Comparing Content Of Phenylethanol Glycosides in Two Different Host Cistanches Herba After Different Drying Treatments Ⅱ
Mar 13, 2024
2.6 Methodological review
2.6.1 Precision test
Take the mixed reference solution and continuously inject and measure 6 times according to the conditions of "2.2" and "2.3". Calculate echinacoside, verbascoside, anthocynoside A, gardeniposide, and 2'-acetyl The relative standard deviation (RSD) of the peak areas of verbascoside, salidroside, motherwort, cistancheside A, and tubulin B were 0.98%, 0.68%, 0.91%, 0.49%, 0.77%, and 0.79 respectively. %, 0.84%, 0.69%, 0.54%, 0.62%, indicating that the instrument has good precision.
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2.6.2 Stability test
Aspirate the same Cistanche deserticola test solution, and inject samples for measurement at 0, 2, 4, 6, 8, 10, 12, and 24 hours according to the conditions under "2.2" and "2.3". Calculate the RSD of the peak area of the above components as follows: 0.28%, 0.43%, 0.75%, 1.14%, 0.89%, 1.07%, 1.23%, and 1.01%, indicating that the instrument has good precision.
2.6.3 Repeatability test
Take 6 portions of the same batch of test sample, weigh each portion at 1.0 g, prepare the test solution according to the method under "2.5", inject and measure according to the conditions under "2.2" and "2.3", and calculate the content of the above 10 ingredients. The RSDs are 0.41%, 0.27%, 0.49%, 0.36%, 0.57%, 0.97%, 0.24%, .61%, 0.32%, and 0.63% respectively, indicating that the method has good repeatability.
2.6.4 Sample Recovery Test
Take the test solution that has been analyzed and tested for the content of 10 phenylethanoid glycosides, and add 50% of the reference substance stock solution that is the same as the 10 phenylethanoid glycosides in the sample (the mass concentration of each component is 100 μg/L). %, 100%, 150%, according to the conditions of "2.2" and "2.3", the sample recovery rate of the above 10 components is calculated to be 100.21%~107.92%, and the RSD is 0.03%~1.09%, see Table 3. It can be seen that the accuracy of this method is good.
Table 3 Recoveries and RSDs of ten phenylethanoid glycosides
Table 6 Standardized processing of test data
3 Results and analysis
3.1 Sample variance analysis
SPSS 20.0.0 was used, specifying 5 drying methods as independent variables, and the contents of 10 phenylethanol glycosides as dependent variables, and conducted a one-factor analysis of variance (see Table 4 and Table 5). The analysis results show that the main chemical components of Cistanches deserticola and Haloxylon ammodendron are the same, but the content differences are large. In addition, except for verbascosides, different drying treatments have different effects on each phenylethanol glycoside detected. There are significant differences (P < 0.05) in the various components. Among them, among the Cistanche deserticola hosts, the sun-dried Cistanche deserticola has the highest content of gardeniposide, and the Cistanche deserticola dried at 80 ℃ has the highest content of gardeniposide and motherwort. The contents of sedum and cistanche A are the highest, and the freeze-dried cistanche has the highest contents of echinacoside, transcode A, verbascoside, isocarbastoside, transcode B and 2'-acetyl verbascoside; in Haloxylon ammo dendron Among the host Cistanche deserticola, the sun-dried Cistanche deserticola has the highest genocide content, the 80°C drying process has the highest echinacoside and cistancheaside A contents, and the freeze-drying process has the highest contents of salidroside and tuberose A., verbascoside, anthophyllite B, isorebascoside and 2'-acetyl verbascoside have the highest contents.
3.2 Sample standardization processing and analysis
To more scientifically and intuitively explain the effects of different drying methods on the contents of 10 phenylethanol glycosides in Cistanches deserticola on different hosts, the Z-score standardized processing data in SPSS 20.0.0 was used (Table 6). It can be seen from the comprehensive score of phenylethanoid glycosides content in Cistanche deserticola, the host of Chenopodium, was the highest after freeze-drying, followed by drying at 80°C. The comprehensive score of phenylethanoid glycosides in Cistanche deserticola, the host of Haloxylon ammodendron, was the highest after freeze-drying, followed by drying at 80°C. Dry processing. This also reflects that the total content of 10 phenylethanoid glycosides in the Cistanche deserticola of the two hosts was the highest after freeze-drying, followed by drying at 80°C.
