Biomedical Chromatography

Ma, Yanhong; Gao, Bixing; Qi, Jingliang; Gou, Yan; Li, Zhi; Chen, Yingxin; Li, Min; Zhou, Juan; Zhong, Lian; Cai, Xiaoyang

doi: 10.1002/bmc.5526pmid: 36250730

Because of the immense difficulty in identifying Cyathulae Capitatae Radix adulteration in Cyathulae Radix, this research aims at fortifying the quality control of Cyathulae Radix and its decoction pieces to guarantee the effectiveness and safety of its clinical use in terms of source material. A method was devised to identify Cyathulae Capitatae Radix adulteration in Cyathulae Radix and its decoction pieces. This research takes achybidensaponin I, that is, the characteristic component of Cyathulae Capitatae Radix, as reference substance and adopts HPLC for detection. The results revealed that, among all samples collected, no trace of achybidensaponin I was found in the 21 batches of Cyathulae Radix, whereas achybidensaponin I was found in all the 14 batches of Cyathulae Capitatae Radix. The research sets 5% as the adulteration limit, that is, 1.45 mg/g Cyathulae Capitatae Radix was detected in 57.14% of the 49 batches of market samples collected and the ratio was 51.02% in the case of 5% adulteration limit. The method is not only precise and reliable but can also be used as a supplement for provisions regarding quality control of Cyathulae Radix and its decoction pieces in Pharmacopoeia of the People's Republic of China, to effectively crack down on Cyathulae Capitatae Radix adulteration in the market.

Liu, Hao; Guo, Siyuan; Xi, Shuyi

doi: 10.1002/bmc.5511pmid: 36100977

Graveoline is a biologically active ingredient extracted from Ruta graveolens. Current work aimed at investigating in vitro metabolism of graveoline using rat or human liver microsomes and hepatocytes. Graveoline (20 μM) was incubated with nicotinamide adenine dinucleotide phosphate–supplemented rat and human liver microsomes as well as hepatocytes. LC coupled to a photo diode array detector and quadrupole/time‐of‐flight tandem mass spectrometry was used to detect and identify the metabolites. The structures of the metabolites were identified by accurate mass, elemental composition, and indicative fragment ions. A total of 12 metabolites, comprising 6 phase I and 6 phase II metabolites, were obtained. The metabolic pathways included demethylenation, demethylation, hydroxylation, glucuronidation, and glutathion conjugation. The metabolite (M10) produced by opening the ring of the methylenedioxyphenyl moiety was detected as the most abundant in both liver microsomes and hepatocytes, mainly catalyzed by CYP1A2, 2C8, 2C9, 2C19, 2D6, 3A4, and 3A5. This study provides valuable information on the in vitro metabolism of graveoline, which is indispensable for further development and safety evaluation of this compound.

Wang, Fusheng; Wang, Guanghuan; Li, Weizhong; Xu, Chenbin; Zeng, Zailin; Zhou, Yongcui

doi: 10.1002/bmc.5505pmid: 36093571

Preterm birth and enteral feeding are two main factors leading to necrotizing enterocolitis (NEC). The metabolomics of preterm infants before and after feeding can provide a basis for the prediction of NEC. Using the method of cross‐sectional study, the mode was established with the serum samples of 19 premature infants at birth and after feeding as the control group. The serum was analyzed using GC–MS. Chemometric analysis includes principal component analysis, partial least squares‐discriminant analysis, and orthogonal partial least squares‐discriminant analysis. Spectral separation of serum metabolites occurred in premature infants before and after feeding. The levels of xylose, d‐talose, phosphoglycolic acid, maleimide, l‐gulonolactone, maleic acid, β‐hydroxypyruvate, itaconic acid, and pantothenic acid in the serum of premature infants after feeding were significant in both multidimensional and single‐dimensional modes (variable importance in projection >2, P < 0.01). There was a moderate correlation between total bilirubin and l‐gulonolactone and β‐hydroxypyruvate (0.8 > r > 0.5). Maleimide, maleic acid, and itaconic acid have diagnostic value (area under the curve >0.9). The results indicated that serum metabolism of preterm infants changes significantly after feeding. Some metabolites have potential value in predicting NEC.

