Neopterygosoma Fajfer, 2019 (Acariformes: Pterygosomatidae) formerly placed in the genus Pterygosoma Peters, 1849, is presented. Two new natural species groups are established: the chilensis group for species found on Chilean liolaemid lizards (Sauria: Liolaemidae) and the patagonica group for N. patagonica  found on several Liolaemus spp. from Argentina. A neotype of N. patagonica is designated. A leg chaetotaxy model for tarsi-coxae I–IV is proposed for the genus. A key to all species of Neopterygosoma is provided and a full list of Neopterygosoma spp. with their updated host associations and distribution data is compiled. Additionally, a new species, N. schroederi n. sp. found on Liolaemus schroederi Müller & Hellmich, is described, including for the first time, description and illustrations of the immature stages of a species of Neopterygosoma.A systematic revision of the scale mites of the genus  Pimeliaphilus Trägårdh, 1905 are only found on terrestrial arthropods   distributed throughout the world, except for Antarctica. Mites of the genus Neopterygosoma Fajfer, 2019 are associated with lizards of the genus Liolaemus (Sauria: Liolaemidae), and until recently they were placed in the genus Pterygosoma Peters, 1849  . Currently, the genus includes six monoxenous species found on Chilean liolaemids, and one oligoxenous species, N. patagonica Dittmar de la Cruz, Morando & Avila, 2004, observed on several Liolaemus spp. . The deutonymph, protonymph and larva are described for the first time for a species of the genus Neopterygosoma. The diagnosis for the genus is provided in Fajfer , ZISP  and ZMUC . The new material used in this study was taken from dead lizards kept in jars with 70% ethanol in the ZSM  and the NHM . Each lizard’s body was completely and carefully checked for mites under a dissecting microscope  than long. Second pair of legs discernible shorter than others. Postero-medial part of idiosoma with several pairs of dorso-median setae Microhabitat: Under the ventral and lateral scales of the head, belly, and tail.Distribution and host range: This group is associated with tree lizards of the genus Liolaemus (Sauria: Liolaemidae) from Chile.Species included: Neopterygosoma chilensis , N. cyanogasteri , N. formosus , N. levissima , N. ligare , N. ovata , N. schroederi n. sp.Neopterygosoma chilensisPterygosoma chilensis Fajfer & González-Acuña, 2013Syn. Type-host: Liolaemus chilensis (Lesson) (Sauria: Liolaemidae).Type-locality: Chile: Río Ñuble .Type-material: The holotype female is deposited in the ZISP (Reg. No. ZISP T-Pt-8), 1 female paratype is deposited in the AMU (Reg. No. AMU-PTE5.1).Other material examined: Five females from Liolaemus chilensis (NHM no. 1904.10.26.103–106), Chile: Concepción Province, Bío Bío Region, Penco , 26.x.1904, coll. S. C. Reed. All mite specimens are deposited in the UKSW (Reg. no. UKSW-PTE1.1).Records: Fajfer & González-Acuña Pterygosoma cyanogasteri Fajfer & González-Acuña, 2013Syn. Type-host: Liolaemus cyanogaster Duméril & Bibron (Sauria: Liolaemidae).Type-locality: Chile (ZMUC-R37901) .Type-material: The holotype female is deposited in the ZMUC (Reg. no. ZMUC-R37901). Paratypes: 3 females in the ZMUC (Reg. no. ZMUC-R37901), 3 paratypes in the AMU (Reg. no. AMU-PTE6.1) and 3 paratypes in the ZISP (Reg no. ZISP T-Pt-9).Records: Fajfer & González-Acuña Pterygosoma formosus Fajfer & González-Acuña, 2013Syn. Type-host: Liolaemus pictus (Duméril & Bibron) (Sauria: Liolaemidae).Type-locality: Chile: Arauco Province, Isla Mocha .Type material: The holotype female is deposited in the ZISP (Reg no. ZISP T-Pt-5); 1 paratype female is deposited in the AMU (Reg. no. AMU-PTE2.1).Records: Fajfer & González-Acuña Pterygosoma levissima Fajfer & González-Acuña, 2013Syn. Type-host: Liolaemus pictus (Duméril & Bibron) (Sauria: Liolaemidae).Type-locality: Chile: Arauco Province, Isla Mocha .Type-material: The holotype female is deposited in the ZISP (Reg. No. ZISP T-Pt-7), 1 paratype female is deposited in the AMU (Reg. No. AMU-PTE4.1).Records: Fajfer & González-Acuña Pterygosoma ligare Fajfer & González-Acuña, 2013Syn. Type-host: Liolaemus pictus (Duméril & Bibron) (Sauria: Liolaemidae).Type-locality: Chile: Arauco Province, Isla Mocha .Type-material: The holotype female is deposited in the ZISP (Reg No. ZISP T-Pt-4); 3 paratype females are deposited in the ZISP (Reg No. ZISP T-Pt-4) and AMU (Reg. No. AMU-PTE1.1)Records: Fajfer & González-Acuña Pterygosoma ovata Fajfer & González-Acuña, 2013Syn. Type-host: Liolaemus pictus (Duméril & Bibron) (Sauria: Liolaemidae).Type-locality: Chile: Arauco Province, Isla Mocha .Type-material: The holotype female is deposited in the ZISP (Reg. No. ZISP T-Pt-6), one paratype female is deposited in the AMU (Reg. No. AMU-PTE3.1).Records: Fajfer & González-Acuña .Type-locality: Chile, Maule region, Curicó Province, Curicó (Reg. No. 54/1933) .Type-material: The holotype female is deposited in the UKSW (Reg. No. UKSW-PTE3.1). Paratypes: 3 paratypes (1 deutonymph and 2 protonymphs) are deposited in the UKSW (Reg. No. UKSW-PTE3.1) and 2 paratypes (1 deutonymph and 3 protonymphs) are deposited in the ZSM (Reg. No. 54/1933).ZooBank registration: To comply with the regulations set out in Article 8.5 of the amended 2012 version of the International Code of Zoological Nomenclature , U.S. oceanographer and ichthyologist.Female [Holotype; Figs. Gnathosoma. Chelicerae 195 long. Swollen, proximal part of cheliceral base 100 long; slender distal part 95 long. Fixed cheliceral digit spinous, 10 long. Palpal femur with serrate seta dF, 50 long, palpal genu with slightly serrate seta dG, 50 long. Palpal tibia l’Ti and l”Ti with barely discernible serration, seta vTi smooth, tarsi with 5 setae and solenidion . Dorsum, antero-mid cluster with c.60 setae, subequal in length, 15 long; antero-lateral part with c.210 pairs of setae on each side, 80–110 long; dorso-median part with c.16–22 pairs of setae dm, 50–115 long; posterior and postero-lateral parts with 28–35 pairs of peripheral slightly serrate setae, 125–185 long, inserted dorsally and ventrally. Venter, 1 pair of slightly serrate setae vm, c.60 long, present anterior to genital area; 16 and 21 pairs of serrate setae vm, 30–110 long, present laterally to genital area and 15–21 pairs of serrate setae, 35–85 long, present in postero-lateral part of idiosomal venter. Genital series represented by 1 pair of setae g1 with barely discernible serration, 60 long. Pseudanal series represented by 4 pairs of setae ps. Setae ps1-ps4, 90, 120, 120 and 130 long, respectively. Setae ps1 and ps2 situated ventrally, ps3 and ps4 dorsally. Legs, coxal setation 1a, 1b, 2b, 3a, 3b, 3c and 3d arranged in formula 2-1-0-4. Setae 3a situated outside coxal plates. All coxal setae smooth except for slightly serrate setae 3d. Setae of trochanters I-IV: 1-1-1-1, femora I-IV: 5-4-3-3, genua I-IV: 5-4-3-3 and tibiae I-IV: 5-5-5-5. Setae d’FI-IV, d”FI-II, lFIII-IV, dGI-IV, l’GII-IV and dTiI serrate; lFI, v’FII-IV, v”FII, l’GI, l”GI, v’GIII-IV, dTiIII-IV, l’TiI-IV and l”TiI-IV slightly serrate; v’GI-II, v” GI, v’TiI-IV and v”TiI-IV with barely discernible serration.ion Fig. A. HypostMale. Unknown.Deutonymph [Based on 2 specimens, Figs. Gnathosoma as in female. Chelicerae 155 long; swollen cheliceral part 70 long, slender distal part 85 long. Fixed cheliceral digit spinous, 25 long. Palpal femur with serrate seta dF and dG, 45–50 and 45–55 long, respectively. Palpal tibia and tarsi as in female. Hypostome 145 long, with small depression present at apex. Peritremes with clearly visible chambers, c.90–95 long. Subcapitular seta n with barely discernible serration, 65 long. Idiosoma 505 long and 770–930 wide. Dorsum, antero-mid cluster with about 50 setae, subequal in length, 20 long; antero-lateral part with c.175–195 pairs of setae, 20–75 long, on each side present; dorso-mednian part with 22–25 pairs of setae dm, 55–170 long; posterior and postero-lateral parts with 28–35 pairs of peripheral setae, 140–215 long, inserted dorsally and ventrally. Venter with 1 pair of slightly serrate setae vm, 60 long, present anteriorly to genital area; 10–18 pairs of serrate setae vm, 55–95 long, present lateral to genital area and 15–21 pairs of postero-lateral setae, 35–90 long. Genital series represented by 1 pair of setae g1 with barely discernible serration, 50–60 long. Pseudanal series represented by 4 pairs of setae ps. Setae ps1-ps4, 90, 120, 120 and 130 long, respectively. Setae ps1 and ps2 situated ventrally, ps3 and ps4 situated dorsally. Legs, coxal setation 1a, 1b, 2b, 3a, 3b, 3c and 3d arranged in formula 2-1-0-4. Setae 3a situated outside coxal plates. All coxal setae smooth except for slightly serrate setae 3d. Setae of trochanters I-IV (1-1-1-1), femora I-IV (5-4-3-3), genua I-IV (5-4-3-3), tibiae I-IV (5-5-5-5). Setae d’FI-IV, d”FI-II, lFIII-IV, dGI-IV, l’GII-IV, dTiI serrate, lFI, v’FII-IV, v”FII, l’GI, l”GI, v’GIII-IV, dTiIII-IV, l’TiI-IV, l”TiI-IV slightly serrate v’GI-II, v” GI, v’TiI-IV, v”TiI-IV with barely discernible serration.Protonymph [Based on 5 specimens, Figs. Gnathosoma as in female. Swollen, proximal part of cheliceral base and slender, distal part equal in length, 50 long. Fixed cheliceral digits spinous, 5 long. Palpal femur with serrate seta dF, 30 long; palpal genu with filiform setae dG, 30 long. Palpal tibia and tarsi as in female. Hypostome c.90 long, with small depression present at apex. Peritremes with clearly visible chambers, 65 long. Subcapitular seta n 35–45 long. Idiosoma 275–365 long, 485–580 wide. Dorsum. Antero-mid-cluster with 26–28 setae, subequal in length, c.20 long. Antero-lateral part with c.58–79 pairs of setae on each side, 20–45 long; dorso-median part with 8–15 pairs of setae dm, 35–50 long; posterior part with 26–35 pairs of peripheral setae, 40–90 long, inserted dorsally and ventrally. Venter with 10–14 pairs of setae vm, 30–50 long, present lateral to genital area. Genital setae absent. Pseudanal series represented by 4 pairs of setae ps. Setae ps1 50–55 long and situated dorsally, setae ps2 40–45 long and situated terminally; setae ps3 and ps4 75–95 and 95–105 long, respectively, situated dorsally. Legs, coxal setation 1a, 1b, 2b, 3a, 3b, 3c and 3d arranged in formula 2-1-0-4. Setae of trochanters I-IV: 1-1-1-0, femora I-IV: 5-4-3-2, genua I-IV: 5-5-3-3 and tibiae I-IV: 5-5-5-5. Setae d’FI-IV, d”FII, d’GI-IV, d”GI-II and lGII-III serrate; d”FI, lFIII, lGI and lGIV, slightly serrate; vTrI-IV, v’FI-IV, v”FI-II, lFI, v’GI-IV, v”GI-II, d’TiI-IV, l’TiI-IV, l”Ti-IV, v’TI-IV and v”TiI-IV with barely discernible serration. Tarsi I-IV as in female.Larva [Based on 1 larva; Fig. Gnathosoma as in female, except for lack of subcapitular setae n. Idiosoma 240 long, 360 wide. Dorsum with 11 pairs of slightly serrate setae, 10–35 long: 4 pairs situated anteriorly, 4 pairs medio-laterally and 3 pairs posteriorly. Eyes present. Genital slit situated terminally. Genital setae absent. Pseudanal setal series represented by 1 pair of slightly serrate setae ps1. Legs, coxal setation 1a and 3a arranged in formula 1-0-1. All coxal setae filiform. Setae of trochanters I-III: 0-0-0, femora I-III: 2-2-0, genua I-III: 2-3-0 and tibiae I-III: 5-5-5. All setae on each podomere smooth, except for serrate setae dFI and dFII. Setation of tarsi: I 11 setae  and solenidion ω1; II 8 setae  and ω1; III and IV with 8 setae each . Setae tc’, tc”, it’ and it” of leg I represented by eupathidia, all setae p’ and p” fan-like, setae a’ and a” of legs I-III and setae u’ and u” of leg I smooth; setae ft, tc’, vs’, vs”, u’ and u” of legs I-III slightly serrate.Differential diagnosisNeopterygosoma chilensis  in Chile  from Argentina.Species included: Neopterygosoma patagonica .Neopterygosoma patagonicaPterygosoma patagonica Dittmar de la Cruz, Morando & Avila, 2004Syn. Type-host: Liolaemus petrophilus Donoso-Barros & Cei (Sauria: Liolaemidae).Type-locality: Argentina.Type-material (not examined): The type material is lost .Neotype (examined): One female (Reg. no. UKSW-PTE2.1) from Liolaemus petrophilus Donoso-Barros & Cei (Sauria: Liolaemidae), Argentina: Chubut Province, Paso de Indios .Other material examined: Three females from Liolaemus petrophilus Donoso-Barros & Cei (Sauria: Liolaemidae), Argentina: Chubut Province, Paso de Indios ; 3 females from Liolaemus austromendocinus Cei, Argentina: Mendoza, San Rafael ; 2 females from Lioleamus rothi Koslowsky, Argentina: Chubut Province, Telsen . All mites are deposited in the UKSW (UKSW-PTE2.2).Records: Dittmar de la Cruz et al. Body much wider than long (1.5–1.8 times). Second pair of legs discernibly shorter than others. Peripheral setae much longer than dorsal setae situated medially and laterally. Leg setae l’GIV absent ……………………………………………………… N. patagonica (patagonica group)Body only slightly wider than long (1.1–1.3 times). Legs I-IV subegual in length. Peripheral setae subequal with medial and lateral setae on idiosomal dorsum. Leg setae Five setae on genu I and 3 on femur IV ………………………………………………………………. 