Lactation-promoting ingredients of Hemerocallis citrina Borani and the corresponding mechanisms

Hemerocallis citrina Borani is a traditional folk food used to promote the lactation of postpartum mothers in China; however, the active ingredients and corresponding mechanisms are still unknown. In this study, the lactogenic effect of alcoholic and aqueous extracts of H. citrina was primarily evaluated, and the aqueous extract (1,000 and 2,000 mg/kg) displayed significant lactation-promoting effects. Three eluates of the aqueous extract (0%, 30%, and 50%HCW) were further evaluated for their lactogenic effect, and 30% and 50% HCW showed significant lactation-promoting activity. Nineteen ingredients, including those with a high content of rutin and isoquercetin, were then identified from 30% and 50%HCW using the ultra-performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry (UPLC-Q-TOF-MS) method. Finally, the lactogenic effect of rutin and isoquercetin was evaluated, and both compounds displayed significant lactation-promoting activity. The mechanisms relative to the lactation-promoting active ingredients for H. citrina extracts and compounds are to stimulate the release of prolactin (PRL) and progesterone (P), as well as to induce the expression of prolactin receptor (PRLR) and improve the morphology of mammary tissue. This study first clarified the lactation-promoting active ingredients of H. citrina and the corresponding mechanisms, which provide a new insight into the new lactation-promoting drug and promote the high-value utilization of H. citrina resources

Macleaya cordata R. Br.

<i> 2.3. Screening and identifying [ring- 13 C]-labeled alkaloids from</i> <i>M.</i> 6 <i>cordata</i> <i>seedlings</i> <p> The [<i>ring</i> - 13 C 6]-labeled isoquinoline alkaloids were screened from the TICs of <i>M. cordata</i> seedlings by EIC of the theoretical [M+6.0201] + values (M represents the <i>m/z</i> values of identified alkaloids). These labeled alkaloids were further identified by comparing their retention times, MS or MS/MS data with those of their unlabeled alkaloids. Take compound <b>2</b> as an example. An obvious peak was formed via EIC of <i>m/z</i> 320.1952 ([314.1751 + 6.0201] +) on the TICs of <i>M. cordata</i> seedlings that were fed [<i>ring</i> - 13 C 6]-tyrosine. In the MS spectrum of alkaloid <b>2</b> (Fig. S5 A), unlabeled (<i>m/z</i> 314.1747) and labeled (<i>m/z</i> 320.1952) mass-tocharge ratio values were observed. In the MS/MS spectrum of unlabeled <i>m/z</i> 314.1747 (Fig.S5 B), the base peak ion at <i>m/z</i> 107.0483 was formed by β- cleavage of the skeleton. However, the corresponding fragment ion was found at <i>m/z</i> 113.0633 in the MS/MS spectrum of labeled <i>m/z</i> 320.1952, which indicated that the C-ring in the structure of alkaloid <b>2</b> was labeled (Fig.S5 C). In addition, the unlabeled (<i>m/z</i> 175.0605) and labeled (<i>m/z</i> 275.1370) fragment ions were also generated from the MS/MS spectrum of labeled parent ion, which further identified alkaloid <b>2</b> as demethylcolletine (Table S2). When we used a similar method, a total of 120 [<i>ring</i> - 13 C 6]-labeled isoquinoline alkaloids were screened and identified from the TICs of <i>M. cordata</i> seedlings (Table S2). However, some of the identified isoquinoline alkaloids, such as compounds <b>52</b> and <b>193</b>, were not found to be labeled by [<i>ring</i> - 13 C 6]. One reason is that the content of [<i>ring</i> - 13 C 6]-isoquinoline alkaloids was low and hard to detect by HPLC-Q-TOF-MS. Another reason is that some unlabeled isoquinoline alkaloids, such as compounds <b>174</b> and <b>180</b>, were found from only <i>M. microcarpa</i> seedlings and not from <i>M. cordata</i> seedlings.</p>Published as part of <i>Qing, Zhixing, Yan, Fangqin, Huang, Peng & Zeng, Jianguo, 2021, Establishing the metabolic network of isoquinoline alkaloids from the Macleaya genus, pp. 1-8 in Phytochemistry (112696) (112696) 185</i> on page 3, DOI: 10.1016/j.phytochem.2021.112696, <a href="http://zenodo.org/record/8259854">http://zenodo.org/record/8259854</a&gt

Fluid Dynamics Studies on Bottom Liquid Detachment from a Rising Bubble Crossing a Liquid–Liquid Interface

