Simultaneous quantification of ten key Kratom alkaloids in Mitragyna speciosa leaf extracts and commercial products by ultra‐performance liquid chromatography−tandem mass spectrometry

Kratom (Mitragyna speciosa) is a psychoactive plant popular in the United States for the self‐treatment of pain and opioid addiction. For standardization and quality control of raw and commercial kratom products, an ultra‐performance liquid chromatography?tandem mass spectrometry (UPLC?MS/MS) method was developed and validated for the quantification of ten key alkaloids, namely: corynantheidine, corynoxine, corynoxine B, 7‐hydroxymitragynine, isocorynantheidine, mitragynine, mitraphylline, paynantheine, speciociliatine, and speciogynine. Chromatographic separation of diastereomers, or alkaloids sharing same ion transitions, was achieved on an Acquity BEH C18 column with a gradient elution using a mobile phase containing acetonitrile and aqueous ammonium acetate buffer (10mM, pH 3.5). The developed method was linear over a concentration range of 1–200 ng/mL for each alkaloid. The total analysis time per sample was 22.5 minutes. The analytical method was validated for accuracy, precision, robustness, and stability. After successful validation, the method was applied for the quantification of kratom alkaloids in alkaloid‐rich fractions, ethanolic extracts, lyophilized teas, and commercial products. Mitragynine (0.7%–38.7% w/w), paynantheine (0.3%–12.8% w/w), speciociliatine (0.4%–12.3% w/w), and speciogynine (0.1%–5.3% w/w) were the major alkaloids in the analyzed kratom products/extracts. Minor kratom alkaloids (corynantheidine, corynoxine, corynoxine B, 7‐hydroxymitragynine, isocorynantheidine) were also quantified (0.01%–2.8% w/w) in the analyzed products; however mitraphylline was below the lower limit of quantification in all analyses.     

液相色谱-质谱联用法分析木瓜中绿原酸与抗癌活性成分的相关性研究

目的：高效液相色谱-质谱联用法（HPLC-MS）分析木瓜中绿原酸与抗癌活性成分的相关性。方法：采用HPLC-MS联用法,在Diamonsil C18色谱柱（4.6 mm×250 mm,5μm）,乙腈-0.05%磷酸水溶液梯度洗脱;流速为0.8 ml/min;检测波长为325 nm;柱温为25℃。分离测定木瓜中绿原酸的含量。结果：木瓜中绿原酸的含量为1.3 mg/g,相对标准偏差（RSD）为0.4%,线性关系良好。结论：木瓜中含有绿原酸,其主要成分与木瓜对脂肪酸合酶（FAS）的抑制及抗肿瘤作用有潜在的相关性。     

木瓜苷对佐剂性关节炎大鼠血清抗体水平的下调作用

目的探讨木瓜苷(glucosides of cheanomeles speciosa,GCS)对AA大鼠血清抗体水平的影响。方法SD大鼠随机分为6组:正常对照组、模型组(AA组)、GCS3个剂量组和雷公藤多苷组(GTW)。大鼠足跖皮内注射弗氏完全佐剂(FCA)诱发大鼠AA模型,聚乙二醇(PEG)6000法检测血清循环免疫复合物(circulating immune complexes,CIC);酶联免疫吸附(ELISA)法检测血清中抗Ⅱ型胶原抗体(anti-CⅡan-tibody)水平、抗结核杆菌抗体(anti-TB antibody)水平。结果大鼠免疫后d17开始分别灌胃GCS(30、60、120mg·kg-1)和GTW(40mg·kg-1)对照药,连续给药8d。GCS(60、120mg·kg-1)体内给药可明显降低AA大鼠血清中升高的CIC水平、抗CⅡ抗体水平和抗结核菌素抗体水平。结论GCS具有明显的下调AA大鼠血清抗体水平的作用,可能是其发挥治疗AA作用的重要机制之一。     

宣木瓜总有机酸的纯化及镇痛抗炎作用

目的:筛选适合分离纯化宣木瓜总有机酸的阴离子交换树脂,并对其进行分离纯化;研究纯化后总有机酸的镇痛抗炎作用。方法:以有机酸的收率为指标,筛选分离纯化宣木瓜有机酸最佳的离子交换树脂、最佳洗脱剂的浓度;通过ig给药,采用热板法和醋酸扭体法研究并比较宣木瓜总有机酸与醇提物的镇痛作用;以二甲苯诱导小鼠耳肿胀,观察并比较宣木瓜总有机酸与醇提物的抗炎作用。结果:717型强碱性阴离子交换树脂对宣木瓜总有机酸的交换能力最强,最佳洗脱剂为0.3 mol·L-1的盐酸,纯化后的有机酸纯度可达65.15%;宣木瓜总有机酸(0.15,0.3,0.6 g·kg-1)对醋酸和温度引起的小白鼠疼痛有明显的镇痛作用,对二甲苯致小鼠耳肿胀有一定的抑制作用,且与醇提物(1.25,2.5,5 g·kg-1)无显著性差异。结论:宣木瓜总有机酸可能是宣木瓜镇痛抗炎作用的有效组分之一。     

不同产地木瓜多糖含量的测定

目的:探讨不同产地木瓜多糖的含量,为临床合理应用和质量评价提供科学依据.方法:采用蒽酮-浓硫酸法显色,利用紫外分光光度法测定不同产地木瓜多糖的含量.结果:不同产地木瓜多糖的含量具有明显差异.木瓜多糖的含量与产区的平均日照、温度、气候、地理条件等有关.结论:本测定方法操作简单,重复性较好,结果准确可靠,可用于木瓜多糖的含量测定.     