3N. formosusFour setae on genu I and 2 setae on femur IV ………………………………………. Four setae on femur II ………………………………………………………………………………………. 4Five setae on femur II ……………………………………………………………………………………….. 5N. chilensisPseudanal setae 5 ………………………………………………………………………………. N. schroederi n. sp.Pseudanal setae 4 …………………………………………………………………… N. cyanogasteriFixed cheliceral digit spinous ……………………………………………………….. Fixed cheliceral digit reduced to rounded structure ………………………………………………. 6Coxal fields I with 2 setae. Gnathosoma situated apically ……………………………………… 7N. ovataCoxal fields I with 3 setae. Gnathosoma displaced on dorsal side ………………… a’ and a” of tarsi I slightly serrate. Setae v’TrI-IV serrate. Setae 3a smooth …………………………………………………………………………………………….. N. levissimaAntero-median setae increase in length from the anterior to the posterior part of setal cluster. Setae a’ and a” of tarsi I smooth. Setae v’TrI-IV with barely discernible serration. Setae 3a serrate …………………………………. N. ligareAntero-median setae subequal in length. Setae


SOS was mixed with LLL at several mass fractions of LLL (wLLL); the mixtures obtained were analyzed for polymorphic phase behavior using differential scanning calorimetry (DSC) and synchrotron radiation X-ray diffractometry (SR-XRD). In melt crystallization with constant-rate cooling, SOS and LLL formed eutectics in their metastable polymorphs, allowing the occurrence of a compatible solid solution at wLLL ≥ 0.925. With subsequent heating, the resultant crystals transformed toward more stable polymorphs, then melted in a eutectic manner. For mixtures aged at 25 °C after melt crystallization, eutectics were found in the extended wLLL region, even at wLLL = 0.975. These results indicate that phase separation between SOS and LLL progressed in their solid solution under stabilization. The crystal growth of the separated SOS fraction may cause fat-bloom formation in compound chocolate containing CB and CBS. To solve this problem, the development of retardation techniques against phase separation is expected.This paper reports the precise analysis of the eutectic mixing behavior of 1,3-distearoyl-2-oleoyl- Theobroma cacao L.) by press, expeller, or solvent extraction [sn-glycerols of 1(3)-palmitoyl-2-oleoyl-3(1)-stearoyl-sn-glycerol (POS), 1,3-distearoyl-2-oleoyl-sn-glycerol (SOS), and 1,3-dipalmitoyl-2-oleoyl-sn-glycerol (POP) comprise at least 70% of the total TAGs in CB [2-2), form IV (β′1-2), form V (β2-3), and form VI (β1-3) in ascending order of thermal stability [Cocoa butter (CB), a material fat for chocolate products, is obtained from the beans of the cacao plant , and 1(3)-lauroyl-2,3-dimyristoyl-sn-glycerol (LMM) [The unstable supply, high price, and uncertain quality of CB have resulted in the production of alternatives derived from other vegetable oils ,19,20,21ol (LMM) ,18,22. Tol (LMM) . Howeverol (LMM) ,26. Indeol (LMM) ,28,29.Despite their similar physical properties, CB and CBS incompatibly form eutectics in their mixtures due to the significant difference in TAG composition ,22,23,24Phase diagrams are useful to clarify the interactions between two components of binary mixtures ,37. The At present, the mechanisms of eutectic formation between CB and CBS have not yet been determined, possibly because few fundamental studies on the mixing phase behavior of the component TAGs have been conducted. This study aimed to clarify the mixing phase behavior of SOS and LLL as a typical model case of the mixture of CB and CBS. For this purpose, mixtures of SOS and LLL (SOS/LLL) were precisely analyzed for crystallization kinetics and polymorphic phase behavior. wLLL). For example, the mixture at wLLL = 0.667 contains SOS and LLL at a mass ratio of SOS/LLL = 0.333/0.667. Differential scanning calorimetry (DSC) and synchrotron radiation XRD (SR-XRD) were employed to analyze the polymorphic crystallization kinetics of the mixtures during constant-rate cooling (2 °C/min) and subsequent heating (5 °C/min). Before cooling, the mixtures were pretreated at 80 °C for 10 min to ensure the molten state, eliminating any crystal memory. The following aging process was inserted as necessary between the cooling and heating: cooling at 15 °C for 30 min, heating to 25 °C at a rate of 5 °C/min, and then maintaining the temperature for two weeks.Samples for all experiments in this study were prepared by mixing SOS with LLL at several mass fractions of LLL  to 17.5 °C (wLLL = 0.500) for SOS and from 17.9 °C (wLLL = 0.111) to 17.0 °C (wLLL = 0.500) for LLL. Separation between the two peaks indicates the formation of incompatible eutectics between SOS and LLL.For SOS-dominant mixtures at wLLL = 0.600 to 1.000 exhibit one broad peak alone in their cooling thermograms. With increasing wLLL value, this peak increases its peak-top temperature from 11.6 °C (wLLL = 0.600) to 18.0 °C (wLLL = 1.000). The similar peak shape between the mixtures at wLLL = 0.600 to 0.975 and pure LLL at wLLL = 1.000 implies the possibility that LLL crystallized compatibly incorporating the minor fraction of SOS to form a solid solution.In contrast, the LLL-dominant mixtures at wLLL = 0.667 exhibits as many as seven peaks: a faint endothermic peak at 19.3 °C, a small exothermic peak at 20.6 °C, an endothermic peak at 29.7 °C, a large exothermic peak at 31.0 °C, an endothermic peak at 33.9 °C, a small exothermic peak at 37.6 °C, and a large endothermic peak at 44.5 °C.Immediately after the completion of cooling to 0 °C, the mixtures were reheated with constant-rate heating. As indicated in the right part of wLLL = 0.000 and pure LLL at wLLL = 1.000; one is a minor exothermic peak at 27.1 to 27.9 °C; the other is a main endothermic peak at the higher temperature. This endothermic peak is ascribed to the melting of SOS or LLL crystals. Corresponding endothermic peaks are also observed for the selected mixtures, as indicated by open and closed triangles in wLLL = 0.000 to 0.900, exhibits a peak-top temperature ranging from 36.2 °C (wLLL = 0.000) to 33.4 °C (wLLL = 0.900). In contrast, the endothermic peak for the melting of LLL crystals, found in mixtures at wLLL = 0.500 to 1.000, exhibits a peak-top temperature ranging from 41.4 °C (wLLL = 0.500) to 45.8 °C (wLLL = 1.000).In contrast, only two peaks are observed in the heating thermograms of pure SOS at wLLL = 0.600 and 0.667 exhibit more peaks than the other aged mixtures. For example, the mixture at wLLL = 0.667 exhibits four endothermic peaks at 33.3, 35.8, 39.5, and 43.5 °C; the former three peak-top temperatures are higher than those of the corresponding endothermic peaks observed for the non-aged mixture at wLLL = 0.667. In contrast with the non-aged mixtures, no marked exothermic peaks are found for the aged mixtures.wLLL = 0.000 exhibits a single endothermic peak at 41.8 °C, and pure LLL at wLLL = 1.000 exhibits a single endothermic peak 46.1 °C. Corresponding peaks due to the melting of SOS and LLL crystals are also observed for the selected mixtures, as indicated by open and closed triangles in wLLL = 0.000 and 0.111, exhibits a peak-top temperature ranging from 41.8 °C (wLLL = 0.000) to 41.1 °C (wLLL = 1.000). In contrast, the endothermic peak for the melting of LLL crystals, found in mixtures at wLLL = 0.600 to 1.000, exhibits a peak-top temperature ranging from 42.7 °C (wLLL = 0.600) to 46.1 °C (wLLL = 1.000). The comparison of the heating thermograms between non-aged ), where θ is the scattering angle and λ is the wavelength of X-rays applied [d values with the corresponding values in For the further analysis of crystallization kinetics from the viewpoint of crystal structure, we performed SR-XRD measurements of the mixtures under the same thermal conditions as the DSC measurements. Because of the high-flux X-ray source and the high-sensitivity detector, SR-XRD enables the time-resolved monitoring of the polymorphic phase transition of TAG crystals ,44. In o applied . The TAGwLLL = 0.950 during the cooling-and-heating. The data are displayed three-dimensionally in topographical plots where the time-dependent variation of the SAXS (the SAXS a and WAXthe SAXS b profiles| = 0.307 nm−1 (d = 3.25 nm) occurs first at ~21 °C in the cooling process, attributable to the crystallization of LLL in the β′-2 polymorph. As the temperature decreases, this peak sharpens, increasing its diffraction intensity until ~14 °C and then plateaus, preserving intensity and position. In subsequent heating, this peak grows again from ~21 °C and rapidly declines from ~34 °C before disappearing at ~39 °C. Meanwhile, the peak shifts its position slightly from |s| = 0.306 nm−1 (d = 3.27 nm) at 0 °C to |s| = 0.309 nm−1 (d = 3.24 nm) at 35 °C. Shortly afterward, a new peak at |s| = 0.318 nm−1 (d = 3.15 nm) appears from ~35 °C, ascribable to the occurrence of LLL β-2 crystals. With further heating, this peak develops until ~43 °C before disappearing at ~55 °C, exhibiting a slight positional shift to |s| = 0.319 nm−1 (d = 3.13 nm) at 50 °C. The extracted SAXS profiles at 0, 35, 40, and 50 °C in the heating process are presented in For the SAXS profiles a, a sings| = 2.37 nm−1 (d = 0.422 nm) and 2.61 nm−1 (d = 0.384 nm) with positional modification depending on the temperature. These peaks, characteristic of the β′ sub-cell structure, occur first at ~21 °C during cooling and start to separate at ~27 °C during subsequent heating.Corresponding to the varying SAXS profiles, the WAXS profiles b presents| = 2.19 nm−1 (d = 0.457 nm), 2.29 nm−1 (d = 0.437 nm), 2.34 nm−1 (d = 0.427 nm), 2.39 nm−1 (d = 0.418 nm), 2.47 nm−1 (d = 0.405 nm), and 2.59 nm−1 (d = 0.386 nm). The occurrence of these peaks was recently reported for LLL β′-2 crystals, which were obtained by melt crystallization with isothermal cooling at 28 °C [s| = 2.19 nm−1 (d = 0.457 nm) is specific to the β sub-cell structure and appears to be carried over to the corresponding peak at |s| = 2.18 nm−1 (d = 0.459 nm), which is observed with the intensified signal at 40 °C. Therefore, we conclusively ascribe the complex peak pattern at 35 °C to the coexistence of the LLL β′-2 crystals with improved perfection and the LLL β-2 crystals that converted from the imperfect LLL β′-2 crystals. The LLL β-2 crystals have potential to act as the subsequent crystallization of LLL in the β-2 polymorph.The second peak pattern appears clearly at 33 to 35 °C during heating. As typically observed at 35 °C, this pattern is complicated with peaks at |at 28 °C . Takegucs| = 2.18 nm−1 (d = 0.459 nm), 2.20 nm−1 (d = 0.454 