The detachment regimes and corresponding detachment height of lower liquid from a coated bubble during the bubble passage through an immiscible liquid–liquid interface were studied. High-speed imaging techniques were used to visualize the lower liquid detachment from a rising bubble near the interface. Analysis of industrial slag samples by a scanning electron microscope (SEM) was also carried out. The results indicate that the detachment height of lower liquid from a rising bubble showed a distinct correlation to penetration regimes. Bubble size and a fluid’s physical properties exerted a significant influence on the detachment height of the lower liquid. The detachment height for medium bubbles (Weber number: 4~4.5; Bond number: 2.5~7.5) varied significantly with increasing bubble size, which contributes to the lower liquid entrainment in the upper phase due, significantly, to the higher detachment height and large entrainment volume. The maximum detachment height for large bubbles is limited to approximately 100 mm due to the early detachment with the liquid column at the interface though large bubbles transporting a larger volume of lower liquid into the upper phase

Macleaya cordata R. Br.

<i>2.2. Screening and identifying isoquinoline alkaloids from M. cordata and</i> <i>M. microcarpa samples</i> <p> Screening and identifying as many compounds as possible are critical steps for establishing metabolic networks of isoquinoline alkaloids in the <i>Macleaya</i> genus. The metabolites were detected by the established screening method, and their structures were identified according to their characteristic MS/MS spectra. In this screening method, 1608 exact theoretical masses were first formed by combining 29 well-known skeletons with five common groups, after which 640 potential candidates were obtained via EIC of the exact theoretical mass on the TICs of 45 different samples. Finally, 392 potential candidates were generated, and 204 of them were further identified by their characteristic MS/MS data (Fig. S3) and the fragmentation pathways of isoquinoline alkaloids that have been systematically investigated in previous studies (Qing et al., 2013, 2020; Jeong et al., 2012). Identifying isoquinoline alkaloids from <i>M. cordata</i> and <i>M. microcarpa</i> fruits was performed in detail in our previous studies (Qing et al., 2014, 2015a, 2015b; Zuo et al., 2017). Therefore, in this study, only screening and identifying benzyltetrahydroisoquinoline alkaloids was taken as an example.</p> <p> Previous studies indicated that benzyltetrahydroisoquinoline (whose theoretical <i>m/z</i> values is 220.1122 [M + H] +), <i>N</i> -methyl-benzyltetrahydroisoquinoline (234.1287) and <i>N</i>, <i>N</i> -methyl-benzyl- tetrahydroisoquinoline (248.1435) are the basic skeletons, and OCH 2 O (46.0055), OCH 3 (31.0184), OH (17.0027), H (1.0078), and glucose (179.0556) are the main substituent groups of this type of alkaloid. In total, 138 exact theoretical masses (Table S1) were generated by adding four substituent groups to the parent skeletons. Using EIC of the formed theoretical masses on the TICs of the 45 different <i>Macleaya</i> genus samples, 80 candidate compounds were generated, and target-MS/ MS analysis was performed for each candidate. Finally, 37 benzyltetrahydroisoquinoline alkaloids (<b>1-37</b>, Table S2) were tentatively identified based on their characteristic MS / MS spectra and the fragmentation pathways of reference alkaloids.