Quality control and efficacy of Thatbunjob remedy in abdominal pain relief of patients with foodborne illness

This comprehensive study delves into the multifaceted aspects of Thatbunjob (TBJ), a Thai traditional medicine enlisted in Thailand's national essential medicines, primarily utilized for mitigating symptoms associated with diarrhea and bloating. Despite its extensive historical application, the lack of rigorous clinical studies has left the efficacy of TBJ unverified. This research aims to bridge this gap by conducting a meticulous examination involving In-vitro study, medicinal quality assessment, chemical analysis, and clinical trials to substantiate the therapeutic potential of Thatbunjob. The antimicrobial findings significantly contribute to reaffirming TBJ's confirmed antibacterial properties. Results align with previous research, demonstrating effectiveness against gastrointestinal pathogens such as S.aureus and S.dysenteriae, with MIC values of 1.25 and 5 mg/mL, respectively. The investigation into the anti-inflammatory properties of the 95% ethanolic extract from Thatbunjob involved a detailed assessment of its impact on nitric oxide, Interleukin 6, and tumor necrosis factor-alpha production in LPS-induced RAW 264.7 cells. The study revealed a substantial inhibitory effect on nitric oxide and IL-6 production, with an IC50 value of 64.13 ± 1.84 and 87.69 ± 0.07 μg/mL. No inhibitory effect was observed in tumor necrosis factor-alpha production. In the anti-foaming assay, Thatbunjob exhibited notable superiority over simethicone, showcasing a rapid onset in the initial phase. It is highlighted that Thatbunjob's potential significance in managing conditions characterized by excessive gastrointestinal gas offers a promising approach to relieve associated symptoms, such as abdominal discomfort. The chemical fingerprint analysis of Thatbunjob and confirmation through UV spectra provided substantial evidence supporting the identification of gallic acid, chebulagic acid, ellagic acid, and eugenol. The HPLC method validation adheres to ICH guidelines, suggesting its potential future use in controlling the amount of Thatbunjob's main chemical. The clinical trial process starts with drug production and quality testing. Once the drug met predefined standards, individuals aged 18 to 60 experiencing frequent diarrhea were evaluated for inclusion criteria. After treatment completion and questionnaire receipt from all 114 participants, statistical analysis of clinical results ensued.In the patients' enrollment and baseline clinical characteristics phase, the clinical trial utilized Thatbunjob and placebo capsules, adhering to defined quality standards. Of 114 initially enrolled participants, 109 subjects (Thatbunjob group = 55, placebo group = 54) completed the trial, with exclusions primarily due to non-compliance. Baseline characteristics analysis revealed no statistically significant differences in sex distribution and mean age between the Thatbunjob and Placebo groups. Both groups exhibited comparable male-to-female ratios and mean ages. Pre-intervention baseline VAS scores showed no significant differences, and an analysis of pain scores from the Applied Thai Short-form McGill Pain Questionnaire revealed no significant differences in various pain descriptors, except for the Sickening category (p-value = 0.042). The study exhibited a significant reduction in VAS scores among abdominal pain patients administered with Thatbunjob compared to the placebo group. This aligns with prior research highlighting herbal interventions' potential in ameliorating pain perception, indicating the potential therapeutic efficacy of Thatbunjob in managing symptoms associated with diarrhea. Results from the clinical trial provided compelling evidence of Thatbunjob's efficacy in alleviating abdominal pain and associated symptoms. Pain reduction within the initial 24 hours was significantly higher in the Thatbunjob group (52.7%) compared to the placebo group (9.3%). VAS scores for the Thatbunjob group were consistently significantly lower across multiple time points (24-, 48-, and 72-hours post-treatment), highlighting sustained efficacy in alleviating pain. In the initial 24-hour period, the Thatbunjob group exhibited significantly lower Buscopan tablet usage (16.4%) compared to the placebo cohort (44.4%), indicating reduced reliance on medication. No individuals in the Thatbunjob group required up to 3 Buscopan tablets per day, while a small percentage in the placebo group did. Although no significant difference in stool frequency was observed at the pretreatment assessment (0 hours), the Thatbunjob group exhibited significantly fewer stool occurrences at 24-48 hours post-treatment compared to the Placebo group, underscoring the effective properties of Thatbunjob in mitigating diarrhea.The quality control assessment of Thatbunjob products in Thailand's market, encompassing TBJ1 as the authentic drug and TBJ2 through TBJ8 as distinct products, involved a meticulous examination of physical and chemical attributes. The evaluation revealed that certain variations were observed among the products. Notably, deviations were found in water extractive value and ash content for some samples. Furthermore, microbial contamination testing identified areas for improvement in total aerobic bacterial testing for three samples. Comparisons of their concentrations of gallic acid, chebulagic acid, ellagic acid, and eugenol to the authentic drug (TBJ1) revealed significant variations. This emphasizes the impact of environmental factors on the secondary metabolites of medicinal plants. The observed differences underscore the necessity for robust quality control measures to maintain consistent levels of active compounds in Thatbunjob products.In summary, the presented findings offer valuable insights into the potential efficacy of Thatbunjob for alleviating gastrointestinal discomfort and associated symptoms. The results underscore the imperative for standardized production processes and emphasize the necessity for continued research in this domain.