光皮木瓜质量标准研究与不同产地药材的质量评价

目的探讨光皮木瓜的质量控制方法,对不同产地光皮木瓜的质量进行评价。方法参照文献对不同产地的光皮木瓜进行薄层色谱鉴别,并依照药典附录要求对不同产地光皮木瓜进行水分、总灰分和醇溶性浸出物等常规检测项目测定。结果薄层色谱条件可以有效对光皮木瓜进行鉴别。此外光皮木瓜10个测定样品中,水分含量最高为13.11%,最低为9.28%;总灰分含量最高为3.12%,最低为2.22%;采用热浸法测定醇溶性浸出物含量最高值为24.51%,最低为14.10%。结论薄层色谱条件可以有效地鉴别光皮木瓜,新增的水分、总灰分、醇溶性浸出物的试验结果可作为建立光皮木瓜药材质量标准的实验依据。     

中药木瓜多糖的提取及含量测定

用苯酚—硫酸法对中药木瓜中多糖含量进行了测定 ,结果显示 ,中药木瓜中多糖平均含量为 7 0 3%。     

Kratom induced severe cholestatic liver injury histologically mimicking primary biliary cholangitis: A case report

BACKGROUND Kratom is a psychoactive substance that is isolated from the plant Mitragyna speciosa. The leaves can be chewed fresh or dried, smoked, or infused similar to herbal teas. The plant leaves have been used by natives of Southeast Asia for centuries. The substance has been used for its stimulant activity at low doses, and as an opium substitute at higher doses due to a morphine like effect.CASE SUMMARY A 37-year-old female with a history of depression and obesity(body mass index: 32) presented to emergency room with a week-long history of nausea, decreased appetite, fatigue, and two days of jaundice. On admission bilirubin was markedly elevated. Her condition was thought to be due to consumption of Kratom 2 wk before onset of symptoms. Liver biopsy showed changes mimicking primary biliary cholangitis. Patient's symptoms and jaundice improved quickly.CONCLUSION The use of Kratom has been on the rise in recent years across the United States and Europe. Several case reports have associated adverse health impact ofKratom-containing products including death due to its ability to alter levels of consciousness. Only a few case reports have highlighted the hepatotoxic effects of Kratom. Even fewer reports exist describing the detailed histopathological changes.     

Acute toxicity study of the standardized methanolic extract of Mitragyna speciosa Korth in Rodent

Ethnopharmacological relevance: Mitragyna speciosa Korth (ketum) is widely used in Malaysia as a medicinal agent for treating diarrhea, worm infestations and also acts as an analgesic and antipyretic.

Aim

The aim of the study is to determine the acute toxicity of Mitragyna speciosa Korth standardized methanol extract in vivo in 4-weeks-old Sprague-Dawley rats.

Methodology

Rats were orally administrated single dose of 100, 500 and 1000 mg/kg Mitragyna speciosa Korth standardized methanol extract and the control group received 430 mg/kg of morphine orally. There were 10 rats in each group. All animals were sacrificed after 14 days of treatment. Eight parameters were tested: cage side observation, body weight measurement, food and water consumption, blood pressure, absolute and relative organ weight, hematology, biochemical analysis and histopathology, to look for evidence of toxicity.

Result

No mortality was noted after 14 days of treatment. In general, behavior, food and water consumption, hematological studies and organ weights showed no significant changes. The standardized methanol extraction of Mitragyna speciosa Korth increased rat blood pressure (systolic: 147.4 ± 1.01, 131.64 ± 4.94 and 137.8 ± 4.46) after an hour of 100, 500 and 1000 mg/kg doses, respectively. Biochemical studies showed significant elevation of ALT, AST, albumin, triglycerides, cholesterol and albumin (p > 0.05), at all levels of doses. But, nephrotoxicity evidenced by elevated creatinine was seen only at a dose of 1000 mg/kg. Histological examination showed congestion of sinusoids, hemorrhage hepatocytes, fatty change, centrilobular necrosis and increased number of Kuppfer cells in the liver of all Mitragyna speciosa Korth standardized methanol extract treated groups.

Conclusion

Oral administration of standardized methanolic extraction of Mitragyna speciosa Korth resulted in increasing rat blood pressure after an hour of drug administration. The highest dose of extract also induced acute severe hepatotoxicity and mild nephrotoxicity. However, Mitragyna speciosa Korth shows no effects on body weight, food and water consumption, absolute and relative organ weight and also hematology parameters.