nm), 2.57 nm−1 (d = 0.390 nm), and 2.64 nm−1 (d = 0.379 nm). These peaks, specific to the β sub-cell structure, and a split of the former two peaks were previously observed for the LLL β-2 crystals that occurred at the initial stage of crystallization under ultrasonication [The third peak pattern appears to be replacing the second one from 35 to 39 °C during heating. As typically observed at 40 °C, this pattern is characterized by major peaks at |nication . Since ps| = 2.17 nm−1 (d = 0.461 nm), 2.19 nm−1 (d = 0.457 nm), 2.59 nm−1 (d = 0.386 nm), and 2.66 nm−1 (d = 0.375 nm). Although the extent of the peak shifts is not as large, the overhead projection of the WAXS profiles . These changes indicate the rearrangement of the acyl chains in the sub-cell structure of the LLL β-2 crystals; the characteristic short spacings before and after the changes have recently been reported for pure LLL [2-2) to the more stable β-2 (β1-2). Two subtypes of the β polymorph were previously reported for LLL β-2 crystals occurring in the mixture of LLL with cholesterol at the 4% molar fraction [The fourth peak pattern is apparently derived from the third one as a result of peak shifts at 45 to 47 °C during heating. As typically observed at 50 °C, this pattern is characterized by major peaks at |pure LLL . Therefofraction . The samwLLL = 0.950 as follows. (1) During cooling, LLL starts to crystallize in the β′-2 polymorph at ~21 °C and grows until ~14 °C. (2) During subsequent heating, LLL β2-2 crystals occur at the partial expense of the LLL β′-2 crystals from ~21 °C, improving the crystal perfection of the residual LLL β′-2 crystals. (3) The remaining LLL β′-2 crystals melt from ~34 to ~39 °C and recrystallize as LLL β2-2 crystals from ~35 to ~43 °C. (4) The LLL β2-2 crystals melt from ~43 °C, partly converting to LLL β1-2 crystals from ~45 to ~47 °C. (5) The LLL β1-2 crystals melt completely at ~55 °C. These phase transitions are in good agreement with the thermal behavior in the DSC measurement . From the position at |s| = 0.266 nm−1 to 0.271 nm−1 (d = 3.76 nm to 3.69 nm), the diffuse peak indicates the occurrence of SOS crystals. Since the sharp peak behaves in the same manner as that observed for the mixture at wLLL = 0.950, this peak is associated with LLL crystals.For the SAXS profiles in s| = 0.304 nm−1 (d = 3.29 nm) occurs first at ~18 °C, indicating the crystallization of LLL in the β′-2 polymorph. This peak sharpens, increasing its diffraction intensity from ~18 to ~10 °C, and then plateaus, preserving intensity and position. Shortly after the occurrence of the peak for the LLL β′-2 crystals, a bulge appearing on the lower |s| side of this peak at ~14 °C grows to a diffuse peak at |s| = 0.266 nm−1 (d = 3.76 nm) with its plateaued maximum intensity at ~10 °C. Considering the fact that the diffuse peak weakens from ~10 °C during subsequent heating, we attribute this peak to the transitional structure of SOS α2-(2 + 3) crystals reorganizing the acyl-chain packing from hexagonal to orthorhombic. The α2-(2 + 3) phase, having mixed double and triple chain-length structure, was defined as the less stable subtype of the α polymorph together with the more stable α1-2 [During cooling, a single peak at |ble α1-2 .s| = 0.271 nm−1 (d = 3.69 nm). The resultant peak, attributed to the 002 reflection of SOS γ-3 crystals, weakens again without a break and disappears at ~35 °C. While the previously occurring diffuse peak attenuates from ~10 to ~24 °C, the sharp peak for LLL β′-2 crystals grows again from ~18 °C and declines from ~24 °C before disappearing at ~35 °C; in the meantime, the peak shifts position slightly from |s| = 0.304 nm−1 (d = 3.29 nm) at 0 °C to |s| = 0.307 nm−1 (d = 3.25 nm) at 30 °C. Shortly afterward, a new peak at |s| = 0.318 nm−1 (d = 3.15 nm) occurs from ~34 °C, ascribable to the occurrence of LLL β-2 crystals. With further heating, this peak develops until ~38 °C before disappearing at ~50 °C, exhibiting a slight positional shift to |s| = 0.319 nm−1 (d = 3.13 nm) at 45 °C. The extracted SAXS profiles at 0, 30, 40, and 45 °C during heating are presented in During heating, the SAXS-profile change begins with the attenuation of the diffuse peak from ~10 to ~24 °C, resulting in a less-intensified diffuse peak at |wLLL = 0.950 (s| = 2.12 nm−1 (d = 0.472 nm), attributable to SOS γ-3 crystals, contaminates from ~11 °C during cooling and intensifies from ~14 to ~30 °C before disappearing at ~35 °C during subsequent heating. No trace for the transitional SOS α2-(2 + 3) crystals is found behind the strong peaks at |s| = 2.37 nm−1 (d = 0.421 nm) and 2.61 nm−1 (d = 0.383 nm), as typically observed at 0 °C  During cooling, LLL starts to crystallize in the β′-2 polymorph at ~18 °C. (2) During the growth of LLL β′-2 crystals from ~18 to ~10 °C, SOS starts to crystallize in the transitional α2-(2 + 3) polymorph at ~14 °C and in the γ-3 polymorph at ~11 °C. (3) During subsequent heating, the SOS α2-(2 + 3) and γ-3 crystals melt successively at ~10 to ~35 °C, accompanied by the recrystallization of SOS γ-3 starting from ~14 °C. (4) LLL β2-2 crystals occur at the partial expense of the LLL β′-2 crystals from ~18 °C, improving the crystal perfection of the residual LLL β′-2 crystals. (5) The remaining LLL β′-2 crystals melt from ~24 to ~35 °C and recrystallize as LLL β2-2 crystals from ~34 to ~38 °C. (6) The LLL β2-2 crystals melt from ~38 °C, partly converting to LLL β1-2 crystals from ~41 to ~44 °C. (7) The LLL β1-2 crystals melt completely at ~50 °C. These phase transitions are in good agreement with the thermal behavior in the DSC measurement  and WAXS peaks at |s| = 2.11 nm−1 to 2.12 nm−1 (d = 0.474 nm to 0.472 nm), 2.56 nm−1 to 2.58 nm−1 (d = 0.391 nm to 0.388 nm), 2.75 nm−1 to 2.79 nm−1 (d = 0.363 nm to 0.358 nm). In addition, weak SAXS peaks at |s| = 0.129 nm−1 to 0.132 nm−1 (d = 7.77 nm to 7.57 nm) and 0.403 nm−1 to 0.413 nm−1 (d = 2.48 nm to 2.42 nm) are concurrently observed as 001 and 003 reflections from the SOS γ-3 crystals. These peaks occur prior to the peaks for LLL β′-2 crystals during cooling. Variation in SAXS-peak intensity implies the temporary formation of transitional SOS α2-(2 + 3) crystals and the following recrystallization as SOS γ-3 crystals. Aside from these observations, the overall evolvement of the SAXS and WAXS profiles is similar for the mixtures at wLLL = 0.417 and 0.667. The representative SAXS and WAXS profiles at 0, 30, 35, and 42 °C during heating are depicted in the SAXS  and WAXSthe SAXS  profileswLLL = 0.417 as follows. (1) During cooling, SOS starts to crystallize in the γ-3 polymorph at ~20 °C. (2) Following the rapid growth of SOS γ-3 crystals from ~20 to ~19 °C, LLL β′-2 crystals grow from ~19 to ~16 °C, and transitional SOS α2-(2 + 3) crystals grow from ~18 to 16 °C. (3) The SOS α2-(2 + 3) crystals gradually dissolve the structure from ~9 to 0 °C. (4) During subsequent heating, the SOS γ-3 crystals grow again from ~3 to ~13 °C before melting from ~28 to ~41 °C. (5) The LLL β′-2 crystals melt from ~28 to ~34 °C and recrystallize as LLL β2-2 crystals from ~32 to ~38 °C; this process includes the occurrence of LLL β′-2 crystals with improved perfection. (6) The LLL β2-2 crystals melt from ~39 °C, partly converting to LLL β1-2 crystals from ~41 to ~43 °C. (7) The LLL β1-2 crystals melt completely at ~46 °C. These phase transitions are in good agreement with the thermal behavior in the DSC measurement , 0.306 nm−1 (d = 3.27 nm), and 0.319 nm−1 (d = 3.13 nm) are observed at the initial stage of heating; the former two peaks are identified as 001 and 002 reflections of SOS β2-3 or β1-3 crystals, and the last intense peak is attributable to the 001 reflection of LLL β2-2 or β1-2 crystals, respectively. During heating, the SOS-specific peaks decrease their intensity from ~39 °C and disappear at ~43 °C. In contrast, the LLL-specific peak intensifies from ~35 to ~39 °C and then declines from ~41 °C before disappearing at ~48 °C.For the SAXS profiles in s| = 2.52 nm−1 (d = 0.398 nm), 2.59 nm−1 (d = 0.386 nm), 2.67 nm−1 (d = 0.374 nm), 2.74 nm−1 (d = 0.365 nm), and 2.82 nm−1 (d = 0.355 nm). The group that disappears at the higher temperature includes representative peaks at |s| = 1.86 nm−1 (d = 0.536 nm), 2.17 nm−1 (d = 0.460 nm), 2.20 nm−1 (d = 0.454 nm), 2.60 nm−1 (d = 0.384 nm), and 2.68 nm−1 (d = 0.373 nm). Because of the two-step decrement in peak intensity, a peak at |s| = 2.17 nm−1 (d = 0.460 nm) is assigned to both groups. Therefore, we presume from these d values that during heating, SOS β2-3 crystals and LLL β2-2 or β1-2 crystals melted in this order. Moreover, a peak at |s| = 2.84 nm−1 (d = 0.353 nm) gradually shifts its position to |s| = 2.82 nm−1 (d = 0.355 nm) until ~38 °C, intensifies from ~38 to ~40 °C, and then disappears at ~48 °C. This is consistent with the fact that the LLL-specific SAXS peak at |s| = 0.319 nm−1 (d = 3.13 nm) intensifies from ~35 to ~39 °C, indicating the occurrence of the polymorphic transformation of LLL crystals from β2-2 to β1-2.For the WAXS profiles in wLLL = 0.667, initially-coexisting SOS β2-3 and LLL β2-2 crystals undergo the polymorphic transformation of LLL crystals from β2-2 to β1-2 from ~35 to ~39 °C, the melting of SOS β2-3 crystals from ~39 to ~43 °C, and the melting of LLL β1-2 crystals from ~41 to ~48 °C.To summarize the SR-XRD results for the 25 °C-aged mixture at wLLL = 0.950, LLL β2-2 crystals convert to LLL β1-2 crystals from ~37 to ~42 °C; SOS β2-3 crystals melt from ~37 to ~41 °C; and LLL β1-2 crystals melt from ~47 to ~52 °C. In the mixture at wLLL = 0.417, LLL β2-2 and SOS β2-3 crystals melt simultaneously from ~40 to ~47 °C. In the mixture at wLLL = 0.111, LLL β2-2 crystals melt from ~40 to ~43 °C, and SOS β2-3 crystals melt from ~40 to ~47 °C.The other aged mixtures were also analyzed for polymorphic phase transitions during heating. Although the SR-SRD data are not presented here, the representative results are summarized as follows. In the mixture at wLLL value at which SAXS peaks or traces for SOS are confirmed. The less intense and denser WAXS peaks are not suitable for this analysis.The SR-XRD data are also analyzed for the determination of the solid-solubility limit of SOS in SOS/LLL. In this study, the solid-solubility limit is defined as the maximum wLLL = 0.925, 0.900, and 0.889, taken during melt crystallization with a constant-rate cooling (2 °C/min). Profiles are displayed every 1 °C, and the low-|s| parts of the profiles with magnified intensity are inserted as insets in this figure.wLLL = 0.925 exhibits an intense peak at |s| = 0.307 nm−1 (d = 3.25 nm). This peak is due to LLL β′-2 crystals growing from the melt from ~21 to 0 °C. No other peaks or shoulders due to the crystallization of SOS are observed. This means that SOS is fully incorporated into the LLL β′-2 crystals as a solid solution.In wLLL = 0.900 also exhibits an intense peak at |s| = 0.306 nm−1 (d = 3.27 nm). Compared to the mixture at wLLL = 0.925, this peak due to LLL β′-2 crystals is located at a slightly lower |s| position, although the first occurrence temperature is unchanged. In addition, two shoulders at |s| = ~0.127 nm−1 (d = ~7.87 nm) and ~0.261 nm−1 (d = ~3.83 nm) are observed from ~8 to 0 °C, as indicated by the closed triangle. These shoulders mean the occurrence of SOS crystals, because the corresponding peaks due to the 001 and 002 reflections from transitional SOS α2-(2 + 3) crystals were previously reported for pure SOS [In pure SOS .wLLL = 0.889 exhibits almost the same profiles as the mixture at wLLL = 0.900. While the shoulders due to SOS crystals become more prominent, the peak due to LLL crystals starts at the lower temperature of ~19 °C.In wLLL = 0.900 and 0.925. The continued data for subsequent