綦江皱皮木瓜果实有机酸特征性成分鉴定与不同发育期变化规律
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摘要
【目的】鉴定重庆綦江皱皮木瓜果实有机酸特征性成分,解析果实不同发育期有机酸变化规律,为皱皮木瓜果实发育期有机酸代谢研究提供基础数据。【方法】以重庆綦江皱皮木瓜(‘大罗’木瓜)为研究对象,采用溶剂提取、甲酯化衍生与气相色谱-质谱联用仪检测,进行不同发育期果实有机酸组成及含量测定,药典委2012版中药色谱指纹图谱相似度分析软件进行共有特征性成分峰匹配,SigmaPlot 10.0进行果实发育过程总有机酸、强酸味和弱酸味成分变化规律分析,Simca-P 11.5与SPSS 20.0结合进行果实发育期共有特征性成分PCA分析与HCA聚类。【结果】经甲醇提取、甲酯化衍生、氯仿萃取和GC-MS检测,从綦江皱皮木瓜8个发育期果实中共分离出共有特征性成分41种,包括低碳羧酸10种、长链脂肪酸21种、芳香族有机酸5种、一元酚酸类3种和氨基酸2种,TIC图基线平稳,成分峰分布均匀且分离度高,分离效果好。綦江木瓜从盛花后90 d至果实完熟(160 d)总有机酸含量呈先下降再上升再下降的倒"之"字型,总有机酸与强酸味(r=0.970)、弱酸味成分(r=0.998)极显著正相关;而强酸味成分与低碳羧酸极显著正相关(r=0.999),与一元酚酸显著正相关(r=0.747);弱酸味成分与长链脂肪酸极显著正相关(r=0.999)。綦江木瓜完熟期检出的强酸味有机酸以苹果酸、乙酰丙酸、柠檬酸为主,累计相对含量占检出总强酸味成分的90%以上;苹果酸在发育期经历了含量下降、略有上升然后再下降的变化过程,呈从盛花后90—120 d逐渐下降,到130 d略有上升,然后再下降的倒"之"字型;柠檬酸与苹果酸变化规律相似,但乙酰丙酸与苹果酸截然相反,总体呈现上升趋势,到盛花后130 d增至最高点,之后略有下降(150 d降至最低),进入完熟期再次升高。相关性分析表明,果实强酸味有机酸总量与苹果酸、柠檬酸呈极显著正相关,但与乙酰丙酸、异柠檬酸和水杨酸弱负相关。弱酸味成分变化规律分析表明,在果实发育期,油酸、亚油酸、棕榈酸和10-羟基-十六烷酸等多数弱酸味成分均经历了迅速下降、缓慢上升再下降的变化过程,而十九烷酸却呈缓慢上升,然后下降再上升的相反过程;相关性分析表明,以油酸、亚油酸等多数弱酸味有机酸与总弱酸味有机酸极显著正相关,与十九烷酸和硬脂酸弱负相关。Simca-P主成分分析表明PC1和PC2分别解释了总变量40.00%和23.20%,样品主成分得分图显示,S1和S2聚为一类,α-酮戊二酸、苹果酸、奎尼酸、莽草酸、棕榈酸、亚油酸对样品此类有机酸组成起决定性作用;S3、S4、S5聚在一起,油酸、10-羟基-十六烷酸对这3个发育期样品起决定性作用;S6和S7聚在一起,丙二酸、乙酰丙酸、异柠檬酸、水杨酸是其主要贡献性成分;S8单独存在,琥珀酸、十九烷酸、二十四烷酸是其主要贡献性成分;该结果与基于SPSS平方欧氏距离的离差平方和聚类分析结果基本一致。【结论】綦江皱皮木瓜属苹果酸型水果,果实发育过程中有机酸的积累模式由盛花后90 d的苹果酸-柠檬酸积累型向完熟期(160 d)乙酰丙酸-苹果酸-柠檬酸积累型转变,酸积累模式的转变在重庆皱皮木瓜果实酸度与风味品质决定中具有重要作用。
Objective】The study was carried out to identify characteristic organic acids and disclosing their dynamic changing law of Chaenomeles speciosa fruit from Qijiang,Chongqing during its fruit developing period,so as to provide the basic data for the C.speciosa fruit organic acids metabolic research.【Method】By methanol extracting,methyl derivating and GC-MS detecting,the organic acids composition and their content of 8 developing period C.speciosa cv.Daluo fruit samples of Qijiang was measured.The 2012 chromatographic similarity analysis software of Chinese Pharmacopoeia Commission was used to match their common constituent peaks.The dynamic law of the total organic acids,strong-sour and weak-sour organic acids were showed in SigmaPlots10.0 software and the PCA and HCA of all the samples was processed separately by Simca-P 11.5 and SPSS 20.0softwares.【Result】The total 41 common characteristic constituents,including 10 short-chain carboxylic acids,21 long-chain fatty acids,5 aromatic-organic acids,3 monobasic-phenol acids and 2 amino-acids,were successfully matched among 8 TICs of different period fruit samples C.speciosa cv.Daluo from Qijiang.The baseline of all the TICs were smooth and stable,the organic acid derivatives peaks'distribution were well-distributed,and the separating-degree of samples’peaks was high.All target components were well separated.The total organic acids content of Qijiang C.speciosa cv.Daluo fruit from 90~(th) day after its full-bloom stage to160~(th) days showed the changing on reversed‘Z’that including the sharply decreasing,slowly increasing and then decreasing