Zhu, Yong‐liang; Ding, Hao‐zhe; Gou, Kai‐feng; Sun, Bo; Chen, Ye; Zhang, He‐wei

doi: 10.1002/bmc.5504pmid: 36094354

A reliable and sensitive ultra‐performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was developed for the determination of zanubrutinib in the plasma of beagle dogs. The column used was an Acquity BEH C18 column (2.1 mm × 50 mm, 1.7 μm), maintained at 40°C with an injection volume of 2 μl. The gradient elution program was as follows: 0–1 min, 10–10% A; 1–1.1 min, 10–90% A; 1.1–2.1 min, 90–90% A; 2.1–2.2 min, 90–10% A; 2.2–3.0 min, 10–10% A. Mobile phase A was 0.1% formic acid, B was acetonitrile, and the total analysis time was 3 min. The mass spectrometry was performed in positive ion mode, and the scanning mode was multi‐reaction monitoring mode with electrospray ionization as the ion source; m/z 472.2 → 455.01 for zanubrutinib and m/z 441.03 → 137.99 for ibrutinib (internal standard). The plasma samples were processed by protein precipitation. The standard curve showed good linearity (r2 = 0.999 8) in the range of 1.0–1,000 ng/ml (zanubrutinib) with a low limit of quantification of 1 ng/ml. Also, the intra‐day and inter‐day precision (RSD) was <5.88% and the accuracy (RE) ranged from −1.56 to 1.08%; the recoveries of zanubrutinib in beagle plasma ranged from 90.12 to 93.53% (RSD 1.67–6.42%) and the ME values of zanubrutinib were 98.70–101.06% (RSD 5.37–8.49%, n = 6). All values meet US Food and Drug Administration requirements. A rapid, highly selective and sensitive method for the determination of zanubrutinib concentration in plasma by UPLC–MS/MS was successfully developed. This method is suitable for pharmacokinetic studies in beagle dogs by following oral administration of zanubrutinib.

Huang, Guochu; Xie, Sheng; Wang, Meng; Mao, Dewen; Huang, Guye; Huang, Jingjing; Liu, Xirong; Zhang, Rongzhen; Xie, Jiacheng; Huang, Liang Jiang; Cheng, Chen; Yao, Fan; Zhong, Yu; Lin, Long; Yao, Chun

Chen, Xingyu; Li, Guoliang; Zhang, Weipeng; Ou, Jiayi; Li, Hecheng; Huang, Yiqi; He, Shuirong; Zhou, Jiazhen; Zhao, Zhiqiang; Chen, Jiabin; Meng, Xiaojing; Liu, Lili

doi: 10.1002/bmc.5523

Han, Jiahong; Fei, Xuan; Sun, Nian; Xing, Junjia; Cai, Enbo; Yang, Limin

doi: 10.1002/bmc.5524pmid: 36241188

In this study, we aimed to demonstrate the therapeutic effect of Ligustri Lucidi Fructus on chemotherapy‐induced myelosuppression and elucidate its mechanism. A pharmacological study was conducted to investigate the mechanism of the inhibiting effects of Ligustri Lucidi Fructus on cyclophosphamide‐induced bone marrow suppression in mice. HPLC was used to measure the chemical components. We demonstrated that medium and high doses of Ligustri Lucidi Fructus increased the amount of white blood cells and bone marrow nucleated cells (p < 0.05) in the cyclophosphamian‐induced mouse model, and at the same time reduced granulocyte–macrophage–colony stimulating factor and thrombopoietin in the serum of myelosuppression mice (p < 0.01). Medium and high doses of Ligustri Lucidi Fructus can also adjust the thymus index and spleen index(p < 0.05). Ligustri Lucidi Fructus regulates the balance of bcl‐2/bax, inhibits the expression of Caspase‐3 and meanwhile stimulates the expression of mitogen‐activated protein (MEK) and phospho extracellular regulated protein kinases (p‐ERK) on the MAPK pathway. Five chemical constituents of Ligustri Lucidi Fructus, which may be related to myelosuppression, were analyzed. The content of specnuezhenide was 0.281%, that of ligustroflavone was 0.004%, that of salidroside was 0.094%, that of hydroxytyrosol was 0.060% and that of tyrosol was 0.069%. The effect of Ligustri Lucidi Fructus on myelosuppression after chemotherapy may be related to its multicomponent and multitarget nature. Ligustri Lucidi Fructus may be a promising potential drug for treatment after chemotherapy.