本综合研究深入探讨了Thatbunjob（TBJ）的多维度特性——这是一种被列入泰国国家基本药物的传统医学制剂，主要用于缓解腹泻与腹胀相关症状。尽管其应用历史悠久，但缺乏严谨的临床研究导致TBJ的疗效尚未得到证实。本研究旨在通过开展涵盖体外研究（In-vitro study）、药用质量评估、化学分析及临床试验的细致考察，填补这一空白，以证实TBJ的治疗潜力。抗菌研究结果为重申TBJ已被确认的抗菌特性做出了重要贡献，其结果与既往研究一致，显示对金黄色葡萄球菌（S.aureus）和痢疾杆菌（S.dysenteriae）等胃肠道病原体具有抑制效果，最低抑菌浓度（MIC）值分别为1.25 mg/mL和5 mg/mL。

关于TBJ 95%乙醇提取物抗炎特性的研究，详细评估了其对脂多糖诱导的RAW 264.7细胞中一氧化氮、白细胞介素6及肿瘤坏死因子-α生成的影响。研究显示，该提取物对一氧化氮和白细胞介素6的生成具有显著抑制作用，半数抑制浓度（IC50）值分别为64.13 ± 1.84 μg/mL和87.69 ± 0.07 μg/mL，但对肿瘤坏死因子-α的生成无抑制效果。在抗发泡试验中，TBJ表现出优于西甲硅油（simethicone）的显著性能，尤其在初始阶段起效迅速。值得强调的是，TBJ在管理胃肠道气体过多相关病症方面的潜在价值，为缓解腹部不适等症状提供了新的思路。

TBJ的化学指纹分析及紫外光谱（UV spectra）验证提供了充分证据，支持对没食子酸、诃子酸、鞣花酸及丁香酚的鉴定。高效液相色谱法（HPLC）验证符合国际人用药品注册技术协调会（ICH）指南，表明其未来有望用于控制TBJ主要化学成分的含量。

临床试验流程始于药物生产与质量检测。当药物达到预设标准后，对18至60岁频繁腹泻患者进行纳入标准评估。在114名参与者均完成治疗并提交问卷后，开展了临床结果的统计分析。在患者招募与基线临床特征阶段，临床试验采用符合既定质量标准的TBJ及安慰剂胶囊。在最初招募的114名参与者中，109名受试者（TBJ组=55人，安慰剂组=54人）完成了试验，排除原因主要为不依从。基线特征分析显示，TBJ组与安慰剂组在性别分布及平均年龄方面无统计学显著差异，两组的男女比例及平均年龄具有可比性。干预前的视觉模拟评分法（Visual Analog Scale, VAS）基线分数无显著差异；通过泰国版简明麦吉尔疼痛问卷进行的疼痛评分分析显示，除恶心类别（p值=0.042）外，各疼痛描述项均无显著差异。

研究显示，与安慰剂组相比，接受TBJ治疗的腹痛患者VAS分数显著降低，这与既往关于草药干预改善疼痛感知的研究一致，表明TBJ在缓解腹泻相关症状方面具有潜在治疗效果。临床试验结果为TBJ缓解腹痛及相关症状的疗效提供了有力证据：TBJ组在初始24小时内的疼痛缓解率（52.7%）显著高于安慰剂组（9.3%）；TBJ组在治疗后24、48及72小时的VAS分数持续显著低于安慰剂组，体现了其缓解疼痛的持续疗效。在初始24小时内，TBJ组的Buscopan片使用率（16.4%）显著低于安慰剂组（44.4%），表明患者对药物的依赖减少；TBJ组无患者需每日服用3片Buscopan，而安慰剂组有少量患者需此剂量。尽管治疗前（0小时）两组排便频率无显著差异，但TBJ组在治疗后24-48小时的排便次数显著少于安慰剂组，突显了TBJ缓解腹泻的有效特性。

针对泰国市场上TBJ产品的质量控制评估涵盖了作为正品药物的TBJ1及TBJ2至TBJ8等不同产品，考察内容包括理化特性。评估结果显示，各产品间存在一定差异：部分样品的水溶性浸出物值及灰分含量偏离标准；此外，微生物污染检测发现3个样品的总需氧菌检测结果存在改进空间；与正品药物TBJ1相比，各产品中没食子酸、诃子酸、鞣花酸及丁香酚的浓度存在显著差异。这强调了环境因素对药用植物次生代谢产物的影响，而观察到的差异也突显了建立严格质量控制措施以维持TBJ产品活性成分含量一致性的必要性。

综上，本研究结果为TBJ缓解胃肠道不适及相关症状的潜在疗效提供了宝贵见解，同时强调了标准化生产流程的必要性及该领域持续研究的重要性。