heating at a rate of 5 °C/min indicate the unchanged solid-solubility limit (data not shown).We conclude from these results that in melt crystallization, the solid-solubility limit of SOS in SOS/LLL lies between wLLL = 0.975, 0.950, and 0.925, taken during constant-rate heating (5 °C/min) after aging at 25 °C for two weeks. Profiles are displayed every 1 °C, and the low-|s| parts of the profiles with magnified intensity are inserted as insets in this figure. It should be noted that the SAXS profiles of the mixture at wLLL = 0.925 ( = 0.925 c are lar = 0.925 a.s| = 0.153 nm−1 (d = 6.54 nm) and a strong peak at |s| = 0.318 nm−1 (d = 3.15 nm). The peak-intensity ratio of the weak peak to the strong peak rises with decreasing wLLL. As stated for the mixture at wLLL = 0.950 in 2-2 crystals convert to LLL β1-2 crystals during heating. Therefore, at the initial stage of heating, the weak SAXS peak is identified as due to SOS β2-3 crystals, and the strong SAXS peak is identified as due to LLL β2-2 crystals. With elevating temperature, the weak peak disappears completely at ~41 °C, and the strong peak disappears completely at ~52 °C; these temperatures are constant among the three mixtures. Behind the strong peak, its shoulder at |s| = ~0.280 nm−1 (d = ~3.57 nm) is observed for only the mixture at wLLL = 0.925.These three mixtures all exhibit a weak peak at |wLLL = 0.975. That is, the solid-solubility limit of SOS in SOS/LLL lies between wLLL = 0.975 and 1.000.These results confirm that after aging, SOS and LLL form eutectics in the mixture even at wLLL were examined for crystallization kinetics, polymorphic phase transitions, and the solubility limit of SOS. Integrating all the results acquired, we construct diagrams for the binary phase behavior of SOS/LLL. In the diagrams, the phase-transition temperatures, defined as average values of the peak-top temperatures in the multiple DSC measurements  to 17.5 °C (wLLL = 0.500). Just below these temperatures, transitional SOS α2-(2 + 3) crystals and LLL β′-2 crystals occur at almost the same time, exhibiting nearly constant phase-transition temperatures from 17.0 to 18.5 °C.In the mixtures at wLLL = 0.600 to 0.900, three solid phases  crystals, SOS γ-3 crystals, and LLL β′-2 crystals) form from the melt at phase-transition temperatures ranging from 11.6 °C (wLLL = 0.600) to 14.8 °C (wLLL = 0.900). Among the three phases, the first-occurring phase is considered LLL β′-2 crystals, as observed for the mixture at wLLL = 0.667 in the SR-XRD result (2-(2 + 3) crystals probably surpass SOS γ-3 crystals in these mixtures.In the mixtures at D result . From thwLLL = 0.925 to 1.000, only LLL β′-2 crystals occur from the melt at phase-transition temperatures ranging from 13.5 °C (wLLL = 0.925) to 18.0 °C (wLLL = 1.000). This indicates that the LLL β′-2 crystals grow incorporating the minor fraction of SOS as a solid solution.In the mixtures at wLLL = 0.000 to 0.500, 0.600 to 0.900, and 0.925 to 1.000. On the behalf of each group, the mixtures at wLLL = 0.417, 0.667, and 0.950 are selectively explained for the phase-transition behavior as follows.In wLLL = 0.417, the transitional SOS α2-(2 + 3) crystals have already dissolved at the start of heating, so that only SOS γ-3 and LLL β′-2 crystals exist at the initial stage of heating. After the growth of the SOS γ-3 crystals without the fluctuating DSC heat flow, LLL β2-2 crystals occur at the partial expense of the LLL β′-2 crystals, giving one exothermic effect at the phase-transition temperature of 26.9 °C. With further heating, the SOS γ-3 crystals and the residual LLL β′-2 crystals with improved perfection melt and the LLL β2-2 crystals grow, resulting in one endothermic effect at the phase-transition temperature of 34.2 °C. The higher phase-transition temperatures, due to the melting of the resultant LLL β2-2 crystals, the conversion of the remaining LLL β2-2 crystals to LLL β1-2 crystals, and the melting of the LLL β1-2 crystals are not available: this is because the corresponding endothermic effect appears as a peak shoulder in the DSC thermogram, as indicated by a red arrow in the right part of In the mixture at wLLL = 0.667, the phase-transition behavior starts from the solid phase consisting of transitional SOS α2-(2 + 3) crystals, SOS γ-3 crystals, and LLL β′-2 crystals. Compared to the mixture at wLLL = 0.417, a larger number of phase-transition points are found in the phase behavior. A pair of endothermic and exothermic effects at phase-transition temperatures of 19.3 and 20.6 °C provide evidence that the transitional SOS α2-(2 + 3) crystals convert to SOS γ-3 crystals via melt-mediated transformation. Likewise, a pair of endothermic and exothermic effects at phase-transition temperatures of 29.7 and 31.0 °C provide evidence that the LLL β′-2 crystals partly convert to LLL β2-2 crystals via melt-mediated transformation. The endothermic effect at the phase-transition temperature of 33.9 °C corresponds to the melting of the SOS γ-3 crystals. The exothermic effect at the phase-transition temperature of 37.8 °C is due to the melt-mediated transformation from the residual LLL β′-2 crystals with improved perfection to LLL β2-2 crystals, where the exothermic heat of crystallization prevails over the endothermic heat of fusion. The endothermic effect at the phase-transition temperature of 43.6 °C includes a sequence of melting of the LLL β2-2 crystals, polymorphic transformation of the LLL crystals from β2-2 to β1-2, and the melting of the resultant LLL β1-2 crystals.In the mixture at wLLL = 0.950, LLL β′-2 crystals incorporating SOS as a solid solution solely exist at the initial stage of heating. The LLL β′-2 crystals partly convert to LLL β2-2 crystals probably via melt-mediated transformation, giving the exothermic effect at the phase-transition temperature of 32.1 °C. The other exothermic effect at the phase-transition temperature of 38.5 °C is due to the melt-mediated transformation from the residual LLL β′-2 crystals with improved perfection to LLL β2-2 crystals. The endothermic effect at the phase-transition temperature of 45.7 °C includes a sequence of melting of LLL β2-2 crystals, the polymorphic transformation of LLL crystals from β2-2 to β1-2, and the melting of resulting LLL β1-2 crystals.In the mixture at 2-3 and LLL β2-2 crystals at the initial stage of heating. The melt crystallization with constant-rate cooling was interrupted at 15 °C by subsequent aging including preliminary isothermal cooling at 15 °C for 30 min. Nevertheless, the aged mixtures exhibit a phase diagram similar to that of the non-aged mixtures. This indicates the possibility that melt-crystallization memory is inherited even after the long-term storage of the mixtures. Accordingly, the phase diagram of the aged mixtures is also divided into three areas of the mixtures at wLLL = 0.000 to 0.500, 0.600 to 0.900, and 0.925 to 1.000, whose phase-transition behavior is explained by representative mixtures at wLLL = 0.417, 0.667, and 0.950.wLLL = 0.417, the initially coexisting SOS β2-3 and LLL β2-2 crystals melt simultaneously, giving only one endothermic effect at the phase-transition temperature of 39.9 °C.In the mixture at wLLL = 0.667, the phase-transition behavior starts from the solid phase consisting mainly of SOS β2-3 and LLL β2-2 crystals. The endothermic effect at the phase-transition temperature of 33.3 °C indicates melting of the contaminated SOS γ-3 crystals, although no corresponding peaks are found in the SR-XRD data  and LLL with ≥ 99% purity was purchased from Sigma-Aldrich . These TAGs were used as received without further purification.wLLL): specifically, wLLL = 0.000, 0.111, 0.200, 0.333, 0.417, 0.500, 0.600, 0.667, 0.800, 0.889, 0.900, 0.925, 0.950, 0.975, and 1.000. For example, the mixture at wLLL = 0.667 contained SOS and LLL at a mass ratio of SOS/LLL = 0.333/0.667. The melt-mixing was conducted with vortex stirring in glass vials immediately after heating the vials above 80 °C.Mixtures of SOS/LLL were obtained by melt-mixing SOS with LLL at several mass fractions of LLL  heating at 80 °C for 10 min, (ii) cooling to 15 °C at a rate of 2 °C/min, (iii) isothermally cooling at 15 °C for 30 min, (iv) heating up to 25 °C at a rate of 5 °C/min, then (v) annealing at 25 °C for two weeks. Steps (i)–(iv) were completed using a thermal control device 10,021 . Step (v) was done in an incubator MIR-152 .During the measurements, the non-aged samples were heated at 80 °C for 10 min, cooled to 0 °C at a rate of 2 °C/min, then reheated to 80 °C at a rate of 5 °C/min; the aged samples were heated from 25 to 80 °C at a rate of 5 °C/min.n-decane standards. Each SOS/LLL mixture (~3 mg) in the molten state was weighed on an aluminum pan and hermetically sealed with the pair lid, giving a sample pan. After being subjected to the aging process as necessary, the sample pan and an empty pan for reference were introduced to the thermally controlled measurement chamber where dry nitrogen gas flowed at a constant rate of 25 mL/min.DSC measurements were conducted using heat-flux DSC Q100 . Temperature and heat flow were calibrated with reference to the melting points and enthalpies of indium and The measurement was repeated at least three times for each mixture. The thermograms obtained were analyzed for the peak-top temperatures using the software included in the DSC apparatus.SR-XRD measurements were carried out at a beamline BL-6A of Photon Factory, a synchrotron radiation facility of the High Energy Accelerator Research Organization (KEK) in Tsukuba, Japan. X-rays with a wavelength of 0.15 nm were applied to a sample cell, and the SAXS (WAXS) from the sample therein were simultaneously detected by PILATUS 1M ). The sample-to-detector distance was calibrated using a silver behenate standard.The sample cell was made with a 1.5 mm-thick aluminum plate, having a 2 mm diameter round-hole to be filled with the SOS/LLL mixtures, and X-ray transmission windows of 25 μm thick polyimide films to seal both sides of the hole. After being aged as necessary, the sample cell was placed on the temperature-controlled stage of a thermal control device 10002L ; in the measurement setup, the stage was vertically installed in front of the detectors. The SAXS and WAXS data were acquired every 6 s with 5 s X-ray exposure time and 1 s interval. Topographic plots of the SAXS and WAXS profiles were performed using OriginPro 2020b software .Under thermodynamically metastable conditions, SOS and LLL form eutectics in their metastable polymorphs, allowing the occurrence of a compatible solid solution;The eutectic nature occurring in metastable conditions is retained even after the polymorphic stabilization of SOS and LLL crystals by aging;The polymorphic stabilization narrows the solid-solution area of SOS in LLL crystals, indicating that SOS incorporated in LLL crystals as a solid solution separates from the LLL crystals and crystallizes to form a new solid phase.This study precisely analyzed the eutectic mixing behavior of SOS and LLL, as a typical model case of the mixture of CB and CBS. Using DSC and SR-XRD, the SOS/LLL mixtures were examined for crystallization kinetics, polymorphic phase-transition, and the solid-solubility limit of SOS. Integrating the acquired results, we obtained diagrams for the binary phase behavior of SOS/LLL that are similar to those of CB/CBS. The major findings on the mixing behavior of SOS/LLL are:Such phase separation and crystallization of the separated phase are considered the crucial cause of fat bloom in CBS-based compound chocolate. To solve this problem, we hope to develop retardation techniques against phase separation by applying new ways of fat blending: for example, molecular compound crystal formation .