again.The total organic acids content was significant positive correlated with the strong sour(r=0.970)and the weak sour taste organic acids(r=0.998).At the same time,the strong sour taste organic acids were significant positive correlated with short-chain carboxylic acids(r=0.999)and positive correlated with monobasic phenol acids(r=0.747).The weak sour taste organic acids were significant positive correlated with long chain fatty acids(r=0.999).The malic acid,laevulic acid and citric acid,whose relative content sum was more than 90%,were the key ingredients of strong sour organic acids.The malic acid content had experienced a reverse‘Z’trend that decreased firstly(90~(th )d-120~(th )d),increased slightly(120~(th )d-130~(th )d)and decreased finally(130~(th )d-160~(th )d).The citric acid had a similar process,but the levulic acid had an obviously reverse process of changing with malic acid.Actually the levulic acid showed the increasing progress gradually.The correlation analysis showed that the strong sour taste organic acids had significant positive correlation with malate and citrate,but weakly negative correlation with levulic acid,isocitric acid,and salicylic acid.The analyzing result showed that most of the weak sour organic acids including 9-octadecenoic acid,9,12-octadecadienoic acid,hexadecanoic acid and 10-hydroxy-hexadecanoic acid experienced the similar accumulating progress which was rapidly decreasing firstly,then slowly increased and decreased in the final stage.However,nonadecanoic acid showed an opposite process that was gradually increasing.Correlation analysis showed that the total weak sour organic acids were positively related to most of them such as 9-Octadecenoic acid and 9,12-Octadecadienoic acid but negative to nonadecanoic acid.The PCA result by Simca-p 11.5 disclosed that PC1 and PC2separately distributed 40.00%and 23.20%of the total variance contribution rate.The main component score of each sample showed that:S1 and S2 clustered together which decided byα-Ketoglutaric acid,malic acid,quinic acid,shikimic acid,hexadecanoic acid and linoleic acid.The S3,S4 and S5 got into one branch that mainly because of oleinic acid and 10-hydroxy-Hexadecanoic acid.The S6 was closer to S7 largely because of malonic acid,levulic acid,isocitric acid,salicyluric acid.The S8 formed a unit alone that mostly decided by succinic acid,nonadecanoic acid,tetracosanoic acid.This result was very similar to the other one that clustered by Ward's method with Squared Euclidean Distance in SPSS.【Conclusion】C.speciosa cv.Daluo in Qijiang,Chongqing is a typical malate-accumulating fruit.During its developing period of C.speciosa fruit,the accumulating pattern of organic acids changed from Malate-Citrate accumulating type on 90~(th) days to Levulinic acid-Malic acid-Citric acid accumulating type on 160~(th) days after its flower-blooming.The change of acid accumulation pattern played a key role in the determination of acidity and flavor quality of C.speciosa fruit in Chongqing.