Yan, Yan; Liu, Jiaxing; Zhang, Min; Zhang, Yinjie; Shi, Biyun; Qin, Xuemei; Du, Chenhui

doi: 10.1002/bmc.5530pmid: 36264602

Ziziphi Spinosae semen (ZSS), the dried and ripe seed of Ziziphus jujube Mill. var. spinosa (Bunge) Hu ex H. F. Chou, has been used as a sedative in China and other Asian countries for over a millennium. However, its quality markers (Q‐markers) are not completely clear. In this study, Q‐markers selected by a metabolic in vivo study combined with network pharmacology are proposed for ZSS quality control. An UHPLC (ultra‐high‐performance liquid chromatography)‐Q‐Orbitrap‐MS method was developed to identify or tentatively assign 48 components including 21 flavonoid C‐glycosides, 2 flavonoid O‐glycosides, 11 dammarane triterpenoid saponins, 13 alkaloids, and 1 other, using a diagnostic product ion filtering strategy in ZSS. Subsequently, 147 metabolites detected from serum, urine, bile, and feces samples of para‐chlorophenylalanine–induced insomnia rats treated with ZSS aqueous extracts could be linked to their respective parent compounds, including 27 prototypes. Meanwhile, three metabolic networks of flavonoids, saponins, and alkaloids are preliminarily established and potential metabolic pathways are investigated under the insomnia condition. Finally, 12 key bioactive components against insomnia including magnoflorine, caaverine, coclaurine, norisocorydine, genkwanin, juzinrine, apigenin, jujubogenin, kaempferol‐3‐O‐rutinoside, jujuboside A, jujuboside B, and spinosin with the highest degree values in component–target–pathways network were selected as Q‐markers for the quality control of ZSS.

Jiang, Hai; Zhang, Jiaxu; Yu, Huan; Hou, Ajiao; Wang, Song; Wang, Xuejiao; Zheng, Senwang; Yang, Liu; Kuang, Haixue

doi: 10.1002/bmc.5520pmid: 36205398

Rheumatoid arthritis is a systemic autoimmune disease characterized by chronic symmetrical multiple arthritis. Current traditional counter‐therapies are expensive and have side effects. Xanthii Fructus has effects in expelling wind and cold, draining the nasal orifice, and removing wind and dampness. However, its mechanism of action against rheumatoid arthritis is unknown. In this paper, the mechanism of the anti‐ rheumatoid arthritis effect of Xanthii Fructus is studied by proteomics. The experimental results show that it could significantly reduce serum inflammatory factor levels, alleviate joint edema, improve vasodilation and congestion, and significantly reduce the number of inflammatory cells. Proteomics results show that the PI3K‐AKT signaling pathway is the key pathway for Xanthii Fructus to treat rheumatoid arthritis. In this study, we obtained a new understanding of the mechanism of Xanthii Fructus in the treatment of rheumatoid arthritis, which provided a theoretical basis for its prevention and treatment and laid the foundation for further research.

Ye, Li‐Hong; Dong, Xin; Cao, Jun

doi: 10.1002/bmc.5514pmid: 36181280

A highly sensitive method was developed for simultaneously separating and identifying multiple compounds in radix curcumae. The determination of these compounds was achieved by combining supercritical fluid chromatography with drift tube ion mobility quadrupole time‐of‐flight MS. Related parameters were optimized: the RX‐SIL column was used as the stationary phase, methanol was selected as the organic modifier, back pressure was 120 bar, back temperature was 60°C, the mobile phase flow rate was 1.75 mL/min, the makeup solvent was 0.2% formic acid/methanol with a flow rate of 0.7 mL/min. Under optimal conditions, multipolar compounds were separated. Furthermore, these compounds were identified by the values of collision sectional areas. The established method was verified by related parameters and exhibited good linearity, sensitivity, precision and accuracy. It could be extended to analyze other curcuminoids and sesquiterpenoids in natural products.