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Objective: To develop the EST-SSR molecular identification system of Lilium lancifolium, Lilium davidii var. willmottiae, Lilium regale, Lilium casa blanca and Lilium brownie var. viridulum, and analyze the development efficiency and identification ability of EST-SSR molecular marker technology for Lilium genus. Methods: The MISA.pl program was used to identify the SSR locus of the EST gene sequence published by NCBI. The EST-SSR primers were generated by Primer3 program module, and the primers were screened by PCR amplification and agarose gel electrophoresis. The primary screening primers were verified by polyacrylamide gel electrophoresis, and the characteristic bands of different germplasms were labeled and analyzed to construct an identification system. Results: A total of 199 pairs of SSR primers were designed. After screening 26 pairs of primers, two pairs of highly efficient primers were obtained. The molecular identification system constructed by primer JZ391 can effectively identify the mixed commercial materials, which had certain practical value. Conclusion: Based on the extreme genetic characteristics of the research materials, this identification marker development is very efficient. The results confirm that the genetic basis of the species is an important factor affecting the development efficiency of its EST-SSR molecular marker. At the same time, this case can be used as a reference for the development of EST-SSR markers for other Chinese medicinal herbs similar to Lilium genus.
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Objective To analyze and discuss the content determination of As and Hg from Lilium Lancifolium in Longshan County; To optimize microwave digestion conditions. Methods Automatic microwave digestion appratus was used. Lilium Lancifolium samples from Longshan County were digested in teflon microwave tube with HNO3-H2O2, and As and Hg were measured with atomic fluorescence spectrometry (AFS). Results The content of As was in the range of 0.031–0.507 mg/kg, and the highest content of Hg was 0.024 mg/kg. The regression equation was Y=221.23X+170.72(r=0.995 9),Y=503.52X-682.43,(r=0.999 2).For the production base,the recoveries of As and Hg were 94.32% and 92.48% in the samples, and RSD were 2.14% and 2.70%; for the breeding base, the recoveries of As and Hg were 94.95% and 93.52% in the samples, and RSD were 1.15% and 1.97%. Conclusion The method is simple and reliable, which can be used to the content determination of As and Hg from Lilium Lancifolium, and provide references for the choice of base of production and breeding of Lilium Lancifolium in Longshan County.
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This study established a rapid UPLC-TQ-MS/MS method for determination of eight active ingredients in Lilium lancifolium. The contents range of regaloside E, F, C and B are as follows: 0.604 0×10⁻¹-18.62×10⁻¹, 0.680 0×10⁻²-44.75×10⁻², 0.700 0×10⁻³-29.65×10⁻¹, 0.170 0×10⁻¹-4.724 mg•g⁻¹; the contents of chlorogenic acid, caffeic acid, protocatechualdehyde and ferulic acid, within the range of 6.827×10⁻³-16.07×10⁻³, 0.011 1×10⁻³-79.71×10⁻³, 0.593 7×10⁻³-2.962×10⁻³, 2.606×10⁻²-45.89×10⁻² mg•g⁻¹, respectively. According to PCA (principal components analysis) plotting, 35 batches can be divided into two categories, namely Anhui Huoshan and Hunan Longshan. The main different elements between these two categories are caffeic acid and ferulic acid according to the VIP (variable importance in the projection) points figure. Based on comprehensive principal component values, there are eight batches of L. lancifolium from Huoshan among the comprehensive ranking of ten. The UPLC-TQ-MS method for simultaneous analysis of eight active ingredients is accurate, efficient and convenient. This result can provide scientific basis for quality control of L. lancifolium.
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Objective: To study the steroidal saponins and phenylic constituents in the bulbs of Lilium lancifolium and their anti-oxidant activities in vitro. Methods: The constituents were isolated and purified by chromatographic technique and recrystallization, and their structures were identified by spectral methods together with physiochemical analyses. The anti-oxidant effects of these constituents on DPPH and ABTS were screened in vitro. Results: Eleven constituents were isolated including seven first-found ones (1-5, 9, and 10) and four known compounds in this plant. They were cis-1-O-p-coumaroylglycerol (1), trans-1-O-p-coumaroylglycerol (2), caffeoyl glyceride (3), 3,4- dihydroxybenzaldehyde (4), salicylic acid (5), (25R,26R)-26-methoxyspirostan-5- ene-3β-O-α-L-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl- (1→6)]-β-D-glucopyranoside (6), (25R,26R)-3β, 17α-dihydroxy-26-methoxyspirostan-5-ene-3β-0-a-Z.-rhamnopyranosyl-(l- >2)-[β-D-glucopyranosyl-(l-"6)]-β-D-glucopyranoside (7), diosgenin 3-O-{O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-xylopyranosyl- (1→3)]-β-D-glucopyranoside} (8), (25R)-3β,17α-dihydroxy- 5α-spirostan-6-one-3-O-α-L-rhamnopyranosyl-(1→2) -β-D-glucopyranoside (9), (25R)-3β-hydroxy-5α-spirostan-6-one-3- O-α-L-rhamnopyranosyl-0 (1→2)-β-D-glucopyranoside (10), and (25R)-spirost-5-ene-3β-O-α-L-rhamnopyranosyl-(1→2) -[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside (11). Compounds 1-5 showed the favorable scavenging effects on DPPH and ABTS. Conclusion: The study suggests that the phenylic constituents from the bulbs of L. lancifolium have the anti-oxidant effects.