</p> <p> Take alkaloid <b>1</b> as an example. The exact theoretical mass at <i>m/z</i> 272.1281 (Table S1) was formed by combining the skeletons with substituent groups. The candidate (alkaloid <b>1</b>, t <i> R</i> 3.39 min) was found from the TICs of 0- to 90-day tissue culture seedlings, <i>M. cordata</i> fruits (III), and <i>M. cordata</i> roots (IV) via EIC of the formed theoretical mass on the TICs of the 45 <i>Macleaya</i> genus samples. The MS/MS spectrum of alkaloid <b>1</b> was obtained using the relatively high-content 0-day-old sample (Fig. S4 A and B). In the MS/MS spectrum of alkaloid <b>1</b> (Fig.S4 C), the neutral loss of the NH 3 moiety from the protonated exact mass at <i>m/z</i> 272.1293 and formation of the fragment ion at <i>m/z</i> 255.1073 were observed. In addition, the relatively highly abundant fragment ions at <i>m/z</i> 107.0486, 143.0483, and 161.0604 were also formed. Both fragmentation behaviors indicated that alkaloid <b>1</b> was a benzyltetrahydroisoquinoline-type alkaloid according to previous studies (Qing et al., 2013, 2020). The structure of alkaloid <b>1</b> was combined by a benzyltetrahydroisoquinoline skeleton, three hydroxyls, and one hydrogen, as shown in Table S1. The base peak ion at <i>m/z</i> 107.0486 indicated that one hydroxyl was connected to the C-ring of the benzyltetrahydroisoquinoline skeleton. The fragmentation ions at <i>m/z</i> 143.0483, 161.0604, and 255.1073 demonstrated that the remaining two hydroxyls were connected to the A-ring. Therefore, alkaloid <b>1</b> was tentatively identified as higenamine (Fig.S4 C). Moreover, the structure of alkaloid <b>1</b> was unambiguously determined by comparing the retention time, exact MS and MS/MS data with the standard. The remaining 36 benzyltetrahydroisoquinoline alkaloids were also screened and identified using a similar method (Table S2).</p> <p> In addition to benzyltetrahydroisoquinoline, 28 other types of isoquinoline alkaloids (Fig. S1) were screened from the <i>Macleaya</i> genus (Fig. 1). However, only 11 types of isoquinoline alkaloids, i.e., benzyltetrahydroisoquinoline (<b>1-37</b>), tetrahydroprotoberberine (<b>38-53</b>), <i>N</i> - methyltetrahydroproto -berberine (<b>54-67</b>), protopine (<b>68-90</b>),</p> <p> berberine (<b>91-107</b>), 7,8-dihydroberberine (<b>108-114</b>), aporphine (<b>115- 117</b>), benzophenanthridine (<b>118-167</b>), dihydrobenzophenanthridine (<b>168-201</b>), benzoquinoline (<b>202-203</b>) and arnottianamide (<b>204</b>), were found and identified (Fig. S3; Table S2). Interestingly, among the 204 identified isoquinoline alkaloids, 40 glycosylated alkaloids (<b>21-26</b>, <b>34- 36</b>, <b>42-44</b>, <b>49</b>, <b>52</b>, <b>53</b>, <b>63</b>, <b>64</b>, <b>67</b>, <b>74</b>, <b>93</b>, <b>94</b>, <b>99</b>, <b>100</b>, <b>103</b>, <b>116</b>, <b>117</b>, <b>130-135</b>, <b>158</b>, <b>159</b>, <b>161</b>, <b>167</b>, <b>170</b>, <b>174</b>, <b>180</b>, and <b>181</b>, Fig. S3; Table S2), which have rarely been reported in the Papaveraceae family and may have excellent bioactivities, were detected and characterized from the <i>Macleaya</i> genus. In addition, some addictive compounds, such as morphine and codeine, which have been found within the Papaveraceae family, were not detected in either plant species in the present study (Liu et al., 2017; Poeaknapo et al., 2004). Finally, a metabolic network of isoquinoline alkaloids in the <i>Macleaya</i> genus was established based on the identified metabolites.</p>Published as part of <i>Qing, Zhixing, Yan, Fangqin, Huang, Peng & Zeng, Jianguo, 2021, Establishing the metabolic network of isoquinoline alkaloids from the Macleaya genus, pp. 1-8 in Phytochemistry (112696) (112696) 185</i> on page 3, DOI: 10.1016/j.phytochem.2021.112696, <a href="http://zenodo.org/record/8259854">http://zenodo.org/record/8259854</a&gt

