Cistanche Bioactive Constituents And Quality Markers: Research Progress And Implications For Ingredient Quality Control

Abstract

Cistanches Herba is a well-known tonic herb in traditional Chinese medicine (TCM). Traditionally, it is used to tonify kidney yang, replenish essence and blood, and moisten the intestines to facilitate bowel movements. Its traditional indications include kidney-yang deficiency, depletion of essence and blood, soreness and weakness of the lower back and knees, weakness of the tendons and bones, and constipation associated with intestinal dryness. It is often referred to as "desert ginseng."

Modern research has identified a diverse range of chemical constituents in Cistanche, including phenylethanoid glycosides, iridoids and iridoid glycosides, lignans, sugar alcohols, oligosaccharides, and polysaccharides. Among these constituents, echinacoside and acteoside have been listed as quality-control indicator compounds for Cistanches Herba in the 2005–2025 editions of the Chinese Pharmacopoeia. However, evaluating quality using only these two phenylethanoid glycosides cannot comprehensively characterize the constituents associated with the herb's different traditional functions or fully ensure the quality of the herbal material. A more systematic, function-associated quality-evaluation framework is therefore needed.

Building on more than 30 years of research by our team, this review systematically examines progress in the study of Cistanche chemistry and pharmacological activities. Based on the "three elements" of efficacy-related substances in TCM and the "five principles" of quality markers, echinacoside, acteoside, 8-epiloganic acid, betaine, and mannitol are proposed as a core group of efficacy-related constituents for evaluating Cistanche quality. Together, these compounds may support a more scientifically grounded and comprehensive set of quality-control indicators.

Keywords: Cistanches Herba; quality markers; echinacoside; betaine; mannitol; acteoside

 

 

Introduction

Cistanches Herba is a traditional tonic herb first recorded in the Shennong Bencao Jing. It is traditionally used to tonify kidney yang, replenish essence and blood, and moisten the intestines to facilitate bowel movements. Its traditional indications include kidney-yang deficiency, depletion of essence and blood, impotence and infertility, soreness and weakness of the lower back and knees, weakness of the tendons and bones, and constipation associated with intestinal dryness. It is widely known as "desert ginseng."

The 2025 edition of the Chinese Pharmacopoeia defines Cistanches Herba as the dried fleshy stem bearing scale leaves of Cistanche deserticola Y. C. Ma or Cistanche tubulosa (Schenk) Wight, both belonging to the family Orobanchaceae [1]. In addition, Cistanche salsa (C. A. Mey.) G. Beck is also used as Cistanches Herba in some parts of northwestern China [2–3].

In recent years, C. deserticola has been included in China's catalogue of substances traditionally used as both foods and Chinese medicinal materials. An application for C. tubulosa extract as a new food ingredient was also accepted for review by the National Health Commission in July 2026. As applications expand from traditional herbal medicine into foods and health products, greater demands are being placed on the consistency and scientific evaluation of raw-material quality.

Modern studies show that Cistanche has a complex chemical composition. Its major constituents include phenylethanoid glycosides, iridoids and iridoid glycosides, lignans, oligosaccharide esters, polysaccharides, mannitol, and betaine. Experimental studies have reported activities related to reproductive function, neuroprotection, aging, osteoporosis, immune regulation, and bowel function.

Different constituent classes do not contribute equally to every activity. Phenylethanoid glycosides are currently the most extensively investigated group of active constituents, whereas sugar alcohols, oligosaccharides, and betaine are closely associated with the herb's bowel-related activity. These findings suggest that the functional profile of Cistanche reflects the combined effects of several constituent classes.

The 2005–2025 editions of the Chinese Pharmacopoeia use echinacoside and acteoside as quantitative assay indicators for Cistanches Herba. Although these compounds are relatively abundant, representative, and readily measurable, they belong to the same chemical class. Other efficacy-related constituents-including iridoid glycosides, sugar alcohols, oligosaccharides, and betaine-are not included in these assay indicators. Consequently, the existing approach cannot fully characterize the material basis associated with the herb's different traditional functions.

 

Moreover, C. deserticola and C. tubulosa differ substantially in the composition and concentrations of phenylethanoid glycosides, iridoid glycosides, betaine, and carbohydrates. Herbal-material quality is also influenced by the host plant, geographical origin, growth duration, harvesting period, processing, and traditional preparation methods. These factors make Cistanche quality control particularly complex.

In recent years, quality evaluation in TCM has gradually shifted from measuring abundant chemical constituents alone toward assessing substances associated with safety, effectiveness, and overall therapeutic function. The quality-marker, or Q-Marker, framework emphasizes efficacy relevance, specificity, transfer and traceability, compatibility within herbal formulas, and measurability. It provides a theoretical framework for developing quality-evaluation systems that better reflect the overall functions of Chinese herbal medicines.

Previous studies have predicted potential Cistanche Q-Markers based on chemical composition, biosynthetic pathways, and traditional functions [4–7]. However, the proposed candidate lists are extensive, and the relationships between individual candidates and the herb's functions require further refinement.

Drawing on more than 30 years of research by our team, this review summarizes progress in Cistanche chemistry and pharmacology, with particular attention to constituents associated with its principal traditional functions. Using the "three elements" of efficacy-related substances in TCM [8] and the "five principles" of Q-Markers [9–10], candidate compounds are assessed in terms of efficacy relevance, specificity, transfer and traceability, and measurability. The aim is to identify Q-Markers that more comprehensively represent the functional profile and chemical characteristics of Cistanches Herba, improve quality standards, and support a multidimensional, function-associated quality-evaluation system.

 

1. Chemical Constituents

Research into the chemical constituents of Cistanches Herba began in the early 1980s, when Kobayashi and colleagues at Yomeishu Seizo Co., Ltd. in Japan carried out substantial pioneering work [11].

Chinese researchers began studying domestically occurring Cistanche in the 1960s. Through systematic resource surveys, Professor Yuquan Ma identified the botanical source of Cistanches Herba described in historical materia medica texts and named it C. deserticola Y. C. Ma [12]. Chemical research in China began mainly in the mid-1980s [13].

Since the 1990s, our team has systematically investigated Cistanche species distributed in China [14–15]. Through extensive isolation, structural identification, and comprehensive analysis, we have characterized the chemical composition of domestically occurring species, including C. deserticola and C. tubulosa [2].

Current research indicates that Cistanche species mainly contain phenylethanoid glycosides, iridoid glycosides, lignans, oligosaccharide esters, polysaccharides, monoterpenes and their glycosides, mannitol, and betaine.

 

1.1. Phenylethanoid Glycosides and Benzyl Alcohol Glycosides

Phenylethanoid glycosides are major constituents of Cistanche species. A total of 70 compounds in this class have been isolated and identified [16].

The sugar substituents in these compounds mainly include glucose, rhamnose, and xylose. The sugar directly attached to the aglycone-the central sugar-is glucose. This central glucose frequently carries an acetyl group at C-2 and a rhamnose unit at C-3. Its C-4 or C-6 position is often esterified with a phenylpropenoyl group, such as caffeoyl, feruloyl, or coumaroyl. In trisaccharide glycosides, an additional glucose, rhamnose, or xylose unit may be attached at C-6 of the central glucose.