There was an error in an author’s name in the published article. The author Dae-Hyuk Kim was incorrectly given as as Dae-Hyun Kim.Corrected author list:1, José R. Castón2, Robert H. A. Coutts3, Bradley I. Hillman4, Daohong Jiang5, Dae-Hyuk Kim6, Hiromitsu Moriyama7 and ICTV Report ConsortiumSaid A. GhabrialAdditionally, in Fig. 2 the two black lines indicating the double-stranded RNA genome of each genome segment were incorrectly merged together at certain magnifications. The correct Fig. 2 is provided below.


Ulcerative colitis (UC) is a chronic intestinal disease with unclear pathogenesis. With an increasing global prevalence over the past two decades, UC poses a serious threat to public health. Baitouweng decoction (BTW), a traditional Chinese medicine, has been shown to have good clinical efficacy for treating intestinal inflammation. Yet, the efficacy of BTW in UC and the underlying mechanism remain unclear. The current study aimed to determine whether BTW suppressed intestinal inflammation in mice and the potential mechanism. We used a dextran sulfate sodium (DSS)-induced murine colitis model to test the anti-inflammatory efficacy of BTW. Clinical symptoms were scored by the disease activity index (DAI), and the colon length and pathological changes in colon tissue were also used to further evaluate the efficacy of BTW. Precisely how BTW affected immune function and the intestinal barrier of UC mice was also examined. BTW significantly reduced DAI score and colonic pathological damage. BTW regulated the balance between T helper (Th)17 and regulatory T (Treg) cells, decreased interleukin (IL)-1β, IL-6, and tumor necrosis factor-α, and increased IL-10 levels. BTW reduced intestinal permeability of UC mice, increased expression of tight junction proteins (occludin and zonula occludens-1), and decreased expression of phospho-nuclear factor (p-NF)-κB and phospho-extracellular signal-regulated kinase (p-ERK) in the colon. BTW inhibited the ERK/p-NF-κB signaling pathway and suppressed expression of cyclo-oxygenase-2 and inducible NO synthase in lipopolysaccharide-activated RAW 264.7 cells. BTW significantly promoted the synthesis of short-chain fatty acids in the gut, particularly acetate, propionate, isobutyric acid, and isovalerate. The results suggest that BTW can protect against DSS-induced UC. The mechanism may be partially attributed to regulating the balance of Th17/Treg cells and restoring the intestinal epithelial barrier. Colonoscopy often reveals congestion, edema, granular changes, and ulceration of the intestinal mucosa. Histologically, UC is characterized by extensive ulcer distribution, with neutrophil infiltration and epithelial necrosis . The pat by 2018 . UC has  by 2018 . Current by 2018 .Pulsatilla chinensis (Bunge) Regel (Baitouweng), Coptis chinensis Franch (Huanglian), Phellodendron chinense C. K. Schneid (Huangbai), and Fraxinus chinensis Roxb (Qinpi). Previous studies have indicated that the extract of Pulsatilla chinensis (Bunge) Regel acts as an anti-inflammatory and immunomodulator in a mouse model of lipopolysaccharide (LPS)-induced systemic inflammation  is a reliable, alternative, and complementary therapy with fewer adverse effects. TCM has attracted worldwide attention for the treatment of many diseases . Baitouwammation . Coptidi in rats . An incrPulsatilla chinensis (Bunge) Regel (20190815), Coptis chinensis Franch (20190723), Phellodendron chinense C. K. Schneid (20190820), and Fraxinus chinensis Roxb (20190901). They were identified by Dr. Yan Hui . Voucher specimens of the above four herbs (No. NJUCM_201909101-201909104) were stored in the Herbarium Center of Nanjing University of Chinese Medicine. Pulsatilla chinensis (Bunge) Regel 15 g, Coptis chinensis Franch 6 g, Phellodendron chinense C. K. Schneid 12 g, and Fraxinus chinensis Roxb 12 g were combined and submerged in distilled water (600 ml) for 30 min. Subsequently, the mixture was extracted twice (1 L first followed by 500 ml) over 1 h. Two rounds of extraction were conducted, and the resulting mixture was filtered. The filtrate was concentrated until the established drug content was 1 g/ml. To prepare the BTW extract used in the in vitro cell study, 10 ml BTW (1 g/ml) was lyophilized by a LAB-1A-50E freeze dryer , yielding 1.84 g powder that was used in the in vitro study.The raw herbs for BTW were purchased from Beijing Tongrentang Co. Ltd, with batch numbers: Concentrated BTW (0.1 ml) was extracted by 1.0 ml methanol. The sample was then mixed and spun at 12,000 g for 5 min. The top layer was collected for LC-Q-TOF/MS analysis. The standard mixture, including esculin, fraxin, pulchinenoside A, coptisine, epiberberine, berberine, and palmatine , was also prepared at the indicated concentration for LC-Q-TOF/MS analysis.A Shimadzu UFLC 20ADXR system , with an online degasser quaternary pump, column temperature controller, and an autosampler, was used. The separation was achieved in a Thermo BDS Hypersil C18 . The column temperature was set at 35 °C. The mobile phase was composed of 0.05% formic acid–water (phase A) and 0.05% formic acid–acetonitrile (phase B) with a flow rate set at 0.35 ml/min. The gradient elution program was as follows: 0–4.5 min, 10% B; 4.5–26 min, 10–30% B; 26–29 min, 30–80% B; 29–30 min, 80–10% B; 30–32 min, 10% B. The injection volume was 5 μL.The raw MS and tandem MS (MS/MS) data were collected by a TripleTOF 5600 mass spectrometer. The analytes were ionized using electrospray ionization (ESI) in positive and negative modes. The m/z scan range for MS1 was 50–1200. For information-dependent acquisition, the m/z scan range for MS/MS was 50–1000. The main parameters for spectrometric analysis were as follows: ion source gases 1 and 2, 50 psi, DP, 55 eV, EP, 15 ev, and the collision energy, 15.Pulsatilla chinensis (Bunge) Regel (BTW) was pulchinenoside A. The chemical markers in Coptis chinensis Franch (Huanglian) and Phellodendron chinense C. K. Schneid (Huangbai) were shared and included epiberberinem, berberine, palmatine, and coptisine. The chemical markers in Fraxinus chinensis Roxb (Qinpi) were esculin and fraxin.The concentrations of the main constituents  in BTW were also determined using a validated HPLC method  with a mMale C57BL/6 mice aged 6–8 weeks were obtained from the Qinglongshan Animal Breeding Farm  (certificate no. SCXK (Su) 2017-0001). Mice were housed in a sterile facility at 25 °C with a 12 h light/dark cycle and given sterile food and water. All studies were conducted under the authorization of the Ethics Committee of Animal Experiments.Sixty mice were randomized into five groups of 12 after 1 week of acclimation: 1) control group; 2) DSS group; 3) sulfasalazine (SASP)-treated group; 4) low-dose BTW-treated (L-BTW) group; and 5) high-dose BTW-treated (H-BTW) group. With the exception of the control group, all mice received 2.5% DSS  in drinking water for 7 days. Subsequently, water was changed back to normal drinking water for an additional 3 days. The SASP group received 100 mg/kg SASP orally (Shanghai Sine Co. Ltd.) daily. The L-BTW and H-BTW groups were given 8 g/kg and 16 g/kg BTW orally, respectively. All doses were diluted with distilled water to ensure that each mouse received an equal volume of drug. The control and DSS groups were orally administered with an equal volume of distilled water.To investigate whether the therapeutic effect of BTW on UC was related to the activation or inhibition of extracellular signal-regulated kinase (ERK), ERK agonist (TBHQ) and ERK inhibitor (U0126) were used alone or in combination with BTW to treat DSS-induced UC. Thirty-six mice were randomly assigned to six groups of six: 1) DSS group; 2) DSS + BTW group; 3) DSS + TBHQ group; 4) DSS + BTW + TBHQ group; 5) DSS + U0126 group; and 6) DSS + U0126 + BTW group. Induction of colitis was the same as above. BTW (16 g/kg) was given orally to the corresponding group. For TBHQ feeding, food pellets were mixed with 1% TBHQ  . For U01The overall performance of each mouse was assessed by specific parameters. Daily body weight was recorded along with any volume loss to bleeding or diarrhea. Body weight loss was recorded on a graded scale, where 0, 1 2, 3, and 4 represented no weight loss, 1–5% loss, 5–10% loss, 10–15% loss, and >15% loss, respectively. Stool consistency was recorded with 0, 1, 2, 3, and 4 representing normal, loose stools, mild diarrhea, diarrhea, and liquid stools, respectively. Fecal blood was quantitated on a scale from 0 to 4, where 0 represented no bleeding and 4 represented gross blood loss per rectum. The average of all scores was calculated and used to record DAI .On day 10, all mice were killed by cervical dislocation. The colon from each mouse was collected and photographed, and the length was determined. A similar procedure was used to collect spleens, which were also weighed. The formula used to determine the spleen coefficient was spleen coefficient (%) = Ws/Wb × 100%, where Ws and Wb represent the spleen weight and body weight on day 10, respectively.Colon tissue was fixed in 4% (wt/vol) paraformaldehyde. Paraffin-embedded samples were processed to obtain 5 μm thick sections. Sections were stained with hematoxylin and eosin (H&E) and examined under light microscopy. The scoring criteria were used to determine the histological score . Data we2. LPS  was dissolved in sterile phosphate-buffered saline, and BTW and MEK-1/-2 inhibitor (U0126) were dissolved in dimethyl sulfoxide (DMSO). The final concentration of DMSO was 0.05% (v/v). RAW 264.7 cells were plated at 105 cells/mL for 24 h and pretreated with BTW for 2 h prior to LPS (1 μg/ml).RAW 264.7 macrophage cell line was purchased from the American Type Culture Collection . The cells were grown in Dulbecco’s modified Eagle’s medium with 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/ml streptomycin and maintained at 37 °C in an environment containing 5% CO4 cells/well in 96-well plates per 100 μL medium. To determine the appropriate concentration that was not cytotoxic, the Cell Counting Kit-8 (CCK-8) was used . CCK-8 solution (10 μL) was added to the cultured cells and the plates were incubated for 1–4 h. Metabolic activity was quantified by measuring light absorbance at 450 nm. Data were collected and analyzed from three independent samples.RAW 264.7 cells were plated at 105 cells/well in 96-well plates and incubated with or without LPS (1 μg/ml) in the absence or presence of BTW for 24 h. Each 50 μL of culture supernatant was mixed with an equal volume of Griess reagent  and incubated at room temperature for 15 min. NaNO2 was used to generate a standard curve, and nitrite production was determined by measuring optical density at 550 nm. Data were collected and analyzed from three independent samples.Nitrite level in the medium was used as an indicator of NO production. RAW 264.7 cells were plated at 2 × 10Whole blood was collected by orbital dislocation from all mice on day 10. The serum was centrifuged at 3500 rpm for 15 min to separate the serum. The serum concentration of interleukin (IL)-6, IL-10, TNF-α, and IL-1β was measured using commercial ELISA systems . Data were collected and analyzed from six independent samples.