Macleaya R. Br.

<i>2.4. Establishing the metabolic network of isoquinoline alkaloids in the</i> <i>Macleaya genus</i> <p> Metabolic networks can provide new insight into the biosynthesis pathways of some important compounds present in plant-derived medicines. Sanguinarine and chelerythrine, which are the main components of the <i>Macleaya</i> genus, are used as animal food additives. We previously characterized the biosynthesis pathways of these two compounds in <i>M. cordata</i> thoroughly (Liu et al., 2017). However, are there any other biosynthesis pathways involving either of these two alkaloids in the <i>Macleaya</i> genus plants? Establishment of the metabolic networks is an effective method to answer this question. In this study, a metabolic framework was established primarily based on the reported biosynthesis pathways. A specific metabolic network of isoquinoline alkaloids was then constructed based on the structural relations and differences for identified compounds’ metabolic levels at various growth stages and isotopically labeled [<i>ring</i> - 13 C 6]-tyrosine feeding experiment.</p> <p> In the <i>Macleaya</i> plants, most isoquinoline alkaloids are synthesized from the benzyltetrahydroisoquinoline alkaloids, which can then be converted to tetrahydroprotoberberine and aporphine alkaloids. <i>N</i> - methyltetrahydroprotoberberine, protoberberine, and 7,8-dihydroprotoberberine are derived from tetrahydroprotoberberine-type alkaloids. Protopine is generated from an <i>N</i> -methyltetrahydroprotoberberine-type alkaloid, and it is continually metabolized to dihydrobenzophenanthridine and benzophenanthridine alkaloids (Liu et al., 2017; Takemura et al., 2013) (Fig. S6(A)). A metabolic framework of isoquinoline alkaloids in the <i>Macleaya</i> genus was established based on the above metabolism of alkaloids. Take alkaloids <b>17</b> (3 ′ -hydroxy- <i>N</i>, <i>N</i> -dimethylcoclaurine) and <b>115</b> (10-demethylmagnoflorine) as an example. Compounds <b>17</b> and <b>115</b> were identified as benzyltetrahydroisoquinoline- and aporphine-type alkaloids, respectively (Fig. S3). Both alkaloids have the same substituent groups, and the benzyltetrahydroisoquinoline can be converted to aporphine in the Papaveraceae family (Sato et al., 2007; Ziegler et al., 2006). Therefore, alkaloid <b>17</b> was regarded as the precursor of <b>115</b>. The relations of substituent groups (Fig. S6(B)) that have been reported in the <i>Macleaya</i> genus were employed for establishing the specific metabolic network of the same skeleton compounds (Liu et al., 2017). Take alkaloids <b>2</b> (4 ′ -demethylcolletine) and <b>28</b> (colletine) as an example. Both of those compounds were identified as benzyltetrahydroisoquinoline-type alkaloids (Fig. S3). Their structures differed in that alkaloid <b>28</b> has an –OCH 3 group at the site of C-4’; however, an –OH group was present in the same position for alkaloid <b>2</b>. In the genus <i>Macleaya</i>, the –OH can be transferred to an –OCH 3 group by the methyl transferase (MT), which is an enzyme encoded by various functional genes, such as 6-OMT, Mc2833, and McSMT (Han et al., 2010; Liu et al., 2017). The structural relation of both compounds indicated that alkaloid <b>2</b> was the precursor of <b>28</b>.</p> <p> The changes in metabolite levels at different growth stages provided further obvious evidence for establishing the metabolic network. The peak area of alkaloid <b>2</b> in the TICs of tissue culture seedlings continually decreased from 0 to 36 days; however, the peak area of alkaloid <b>28</b> successively increased (Fig. 3). Interestingly, the level of change was nearly the same, which further demonstrated that the alkaloid <b>2</b> could be converted to <b>28</b> during plant growth. When we used a similar method, the specific metabolic network of other isoquinoline alkaloids in the <i>Macleaya</i> genus was also established (Fig. 2). Finally, the [<i>ring</i> - 13 C 6]-tyrosine feeding experiment provided visual evidence for establishing the metabolic network of isoquinoline alkaloids in the <i>Macleaya</i> genus. The [<i>ring</i> - 13 C 6]-labeled alkaloids <b>2</b> and <b>28</b> were detected simultaneously in the TICs of <i>M. cordata</i> seedlings, which further indicated that alkaloid <b>2</b> was the precursor of <b>28</b>. All [<i>ring</i> - 13 C 6]- labeled isoquinoline alkaloids were synthesized from [<i>ring</i> - 13 C 6]-tyrosine, and their metabolic relationships could be relatively easily determined (Fig. 2). In addition, four enzymes, i.e., MT, glycosyltransferase (GTA), cyclooxygenase (COA), and oxidase, were proposed for each specific biosynthesis pathway in this metabolic network.</p> <p> Although the isoquinoline alkaloids were screened from 45 TICs of samples, which were collected from various parts of <i>Macleaya</i> plants at different growth stages, some intermediate alkaloids were still no detected or were absent in this metabolic network. Take alkaloids <b>144</b> (sanguinarine) and <b>157</b> (10-methoxy-sanguinarine) as an example (Fig. S3). Sanguinarine can be converted to 10-hydroxy-sanguinarine by an oxidase, and the intermediate alkaloid can be methylated to form alkaloid <b>157</b> in the Papaveraceae family (Fig. S7) (Tanahashi et al., 1990). However, no detectable amount of 10-hydroxy-sanguinarine was found from the TICs of 45 <i>Macleaya</i> genus samples. Therefore, 10-hydroxy-sanguinarine was absent from the metabolic network of isoquinoline alkaloids in the <i>Macleaya</i> genus.</p>Published as part of <i>Qing, Zhixing, Yan, Fangqin, Huang, Peng & Zeng, Jianguo, 2021, Establishing the metabolic network of isoquinoline alkaloids from the Macleaya genus, pp. 1-8 in Phytochemistry (112696) (112696) 185</i> on pages 3-4, DOI: 10.1016/j.phytochem.2021.112696, <a href="http://zenodo.org/record/8259854">http://zenodo.org/record/8259854</a&gt

Establishing the metabolic network of isoquinoline alkaloids from the Macleaya genus

Qing, Zhixing, Yan, Fangqin, Huang, Peng, Zeng, Jianguo (2021): Establishing the metabolic network of isoquinoline alkaloids from the Macleaya genus. Phytochemistry (112696) 185: 1-8, DOI: 10.1016/j.phytochem.2021.112696, URL: http://dx.doi.org/10.1016/j.phytochem.2021.11269