Common phenylethanoid glycosides include:

Echinacoside

Acteoside, also known as verbascoside

Isoacteoside

2′-Acetylacteoside

Poliumoside

Salidroside

Tubuloside A

Tubuloside B

Cistanoside A

Cistanosides C–E

Echinacoside and acteoside are the two principal phenylethanoid glycosides in several Cistanche species. They are also the quantitative assay indicators specified for Cistanches Herba in the 2005–2025 editions of the Chinese Pharmacopoeia.

Our team has additionally isolated six benzyl alcohol glycosides from C. salsa and C. deserticola: salsaside A, salsaside B, salsaside Ca/Cb, 3,4-dimethoxybenzyl-β-D-glucoside, benzyl glucopyranoside, and 4-hydroxybenzyl-β-D-glucoside [16–17].

The aglycones of these compounds are benzyl alcohol derivatives. The central sugar commonly carries rhamnose at C-3 and may be esterified with caffeoyl or coumaroyl groups at C-4 or C-6.

 

1.2. Iridoids and Iridoid Glycosides

Iridoids and their glycosides are another major constituent class in Cistanche. To date, 26 compounds have been isolated and identified from the genus, comprising four iridoids and 22 iridoid glycosides [2].

In iridoid glycosides, glucose is commonly attached at C-1 of the iridoid skeleton, while C-4 often bears a methyl or carboxyl group. The hydrogen atoms at C-5 and C-9 generally have a β configuration. Hydroxyl substitution, double bonds, or a three-membered oxygen-containing ring may occur around C-6–C-8 or C-10.

Representative compounds include:

8-Epiloganic acid

6-Deoxycatalpol

Geniposide

Geniposidic acid

Gluroside

 

1.3. Lignans

Lignans occur at relatively low concentrations in the fleshy stems of Cistanche. However, their concentrations increase markedly as the stems become lignified, at which point lignans become one of the major chemical classes [18]. Lignan content may therefore serve as an indicator of substantial lignification in the herbal material.

Two lignans and 14 lignan glycosides have been isolated from Cistanche species. Among the glycosides, two are neolignan glycosides and one is a tetrahydronaphthalene-type lignan glycoside; the remaining compounds are predominantly bis-tetrahydrofuran-type lignans [2].

Representative compounds include:

(+)-Pinoresinol

(+)-Pinoresinol-O-β-D-glucopyranoside

Liriodendrin

Eucommin A

(+)-Syringaresinol-O-β-D-glucopyranoside

Isoeucommin A

 

1.4. Oligosaccharide Esters and Other Small-Molecule Constituents

Cistanche species also contain sugar alcohols and smaller quantities of sugar esters. Five sugar esters and six sugar or sugar-alcohol-related constituents have been isolated [2].

The five oligosaccharide esters are:

Cistanoside F

Cistanoside I

Cistanche tubulosa sugar ester A1

Cistanche tubulosa sugar ester A2

Kankanose

Structurally, these oligosaccharide esters correspond to phenylethanoid glycosides lacking the hydroxytyrosol structural unit.

The sugars and polyol-related compounds isolated from Cistanche mainly include glucose, fructose, sucrose, mannitol, and (2S,3S,4S)-trihydroxypentanoic acid.

Mannitol is one of the relatively abundant constituents of C. deserticola. It was previously misidentified as galactitol. Our team subsequently used nuclear magnetic resonance spectroscopy and online energy-resolved mass spectrometry to establish that the compound was mannitol rather than galactitol; the study reported that galactitol was not present in Cistanche [19].

Betaine is also a major constituent of C. deserticola and certain other Cistanche materials [2]. These water-soluble constituents occur at substantially higher concentrations in C. deserticola than in C. tubulosa. They have been proposed as important factors contributing to the cold tolerance and comparatively stronger bowel-related activity of C. deserticola.

 

1.5. Monoterpenes and Their Glycosides

Seven monoterpenes and monoterpene glycosides have been isolated from Cistanche species, comprising two monoterpenes and five monoterpene glycosides [2]:

8-Hydroxygeraniol-1-β-D-glucopyranoside

Kankanoside E

(2E,6Z)-8-β-D-Glucopyranosyloxy-2,6-dimethyl-2,6-octadienoic acid

8-Hydroxygeraniol-8-O-β-D-glucopyranoside

(2E,6R)-8-Hydroxy-2,6-dimethyl-2-octenoic acid

Betulalbuside A

8-Hydroxygeraniol

 

1.6. Polysaccharides

Polysaccharides are another major class of active constituents in Cistanche. Their constituent monosaccharides mainly include glucose, galactose, rhamnose, arabinose, and fructose.

Thirteen polysaccharide constituents have been isolated and characterized from Cistanche species [2]:

Pectic polysaccharides P1–P3 [20]

Cistan A [21]

Mannoglucan [22]

Arabinogalactan ACDP-2 [23]

Linear glucan CDP-4 [24]

Glucans 1–3 [25]

SPA [26]

CDA-1A [27]

RG-I polysaccharide CDA-3B [27]

 

1.7. Other Constituents

Other compounds isolated from Cistanche species include phenolic glycosides, sterols, and alkaloids [2]. Volatile constituents have also been widely investigated.

Lu and colleagues [28] used gas chromatography–mass spectrometry (GC-MS) to analyze a petroleum ether extract of C. deserticola. They identified 25 compounds, with esters and alkanes representing the principal constituents.

Hui and colleagues [29] reportedly extracted volatile oil from C. deserticola using a distillation–extraction apparatus and identified 24 constituents by GC-MS. Eugenol accounted for 83.60% of the total volatile oil.

Ma and colleagues [30] used supercritical carbon dioxide extraction to obtain volatile constituents from Cistanches Herba. More than 30 principal components were separated by gas chromatography. These mainly included C16–C18 normal alkanes, esters, and low-molecular-weight oxygen- and nitrogen-containing compounds. Dibutyl phthalate, dibutyl sebacate, and diisooctyl phthalate reportedly accounted for more than 90% of the total ester fraction. Eugenol was a major constituent among the low-molecular-weight compounds.

Zhang and colleagues [31] analyzed the essential oil of C. tubulosa using GC-MS-DS and identified 21 compounds, mainly fatty acids, esters, alcohols, and aliphatic hydrocarbons.

Overall, reported volatile constituents of Cistanche species include fatty acids and related alcohols and esters, sterols, hydrocarbons, and other oxygen- and nitrogen-containing compounds. Eugenol has been reported as a principal volatile phenolic constituent.

 

2. Efficacy-Related Constituents and Mechanisms of Action

Before the 1990s, pharmacological research on Cistanche mainly focused on kidney-yang-tonifying, bowel-related, and antioxidant activities, often using relatively simple in vitro experiments.

With the successful development of artificial cultivation and advances in research methods, investigations into the active constituents and mechanisms of Cistanche have become increasingly detailed.

Drawing on historical materia medica descriptions of the herb's functions and indications, our team has conducted systematic research into efficacy-related constituents, molecular targets, and mechanisms associated with "tonifying the kidney," "replenishing marrow," "strengthening tendons and bones," "nourishing the five zang organs," "promoting longevity," and "facilitating bowel movements."

The total phenylethanoid glycoside fraction associated with the traditional function of "tonifying the kidney and replenishing marrow," and its principal active constituent echinacoside, have been developed separately as drug candidates or medicines for vascular dementia (VD). The total sugar-alcohol fraction associated with bowel function has been developed as a new drug for constipation and, according to the source manuscript, is undergoing Phase III clinical trials.