−ΔΔCt method  was used to extract total RNA. HiScript II qRT SuperMix (Vazyme Biotech) was used to generate 1 µg of first-strand cDNA. PCRs were conducted with ChamQ SYBR qPCR Master Mix (Vazyme Biotech). The primers were purchased from Sangon Biotech (Shanghai) Co. Ltd., and the primer sequences (designed and checked by Primer Premier 6.0) were as follows: GAPDH-F, 5′-AGGTCGGTGTGAACGGATTTG-3′; GAPDH-R, 5′-TGTAGACCATGTAGTTGAGGTCA-3′; TNF-α-F, 5′-CCTCTCTCTAATCAGCCCTCTG-3′; TNF-α-R, 5′-GAGGACCTGGGAGTAGATGAG-3′; IL-1β-F, 5′-CTCGCCAGTGAAATGATGGCT-3′; IL-1β-R, 5′-GTCGGAGATTCGTAGCTGGAT-3′; IL-6-F, 5′-ACAACCACGGCCTTCCCTAC-′; IL-6-R, 5′-TCTCATTTCCACGATTTCCCAG-3′; IL-10-F, 5′-GCTCTTACTGACTGGCATGAG-3′; IL-10-R, 5′-CGCAGCTCTAGGAGCATGTG-3′; ZO-1-F, 5′-ACTCCCACTTCCCCAAAAAC-3′; ZO-1-R, 5′-CCACAGCTGAAGGACTCACA-3′; occludin-F, 5′-ACTGGGTCAGGGAATATCCA-3′; and occludin-R, 5′-TCAGCAGCAGCCATGTACTC-3′. Real-time PCR was conducted using a CFX Connect Real-Time PCR Detection System . Parameters for amplification were as follows: 95 °C for 30 s, followed by 40 cycles at 95 °C for 10 s and 58 °C for 30 s. Relative mRNA expression was calculated using the 2t method . Data weThe proteins from colonic tissues and RAW 264.7 cells were extracted using RIPA buffer (Beyotime) and protease inhibitor cocktail  (Beyotime). Protein was then extracted following centrifugation. A BCA kit (Beyotime) was used to determine protein concentration. Subsequently, polyacrylamide gel electrophoresis (Bio-Rad) was conducted. The resulting membranes were replaced with polyvinylidene difluoride  before being submerged for 2 h in blocking solution. Primary antibodies against ZO-1 (1:1000), occludin (1:1000), cyclo-oxygenase (COX)-2 (1:1000), inducible NO synthase (iNOS) (1:1000), ERK 1/2 (1:1000), phospho (p)-ERK 1/2 (1:2000), nuclear factor (NF)-κB p65 (1:1000), p-NF-κB p65 (1:1000), β-actin (1:1000), and GAPDH (1:1000)  solution were added to the membranes which were stored overnight at 4 °C. On the next day, the membranes were washed, and a secondary antibody was added for 1 h at room temperature. The secondary antibody was then washed away, and chemiluminescence was measured . Data were collected and analyzed from three independent samples.Mesenteric lymph nodes (MLNs) were lysed into single-cell solutions using collagenase type D  (1 mg/ml) and DNase I (Roche) (0.1 mg/ml) in Hanks’ Balanced Salt Solution for 30 min at 37 °C. The red cells in the preparation were lysed. After washing, the cells were processed for FCM (30). To identify T helper (Th)17 cells, plated cells were combined with 5 ng/ml phorbol myristate acetate along with 1 ng/ml ionomycin for 5 h. At 30 min, 10 ng/ml brefeldin A was added. Cells were washed and stained with 1 ml CD4-fluorescein isothiocyanate (FITC) per test, before being permeabilized with Cytofix or Cytoperm. Subsequently, 0.3 ml of IL-17A-phycoerythrin (PE) was added. For the detection of regulatory T (Treg) cells, a similar permeabilization protocol was used, but cells were stained with 0.3 ml forkhead box (Fox)p3-PE. Following washing, cells were then stained with CD4-FITC and CD25-APC, both at 1 ml per test. Finally, cells were rinsed using a buffer solution to remove any additional stain, and analysis was conducted in FCM buffer . Data were collected and analyzed from three independent samples.Concentrations of short-chain fatty acids (SCFAs) in mouse feces were determined using a previously reported method with minor modification . BrieflyA Shimadzu LC-20A system  was coupled with a 5500 QTRAP mass spectrometer . The LC system and MS system were connected with an ESI source. Chromatographic separation was achieved in a Thermo BDS Hypersil C18 column . The column temperature was set at 40 °C, and the mobile phases consisted of 0.1% formic acid (A) and acetonitrile (B). A 19 min gradient elution program (with flow rate set at 0.35 ml/min) was used as follows: 0–2 min, 10% B; 2–9 min, 10–48% B; 9–14 min, 48–90% B; 14–16 min, 90% B; 16–17 min, 90–10% B; 17–19 min, 10% B. Samples were analyzed in negative ion mode. Multiple reaction monitoring mode was used, and the Q1/Q3 pairs were as follows: m/z 194.2 → 137.2 for acetate, m/z 208.1 → 165.1 for propionate, m/z 222.2 → 137.1 for isobutyrate and butyrate, m/z 236.1 → 137.1 for isovalerate and valerate, and m/z 253.2 →137.1 for internal standard (IS). The collision energy was set at −21, −20, −20, −24, and −26 for acetate, propionate, isobutyrate/butyrate, isovalerate/valerate, and IS, respectively. Data were collected and analyzed from eight independent samples.To quantitate the inflammation of colon tissues and function of the intestinal barrier, IF and immunohistochemistry (IHC) were used. Tissues were first dewaxed and rinsed. Subsequently, 0.01 M antigen was added along with sodium citrate buffer (pH 6). An IHC Biotin Block Kit was used to block endogenous biotin. Tissues were incubated with various primary antibodies at 4 °C overnight. Tissue slices were stained using a DAB kit along with hematoxylin after incubation using horseradish-peroxidase-conjugated antibodies for 1 h. For IF staining, Cy-3-labeled goat anti-rabbit IgG was used, and tissues were incubated for 1 h. Subsequently, 4′,6-diamidino-2-phenylindole (DAPI) was used to stain slices at room temperature for 10 min. Confocal microscopy was used to visualize tissues. Data were collected and analyzed from three independent samples.After mice were killed at the indicated time points, FITC–dextran (FD40) (Sigma) was used to assess intestinal permeability. FD40 was first dissolved in normal saline (50 mg/ml) and gavaged with 0.1 ml/10 g blood by body weight for 4 h. Separation of plasma was achieved using centrifugation. Plasma samples (40 μL) were combined with 160 μL normal saline. The concentration of FD40 was determined with fluorospectrometry at 485 nm (excitation wavelength) and 533 nm (emission wavelength). Data were collected and analyzed from four independent samples.p < 0.05 was defined as statistically significant.All results were presented in the form of mean ± SEM from two or three independent experiments as each experiment was conducted at least twice. One-way analysis of variance and Dunnett’s test were used to evaluate statistical significance between control groups and multiple dose groups. An in-house component library of BTW and MS1 and MS/MS data was compared to the literature or standards. More information was acquired under the anion mode when compared to the cation ion mode. The total ion chromatography of the standard mixture and BTW are shown in p < 0.01) (p < 0.01)  . Histolo < 0.01) .p < 0.01)  . DSS tre < 0.01) . These r+ IL-17A+) in the MLNs of BTW- and SASP-treated mice was lower than that in the DSS-treated mice (p < 0.01) (+CD25+Foxp3+), which usually functions to suppress inflammation, was significantly higher compared with that in the DSS-treated mice (p < 0.01) (+ T cells were significantly increased by DSS but decreased significantly after BTW and SASP treatment (p < 0.01)  . At the  < 0.01) . CD4+ T  < 0.01) .p < 0.01 or p < 0.05). Expression of IL-10, an anti-inflammatory cytokine secreted by Treg cells, was significantly upregulated in the H-BTW group compared with the DSS group (p < 0.01) (p < 0.01 or p < 0.05)  . Express < 0.01) . A simil < 0.05) .p < 0.05, p < 0.01) (p < 0.01 or p < 0.05) (p < 0.05)  . We meas < 0.01) . BTW res < 0.05) . To inve < 0.05) .p < 0.01) (p < 0.01 or p < 0.05) (Previous studies have shown that NF-κB and ERK are related to downregulation of tight junction proteins . Therefo < 0.01) . Simulta < 0.05)  and 4A.p < 0.01 or p < 0.05)  .p < 0.01). The combination of U0126 and BTW showed a tendency to improve the colonic histological score compared with BTW alone (p > 0.05). ERK agonist TBHQ increased the activation of ERK compared with that in the DSS group (p < 0.05); however, it did not aggravate the symptoms of UC, compared with those in the DSS group (p > 0.05). The combined use of TBHQ and BTW significantly reduced the inhibition of ERK activity (p < 0.01) and weakened the improvement of body weight (p < 0.05 or p < 0.01).ERK inhibitor U0126 downregulated ERK activity  and showy weight , DAI  (p < 0.01) (Growth inhibition of RAW 264.7 by BTW was established by CCK-8 assay. BTW was not cytotoxic at any of the tested concentrations ( > 0.05) . Based o > 0.05) . The res < 0.01) . We usedp < 0.01) (p < 0.01) (The effect of BTW on the activation of ERK/NF-κB in LPS-induced RAW 264.7 cells was determined by western blotting. LPS (1 μM) significantly increased the expression of p-ERK and p-p65. Pretreatment of cells with 5, 10, or 50 μg/ml BTW significantly decreased the activation of ERK and p65 in a concentration-dependent manner ( < 0.01) . LPS inc < 0.01) .p < 0.01), which was similar to the effect of BTW. The combination of U0126 with BTW augmented the inhibition of the ERK/NF-κB pathway (p < 0.01)  . This suBTW is a commonly used prescription and has been widely used as an adjuvant therapy for UC. Our previous meta-analysis based on clinical evidence showed that the use of BTW can significantly improve the clinical efficiency and endoscopic remission rate of UC . AlthougTo determine the effect of BTW on UC, parameters such as clinical symptoms and histopathology were used as indicators of drug efficacy . In our + T cells in MLNs of UC mice. It is known that the activation of CD4+ T cells requires the involvement of transcription factor NF-κB, which activates immunity -mediated antioxidant response, which may partially neutralize the colonic damage caused by the activation of the ERK pathway ; Jiangsu University Advantageous Discipline Construction Project (No. ZYX03KF058); Jiangsu Province Traditional Chinese Medicine Science and Technology Development Plan Project (No. RC201903); Suzhou Science and Technology Development Plan Project (No. SYSD2017007); and Zhangjiagang Science and Technology Project (No. ZKS1839).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


