Analyses of Physical and Chemical Compositions of Different Medicinal Specifications of CRPV by Use of Multiple Instrumental Techniques Combined with Multivariate Statistical Analysis

Citri Reticulatae Pericarpium Viride (CRPV) is the processed product of Citrus reticulata Blanco. We systematically analyzed two CRPV types, Geqingpi (GQP) and Sihuaqingpi (SHQP), based on powder color, microscopic characteristics, and chemical composition. In addition, we characterized their constituents via ultra-high-performance liquid chromatography with hybrid quadrupole-orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap-MS). Both showed significant differences in their powder color and microscopic characteristics. Fourier-transform infrared (FT-IR) spectroscopic analysis results showed that the C=O peak absorption of carboxylic acids and their carbonyl esters in SHQP was higher than that of GQP, while the C-OH and C-H plane bending peaks of polysaccharides were lower than those of GQP. We analyzed these data via similarity analysis, PCA, and OPLS-DA. GQP and SHQP had large distinct differences. Based on the mass measurements for molecular and characteristic fragment ions, we identified 44 main constituents from CRPV, including different flavonoid glycosides and flavonoid aglycones in SHQP and GQP, respectively. We found luteolin-6-C-glucoside, orientin, rhoifolin, and pilloin solely in SHQP, and naringenin and hesperetin only in GQP. The peak area measurements showed GQP having a higher flavonoid glycoside (narirutin, hesperidin, etc.) content, whereas SHQP had a higher polymethoxyflavone (nobiletin, tangeretin, etc.) content. Since we holistically analyzed two CRPV types, the results can not only support future pharmacological research, but also provide a scientific basis for formulating more reasonable CRPV quality standards and guide its clinical potential as a precision medicine.


Introduction
CRPV is a traditional Chinese medicine (TCM) commonly used in China. According to the "Pharmacopoeia of the People's Republic of China", CRPV is the processed product of the peel of the dried young or immature fruit of Citrus reticulata Blanco and its cultivars. The earliest records of its medicinal application can be traced back to the Tang Dynasty in China [1]. As per the TCM theory, CRPV soothes the liver and breaks Qi, eliminates accumulation, and resolves stagnation [2]. According to its harvesting time, CRPV can be divided into two types: GQP and SHQP. GQP is the young citrus fruit and is often

Powder Color Analysis
Upon analyzing the pictures of both GQP and SHQP powder, we found the GQP powder color was darker than that of SHQP ( Figure 1). Based on the L*, a*, and b* values of 22 samples shown in Table 1, we can see that SHQP had a higher brightness L* value than GQP, thereby indicating that the SHQP powder color was brighter. Although the red-green degree value a*: GQP > SHQP > 0, indicated that both their powder colors were red, the SHQP color was redder. Furthermore, the yellow-blue degree value b*: SHQP > GQP > 0, indicated that both their powder colors were yellow, but the color of the SHQP was yellower.
Molecules 2022, 27, x FOR PEER REVIEW

Powder Color Analysis
Upon analyzing the pictures of both GQP and SHQP powder, we foun powder color was darker than that of SHQP ( Figure 1). Based on the L*, a*, an of 22 samples shown in Table 1, we can see that SHQP had a higher brightne than GQP, thereby indicating that the SHQP powder color was brighter. Al red-green degree value a*: GQP > SHQP > 0, indicated that both their powder red, the SHQP color was redder. Furthermore, the yellow-blue degree value GQP > 0, indicated that both their powder colors were yellow, but the color o was yellower.  Table 1. The value of L*, a*, and b* of GQP and SHQP.

Microscopic Cell Analysis
According to the microscopic observation, the color of the microscopic c was darker, while that of SHQP was lighter, thus being consistent with the po results. According to the Chinese Pharmacopoeia 2020 edition, while the GQP comprise calcium oxalate crystallization, epidermal cells of the pulp capsule, a idin crystal (a), those of SHQP mainly comprise tracheal, calcium oxalate cry exocarp, mesocarp parenchyma, hesperidin crystal, and stoma (b). The result Figure 2. However, while observing, we found that the characteristic cells of existed in individual GQP batches.  Table 1. The value of L*, a*, and b* of GQP and SHQP.