These developments have contributed to the transformation of Cistanches Herba from an endangered plant resource into a major Chinese medicinal product category.

 

2.1. Investigation of Traditional Functions

To explore the clinical value of Cistanche and guide research into its efficacy-related constituents and applications, our team comprehensively examined its functions and indications in historical materia medica texts and modern works, including the Chinese Pharmacopoeia and Zhonghua Bencao [32–33].

The recorded traditional functions and indications can be summarized as follows:

Tonifying the kidney and replenishing essence.
Traditional indications in men include kidney deficiency, insufficient essence and blood, reduced fertility attributed to deficient essence, impotence, premature ejaculation, hematuria, urinary dribbling, and cold, heat, or pain affecting the penis. Traditional indications in women include kidney-yin deficiency, abdominal masses, excessive uterine bleeding, abnormal vaginal discharge, infertility associated with coldness, and a darkened complexion attributed to depletion of essence. Other recorded indications include hernia, soreness and weakness of the lower back and knees, weakness of the tendons and bones, and physical and mental fatigue.

Replenishing marrow and supporting cognition.
Recorded indications include deficiency of the "sea of marrow," neurasthenia, insomnia, forgetfulness, senile dementia, and paralysis agitans, corresponding to Parkinson's disease (PD).

Moistening the intestines and facilitating bowel movements.
Traditionally used for constipation associated with intestinal dryness, particularly in older adults, individuals with constitutional weakness, and pregnant women.

Maintaining appearance and complexion.
Recorded uses include aging or dry skin and poor complexion.

Nourishing the five zang organs and supporting longevity.
Traditional descriptions include anti-aging and life-prolonging functions.

These are historical TCM descriptions and should not be interpreted as equivalent to modern, clinically validated indications.

 

2.2. Constituents and Mechanisms Associated with "Tonifying the Kidney and Replenishing Essence"

The TCM concept of the "kidney" differs substantially from the kidney as defined in modern anatomy and physiology.

In TCM, the kidney stores essence, governs water metabolism, and receives qi. It stores congenital essence and is associated with reproduction and the fundamental basis of life, hence its description as the "root of the congenital constitution." Kidney essence is understood to generate kidney qi, which encompasses yin and yang. Kidney yin and kidney yang are believed to support and coordinate the yin–yang balance of the other organs, leading to the description of the kidney as the "root of yin and yang of the five zang organs."

Within this framework, kidney function is associated with the bones, bone marrow, spinal cord, brain, thinking, memory, teeth, hair, hearing, reproduction, excretion, and emotional activity. "Kidney deficiency" is therefore understood to affect a wide range of organs and tissues.

Cistanche is a well-known tonic herb with a particularly prominent role in traditional kidney-tonifying formulas. The Rihuazi Bencao describes its use for male sexual dysfunction and female infertility, as well as for nourishing the internal organs, supporting muscle growth, warming the lower back and knees, and addressing seminal leakage, hematuria, urinary dribbling, abnormal vaginal discharge, and genital pain.

In the Bencao Gangmu, Li Shizhen described Cistanche as tonifying without being excessively forceful, reflecting its traditional reputation as a gentle warming tonic.

During the twentieth century, several research teams used animal models of kidney-yang deficiency to investigate these traditional functions [34–35].

To clarify the associated constituents, our team established a hydrocortisone-induced rat model of reproductive impairment and kidney-yang deficiency. Measures included mounting latency, mounting frequency, ejaculation latency, ejaculation frequency, and sperm motility.

Low-molecular-weight water-soluble constituents-mainly mannitol, betaine, and oligosaccharides-polysaccharides, and phenylethanoid glycosides all improved erectile and mating-related measures in the model. They also improved characteristic model features such as reduced food intake, slower body-weight gain, sparse and disordered fur, reduced activity, and cold-seeking avoidance behaviors. Phenylethanoid glycosides showed the strongest effects [36].

The hypothalamic–pituitary–gonadal (HPG) axis helps maintain reproductive endocrine function. Disturbances in this axis can lead to reproductive dysfunction. HPG-axis impairment has been proposed as one of the biological features associated with experimental kidney-yang-deficiency models, contributing to reduced testosterone secretion and impaired sexual function.

A cited clinical study reported that kidney-tonifying medicines, including Cistanche, could influence HPG-axis function and the release of related signaling molecules and hormones [37].

Our team found that Cistanche oligosaccharides, polysaccharides, and phenylethanoid glycosides could regulate hydrocortisone-induced disturbances in sex-hormone levels and improve reproductive-function measures in rats [36]. Phenylethanoid glycosides also improved atrophy of the testes, epididymides, seminal vesicles, and penis, preserved testicular morphology, and moderately increased testosterone levels [38].

More recently, our team reported that total glycosides of C. deserticola (TGCD) dose-dependently regulated the HPG axis in an alcohol-induced reproductive-injury model. TGCD improved sperm-quality measures, alleviated pathological damage to the testes and small intestine, increased testosterone levels, and reduced oxidative stress and inflammation.

The proposed mechanism involved remodeling the gut microbiota, with Lactobacillus reuteri (LR) identified as a key organism. TGCD acted as a prebiotic substrate for LR, increased its abundance, and raised concentrations of the microbial metabolite spermidine. This activated the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway in Leydig cells, reduced oxidative stress, restored the expression of steroidogenic enzymes, and enhanced testosterone biosynthesis.

The study also reported lower fecal LR abundance in clinical patients. Analysis of human testicular single-cell transcriptomic data showed reduced expression of genes associated with spermidine biosynthesis and the Nrf2 pathway in infertile patients. These findings provide a rationale for further investigation of microbiota-targeted TGCD interventions for oligoasthenozoospermia [39].

In summary, experimental studies support reproductive-function-related activities of Cistanche. Phenylethanoid glycosides, low-molecular-weight water-soluble constituents-including mannitol, betaine, and oligosaccharides-and polysaccharides have all shown activity in relevant models, with phenylethanoid glycosides generally producing the strongest effects.

 

 

Cistanche Benefits in Modern Medical Terms

2.3. Constituents and Mechanisms Associated with "Replenishing Marrow and Supporting Cognition"

As China's population ages, neurodegenerative diseases have attracted increasing attention. Experimental pharmacological research has investigated Cistanche in models relevant to Alzheimer's disease (AD), vascular dementia, and Parkinson's disease.

2.3.1. Constituents and Mechanisms Related to Learning and Memory

Researchers have used a range of experimental models, including:

Scopolamine-induced impairment of learning and memory acquisition in mice

Sodium nitrite-induced learning and memory impairment in mice

Ischemia–reperfusion-induced memory impairment in rat models of vascular dementia

Ethanol-induced impairment of memory retrieval in mice

Hydrocortisone-induced learning and memory impairment in kidney-yang-deficiency mouse models

Behavioral testing indicated that total Cistanche glycosides reduced errors in water-maze tests, prolonged latency in step-down tests, increased the proportion of correct responses, and shortened the time required to reach the endpoint [40–44].

Our team reported that total Cistanche glycosides reduced cerebral infarct area, increased blood flow on the ischemic side, improved blood–brain barrier integrity, promoted angiogenesis and neurovascular-unit remodeling, and increased the activities of antioxidant enzymes such as glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in a middle cerebral artery occlusion/reperfusion model.