In the course of these investigations, the compound [Fe(C6H8BN4)2(C12H7BrN2)] was synthesized, for which magnetic measurements revealed an incomplete spin-crossover behaviour. Crystallization of this compound from toluene led to the formation of crystals of the toluene disolvate, [Fe(C6H8N4B)2(C12H7N2Br)]·2C7H8. Its asymmetric unit comprises two discrete metal complex molecules and two toluene solvent molecules. One of the latter is severely disordered and its contribution to the diffracted intensities was removed using the SQUEEZE routine [Spek borate ligands in the form of a slightly distorted octahedron. The discrete complexes are arranged in columns along the a-axis direction with the toluene solvate molecules located between the columns. The 5-bromo-1,10-phenanthroline ligands of neighbouring columns are approximately parallel and are slightly shifted relative to each other, indicating π–π interactions.The structure determination of the title compound was undertaken as part of a project on the modification and synthesis of new spin-crossover (SCO) compounds based on octahedral FeSpek 2015. Acta Cr Such materials are also of interest because of their potential for future applications as molecular switches, in data storage or in spintronics  and the diamagnetic low-spin state  by external stimuli such as temperature or light  bond lengths and also the unit-cell volume change significantly. Therefore, cooperativity effects are of importance, which frequently lead to abrupt spin transitions, very often associated with a hysteresis or a more complicated SCO behaviour 2)2(L)] (with pz = pyrazole and L = diimine co-ligand) have become of particular interest, because they can be evaporated in vacuo and therefore allow a facile preparation of thin films  occurs in octahedrally coordinated transition-metal complexes with an electron configuration of 3L and the pyrazole ligand in iron(II) complexes with general composition [Fe(H2B(pz)2)2(L)], which leads to characteristic changes in the spin-transition behaviour in the solid state 2)2]·0.5C7H8 exhibits a short π–π intra-dimer distance (3.507 Å at 293 K and 3.483 Å at 200 K) and consequently the complexes are locked in the high-spin state, whereas for [Fe(H2B(pz)2)2] with a long π–π distance (3.753 Å at 293 K and 3.736 Å at 200 K) complete thermal SCO is observed 2)2(bipy)] an incomplete spin-crossover is observed 2)2(5-Br-phen)] . Magnetic measurements of this new complex revealed an incomplete SCO in the temperature range from 2 to 300 K with only one step during the spin transition . This compound can easily be crystallized from toluene whereby a toluene disolvate, [Fe(H2B(pz)2)2(5-Br-phen)]·2C7H8, is formed. The crystal structure of this solvate shows dimers that are linked by phenanthroline ligands with an intra-dimer distance of 3.465 (6) Å at 200 K, indicating strong intermolecular π–π interactions (see above). Therefore, the system may be locked in the HS, which would concur with the observed bond lengths at 200 K, reflecting a HS configuration, see Structural commentary. Unfortunately, we were not able to prepare large amounts of pure samples of the title compound for magnetic measurements. On the other hand, a comparison of the experimental XRPD pattern of the ansolvate with the simulated pattern of the title complex based on single-crystal data  reveals that the crystal structure of the disolvate is entirely different from that of the ansolvate. Therefore, we have no information as to whether π–π interactions are also present in the ansolvate and, if so, how strong these are. Nevertheless, from the observation of thermal spin-crossover in the latter , we can conclude that the strength of the π–π interactions must be weaker in the ansolvate than in the solvate.In our own systematic investigations we are interested how a chemical modification of such Fe organoborate complexes influences the SCO behavior in the bulk material and in thin films. This includes functionalization of the neutral diimine ligand 2B(pz)2)2] and two toluene molecules . The FeII cation of each independent complex is distorted octahedrally coordinated  of neighbouring 5-bromo-1,10-phenanthroline planes is 3.465 (6) Å, indicating strong π–π interactions.In the crystal structure, the discrete complexes are arranged into columns that extend along the on Fig. 2. Betweenon Fig. 2. The sho2B(pz)2)2(phen)] and [Fe(H2B(pz)2)2] /(S)-4,5-pinenepyridyl-2-pyrazine ligands 2)2(MeOH)2], has also been published borate and different co-ligands reported in the literature, which include  was synthesized according to previously reported procedures 2)22  in methanol (3 ml), a solution of K(H2B(pz)2)  in methanol (5 ml) was added, leading to the formation of a slightly yellow-coloured solution, which was stirred for 15 min at room temperature. A solution of 5-bromo-1,10-phenanthroline  in methanol (3 ml) was added dropwise to the reaction mixture. Immediately, the solution turned purple and a purple-coloured precipitate was formed. The solution was stirred for 1 h at room temperature and then the precipitate was filtered off, washed with methanol (7 ml), and dried under reduced pressure. Yield: 373mg .Elemental analysis calculated for C24H23B2BrFeN10: C 47.34, H 3.81, N 23.00, Br 13.12%, found C 47.11, H 3.92, N 22.97, Br 13.41%.HRESI–MS (+) (MeOH):m/z (%) = [H2B(Hpz)2]+ calculated 149.09930, found 149.09927 (33), [co-ligand + H]+ calculated 258.98654, found 258.98631 (3), [M – H2B(pz)2]+ calculated 460.99786, found 460.99758 (25), [M + H]+ calculated 609.08988, found 609.08946 (6), [M – H2B(pz)2 + co-ligand]+ calculated 720.97507, found 720.97455 (100).IR : ν/cm−1 = 3105, 3058 , 2401, 2352 , 2285 , 1599 (w), 1574 (w), 1501 (m), 1482 (w), 1421 (m), 1398 (m), 1372 (w), 1339 (w), 1292 (m), 1258 (w), 1203 (m), 1184 (m), 1154 (s), 1092 (m), 1064 (m), 1049 (s), 996 (w), 978 (m), 933 (m), 921 (w), 880 (s), 845 (w), 823 (w), 804 (m), 766 (s), 747 (s), 731 (s), 719 (m), 674 (m), 637 (s), 621 (m), 612 (m), 421 (m).Raman : ν/cm−1 = 2781 , 1447 (w), 1410 (w), 1366 (w), 1339 (w), 1312 (w), 1290 (w), 1205 (w), 1092 (w), 1051 (w), 758 (w), 719 (w).UV/Vis : λmax/nm = 212, 234, 279, 314, 425–702 (br).Crystallization: Single crystals of  between 2 and 300 K. The curves resulting from cooling and heating the sample almost coincide. DOI: Click here for additional data file.10.1107/S2056989020010361/wm5576sup4.pngFigure S2. Experimental XRPD pattern of [Fe(H2B(pz)2)2(5-Br-phen)] measuered at rt (a) as well as simulated XRPD pattern calculated from the single crystal structure of [Fe(H2B(pz)2)2(5-Br-phen)]*2 toluene (b) (200 K). DOI: 2019452CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Correction to: Neurocrit Care https://doi.org/10.1007/s12028-023-01707-3The original article has been updated to correct Row 2 of Table 4 “04-Jul” to “4–7”.


Two new single-enantiomer phosphorus structures were studied. Their geometries, conformations and NMR features are discussed. i.e. diphenyl [(R)-(+)-α-ethylbenzylamido]phosphate, C21H22NO3P or (C6H5O)2P(O)[NH-(R)-(+)CH(C2H5)(C6H5)] (I), and N-[(R)-(+)-α-ethylbenzyl]-P,P-diphenylphosphinic amide, C21H22NOP or (C6H5)2P(O)[NH-R-(+)CH(C2H5)(C6H5)] (II), were studied. The different environments at the phosphorus atoms, (O)2P(O)(N) and (C)2P(O)(N), allow the P=O/P—N bond strengths to be compared, as well as the N—H⋯O=P hydrogen-bond strengths, and P=O/N—H vibrations. The following characteristics related to diastereotopic C6H5O/C6H5 groups in I/II were considered: geometry parameters, contributions to the crystal packing, solution 13C/1H NMR chemical shifts, conformations, and NMR coupling constants. The phosphorus-carbon coupling constants nJPC (n = 2 and 3) in I and mJPC  in II were evaluated. For a comparative study, chiral analogous structures were retrieved from the Cambridge Structural Database (CSD) and their geometries and conformations are discussed.The crystal structures of two single-enantiomer compounds,  The chirality may also be essential for some particular applications, such as the manufacture of drugs and the planning of some reactions related to different reactivities of diastereotopic groups 2P(O)[NH-(+)CH(C2H5)(C6H5)] phosphoramide, (I), and (C6H5)2P(O)[NH-(+)CH(C2H5)(C6H5)] phosphinamide, (II) to study structural differences of two diastereotopic C6H5O/C6H5 groups, caused by the same chiral amine. Structure I is the enantiomer of the previously reported (C6H5O)2P(O)[NH-(–)CH(C2H5)(C6H5)] (N) for I and (C)2P(O)(N) for II, and the maximum/minimum bond angles at phosphorus are related to O=P—O/O—P—O and O=P—N/N—P—C. The differences between maximum and minimum values are about 16.8° for I and 17.5°/16.9° for the two symmetry-independent molecules of II. The P—N—C angles in I and II, for example, P1—N3—C4 angle in II of 120.91 (14)° °/121.91 (16)°, similarly show the hybridization state of the ester oxygen atoms is close to sp2.The P atoms display a distorted tetrahedral environment, (O)° Table 2, demonstI is similar to its S-enantiomer  of 0.0089 Å and a maximum deviation of 0.0153 Å.The structure I, 1.469 (2) Å, is shorter than the P=O bonds in II, 1.4846 (15)/1.4933 (15) Å, and the same is true about the P—N bonds . The differences result from the effect of electronegative oxygen atoms of two C6H5O groups in I attached to phosphorus, while in II, there are two C6H5 groups. The longer P—N bond in II is also caused by the steric effects of two phenyl groups directly attached to phosphorus. Minor differences are observed for the bond lengths related to the diastereotopic pairs. Typically, the P—O and P—C bonds in I and two independent molecules of II are 1.581 (2)/1.592 (2) Å, 1.802 (2)/1.808 (2) Å and 1.808 (2)/1.797 (2) Å.The P=O bond in I, the N—H unit adopts an antiperiplanar (–ap) orientation with respect to the P=O group (based on the O=P—N—H torsion angle of −157.18°), and in two symmetry-independent molecules of II, the same units adopt synclinal (–sc and +sc) conformations (the torsion angles are −80.63° and +84.78°). The different conformation of II (in comparison to I) results from intramolecular rotations of the chiral amine, and the two independent molecules feature different rotations, but with a similar O=P—N—H conformation.In compound II, the symmetry-independent molecules are similar concerning the bond lengths and angles °/−98.2 (2)°/60.7 (2)° correspond to +ap/−ac/+sc conformations . Similar torsion angles in the other molecule, −178.3 (2)°/−158.6 (2)°/−62.0 (2)°, define −ap/–ap/−sc conformations. The other notable difference between the two molecules is reflected in the direction of the phenyl ring of the chiral segment with respect to the P=O group (an opposite direction in the molecule P1 and the same direction in the second molecule). Fig. 3b shows the overlay of two molecules, and the root-mean-square deviation (r.m.s.d.) of the fit of them is 1.3533 Å with a maximum deviation of 4.6684 Å. The noted difference is reflected in the spatial distances of phenyl groups bonded to P and the phenyl group of chiral amine in the two molecules. The differences in diastereotopic