Microscopic Cell Analysis
According to the microscopic observation, the color of the microscopic cells of GQP was darker, while that of SHQP was lighter, thus being consistent with the powder color results. According to the Chinese Pharmacopoeia 2020 edition, while the GQP microcells comprise calcium oxalate crystallization, epidermal cells of the pulp capsule, and hesperidin crystal (a), those of SHQP mainly comprise tracheal, calcium oxalate crystallization, exocarp, mesocarp parenchyma, hesperidin crystal, and stoma (b). The result is shown in Figure 2. However, while observing, we found that the characteristic cells of SHQP also existed in individual GQP batches.

FT-IR Analysis
The FT-IR spectrum of CRPV is shown in Figure 3, and the location of the most relevant features of the CRPV are shown in Table 2. We divided the infrared spectral absorption peaks of GQP and SHQP into five sections: the first was 3450-3350 cm −1 ; the second was 3000-2800 cm −1 ; the third was 1800-1350 cm −1 ; the fourth was 1300-1000 cm −1 ; and the fifth section was 900-600 cm −1 . There were no significant differences between GQP and SHQP in the first and second sections. Affected by volatile oils, GQP and SHQP showed characteristic peaks near 3419 cm −1 , which were broad and strong, resulting from the hydrogen bonding of free O-H. Due to flavonoids, GQP and SHQP showed characteristic peaks near 2923 cm −1 , generated by the asymmetric stretching vibration of methylene C-H. There was a significant difference between GQP and SHQP in the third sections, affected by carboxylic acids and their esters. GQP and SHQP showed characteristic peaks near 1746 cm −1 , which were carbonyl C=O stretching vibration peaks, with the absorption peak of SHQP in this band being more noticeable than that of GQP, leading to speculation that the content of carboxylic acids and its esters in SHQP was higher than that in GQP. GQP and SHQP showed characteristic peaks near 1639 cm −1 , which were C=C or aromatic ring skeleton vibration superimposed peaks. However, GQP showed two absorption peaks at 1646 cm −1 and 1609 cm −1 , due to asymmetric stretching vibrations. In the fourth and fifth sections, the absorption peaks of GQP were more noticeable than those of SHQP, with the fourth section being 1300-1000 cm −1 , due mostly to C-O stretching vibration peaks, while the C-OH stretching vibration peaks of polysaccharides were mostly around 1070 cm −1 . The fifth segment peak of 900-600 cm −1 was mostly the vibration absorption peak of carbohydrates, affected by polysaccharides. GQP and SHQP showed characteristic peaks around 800 cm −1 , which were C-H plane bending vibration absorption peaks. We speculated that GQP had a higher polysaccharide content than SHQP.

FT-IR Analysis
The FT-IR spectrum of CRPV is shown in Figure 3, and the location of the most relevant features of the CRPV are shown in Table 2. We divided the infrared spectral absorption peaks of GQP and SHQP into five sections: the first was 3450-3350 cm −1 ; the second was 3000-2800 cm −1 ; the third was 1800-1350 cm −1 ; the fourth was 1300-1000 cm −1 ; and the fifth section was 900-600 cm −1 . There were no significant differences between GQP and SHQP in the first and second sections. Affected by volatile oils, GQP and SHQP showed characteristic peaks near 3419 cm −1 , which were broad and strong, resulting from the hydrogen bonding of free O-H. Due to flavonoids, GQP and SHQP showed characteristic peaks near 2923 cm −1 , generated by the asymmetric stretching vibration of methylene C-H. There was a significant difference between GQP and SHQP in the third sections, affected by carboxylic acids and their esters. GQP and SHQP showed characteristic peaks near 1746 cm −1 , which were carbonyl C=O stretching vibration peaks, with the absorption peak of SHQP in this band being more noticeable than that of GQP, leading to speculation that the content of carboxylic acids and its esters in SHQP was higher than that in GQP. GQP and SHQP showed characteristic peaks near 1639 cm −1 , which were C=C or aromatic ring skeleton vibration superimposed peaks. However, GQP showed two absorption peaks at 1646 cm −1 and 1609 cm −1 , due to asymmetric stretching vibrations. In the fourth and fifth sections, the absorption peaks of GQP were more noticeable than those of SHQP, with the fourth section being 1300-1000 cm −1 , due mostly to C-O stretching vibration peaks, while the C-OH stretching vibration peaks of polysaccharides were mostly around 1070 cm −1 . The fifth segment peak of 900-600 cm −1 was mostly the vibration absorption peak of carbohydrates, affected by polysaccharides. GQP and SHQP showed characteristic peaks around 800 cm −1 , which were C-H plane bending vibration absorption peaks. We speculated that GQP had a higher polysaccharide content than SHQP.