The treatment also reduced malondialdehyde levels and brain-cell apoptosis, inhibited acetylcholinesterase (AChE) and nitric oxide synthase (NOS) activities, and regulated excitatory amino-acid metabolism [45].

Echinacoside, a representative active constituent, also showed protective effects in cerebral ischemia models, reducing neuronal apoptosis in the hippocampus [46]. It additionally influenced striatal monoamine neurotransmitters and their metabolites after ischemia, suggesting possible roles in limiting neuronal apoptosis and maintaining neurotransmitter homeostasis.

2.3.2. Constituents and Mechanisms Investigated in Alzheimer's Disease Models

Alzheimer's disease is a neurodegenerative disorder characterized clinically by progressive cognitive and memory impairment, with major pathological changes affecting the cerebral cortex and other brain regions.

Researchers have investigated total Cistanche glycosides using several models, including:

Intracerebroventricular administration of quinolinic acid [47]

Intracerebroventricular administration of β-amyloid peptide 25–35 [48]

Subcutaneous administration of aluminum chloride [49]

D-galactose-induced brain-aging models [50]

These studies reported improvements in learning and memory, reductions in brain malondialdehyde content, changes in GSH-Px and SOD activities, lower AChE activity, reduced brain-cell apoptosis, and reduced calcium accumulation in brain tissue.

They also reported increased expression of the anti-apoptotic protein B-cell lymphoma-2 (Bcl-2) and reduced expression of Bcl-2-associated X protein (Bax).

The proposed mechanisms include enhanced free-radical defense, reduced lipid peroxidation, reduced β-amyloid deposition, and inhibition of neuronal apoptosis. The source manuscript contains an inconsistency concerning the direction of changes in GSH-Px and SOD activities; this is addressed in the editorial notes below.

Cell-based studies have further explored these mechanisms.

Qi and colleagues [51] used human SH-SY5Y neuroblastoma cells and reported that Cistanche increased the expression of α3 and α7 nicotinic acetylcholine receptor subunits. It counteracted the reductions induced by β-amyloid peptide 25–35 and attenuated β-amyloid-associated cellular injury and lipid peroxidation.

Choi and colleagues [52] reported that C. deserticola promoted neurite growth in rat pheochromocytoma cells and increased nerve growth factor (NGF) levels in rat glioma cells. The study also reported enhanced hippocampal neurite growth, neuronal differentiation, and synapse formation in mice, together with increased NGF secretion in the cortex and hippocampus and improved learning and memory.

Phenylethanoid glycosides are the principal constituents investigated for these activities.

Tian and colleagues [53] found that echinacoside increased total brain protein content and total antioxidant capacity, reduced AChE activity and serum interleukin-2 (IL-2), and improved learning and memory in SAMP8 mice.

Wang and colleagues [54] reported that acteoside protected SH-SY5Y cells against β-amyloid peptide 25–35-induced toxicity. It improved cell viability, reduced apoptosis and reactive oxygen species production, and inhibited cytochrome c release, mitochondrial membrane-potential loss, and changes associated with caspase-3.

Our team also found that tubuloside B and echinacoside protected neuronal cells against tumor necrosis factor-α (TNF-α)-induced apoptosis [55–56].

Building on research into the neuroprotective activities of Cistanche phenylethanoid glycosides, our team developed Congrong Zonggan Capsules, a total Cistanche glycoside medicine for vascular dementia that was approved for marketing in 2005.

A cited clinical report involving 124 patients with Alzheimer's disease described delayed disease progression and effects comparable to Aricept, with fewer reported adverse reactions [57]. This finding should be interpreted in the context of the study's design and should not be generalized to all Cistanche ingredients or extracts.

Echinacoside was subsequently developed as a Class I new-drug candidate. According to the source manuscript, it received clinical-trial authorization in 2017 and has completed Phase I trials.

To investigate its molecular targets in cerebral ischemic injury, our team used echinacoside as a molecular probe for target identification. The study identified a catalytic subunit of casein kinase 2 (CK2) as a direct target and further investigated the associated neuroprotective mechanism [58]. This work was included among the "Top Ten Academic Advances in Traditional Chinese Medicine in 2022" [59].

2.3.3. Constituents and Mechanisms Investigated in Parkinson's Disease Models

Parkinson's disease is a chronic, progressive disorder involving extrapyramidal motor function. A major pathological feature is degeneration of dopaminergic neurons in the substantia nigra, leading to reduced dopamine availability.

Changes in acetylcholine, norepinephrine, serotonin, γ-aminobutyric acid, and neuropeptides may also contribute to disturbances in neurotransmitter balance and clinical symptoms such as rigidity, movement difficulties, and postural impairment [60].

A commonly used experimental model is the mouse model induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP).

Studies of echinacoside [61–62], acteoside [63–64], and campneoside II [65] have reported protective effects in Parkinson's-related experimental systems.

Reported findings across animal and cellular models include improved behavioral measures, reduced loss of dopaminergic neurons and dopamine transporters, increased dopamine-related metabolites such as 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid, inhibition of caspase-3- and caspase-8-associated apoptosis, changes in neurotrophic-factor activity and expression, and regulation of apoptosis-related proteins and transcripts.

Cell-based studies have provided additional mechanistic information:

Tubuloside B inhibited apoptosis and oxidative stress induced by 1-methyl-4-phenylpyridinium (MPP+) in rat PC12 cells [66].

Acteoside protected SH-SY5Y cells against rotenone-associated injury, with proposed effects on Parkinson's-related protein degradation and α-synuclein dimer formation [67].

Acteoside protected SH-SY5Y cells against MPP+-induced injury by reducing intracellular reactive oxygen species, increasing Bcl-2 expression, inhibiting caspase-3 activity, and preserving mitochondrial membrane potential [68].

2.3.4. Constituents and Mechanisms Associated with Antidepressant-Like Activity

Depression is characterized by disturbances in mood and may involve sadness, reduced appetite, feelings of guilt, low self-worth, sleep disturbances, and, in severe cases, suicidal thoughts.

Its pathogenesis is complex. Proposed mechanisms include disturbances in monoamine neurotransmitters and receptors, hyperactivity of the hypothalamic–pituitary–adrenal (HPA) axis, altered neuroplasticity, abnormal cytokine signaling, and gut-microbiota imbalance.

Modern clinical management commonly combines pharmacotherapy with psychological interventions. In TCM, depression is generally discussed within the category of "yu zheng," or constraint disorders.

The Shennong Bencao Jing records Cistanche as addressing "five types of taxation and seven injuries," a historical category that the source manuscript interprets as including injury associated with emotional disturbances. The Bencao Gaiyao also describes uses related to neurasthenia.

Experimental models used to investigate antidepressant-like activity include learned helplessness, chronic unpredictable mild stress, olfactory bulbectomy, and glucocorticoid- or corticosterone-induced models.

Studies reported reduced immobility in mouse tail-suspension tests [69]. Repeated administration of Cistanche extracts also improved depression-related behavioral measures in forced-swim, sucrose-preference, open-field, and novelty-suppressed-feeding tests [70].

Fan and colleagues [71] reported that C. tubulosa glycosides alleviated depression-like behaviors in experimental models.

Phenylethanoid glycosides have been reported to increase serum estradiol and testosterone and raise brain serotonin and dopamine levels in experimental perimenopausal models, with improvements in depression-like behavior.