4 Conclusion and discussion
This study established a method for the simultaneous analysis and detection of motherwort, salidroside, gardeniposide, echinaceaside, cistanche glycoside A, and tube flower using ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QQQ-MS). Methods for glycoside A, verbascoside, isorbascoside, anorthosite B, and 2'-acetyl verbascoside. After adding methanol, ultrasonic extraction was performed, and multiple reaction analysis was performed with a triple quadrupole mass spectrometer in electron sprays negative ion mode. This method can quickly, accurately, and conveniently analyze 10 phenylethanoid glycoside components of Cistanche deserticola in different hosts, and has high sensitivity and resolution.
After analyzing the results using different statistical methods, it can be seen that different drying treatment methods have a significant impact on the contents of 10 phenylethanol glycosides in Cistanche deserticola on different hosts. The total content of the 10 phenylethanol glycosides detected in this experiment was highest in the freeze-dried samples of the two host Cistanche deserticola, followed by the samples dried at 80 °C, but the phenylethanol content in the samples obtained by drying the two host Cistanche deserticola under the same conditions was the highest. There are also significant differences in the content of glycosides. From high to low, the total content is as follows: Freeze-dried sample of Atriplex truncatula host Cistanche deserticola > 80 ℃ drying sample of Atriplex truncatula host Cistanche deserticola > Haloxylon ammodendron host Cistanche deserticola freeze-dried sample > Haloxylon ammodendron Samples of host Cistanche deserticola were dried at 80°C. Huang Xiang[17],
Wang Limin [18], Feng Jie [19] and other studies have shown that the Haloxylon ammodendron host Cistanche deserticola contains the largest amount of active ingredients retained after freeze-drying and drying at 80°C, and the content of active ingredients in different hosts of Cistanche deserticola has significant differences, among which four The content of phenylethanol glycosides in the medicinal material of Atriplexa parasitica and Cistanche deserticola is generally higher than that of Haloxylon ammodendron. However, the components detected in the literature are concentrated in echinaceaside, verbascoside, cistancheoside A, isomurabasin, and 2' -Acetyl verbascoside, the results obtained in this experiment are consistent with the literature, and more detection of the content of phenylethanol glycosides has been added, further strengthening the accuracy and theoretical basis of the experimental conclusions.
In the "Chinese Pharmacopoeia" (2020 edition), verbascoside and echinaceaside are listed as index components of Cistanche deserticola. Based on all the analysis results, it is shown that the samples of Cistanche deserticola from the two hosts after freeze-drying and 80 ℃ drying are all the same. It has obvious advantages over other drying methods. Freeze drying takes 2 to 3 times longer than drying at 80°C. In the experiment, Cistanche deserticola was cut into 3 mm thick slices for drying, which is often the case during industrial processing in factories. The whole Cistanche deserticola is dried, so the difference in freeze-drying and drying time may be greater than the experimental time. In addition, there is a curing process before drying, which slows down the loss of medicinal ingredients by destroying enzyme activity [11]. Before freeze-drying, it is necessary to
Fresh samples are pre-cooled in a -80°C refrigerator for some time without killing or destroying enzyme activity. When Cistanche deserticola is stored in large quantities during the harvest period, the problem of low freeze-drying efficiency will greatly affect the quality of the medicinal materials. Therefore, in terms of time consumption and ingredient retention, mechanical cost and operation difficulty are not conducive to large-scale production, so drying at 80 ℃ is more economical. This experiment only provides suggestions for drying methods of two host Cistanche deserticola from the perspective of the content of phenylethanol glycosides and provides a certain theoretical reference for the processing of Cistanche deserticola medicinal materials and the quality control of related foods. Subsequent updates will be made on the aspects of pharmacology and efficacy. Deep research.
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