Qualitative Analysis of Constituents
We analyzed the CRPV samples using UHPLC-Q-Exactive Orbitrap-MS in both positive and negative ion modes ( Figure 4). Considering the chromatographic peaks, the MS spectra obtained in positive ion mode were better than those obtained in negative ion mode. Most of the PMFs showed abundant peaks in the positive ion mode rather than in negative ion mode. Details of the identified compounds are presented in Table 3. Finally, 44 major components were identified or preliminarily identified, including 5 flavone-C-glycosides, 4 flavone-O-glycosides, 4 flavanone-O-glycosides, 4 flavanone aglycones, 21 PMFs, 3 alkaloids, 2 limonoids, and 1 other compound (Table 3). Chemical structures of the major constituents are shown in Figures 5-7 and Tables 4 and 5. Among them, luteolin-6-C-glucoside, orientin, rhoifolin, and pilloin were unique to SHQP, most of which were flavonoid glycosides. However, flavanone aglycones like naringenin and hesperetin were unique to GQP.

Qualitative Analysis of Constituents
We analyzed the CRPV samples using UHPLC-Q-Exactive Orbitrap-MS in both positive and negative ion modes ( Figure 4). Considering the chromatographic peaks, the MS spectra obtained in positive ion mode were better than those obtained in negative ion mode. Most of the PMFs showed abundant peaks in the positive ion mode rather than in negative ion mode. Details of the identified compounds are presented in Table 3. Finally, 44 major components were identified or preliminarily identified, including 5 flavone-Cglycosides, 4 flavone-O-glycosides, 4 flavanone-O-glycosides, 4 flavanone aglycones, 21 PMFs, 3 alkaloids, 2 limonoids, and 1 other compound (Table 3). Chemical structures of the major constituents are shown in Figures 5-7 and Tables 4 and 5. Among them, luteolin-6-C-glucoside, orientin, rhoifolin, and pilloin were unique to SHQP, most of which were flavonoid glycosides. However, flavanone aglycones like naringenin and hesperetin were unique to GQP. (a)

Analysis of Flavonoid-C-Glycosides
Fragmentations in most flavonoid-C-glycosides were based on the aglycones of flavones [38]. In the MS/MS spectra, the main product ion of flavone-C-glycosides usually occurred in the glycosyl moiety, which was generated by the loss of water molecule(s) and the glycosidic methylol group as formaldehyde [39]. According to previous reports [34][35][36][37], the product ions [M + H-n18Da] + are diagnostic ions, while the other two fragments,

Analysis of PMFs
PMFs are almost exclusively found in citrus species [40]. They have the same aglycon structure; the difference lies in the number and position of the hydroxyl groups (-OH) and/or methoxy groups (-OCH3) connected to the A, B, and C rings of the aglycon.