Iridoid glycoside fractions, described in the source manuscript as containing mainly 8-epiloganic acid and geniposidic acid, have also been associated with antidepressant-like effects.

Further research cited in the manuscript examined changes in hippocampal signaling and neuronal morphology, proposing that iridoid-related activity may involve restoration of HPA-axis function [72].

Gut microbiota may also contribute to Cistanche-associated antidepressant-like effects. Studies suggest that Cistanche can influence central nervous system function, emotional responses, and behavior through the gut–brain axis [73–74].

Overall, phenylethanoid glycosides are the main constituent class associated with the traditional function of "replenishing marrow and supporting cognition." Iridoid glycosides may also contribute to selected neurological activities, although their specific roles require further clarification.

 

2.4. Constituents and Mechanisms Associated with "Strengthening Tendons and Bones"

Within TCM theory, kidney function is closely associated with the bones, bone marrow, and spinal cord. Experimental research has therefore investigated Cistanche for bone-related and anti-fatigue activities.

Li and colleagues [75] reported that C. deserticola extract increased alkaline phosphatase (ALP), bone morphogenetic protein-2 (BMP-2), osteopontin mRNA, and mineral deposition in bone-related experimental systems.

Zeng and colleagues [76] found that serum obtained after Cistanche administration promoted the differentiation of bone-marrow mesenchymal stem cells toward osteoblasts, suggesting a basis for further research into bone formation.

Our team compared different C. deserticola fractions in SAMP6 mice. TGCD and Cistanche polysaccharides improved pathological changes in the femur, increased trabecular number, promoted bone formation, and reduced bone resorption. The sugar-alcohol fraction did not show activity in this model [77].

Mechanistic investigations indicated that TGCD and polysaccharides increased the expression of BMP-2, osteocalcin, and osteoprotegerin, reduced receptor activator of nuclear factor-κB ligand (RANKL) expression, and activated proteins associated with Wnt/β-catenin signaling.

Echinacoside also improved bone-quality measures in ovariectomized rats, an experimental model of estrogen-deficiency-associated osteoporosis [78].

In MC3T3-E1 osteoblast precursor cells, echinacoside promoted proliferation, increased ALP activity and collagen content, improved osteocalcin and mineralization-related measures, and increased the osteoprotegerin/RANKL ratio [79].

Anti-fatigue-related studies have reported additional effects:

Yang and colleagues [80] found that a C. deserticola decoction reduced liver-cell and endothelial-cell injury in mice subjected to loaded swimming, increased NOS3 expression, and promoted liver-glycogen synthesis.

Cai and colleagues [81] reported lower serum creatine kinase, lactate dehydrogenase, and lactate, together with higher hemoglobin and glucose levels.

C. deserticola prolonged swimming time and improved hypoxia tolerance and cold tolerance in aging-model mice [34].

It reduced the post-exercise rise in serum creatine kinase and helped preserve skeletal-muscle ultrastructure [82].

Xu and colleagues [83] reported increased body weight, improved hypoxia tolerance, and longer sustained swimming time in mice.

These findings support further investigation of Cistanche for bone-related and anti-fatigue applications. Echinacoside and other constituents are proposed as important contributors.

 

2.5. Constituents and Mechanisms Associated with "Nourishing the Five Zang Organs"

The research reviewed in this section mainly concerns liver-protective activity.

Alcohol-induced injury in primary hepatocytes reduces viability and causes apoptotic and necrotic changes. It is associated with reduced expression of the anti-apoptotic gene Bcl-2 and increased expression of c-fos.

Zhao and colleagues [84] reported that C. deserticola extract improved liver-tissue structure, increased liver glycogen, and reduced lactate dehydrogenase isoenzyme activity in mice subjected to strenuous exercise.

Our team compared different C. deserticola fractions in a mouse model of acute alcohol-induced liver injury and identified TGCD as an active fraction.

TGCD reduced hepatic lipid accumulation, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and malondialdehyde. It increased SOD, catalase, and GSH-Px activities and improved liver histology and intestinal-barrier-related measures.

It also reduced serum lipopolysaccharide, diamine oxidase, and D-lactate, limited the transfer of bacterial metabolites toward the liver, promoted Nrf2 nuclear translocation, and inhibited the expression of Kelch-like ECH-associated protein 1 and plasmalemma vesicle-associated protein 1 in the intestinal barrier [85].

Our team subsequently combined a recognized chronic alcohol-induced liver-injury model with multi-omics analysis. TGCD improved pathological changes associated with chronic injury.

The proposed mechanism involved inhibition of bile salt hydrolase activity, remodeling of bile-acid homeostasis, alleviation of cholestasis, regulation of the stearoyl-CoA desaturase 1/AMP-activated protein kinase/mechanistic target of rapamycin signaling pathway, improved lipid-metabolism balance, and reduced inflammation [86].

Luo and colleagues [87] also reported that total Cistanche glycosides dose-dependently improved apoptosis and necrosis-related changes, increased cell viability, enhanced anti-apoptotic gene expression, and reduced pro-apoptotic gene expression in primary hepatocyte cultures.

Houghton and colleagues [88] reported hepatoprotective activity associated with echinacoside.

Wu and colleagues [89] found that echinacoside reduced serum ALT and AST and tissue malondialdehyde in rats with carbon tetrachloride-induced acute liver injury. It also reduced caspase-3 activity and TNF-α, increased SOD activity, and improved liver histology.

Our recent research reported that echinacoside could promote the growth of Lactobacillus reuteri as a prebiotic substrate while also activating hepatic farnesoid X receptor signaling. These complementary pathways were associated with lower harmful bile-acid levels and improved liver injury in mice with chronic alcohol-associated liver disease [90].

Other phenylethanoid glycosides with reported hepatoprotective activity include acteoside [91], isoacteoside [92], 2′-acetylacteoside [92], tubuloside B [92], kankanoside K1 [93], kankanoside K2 [93], and isocampneoside I [93].

Proposed mechanisms include superoxide-radical scavenging, inhibition of lipid peroxidation in hepatic microsomes, and reduced AST release from injured cells.

Overall, echinacoside, acteoside, and related phenylethanoid glycosides are the principal constituents investigated for Cistanche-associated liver-protective activity.

 

2.6. Constituents and Mechanisms Associated with "Moistening the Intestines and Facilitating Bowel Movements"

In TCM, Cistanche is associated with the kidney and large-intestine meridians. It is traditionally understood to support kidney yang while moistening dryness and facilitating bowel movements.

Its bowel-related effects were initially recorded as cautions concerning use. The Bencao Yanyi Buyi described loose bowel movements following abrupt use, while the Shennong Bencao Jing Shu advised against use in diarrhea.

The Yizong Bidu: Bencao Zhengyao subsequently identified moistening the large intestine as a function of the herb. The Ming-dynasty Jingyue Quanshu: Bencao Zheng described its use for constipation in individuals considered too weak for purgative treatment.

Bowel-related applications subsequently became a distinctive traditional use of Cistanche.

Zhang and colleagues [94] reported that C. deserticola increased small-intestinal transit, shortened defecation time, counteracted atropine-associated inhibition of defecation, and reduced water absorption in the large intestine. The authors initially proposed inorganic salts and hydrophilic colloidal polysaccharides as contributors to its mild laxative activity.

