| Part one Qualitative analysis of the extract of Opuntia Milpa Alta based on the UHPLC-QE-MS.Objective:To identify the main components of the Opuntia Milpa Alta（cladodes peels and pulps）and the fruits（peels and juice）by using of ultra high-performance liquid chromatography-quadrupole-static field orbital trap mass spectrometry（UHPLC-QE-MS）.Methods:In this experiment,the extraction method of heated reflux was used to determine the best extraction solvent of all parts of Opuntia Milpa Alta,and the chemical composition of four parts of cladodes peels and pulps,the fruits peels and juice was systematically analyzed by high resolution and sensitive UHPLC-QE-MS technology in positive and negative ion mode.Results:The Opuntia Milpa Alta extraction solvents were water,70%ethanol and 50%ethanol,respectively,and the juice was directly applied.A total of 66 compounds were identified in the Opuntia Milpa Alta,41 in negative ion detection mode and 26 in positive ion detection mode were identified,among which rutin was detected in positive and negative ion mode,mainly organic acid and flavonoids.Piscidic acid was found in the cladode peels and pulps,the fruits peels and juice,and flavonoids were found mainly in the cladode peels and fruits peels.Conclusion:In this experiment,the chemical composition of Opuntia Milpa Alta was systematically analyzed for the first time,which provided scientific basis for its future development and utilization.Part two Quantitative analysis of the extracts from Opuntia Milpa Alta by spectrophotometryObjective:To determine the content of flavonoids,polyphenols,polysaccharides and proteins in the Opuntia Milpa AltaMethods:Flavonoids,polyphenols,polysaccharides and proteins in crude extracts of Opuntia Milpa Alta were quantitatively analyzed by spectrophotometry.Results:The contents of polysaccharides in cladodes peels and pulps,the fruits peels and juice were 37.31 mg·g-1,24.24 mg·g-1,57.15 mg·g-1,5.08mg·m L-1,protein of each extracts were141.45 mg·g-1,78.92 mg·g-1,193.09mg·g-1,7.77 mg·m L-1,and polyphenols were 23.72 mg·g-1,11.21 mg·g-1,29.70 mg·g-1,0.76 mg·m L-1,and flavonoids were 0.01 mg·g-1,0.28 mg·g-1,3.33 mg·g-1 and no flavonoids were detected in juice.Conclusion:Opuntia Milpa Alta is rich in polysaccharides and protein compounds,which indicates that it has certain nutritional value and provides a reference for future development and utilization in industry.Part three Quantitative analysis and bioactivity evaluation of two main components in Opuntia Milpa Alta based on HPLCObjective:To quantitatively analyze the two main chemical constituents in Opuntia Milpa Alta and biological activities of piscidic acid.Methods:The two compounds were isolated and identified by liquid chromatography and nuclear magnetic resonance,analyzed quantitatively by HPLC,and the antioxidant activity of piscidic acid was evaluated by scavenging DPPH free radicals and scavenging hydroxyl free radicals in vitro.The anti-sensitivity activity was evaluated by inhibiting hyaluronidase activity in vitro.Results:5-hydroxymethyl-2-furanformaldehyde（1）and piscidic acid（2）were isolated from the Opuntia Milpa Alta,and piscidic acid was isolated from the plant for the first time.Content of 5-hydroxymethyl-2-furanformaldehyde in crude extract of cladodes pulps and peels,the fruits peels were 2.04 mg·g-1,0.67 mg·g-1,0.32 mg·g-1,and piscidic acid of crude extract of cladodes pulps and peels,the fruits peels were 18.53 mg·g-1,45.71mg·g-1,29.01 mg·g-1,and no 5-hydroxymethyl-2-furanformaldehyde detected in juice,piscidic acid was 0.44 mg·m L-1.The IC50 value of scavenging DPPH free radicals of piscidic acid was 3692.04μg·m L-1;and the IC50 value of scavenging hydroxyl radical was 66.79μg·m L-1,which was stronger than that of positive control vitamin C and inhibited hyaluronidase activity was 3662.60μg·m L-1,which was slightly lower than that