Statistical Analysis
We applied different statistical analyses like the similarity analysis, PCA analysis, and OPLS-DA analysis, based on the FT-IR information. Similarity analysis between samples was calculated as Euclidean distance, and the final result is shown in Table 6. The larger the value, the higher the similarity between the samples, while the smaller the value, the lower the similarity between the samples. The similarity value between the GQP varieties is 1.00, and the similarity between the SHQP varieties is also 1.00. However, the similarity value between GQP and SHQP is 0.65~0.66. This means that there are large differences between GQP and SHQP.
We established a PCA model and an unsupervised pattern recognition technique with R 2 (X) (cum) = 0.748 and Q 2 (cum) = 0.588. The final PCA score scatter plot and bi-plot are shown in Figure 8a,b. As shown in Figure 8a, we separated the samples into distinct groups. GQP and SHQP were clustered together; as well as being visible to the naked eye in Figure 8b, this feature indicates that samples fell into two classes; different infrared peaks had different effects between GQP and SHQP. The infrared peaks numbered 7, 16, 2, 8, and 3 had a greater impact on the clustering of SHQP, while the rest had a greater impact on the clustering of GQP. A clear separation between GQP and SHQP was achieved, indicating the significant differences between these two species.  Sample  Number  G1  G2 G3  G4  G5 G6  G7  G8  G9 G10 G11 G12 S13 S14 S15 S16 S17 S18 S19 S20 S21 S22 G1 1.00 G2 1.00 1.00 G3 1.00 1.00 1.00 G4 1.00 1.00 1.00 1.00 G5 1.00 1.00 1.00 1.00 1.00 G6 1.00 1.00 1.00 1.00 1.00 1.00 G7 1.00 1.00 1.00 1.00 1.00 1.00 1.00 G8 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 G9 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 G10 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 G11 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 G12 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 S13 0 We established the OPLS-DA model by setting GQP and SHQP as groups I and II, respectively. The OPLS-DA scores plot was established with the model parameters of R 2 (Y) = 0.999 and Q 2 (cum) = 0.997 and is shown in Figure 8c. Therefore, we confirmed the OPLS-DA model based on the FT-IR data used to separate GQP and SHQP. Since the variable importance in the projection (VIP) value reflected the contribution of each variable to the grouping, we used VIP ≥ 1 as the threshold to filter and obtain eleven different infrared peaks related to species classification (Figure 8d). The VIP values of the infrared peaks numbered 13,12,11,21,15,9,17,14,3, and 5 were > 1, and all these were important for distinguishing between GQP and SHQP. The permutated R 2 and Q 2 values on the left were lower than the original point on the right (Figure 8e), thus indicating that the established OPLS-DA mode has high goodness of fit and predictability. We established a PCA model and an unsupervised pattern recognition technique with R 2 (X) (cum) = 0.748 and Q 2 (cum) = 0.588. The final PCA score scatter plot and biplot are shown in Figure 8a,b. As shown in Figure 8a, we separated the samples into distinct groups. GQP and SHQP were clustered together; as well as being visible to the naked eye in Figure 8b, this feature indicates that samples fell into two classes; different infrared peaks had different effects between GQP and SHQP. The infrared peaks numbered 7, 16, 2, 8, and 3 had a greater impact on the clustering of SHQP, while the rest had a greater impact on the clustering of GQP. A clear separation between GQP and SHQP was achieved, indicating the significant differences between these two species.
We established the OPLS-DA model by setting GQP and SHQP as groups I and II, respectively. The OPLS-DA scores plot was established with the model parameters of R 2 (Y) = 0.999 and Q 2 (cum) = 0.997 and is shown in Figure 8c. Therefore, we confirmed the OPLS-DA model based on the FT-IR data used to separate GQP and SHQP. Since the variable importance in the projection (VIP) value reflected the contribution of each variable to the grouping, we used VIP ≥ 1 as the threshold to filter and obtain eleven different infrared peaks related to species classification (Figure 8d). The VIP values of the infrared peaks numbered 13,12,11,21,15,9,17,14,3, and 5 were > 1, and all these were important for distinguishing between GQP and SHQP. The permutated R 2 and Q 2 values on the left were lower than the original point on the right (Figure 8e), thus indicating that the established OPLS-DA mode has high goodness of fit and predictability.
Twenty-two batches of CRPV samples, comprising the GQP and SHQP, from different provinces of China, were purchased from various pharmacies and markets (Figure 9,  Twenty-two batches of CRPV samples, comprising the GQP and SHQP, from different provinces of China, were purchased from various pharmacies and markets (Figure 9, Supporting Information Table S1). They were authenticated by Professor Feng Li (Shandong University of Traditional Chinese Medicine). According to morphological characteristics, all the samples were dried fruit or immature fruit skins of Citrus reticulata Blanco and its cultivars. G1-G12 were GQP, and these types of GQP were round thick slices, the surface was grayish-green or black-green, cut surface was yellow-white or light yellow-brown, densely living with most oil chambers, gas fragrance, bitter taste, and spicy. S13-S22 were SHQP, and these types of SHQP were irregular filamentous, surface grayish-green or black-green, cut surface was yellow-white or light yellow-brown, fragrance, bitter taste, and spicy.