Zhang and colleagues [95] compared C. deserticola, C. tubulosa, C. salsa, C. sinensis G. Beck, and their processed products. All four species increased small-intestinal transit and shortened defecation time in mice. C. deserticola produced the strongest effect, while processing reduced bowel-related activity.

Our team reported that C. deserticola, C. tubulosa, and C. salsa reduced water absorption in the mouse large intestine, promoted colonic movement, and shortened defecation time [96].

Further experiments showed that the total sugar-alcohol fraction, mannitol, and the total sugar-alcohol fraction after mannitol removal all shortened defecation time in constipated mice, increased small-intestinal transit, and raised fecal water content.

The total sugar-alcohol fraction was therefore identified as a principal active fraction associated with bowel function. Proposed mechanisms include increased intestinal osmotic pressure, reduced water absorption, and enhanced intestinal motility.

Our team subsequently developed this fraction as a new drug for constipation. According to the source manuscript, Phase III clinical trials are ongoing.

2.7. Constituents and Mechanisms Associated with Other Activities

Immune-Related Activity

Zhang and colleagues [97–98] reported that Cistanche polysaccharides or acteoside reduced malondialdehyde in the brain, heart, and liver of D-galactose-induced aging-model mice.

They also increased telomerase activity in the heart and brain, enhanced peritoneal-macrophage phagocytosis and lymphocyte proliferation, and increased circulating IL-2.

Wang and colleagues [99] used cyclophosphamide-induced immunosuppression models. Cistanche polysaccharides promoted lymphocyte IL-2 secretion and proliferation in both activated and nonactivated cells.

Intraperitoneal administration increased the spleen index in normal and immunosuppressed mice and counteracted cyclophosphamide-associated reductions in the thymus index.

Zeng and colleagues [100] reported that C. deserticola polysaccharides counteracted lymphocyte-proliferation inhibition associated with dexamethasone, isoproterenol, and high concentrations of TNF-β. They also enhanced the proliferation-promoting effects of low concentrations of TNF-β. Intracellular calcium release was proposed as a contributing mechanism.

Shang and colleagues [101] reported protective effects of total Cistanche glycosides on immune-function measures in mice exposed to cobalt-60 gamma radiation.

Antioxidant Activity

C. deserticola polysaccharides increased SOD and GSH-Px activities in erythrocytes and lung tissue of aging-model mice. They reduced malondialdehyde and collagen in lung tissue and plasma and increased elastin, suggesting activity against oxidative damage in these experimental systems [102].

Teng and colleagues [103] reported that crude C. tubulosa polysaccharides scavenged 1,1-diphenyl-2-picrylhydrazyl radicals.

Sui and colleagues [104] isolated a water-soluble carbohydrate fraction with antioxidant activity and effects in skin-aging-related experiments.

Xiong and colleagues [105] reported free-radical-scavenging and anti-lipid-peroxidation activities among C. deserticola phenylethanoid glycosides. Activity increased with the number of phenolic hydroxyl groups.

Yang and colleagues [106] compared six phenylethanoid glycosides and investigated structure–activity relationships. Antioxidant activity was associated with the number and positions of phenolic hydroxyl groups on the aglycone and phenylpropenoyl group, steric effects, and acetyl substitution at C-2 of the central glucose. The source also proposed a possible contribution from an α,β-unsaturated carbonyl-containing structural feature.

 

Aging-Related Activity

Phenylethanoid glycosides, polysaccharides, and mannitol have shown activity in experimental aging models.

Xue and colleagues [107] reported that C. deserticola polysaccharides and mannitol influenced skin-aging-related measures, increased SOD activity, and reduced lipofuscin accumulation. Mannitol also shifted the lipid-peroxide/SOD relationship in aged animals toward that observed in adult animals.

Xuan and colleagues [108] investigated Cistanche phenylethanoid glycosides in D-galactose-induced aging-model mice, using vitamin E as a positive control.

The glycoside-treated group showed fewer learning and memory errors, shorter learning-response times, longer memory latency, increased SOD activity in serum and brain, lower malondialdehyde in liver and serum, and a higher spleen index.

Anti-Inflammatory Activity

Lin and colleagues [109] reported anti-inflammatory activity of C. deserticola in rodents.

Jia and colleagues [110] reported that an echinacoside-enriched C. tubulosa extract improved disease-related measures in mice with dextran sulfate sodium-induced colitis.

Xiong and colleagues [111] reported inhibition of nitric oxide-related activity by phenylethanoid compounds in activated macrophages.

 

3. Identification of Cistanche Q-Markers and Development of a Quality-Standard System

Cistanches Herba is a tonic Chinese medicinal material widely used in traditional kidney-tonifying applications. The 2025 edition of the Chinese Pharmacopoeia specifies echinacoside and acteoside as quality-control indicator compounds.

However, its chemical composition also includes iridoid glycosides, sugar alcohols, and polysaccharides. Measuring only echinacoside and acteoside therefore provides an incomplete assessment of the material's broader chemical and functional profile.

Based on the "three elements" of efficacy-related substances proposed by Academician Wei Xiao [8] and the Q-Marker concept proposed by Academician Changxiao Liu [9], our team has evaluated Cistanche efficacy-related constituents and potential Q-Markers using systematic chemical and pharmacological research.

This work aims to improve quality stability, consistency, controllability, and traceability.

3.1. Identification of Q-Markers

3.1.1. Botanical Relationships and Chemical Specificity

Most Chinese medicinal materials are derived from plants, and many active constituents are secondary metabolites.

Differences in genetic characteristics and biosynthetic pathways allow different plants to produce distinct metabolite profiles. These differences contribute to the chemical basis of traditional properties and functions and provide important information for quality evaluation.

Chemotaxonomy suggests that closely related plants often share similar chemical classes. Studying chemical characteristics within a family or genus can therefore help predict potential Q-Markers.

Cistanche species are perennial parasitic herbs and constitute one of the more extensively studied genera in Orobanchaceae.

Approximately 22 species have been reported worldwide, distributed mainly in warm deserts and other arid or semi-arid regions of the Northern Hemisphere, including China, Iran, India, Pakistan, and Mongolia [112].

The Key to the Families and Genera of Higher Plants of China recorded six species in China [113]. Subsequent investigations identified four principal species: C. deserticola, C. tubulosa, C. salsa, and C. sinensis. These occur in regions including Inner Mongolia, Xinjiang, Qinghai, Gansu, and Ningxia and have traditional uses related to kidney-tonifying and tendon-strengthening functions [15].

The 2025 Chinese Pharmacopoeia recognizes the dried fleshy stems bearing scale leaves of C. deserticola and C. tubulosa as Cistanches Herba.

Parasitic members of Orobanchaceae frequently contain characteristic glycosides, including phenylpropanoid and phenylethanoid glycosides, with benzyl alcohol glycosides occurring in some taxa.

Plants in the genus Boschniakia contain substantial quantities of phenylpropanoid glycosides, such as boschnialoside B [114]. Cistanche species primarily contain phenylethanoid glycosides such as echinacoside and reportedly contain few phenylpropanoid glycosides. Echinacoside is largely absent from the examined Orobanche species [115].

Among Cistanche species subjected to quantitative analysis, C. tubulosa generally has the highest phenylethanoid-glycoside concentrations, principally echinacoside and acteoside. C. sinensis, by contrast, contains little or no echinacoside.