of positive control potassium glycyrrhizinate.Conclusion:The content of piscidic acid is abundant and has good biological activity.This paper provides a reference for the quality control of Opuntia Milpa Alta.Part four Analysis of Biological Activity of Opuntia Milpa Alta.Objective:To investigate the antioxidant activity and anti-sensitivity of the Opuntia Milpa Alta（cladodes peels and pulps）and the fruits（peels and juice）.Methods:Different elution sites of cladodes peels and pulps,the fruits peels and juice were prepared with macroporous adsorption resin.Antioxidant activity of crude extracts from leaves of Opuntia Milpa Alta were evaluated by removing 1,1-diphenyl-2-picrylhydrazyl radical and hydroxyl radical in vitro.To evaluate the anti-sensitivity of crude extracts from different parts by inhibiting hyaluronidase activity in vitro.Results:Opuntia Milpa Alta juice had the strongest ability to scavenge DPPH free radicals and hydroxyl free radicals,and its IC50 values were 25.21μL·m L-1 and 16.13μL·m L-1 respectively.The DPPH free radical scavenging effect of 30%ethanol elution was better after separation by macroporous adsorption resin,and its IC50 was 62.26μg·m L-1,the removal of hydroxyl radicals by washing and removing crude extracts from leaf epidermis was better,and its IC50was 81.86μg·m L-1.Fruit juice,crude extract of fruits peels and cladodes peels crude extract showed stronger inhibition rate of hyaluronidase than that of potassium glycyrrhizinate4.31μL·m L-1,387.26μg·m L-1,2337.17μg·m L-1,3445.08μg·m L-1respectively.Conclusion:In this paper,the antioxidant activity and anti-sensitivity of the Opuntia Milpa Alta（cladodes peels and pulps）and the fruits（peels and juice）were investigated for the first time,which provided a reference for the development of Opuntia Milpa Alta as a functional food and cosmetics raw material.Objective: To establish a HPLC method for the determination of ten kinds of plant raw materials declared in cosmeticsMethods: Adding appropriate amount of extraction solvent,dissolving sample,vortex mixing,adding some emulsion samples to Na Cl demulsification,ultrasonic extraction of more than 30 min,samples were treated with 12000 r·min-1,centrifugation for 5 min,and the supernatant was injected into the HPLC.Results: Forty-three items of cosmetics declared to contain ten kinds of plant raw materials were tested.Of the 10 different types of cosmetics claiming to contain Paeonia lactiflora Pall.,9 detected Paeonia lactiflora Pall.,1 was undetected.of the 4 different types of cosmetics claimed to contain Scutellaria baicalensis Georgi,2 were detected with S Scutellaria baicalensis Georgi and 2 were not detected.Of the 3 different types of cosmetics claimed to contain Centella asiatica（L.）Urb.,3 detected Centella asiatica（L.）Urb..Of 5 different types of cosmetics claimed to contain Sophora flavescens Ait.,2 detected Sophora flavescens Ait.and 3 did not.No Angelica sinensis（Oliv.）Diels was detected in 7 different types of cosmetics claiming to contain Angelica sinensis（Oliv.）Diels.Of the 7 different types of cosmetics that claimed to contain Selaginella tamariscina（Beauv.）Spring,3 were found to contain Selaginella tamariscina（Beauv.）Spring and 4 were not detected.Two pieces of Polygonum multiflorum Thunb.shampoo did not detect Polygonum multiflorum Thunb.Two different types of cosmetics claimed to contain Carthamus tinctorius L.did not detect the content of Carthamus tinctorius L..1 piece Pueraria lobata（Willd.）Ohwi facial mask cosmetics,did not detect Pueraria lobata（Willd.）Ohwi ingredients.Of the 3 shampoo samples of different types that claimed to contain Platycladus orientalis（L.）Franco,2 were detected and 1 was not.Conclusion: A convenient and rapid method for the determination of plant raw material components in cosmetics was established. |
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