Chemicals and Materials
teristics, all the samples were dried fruit or immature fruit skins of Citrus reticulata Blanco and its cultivars. G1-G12 were GQP, and these types of GQP were round thick slices, the surface was grayish-green or black-green, cut surface was yellow-white or light yellowbrown, densely living with most oil chambers, gas fragrance, bitter taste, and spicy. S13-S22 were SHQP, and these types of SHQP were irregular filamentous, surface grayishgreen or black-green, cut surface was yellow-white or light yellow-brown, fragrance, bitter taste, and spicy.

Sample Preparation
Appropriate amounts of the reference standards were dissolved in methanol. The reference solutions were stored at 4 °C before use. All samples were ground into a thin powder through an 80-mesh sieve.

Sample Preparation for Microscopic Observation
A small amount of homogenized sample powder was accurately weighed, and an appropriate amount of chloral hydrate test solution (chloral hydrate:distilled water:glycerin 50:15:10) was added to it. The mixture was heated and permeated, then covered with the cover glass to make microscopic observations.

Sample Preparation for FT-IR
An appropriate amount of homogenized sample powder and potassium bromide powder were accurately weighed and dried separately in a constant temperature oven at 65 °C and 115 °C for 1.5 h, privately. Sample powder and potassium bromide (1:100) were mixed and ground under an infrared baking lamp, then pressed into flakes by a powder tablet press (Graseby Specac) for FT-IR analysis.

Sample Preparation for UHPLC-Q-Exactive Orbitrap-MS
An appropriate amount of homogenized sample powder (0.2 g) was accurately weighed and ultrasonically extracted with 25 mL methanol/water (50:50, v/v) for 45 min at room temperature. The suspension was centrifuged at 5000 rpm for 5 min to remove residue. Then, the solution was filtered through a 0.22 μm filter for analysis.

Sample Preparation
Appropriate amounts of the reference standards were dissolved in methanol. The reference solutions were stored at 4 • C before use. All samples were ground into a thin powder through an 80-mesh sieve.

Sample Preparation for Microscopic Observation
A small amount of homogenized sample powder was accurately weighed, and an appropriate amount of chloral hydrate test solution (chloral hydrate:distilled water:glycerin 50:15:10) was added to it. The mixture was heated and permeated, then covered with the cover glass to make microscopic observations.

Sample Preparation for FT-IR
An appropriate amount of homogenized sample powder and potassium bromide powder were accurately weighed and dried separately in a constant temperature oven at 65 • C and 115 • C for 1.5 h, privately. Sample powder and potassium bromide (1:100) were mixed and ground under an infrared baking lamp, then pressed into flakes by a powder tablet press (Graseby Specac) for FT-IR analysis.

Sample Preparation for UHPLC-Q-Exactive Orbitrap-MS
An appropriate amount of homogenized sample powder (0.2 g) was accurately weighed and ultrasonically extracted with 25 mL methanol/water (50:50, v/v) for 45 min at room temperature. The suspension was centrifuged at 5000 rpm for 5 min to remove residue. Then, the solution was filtered through a 0.22 µm filter for analysis.

Power Color Determination Analysis
A high-quality COLORIMETER (NH300, ThreeNH Technology Co., Ltd., Shenzhen, China) was used to measure the powder color and obtain L* (brightness), a* (red-green), b* (yellow-blue), and the total color value E*ab, which was obtained by the following formula: E*ab = (L 2 + a 2 + b 2 ) 1/2 . The chromaticity meter used the international universal light source D65 with a standard deviation of ∆E* ab < 0.07 (the average of 30 times of interval measurement after calibration of the standard whiteboard).

Microscopic Observation Analysis
An Olympus BX53F microscope (Olympus Life Sciences, Tokyo, Japan) with MC50 lens (Mshot, Mingmei Technology Co., Ltd., Guangzhou, China) was used to observe the characteristic cells in CRPV. The microscope preview resolution was set as 2560 × 1944; the capture resolution was 2560 × 1944; no color enhancement (saturation default 100); no single color; no automatic white balance; color correction OFF.