phenyl rings in each molecule can also be described by their distances from the phenyl ring of the chiral amine.In I, the distances between the centroid of the phenyl ring of chiral amine and the centroids of two diastereotopic phenyl groups are 5.0848 (1) and 7.9514 (1) Å. For the two symmetry-independent molecules of II, equivalent distances are 5.5767 (5)/7.0325 (6) Å and 7.1614 (6)/6.4951 (3) Å. These spatial distances show that one of the diastereotopic phenyl rings is significantly closer to the phenyl of the chiral amine. The differences in these spatial distances are pronounced in I, where the flexibility is greater (because of the existence of the P—O—C segment and the possibility of rotation).For I, the conformations of phenyl rings can be introduced by the C—C—O—P torsion angles, which are 32.1 (3)°/−149.9 (2)° and 86.7 (3)°/−98.0 (3)° according to the +sc−ac conformations for both phenyl rings. In the structure of II, the C—C—P=O torsion angles were considered for checking the conformations of the phenyl rings. The values are 173.8 (2)°/−10.4 (2)° and 25.4 (2)°/−158.0 (2)° (+ap−sp and +sp−ap) in one molecule and 10.8 (2)°/−169.6 (2)° and −18.4 (2)°/163.0 (2)° (+sp−ap and −sp+ap) in the other molecule, which also show similar conformations.In 3.I and II, the molecules are assembled in a chain arrangement through N—H⋯O(P) hydrogen bonds along [100] . This weakness is the result of the lower hydrogen-bond acceptor capability expected for the phosphoryl group of an (O)2(N)P(O)-based structure, compared to the phosphoryl group of a (C)2(N)P(O)-based structure, and is due to the two atoms with a higher electronegativity bonded to the phosphorus atom. The effect of different electronegativities was previously noted (see above) for different P=O bond lengths.In the crystal structures of 0] Fig. 4 and 4 ▸.I assemble the molecules in a two-dimensional array in the ab plane. Fig. 5Cg = 2.81 Å). The magenta ring takes part in a C—H⋯π interaction with an adjacent symmetry-related magenta ring (H⋯Cg = 3.23 Å) and also in a C—H⋯OP interaction (H⋯O = 2.55 Å). The formed two-dimensional assembly is double-layered and has a thickness of 18.057 Å in the c-axis direction.C—H⋯π interactions in II, two possible C—H⋯π interactions exist (H⋯Cg distances of 3.41 and 3.49 Å), which do not change the dimensionality made by the N—H⋯O hydrogen bonds. In both C—H⋯π interactions, the H donors are chiral amines of two symmetry-independent molecules —N—C(H)(C)(C—C) fragment  were retrieved from the CSD to study possible structural differences for diastereotopic R groups; the metal complexes were not considered. The CSD  bonds, and the structures have different skeletons, (C)2P(O)(N), (C)2P(S)(N), (C)2P(N)(N), (O)2P(O)(N) and (N)2P(O)(N). The related bond lengths are given in Table S1 of the supporting information. The largest difference for the P—C bond lengths made by diastereotopic groups (0.025 Å) exceeds the largest differences for the P—O (0.017 Å) and P—N bond lengths (0.015 Å). The P—C, P—N and P—O bond lengths in these structures vary from 1.773 to 1.837 Å, 1.629 to 1.652 Å and 1.555 to 1.607 Å, respectively, with averages of 1.805, 1.643 and 1.580 Å.The chiral structures with an I and II, i.e. with the O2P and C2P skeletons. Only three O2P-based structures (with the oxygen atom attached to an arene ring) were found in the CSD. For the C2P skeleton, 36 structures, including 64 R2PX fragments, were checked, and the C—C—P=X  torsion angles were evaluated.The conformations of diastereotopic groups attached to phosphorus were analysed in the structures analogous to 2P-based structures mainly include Ph2P(O) fragment (28 structures), similar to compound II; however, structures with Ph2P(S) (seven structures), and (C6H11)2P(=N) (one structure) fragments were also found. Both similar and different conformations were observed for diastereotopic groups. Details of the analysis are given in Table S2 and Fig. S1 of the supporting information. The torsion angles such as C—C—P=O of 0.04° in the structure with refcode MEFCIK  are ±sp±ap/±sp±ap (26 entries) and ±sp±ap/±sc±ac (23 entries). In the systems with phenyl rings directly attached to the phosphorus atom, as a result of crowding, the simultaneous torsion angles around ±90° (a perpendicular conformation) for both phenyl rings were not found for any structure. In some cases, like in the structure with refcode VUGSOG  mapped with dnorm and two-dimensional fingerprint plots , H⋯C (28.8%), H⋯O (12.7%) and O⋯C (1.2%). The O⋯C contact results from the near distance of two symmetry-related phenoxy groups [O2(C3–C8)], through the ester oxygen atom and π-system.To visualize and compare the intermolecular contacts of 3I Fig. 7, the redII, the HS map was generated around two symmetry-independent molecules step by step. Besides N—H⋯O hydrogen bonds, a significant H⋯H contact develops a red area, as seen in Fig. 8For I, the structure of II shows fewer H⋯O, H⋯C and O⋯C contacts , which were compensated with remarkable H⋯H (64.2%/64.8%), and C⋯C contacts (2.5% for both). The smaller volume/Z ratio in II is reflected by the crowding, manifested in increased H⋯H contacts and the observation of C⋯C contacts.The contribution percentages of various contacts were obtained for the two symmetry-independent molecules. Compared with 6.−1 for I and 3152 cm−1 for II. The lower NH stretching wave number of II is attributed to stronger N—H⋯OP hydrogen bonds as discussed in the X-ray crystallography section. The bands at 1244 cm−1 for I and 1192 cm−1 for II are assigned to the P=O vibrations, and the higher wave number for I is in accordance with the presence of more electronegative atoms in the (O)2P(O)N skeleton [versus (C)2P(O)N for II].In the IR spectra, the N—H stretching bands are centred at 3268 cm13C NMR spectra, the doublet signals at 31.80 p.p.m. (3J = 8.1 Hz) for I and at 32.62 p.p.m. (3J = 4.7 Hz) for II correspond to the CH2 group. The dissymmetric carbon atom does not show coupling with phosphorus, and the ipso-C atom attached to it, i.e. with a three-bond separation from phosphorus, shows a doublet at 143.04 p.p.m. (3J = 3.0 Hz) in I and at 145.50 p.p.m. (3J = 4.6 Hz) in II.In the 6H5O groups in I, two sets of carbon signals are observed. For example, the doublets at 150.74/150.92 p.p.m. and 120.12/120.23 p.p.m., with 2J = 7.0 Hz for the first pair and 3J = 4.0 Hz for the second pair, are associated with the diastereotopic ipso-C atoms and diastereotopic ortho-C atoms, respectively. All carbon atoms of diastereotopic phenyl groups in compound II show couplings with phosphorus .For the two diastereotopic CJ = 1.9/2.1 Hz) are assigned to the para-carbon atoms of the phenyl rings with four bonds separation from the phosphorus atom. The doublets at 132.20/132.38 p.p.m. (J = 9.4/9.5 Hz) and at 128.62/128.82 p.p.m. (J = 12.2/12.1 Hz) are assigned to the diastereotopic ortho- and meta-carbon atoms. The doublets centred at 134.44 and 134.77 p.p.m. (J = 127.4 and 126.4 Hz) are related to the diastereotopic ipso-carbon atoms. The separation of these signals is comparable with previously investigated 1J coupling constants for analogous compounds, typically in (C6H5)2P(O)(NH-cyclo-C7H13) with 1J = 129.4 Hz .A brief discussion of 7.6H5O/2×C6H5, were discussed for two new single-enantiomer structures, (C6H5O)2P(O)[NH-(+)CH(C2H5)(C6H5)] (I), and (C6H5)2P(O)[NH-(+)CH(C2H5)(C6H5)] (II). The pronounced differences are related to the contributions in the crystal packing by diastereotopic groups, especially in the C—H⋯π interactions, and the NMR chemical shifts of corresponding 13C signals. The geometry parameters, conformations and NMR coupling constants of diastereotopic groups show minor differences (and/or similarities in some cases). In I with the O2P(O)N skeleton, the shorter P=O/P—N bonds and weaker N—H⋯O=P hydrogen bond are observed with respect to the structure II with the C2P(O)N skeleton. These structural features, resulting from different electronegativities of atoms, are reflected in the higher stretching frequencies of P=O and N—H bonds in the structure I (the latter because of a weaker N—H⋯O=P hydrogen bond). The lower volume/Z ratio of II is reflected by the crowding and observation of C⋯C contacts and raising H⋯H contacts, while I includes more H⋯O and O⋯C contacts. The study of analogous chiral structures retrieved from the CSD shows minor differences in bond lengths for diastereotopic P—C, P—O, and P—N bonds and more significant differences in torsion angles of diastereotopic groups.The differences/similarities of diastereotopic pairs, 2×C8.6H5O)2P(O)[NH-(Preparation of (CR2H5)(C6H5)], (I))-(+)CH(C To a solution of (C6H5O)2P(O)Cl in dry chloroform, a solution of R-(+)-1-phenylpropylamine and triethylamine (1:1:1 molar ratio) in the same solvent was added at 273 K. After stirring for 4 h, the solvent was removed in a vacuum, and the obtained solid was washed with distilled water to remove (C2H5)3NHCl. Colourless crystals were obtained from a solution of the title compound in CHCl3/CH3CN (1:2 v/v) after slow evaporation at room temperature.−1): 3268, 3063, 3029, 2970, 2929, 2854, 1592, 1492, 1453, 1420, 1244, 1200, 1167, 1058, 1020, 949, 900, 750, 689, 634, 579, 552, 520, 496, 457. 1H NMR : δ = 0.84 , 1.81 , 3.84 , 4.32 , 6.99 , 7.10 , 7.16 – 7.35 ; 13C{1H} NMR : δ = 10.59, 31.80 , 58.20, 120.12 , 120.23 , 124.63, 124.83, 126.50, 127.21, 128.44, 129.43, 129.64, 143.04 , 150.74 , 150.92 ; 31P{1H} NMR : δ = −2.16.Analytical data: IR 2P(O)[NH-(Preparation of (CR2H5)(C6H5)], (II))-(+)CH(C To a solution of (C6H5)2P(O)Cl in dry chloroform, a solution of R-(+)-1-phenylpropylamine and triethylamine (1:1:1 mole ratio) in the same solvent was added at 273 K. After stirring for 4 h, the solvent was removed in a vacuum, and the obtained solid was washed with distilled water to remove (C2H5)3NHCl. Colourless crystals were obtained from a solution of the title compound in CHCl3/CH3CN (1:2 v/v) after slow evaporation at room temperature.−1): 3152, 3057, 3027, 2962, 2928, 2868, 1488, 1440, 1383, 1337, 1305, 1192, 1117, 1056, 1017, 929, 904, 838, 751, 721, 695, 603, 564, 533. 1H NMR : δ = 0.79 , 1.69 , 1.82 , 3.84 , 5.91 , 7.26 , 7.37 , 7.50 , 7.64 , 7.79 ; 13C{1H} NMR : δ = 11.60, 32.62 , 57.10, 126.87, 126.97, 128.45, 128.62 , 128.82 , 131.74 , 131.86 , 132.20 , 132.38 , 134.44 , 134.77 , 145.50 ; 31P{1H} NMR : δ = 21.13.Analytical data: IR  values in the range 1.2–1.5×Ueq of the parent atom, after which the positions were refined with riding constraints  global, I, II. DOI: Click here for additional data file.10.1107/S2056989023006278/jq2028sup3.docxFigures  and tables (CSD) and a brief discussion about NMR. DOI: 2283048, 2283049CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information: