in low temperature and low humidity can effectively reduce degradation of some amino acids and carnitines in DBS, improve the accuracy and reliability of detection, and thus ensures the quality of screening for neonatal metabolic diseases.
Amino Acids/analysis* ; Carnitine/analysis* ; Dried Blood Spot Testing/standards* ; Humans ; Humidity ; Infant, Newborn ; Neonatal Screening ; Reproducibility of Results ; Specimen Handling/standards* ; Tandem Mass Spectrometry ; Temperature ; Time Factors

In order to reveal the main nutrients and functional ingredients in the shoots of Polygonatum cyrtonema, the polysaccharides, proteins, amino acids, and total phenols were determined. The tested samples cultured in Ma'nijiaonong, Hengtang village, Tianmushan town, Lin'an, Zhejiang, which were collected from three provenances(Pan'an and Longquan in Zhejiang and Qingyang in Anhui). The results showed that the polysaccharide content of the shoots varied from 2.34% to 12.73%, roughly one-third of rhizomes. The protein content varied from 107.75 to 192.49 mg·g~(-1), nearly 5.50 times more than rhizomes. Moreover, the average of total amino acid content was 193.13-248.74 mg·g~(-1), approximately 4.16 times of rhizomes. And the essential amino acids account for 35.57%-39.44% of the total amino acids content, which was close to the standard of the ideal protein proposed by FAO/WHO(the essential amino acid/total amino acid is about 40%). In addition, the taste amino acids(TaAA) changed from 160.12 to 208.29 mg·g~(-1), revealing the material basis of "shoots were extremely delicious" in Chinese ancient herbal medicine. Additionally, the total phenols varied from 51.21-58.76 mg·g~(-1), about 2.96 times of rhizomes. The DPPH free radical scavenging rate of tested shoots was over 95%, which obviously superior to rhizomes. Therefore, the shoots of P. cyrtonema is a very high-quality vegetable and functional food with good development potential. Furthermore, the main nutrients and functional substances in P. cyrtonema shoots are closely related to the provenances and harvesting seasons. It is important to improve the quality and yield of the shoots by strengthening the variety of breeding and cultivation techniques.
Amino Acids, Essential/analysis* ; Functional Food ; Nutrients/analysis* ; Plant Proteins, Dietary/analysis* ; Plant Shoots/chemistry* ; Polygonatum/chemistry* ; Polysaccharides/analysis* ; Rhizome

The roots of Chuanmingshen violaceum is a commonly used Chinese herb and food, which contains rich amino acids. However, the kinds and amounts of amino acids are variety in this herb among the geographical location and ecological environment. Therefore, this study firstly developed a new pre-column derived HPLC method to quantify the levels of 18 amino acids in Ch. violaceum roots. Then 24 Ch. violaceum samples were harvested from its main cultivating areas in Sichuan, China. These samples were divided into 4 producing areas based on their geographical sites. The 18 kinds of amino acids were quantified in these sample by the developed method. The differences of these amino acids were further analyzed among these herbal samples and the 4 producing areas by t-test and principal component analysis(PCA). The result indicated the peaks of the 18 kinds of amino acids were separated well in 70 min.The correlation coefficients between peak areas and concentration of these amino acids were more than 0.999 1(n=6). All of their recoveries were in the range of 97.38%-101.3%(n=6).Their detection limit was in the range of 0.003-0.379 μg·mL~(-1).It demonstrates that the developed HPLC method can accurately quantify the amounts of multi-amino acids in this herb. The results of t-test analysis showed the contents of histidine, cystine, leucine, valine, tryptophan, phenylalanine and threonine were significantly different(P<0.05) among the 4 producing areas. But the differences of other amino acids were not significant.The first five factors were extracted by PCA to calculate the comprehensive score. The order of comprehensive score for the 4 producing areas was B(0.603, n=10), C(0.206, n=3), A(-0.283, n=7) and D(-1.167, n=4). The total content of amino acids in Ch. violaceum collected in B producing area was largest(12.5 mg·g~(-1)). It is concluded the Ch. violaceum contains multi-kinds of amino acids. On the basis of amino acid amount, Langzhong city and Cangxi county in Sichuan province(producing area B) is the suitable areas for cultivating Ch. violaceum.
Amino Acids/analysis* ; Apiaceae/chemistry* ; China ; Chromatography, High Pressure Liquid ; Plant Roots/chemistry*

To reveal the main nutrients and functional ingredients in the flowers of Polygonatum cyrtonema and P. filipes, the content of the polysaccharides, saponins, amino acids, total phenols, mineral elements, and the DPPH free radical scavenging rates were determined. The flowers and rhizomes of P. cyrtonema were collected from Qingyang in Anhui and Qingyuan in Zhejiang, while the flowers and rhizomes of P. filipes were collected from Longyou in Zhejiang, respectively. The results showed that the polysaccharides content in flowers varied from 60.88 to 97.00 mg·g~(-1), about half of that in rhizomes. The saponins content in flowers varied from 32.55 to 40.93 mg·g~(-1), which was close to the content in rhizomes. The content of total phenols ranged from 40.79 to 50.95 mg·g~(-1), approximately 4.5 times of that in rhizomes. The total amino acids content in flowers was 111.85 to 131.03 mg·g~(-1), about 2.3 times of the content in rhizomes. The essential trace element content was abundant in flowers. The contents of heavy metal elements were all within the limits set by the standards. The DPPH free radical scavenging rate IC_(50) varied from 1.77 to 3.25 mg·mL~(-1), less than one-fifth of that in rhizomes, showing a significant superiority of antioxidant activity compared to rhizomes. The results initially revealed the fundamental of "the flowers exceed the rhizomes in effect", the common saying about the traditional Chinese medicinal herbs over the years, indicating a great developing potential of the flowers. Besides, as polysaccharides, saponins, amino acids, total phenols and other nutritive substances in flowers differ widely among species and provenances, it's important to develop variety breeding to improve the quality and yield of flowers.
Amino Acids/analysis* ; Antioxidants/analysis* ; China ; Flowers/chemistry* ; Nutrients/analysis* ; Nutritive Value ; Plant Extracts ; Polygonatum/chemistry* ; Rhizome/chemistry* ; Trace Elements/analysis*

A method, for determination of saponins, amino acids and nucleosides in Panacis Japonici Rhizoma of ultra fast liquid chromatography with triple quadrupole linear ion trap mass spectrometry (UFLC-QTRAP-MS/MS), was established to investigate the effect of different processing methods on the target components of Panacis Japonici Rhizoma. The chromatographic separation was performed on a XBridgeC₁₈(4.6 mm×100 mm, 3.5 μm) at 30 °C with a gradient elution of 0.1% formic acid solution-0.1% formic acid acetonitrile, and the flow rate was 0.8 mL·min⁻¹, using multiple-reaction monitoring (MRM) mode. The grey relational analysis was adopted for the analysis of different processing samples. The results showed that the thirty-three constituents were in a good linear range and the correlation coefficient was greater than 0.999 0; the precision, repeatability and stability were good; the average recovery rates were between 95.33% and 101.8%, and the relative standard deviations were less than 5%. The result of grey relational analysis showed that the complete rhizomes without peeling, which were adopted for the microwave dried method, had the best quality. The established method was accurate and reliable, which could be used to appraise the quality of Panacis Japonici Rhizoma. Our study may lay the way for the processing method of Panacis Japonici Rhizoma in optimization,normalization and standardization.
Amino Acids ; analysis ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Nucleosides ; analysis ; Panax ; chemistry ; Phytochemicals ; analysis ; Rhizome ; chemistry ; Saponins ; analysis ; Tandem Mass Spectrometry