Objective】The study was carried out to identify characteristic organic acids and disclosing their dynamic changing law of Chaenomeles speciosa fruit from Qijiang,Chongqing during its fruit developing period,so as to provide the basic data for the C.speciosa fruit organic acids metabolic research.【Method】By methanol extracting,methyl derivating and GC-MS detecting,the organic acids composition and their content of 8 developing period C.speciosa cv.Daluo fruit samples of Qijiang was measured.The 2012 chromatographic similarity analysis software of Chinese Pharmacopoeia Commission was used to match their common constituent peaks.The dynamic law of the total organic acids,strong-sour and weak-sour organic acids were showed in SigmaPlots10.0 software and the PCA and HCA of all the samples was processed separately by Simca-P 11.5 and SPSS 20.0softwares.【Result】The total 41 common characteristic constituents,including 10 short-chain carboxylic acids,21 long-chain fatty acids,5 aromatic-organic acids,3 monobasic-phenol acids and 2 amino-acids,were successfully matched among 8 TICs of different period fruit samples C.speciosa cv.Daluo from Qijiang.The baseline of all the TICs were smooth and stable,the organic acid derivatives peaks'distribution were well-distributed,and the separating-degree of samples’peaks was high.All target components were well separated.The total organic acids content of Qijiang C.speciosa cv.Daluo fruit from 90~(th) day after its full-bloom stage to160~(th) days showed the changing on reversed‘Z’that including the sharply decreasing,slowly increasing and then decreasing again.The total organic acids content was significant positive correlated with the strong sour(r=0.970)and the weak sour taste organic acids(r=0.998).At the same time,the strong sour taste organic acids were significant positive correlated with short-chain carboxylic acids(r=0.999)and positive correlated with monobasic phenol acids(r=0.747).The weak sour taste organic acids were significant positive correlated with long chain fatty acids(r=0.999).The malic acid,laevulic acid and citric acid,whose relative content sum was more than 90%,were the key ingredients of strong sour organic acids.The malic acid content had experienced a reverse‘Z’trend that decreased firstly(90~(th )d-120~(th )d),increased slightly(120~(th )d-130~(th )d)and decreased finally(130~(th )d-160~(th )d).The citric acid had a similar process,but the levulic acid had an obviously reverse process of changing with malic acid.Actually the levulic acid showed the increasing progress gradually.The correlation analysis showed that the strong sour taste organic acids had significant positive correlation with malate and citrate,but weakly negative correlation with levulic acid,isocitric acid,and salicylic acid.The analyzing result showed that most of the weak sour organic acids including 9-octadecenoic acid,9,12-octadecadienoic acid,hexadecanoic acid and 10-hydroxy-hexadecanoic acid experienced the similar accumulating progress which was rapidly decreasing firstly,then slowly increased and decreased in the final stage.However,nonadecanoic acid showed an opposite process that was gradually increasing.Correlation analysis showed that the total weak sour organic acids were positively related to most of them such as 9-Octadecenoic acid and 9,12-Octadecadienoic acid but negative to nonadecanoic acid.The PCA result by Simca-p 11.5 disclosed that PC1 and PC2separately distributed 40.00%and 23.20%of the total variance contribution rate.The main component score of each sample showed that:S1 and S2 clustered together which decided byα-Ketoglutaric acid,malic acid,quinic acid,shikimic acid,hexadecanoic acid and linoleic acid.The S3,S4 and S5 got into one branch that mainly because of oleinic acid and 10-hydroxy-Hexadecanoic acid.The S6 was closer to S7 largely because of malonic acid,levulic acid,isocitric acid,salicyluric acid.The S8 formed a unit alone that mostly decided by succinic acid,nonadecanoic acid,tetracosanoic acid.This result was very similar to the other one that clustered by Ward's method with Squared Euclidean Distance in SPSS.【Conclusion】C.speciosa cv.Daluo in Qijiang,Chongqing is a typical malate-accumulating fruit.During its developing period of C.speciosa fruit,the accumulating pattern of organic acids changed from Malate-Citrate accumulating type on 90~(th) days to Levulinic acid-Malic acid-Citric acid accumulating type on 160~(th) days after its flower-blooming.The change of acid accumulation pattern played a key role in the determination of acidity and flavor quality of C.speciosa fruit in Chongqing.
引文
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