Our team's biosynthetic research showed relationships among the pathways producing these phenylethanoid glycosides [116]. Echinacoside is formed through multiple biosynthetic steps, including glycosylation.

After oral administration, Cistanche phenylethanoid glycosides also follow related metabolic pathways. They can be converted into smaller phenolic-acid metabolites, including DOPAC and homovanillic acid, which are also associated with dopamine metabolism [45,117–119]. Their structural relationship to neurotransmitter metabolites has been discussed as a possible contributor to biological activity, although structural similarity alone does not establish equivalent neurotransmitter action.

Accordingly, phenylethanoid glycosides-particularly the relatively abundant and representative echinacoside and acteoside-are suitable candidates for Cistanche Q-Markers.

3.1.2. Prediction Based on Efficacy-Related Constituents

A review of pharmacological studies indicates that polysaccharides, sugar alcohols-particularly mannitol-betaine, and phenylethanoid glycosides have shown activity in experimental models associated with the traditional function of tonifying kidney yang.

Echinacoside, acteoside, and related phenylethanoid glycosides are also the principal constituents investigated for learning and memory, Alzheimer's- and Parkinson's-related experimental models, and antidepressant-like activity.

Iridoid glycosides such as 8-epiloganic acid and geniposidic acid may also contribute to antidepressant-like activity.

Constituents proposed to contribute to bowel-related activity include mannitol, betaine, and fructose.

Polysaccharides, betaine, mannitol, and phenylethanoid glycosides have been associated with activity in aging-related models, while echinacoside and acteoside have been investigated for hepatoprotective effects.

Polysaccharides also contribute to immune-related activities, including lymphocyte proliferation and differentiation and recovery of immune-function measures. Antioxidant activity is mainly attributed to phenylethanoid glycosides and polysaccharides.

Lignans increase substantially as the fleshy stems become lignified, but their relationship to the herb's principal functions has received less attention.

Based on traditional functions and experimental pharmacology, candidate Q-Markers therefore include:

Echinacoside and acteoside

Iridoid glycosides such as 8-epiloganic acid and geniposidic acid

Betaine

Mannitol

Polysaccharide constituents, subject to analytical limitations

3.1.3. Measurability

Measurability is a central requirement in Q-Marker evaluation.

Our team used multidimensional chromatography–mass spectrometry approaches to analyze C. deserticola and C. tubulosa. The work reported the identification of 513 compounds and enabled broad characterization of phenylethanoid glycosides, iridoid glycosides, lignans, sugar alcohols, betaine, and other constituents [120].

This analytical work helped characterize the chemical basis of Cistanche and address challenges in measuring diverse constituent classes.

For polysaccharides, the Chinese Pharmacopoeia describes phenol–sulfuric acid colorimetry as an assay method. However, limitations in specificity and accuracy restrict its suitability for defining precise Cistanche Q-Markers. Polysaccharides are therefore not included in the proposed marker set at this stage, pending improved analytical methods.

From the perspective of measurability, echinacoside, acteoside, 8-epiloganic acid, geniposidic acid, betaine, and mannitol remain suitable candidate compounds.

3.1.4. Compatibility Within Herbal Formulas

Chinese herbal medicines are frequently used in formulas, making compatibility an important consideration in Q-Marker research. Candidate markers should therefore be evaluated within relevant formula environments rather than only in isolated herbs.

Cistanche is also commonly used in compound prescriptions.

The Bencao Xinbian describes its use alongside herbs such as ginseng, atractylodes, prepared rehmannia, and cornus fruit. The Bencao Jing Jie Yao records combinations with a range of other medicinal materials for traditional reproductive, urinary, kidney-deficiency, and bowel-related indications.

Examples include combinations with gelatin, eucommia, rehmannia, angelica, and ophiopogon for infertility; with ginseng, deer antler, eucommia, and psoralea for impotence and age-related yang deficiency; with astragalus for kidney-qi deficiency; and with agarwood and sesame for constipation associated with sweating.

Other historical combinations include cornus fruit, schisandra, Chinese yam, goji fruit, astragalus, angelica, peony, licorice, scutellaria, and red yeast rice for different traditional indications.

Modern formulas containing Cistanche mainly address:

Traditional kidney-tonifying functions, as in Dihuang Yinzi, Roucongrong Wan, and Guilingji

Bowel function, as in Jichuan Jian

Cognitive functions, as in Compound Congrong Yizhi Capsules

Commonly combined herbs include dodder seed, morinda root, prepared rehmannia, cornus fruit, cynomorium, and epimedium.

Li [121] divided Dihuang Yinzi into a kidney-tonifying component, including Cistanche, and a phlegm-resolving component. The research examined complementary effects on neurological injury, infarct size, neuronal apoptosis, and neural-cell proliferation and differentiation in ischemia–reperfusion models.

Chang and colleagues [122] reported that Dihuang Yinzi improved post-stroke depression-related measures by promoting Miro1 ubiquitination-dependent mitophagy and reducing nuclear factor-κB-mediated neuroinflammation. Echinacoside showed strong affinity for the investigated target, suggesting a role as an important active constituent.

Cistanche is the principal herb in Jichuan Jian. Su [123] used formula-disassembly experiments and found that the kidney-warming and intestine-moistening group-Cistanche, achyranthes, and angelica-increased intestinal water content. Other formula groups contributed through different mechanisms.

Wei and colleagues [124] and Gao and colleagues [125] applied the Q-Marker "five principles" and proposed echinacoside and acteoside as potential markers for Jichuan Jian.

Echinacoside, acteoside, and polysaccharides were also proposed as potential Q-Markers in the Tiaoli Buxu formula, in which Cistanche serves as an assisting herb [126].

Overall, Cistanche's principal functions in formulas are traditionally described as tonifying the kidney and replenishing essence, replenishing marrow and supporting cognition, and moistening the intestines. Associated constituent classes include phenylethanoid glycosides, sugar alcohols such as mannitol, and betaine.

3.1.5. Quality Transfer and Traceability

Chinese medicinal materials originate from biological sources. During processing, traditional preparation, extraction, formulation, compatibility with other herbs, absorption, and metabolism, their constituents may be transferred, transformed, concentrated, or depleted.

Identifying compounds that can be tracked through these stages provides an important basis for Q-Marker selection [127].

Most Cistanche studies have focused on processing and absorption or metabolism. Comparatively few have directly compared constituent transfer and composition before and after formula compatibility. This section therefore focuses on processing and in vivo metabolism.

Effects of processing

Processing can alter both the concentrations and identities of constituents.

The earliest description of Cistanche processing appears in the Leigong Paozhi Lun, which instructs that the material should be soaked overnight in clear wine and then steamed. Later materia medica texts generally retained wine-based processing, with differences in subsequent heating or roasting methods.

The Bencao Zhengyi criticized prolonged salt preservation and subsequent washing, arguing that these practices could diminish the herb's traditional properties. This is a historical interpretation rather than a modern quantitative demonstration that all pharmacological activity is lost.

In contemporary TCM practice, unprocessed Cistanche is most common, although wine-processed, salt-processed, and black-bean-processed preparations are also used. Wine processing is the most common of these alternatives and is used in Dihuang Yinzi.

Ma and colleagues [128] reported that phenylethanoid glycosides underwent hydrolysis and isomerization during wine processing. The source manuscript also describes sugar degradation and the formation of products including 5-hydroxymaltol and 5-hydroxymethylfurfural.