FT-IR Analysis
The infrared spectrum of CRPV was scanned using a Frontier FT-IR spectrometer (Perkin Elmer, Waltham, MA, USA) along with an FR-DTGS detector. Scanning parameters were as follows: scanning range of 4000-400 cm −1 , spectral resolution of 4 cm −1 , wavelength reactivity of ±0.02 cm −1 , wavelength accuracy of ±0.1 cm −1 . The interference of H 2 O and CO 2 was deducted during scanning. After the processing of peak position and baseline correction, the maps and data were corrected using OMNIC 9.2 software (Thermo Nicolet Corporation, Madison, WI, USA).
MS/MS identification was performed using a high-resolution mass spectrometer equipped with an electrospray ionization (ESI) source using a quadrupole tandem electrostatic field track well. Ion mode: positive and negative ion mode; auxiliary temperature: 350 • C; auxiliary gas flow rate: 10; sheath gas flow rate: 35; atomization voltage: 3.0 kV; capillary temperature: 350 • C; scan mode: full mass-DD MS2; scanning range: 100-1500. Sheath gas pressure auxiliary gas pressure: 30, 40, 50 arb. Nitrogen was used as an atomizer and auxiliary gas. Data were obtained using Thermo Scientific Xcalibur.

Statistical Analysis
Data obtained from the study are presented as mean ± standard deviation (SD). The one-way ANOVA and similarity analyses were carried out using SPSS 22.0 statistical software (SPSS, Inc., Chicago, IL, USA). Principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were performed using SIMCA 14.0 software (Umetrics, Inc., San Jose, CA, USA).

Results and Discussion
In this experiment, we studied 22 batches of CRPV (12 kinds of GQP and 10 kinds of SHQP) based on their powder color, microscopic cells, FT-IR analysis, and composition analysis. The color of SHQP powder was comparatively brighter, with the color of GQP and SHQP being comparatively redder and yellower, respectively. The microscopic characteristic cells of GQP mainly comprised calcium oxalate crystallization, epidermal cells of the pulp capsule, and hesperidin crystal, while those of SHQP mainly comprised tracheal, calcium oxalate crystallization, exocarp, mesocarp parenchyma, hesperidin crystal, and stoma. FT-IR analysis showed that SHQP had a higher content of carboxylic acids and its esters than GQP, whereas GQP had a higher content of polysaccharides than SHQP.
We detected 44 main components using the UHPLC-Q-Exactive Orbitrap-MS. Among them, the flavanone aglycones, naringenin and hesperetin, were the unique components of GQP. Naringenin can reduce the phosphorylation of STAT3 in the hypothalamus by regulating adipocytokines, to achieve weight loss in obese rats and treat hypertension [42]. In vitro and in vivo experiments have confirmed that naringenin can reduce hepatic lipid accumulation and attenuate the inflammation in mice by downregulating the expression of the NLRP3 /NF-κB signaling pathway both in Kupffer cells and hepatocytes [43]. Moreover, it can also decrease urea, creatine, and uric acid levels, thereby protecting against rat liver and kidney damage [44]. Hesperetin reportedly has both neuroprotective and memoryimproving effects, and it works by reducing both the inflammatory mediators' expression and neuronal apoptosis [45,46].
Luteolin-6-C-glucoside, orientin, rhoifolin, and pilloin were unique components in SHQP. Previous literature has shown that orientin exhibits antibacterial effects and can inhibit the growth of Staphylococcus aureus [47]. Although both rhoifolin and pillion have shown anti-inflammatory effects, their mechanisms are different. Rhoifolin inhibits the secretion of inflammatory factors and inhibits the expression of IKKβ and IκBα in the NF-κB signaling pathway [48,49]. Additionally, it can repair liver and kidney damage in mice with acute inflammation, treat rheumatoid arthritis, and exert anti-pancreatic cancer effects [50]. Pilloin inhibits the production of inflammatory molecules in macrophages and downregulates inflammatory cytokines, thus showing good anti-inflammatory activity both in vitro and in vivo [51]. However, we found no pharmacological studies related to luteolin-6-C-glucoside.
GQP has been reported to be mainly used for breaking Qi and resolving stagnation, while SHQP is mainly used for regulating both the liver and Qi [28]. The above studies prove that the pharmacological activities and action mechanisms of the different components in both GQP and SHQP are different. These may be the reasons for the differences in the clinical efficacy of GQP and SHQP. This research can provide a reference for the establishment of different grade standards of CRPV or establish identification methods for Chinese patent medicines with different CRPV as raw materials, while also helping with their clinical development as a precision medicine. More studies on the pharmacological efficacy should be carried out in the future, thus helping to formulate more reasonable quality standards for CRPV and guide its clinical use as a precision drug.