Zhang and colleagues [129] reported an increase in measured betaine content after wine processing.

Wang and colleagues [130] compared unprocessed and wine-processed extracts and reported generally stronger effects for wine-processed material in the investigated model. Echinacoside, acteoside, isoacteoside, and tubuloside B were proposed as constituents potentially associated with changes in kidney-tonifying activity after processing.

Phenylethanoid glycosides, betaine, and mannitol are therefore considered candidates for monitoring processing-associated quality changes.

Absorption and metabolism

Yan [131] used liquid chromatography–mass spectrometry to analyze blood-associated constituents after oral administration of Cistanche to rats.

Echinacoside, acteoside, cistanoside A, 8-epiloganic acid, and geniposidic acid were detected as parent compounds.

Wang [132] reported that echinacoside and geniposidic acid could be detected as parent compounds in rat serum after administration of unprocessed, steamed, or wine-processed C. tubulosa.

These findings support consideration of phenylethanoid glycosides and selected iridoid glycosides in marker evaluation involving systemic exposure.

Species-specific marker selection

Taken together, the proposed candidate Q-Markers for C. deserticola include echinacoside, acteoside, 8-epiloganic acid, geniposidic acid, betaine, and mannitol.

The source manuscript reports that C. tubulosa contains relatively little mannitol and no detectable betaine in the analyzed materials. It therefore recommends excluding betaine and mannitol from the proposed candidate set for that botanical source.

Although polysaccharides have shown reproductive-function-related, antioxidant, immune-related, and aging-related activities, current analytical limitations prevent their inclusion in the proposed Q-Marker set.

3.2. Development of a Q-Marker-Based Quality-Standard System

Phenylethanoid glycosides have long been used as Cistanche quality-control indicators because they are relatively abundant, readily detectable, and supported by pharmacological research.

Research teams have developed simultaneous quantitative methods using:

HPLC-UV [133]

LC-MS [130,134]

Proton nuclear magnetic resonance spectroscopy [135–136]

These methods have been applied to materials differing in geographical origin [134–135], processing method [137], harvesting season [133], plant part [136,138], and host plant [134,138–139].

Studies have also examined C. salsa [140] and C. sinensis [141].

The results show substantial differences in phenylethanoid-glycoside concentrations between species and between batches.

However, Cistanche quality is also associated with sugar alcohols, polysaccharides, and iridoid glycosides. Existing indicator compounds and evaluation methods do not fully capture this broader composition.

Over more than 30 years, our team has focused on Cistanche research and industrial development. Based on systematic chemical and pharmacological work, we have contributed to quality standards for both crude herbal materials and prepared decoction pieces.

Using echinacoside and acteoside as indicator compounds, our team drafted and revised Cistanches Herba quality standards in the 2005 and 2010 editions of the Chinese Pharmacopoeia.

Comprehensive LC-MS and quantitative NMR analyses showed that:

C. deserticola contained relatively high concentrations of oligosaccharides, mannitol, and betaine.

C. tubulosa contained relatively high concentrations of phenylethanoid glycosides and lower concentrations of oligosaccharides.

Betaine was not detected in the analyzed C. tubulosa materials.

Lignan glycosides occurred at relatively low concentrations in both species [120].

LC-UV fingerprinting showed that echinacoside and acteoside were major characteristic peaks in C. deserticola, C. tubulosa, and C. salsa. C. sinensis contained little or no echinacoside [142].

A quantitative analysis of multiple components by a single marker method was also developed for simultaneous measurement of five phenylethanoid glycosides:

Echinacoside

Tubuloside A

Acteoside

Isoacteoside

2′-Acetylacteoside

This approach provides an economical analytical option for Cistanche quality control [143].

To more broadly characterize constituents associated with traditional reproductive, cognitive, and bowel functions, complementary HPLC–evaporative light-scattering detection and HPLC-UV methods were used to quantify five phenylethanoid glycosides together with betaine, mannitol, fructose, glucose, and sucrose [144–145].

These methods provide analytical tools for Q-Marker-based quality evaluation.

Our team also compared the principal active constituents of C. deserticola grown on different host plants, including the recently introduced host Atriplex canescens, commonly known as fourwing saltbush [145].

According to the source manuscript, our team is participating in the development of Cistanche quality standards for the United States Pharmacopeia and the European Pharmacopoeia, with the aim of supporting international standardization.

The manuscript further reports that these findings formed part of the project "Key Technologies for the Full-Industry-Chain Development and Application of the Endangered Desert Parasitic Medicinal Plant Cistanche," which received a Second-Class National Science and Technology Progress Award for 2025.

Related work also received a First-Class Science and Technology Progress Award in the promotion category of the 2016 Higher Education Outstanding Scientific Research Achievement Awards.

4. Conclusions and Future Perspectives

Cistanches Herba has a long history of medicinal use and a broad range of applications in TCM.

Based on our team's research and a substantial body of domestic and international literature, this review summarizes its chemical constituents and pharmacological activities.

Using the "three elements" of efficacy-related substances and the Q-Marker "five principles," echinacoside, acteoside, 8-epiloganic acid, betaine, and mannitol are proposed as a core marker group associated with Cistanche quality.

These constituents may support a more comprehensive and accurate analytical approach than reliance on echinacoside and acteoside alone. Their application must nevertheless account for botanical-source differences, particularly the differences between C. deserticola and C. tubulosa.

A function-associated quality-evaluation framework may help improve resource utilization, raw-material consistency, and the development of the Cistanche industry.

Several research gaps remain:

Iridoid glycosides: Although they are a major constituent class, their relationships with specific functions and overall quality are not yet fully understood.

Polysaccharides: Their activities and mechanisms have not been systematically characterized to the same extent as those of phenylethanoid glycosides, making their contribution to overall quality difficult to define.

Processing and formula compatibility: More research is needed on how these factors alter chemical composition and biological activity.

Future studies can use metabolomics, polysaccharide-focused omics, and other modern approaches to establish more complete chemical profiles and investigate multicomponent, multitarget interactions.

Mapping constituent–target–function relationships, while integrating processing and formula-compatibility research, may support quality-evaluation models more closely associated with clinical performance.

Practical Implications for Cistanche Ingredient Buyers

For supplement-ingredient procurement, the central implication of this review is that two marker assays cannot fully describe every Cistanche ingredient.

Buyers can use the research framework to ask more specific questions:

Which botanical species is supplied? C. deserticola and C. tubulosa should not be treated as chemically interchangeable.

What exactly is being standardized? An echinacoside specification, an acteoside specification, and a total phenylethanoid-glycoside specification describe different analytical attributes.

Which analytical method supports the specification? Method suitability matters when comparing results from HPLC-UV, LC-MS, ELSD, or other approaches.

Does the constituent profile match the intended application? A glycoside-focused extract does not necessarily represent the sugar-alcohol or polysaccharide profile of the starting herb.

How are origin, host plant, harvest, and processing controlled? These factors can influence composition and batch consistency.

The proposed Q-Markers are a research-based quality-evaluation framework, not a substitute for applicable regulatory requirements, validated product specifications, or evidence supporting a finished product's claims.

Conflict of Interest

All authors declare that they have no conflicts of interest.

References

Chinese-language reference titles are translated into English for accessibility. Author names, publication years, volumes, issues, and page ranges are retained from the supplied manuscript. Abbreviated author lists remain abbreviated as in the original.

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